研究者詳細 - 沢　禎彦

論文 - 沢　禎彦

分割表示 >> ／全件表示 95 件中 1 - 95 件目

Study of Podoplanin-Deficient Mouse Bone with Mechanical Stress. 査読国際誌

Takenori Kanai, Kyoko Osawa, Koichiro Kajiwara, Yoshiaki Sato, Yoshihiko Sawa

Dentistry journal 13 ( 2 ) 2025年1月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Objective: We investigated morphological differences in osteocyte processes between aged mice and our original podoplanin-conditional knockout (cKO) mice in which the floxed exon 3 of podoplanin was deleted by Dmp-1-driven Cre (Dmp1-Cre;PdpnΔ/Δ). Methods: SEM observation on osteocyte cell process, histochemistry for bone remodeling with mechanostress, and RT-PCR for RANKL and M-CSF in podoplanin cKO mouse bone with mechanostress was investigated. Results: SEM observations showed fewer and thinner osteocyte processes in femurs from 23-week-old Dmp1-Cre;PdpnΔ/Δ mice than from 23-week-old wild-type mice, while the numbers of osteocyte processes in femurs and calvarias were similar in 23-week-old Dmp1-Cre;PdpnΔ/Δ mice and 48-week-old wild-type mice. Furthermore, cell process numbers in femurs and calvarias were significantly smaller in 23-week-old Dmp1-Cre;PdpnΔ/Δ mice than in 48-week-old wild-type mice. In the test for differences in alveolar bone resorption under mechanical stress between Dmp1-Cre;PdpnΔ/Δ and wild-type mice, the area of TRAP-positive resorption pits was larger in wild-type mice than in Dmp1-Cre;PdpnΔ/Δ mice. In a quantitative tissue PCR analysis, the mRNA expression levels of RANKL and M-CSF in alveolar bone under mechanical stress were significantly lower in Dmp1-Cre;PdpnΔ/Δ mice than in wild-type mice. These results suggest that a reduction in cell process formation in osteocytes with podoplanin cKO affected the absorption of alveolar bone under mechanical stress in Dmp1-Cre;PdpnΔ/Δ mice. Conclusions: In podoplanin-deficient bone, the deformation of osteocyte processes by mechanical stimuli is not recognized as a stress due to the lower number of cell processes with podoplanin deficiency; therefore, the production of osteoclast migration/differentiation factors by activated osteocytes is not fully induced and macrophage migration to alveolar bone with mechanical stress appeared to be suppressed. These results indicate that podoplanin-dependent osteocyte process formation indirectly plays a key role in sensing mechanical stress in bone.

DOI： 10.3390/dj13020061

Exacerbation of diabetes due to F. Nucleatum LPS-induced SGLT2 overexpression in the renal proximal tubular epithelial cells. 査読国際誌

Aiko Seki, Koichiro Kajiwara, Jumpei Teramachi, Masahiko Egusa, Takuya Miyawaki, Yoshihiko Sawa

BMC nephrology 26 ( 1 ) 38 - 38 2025年1月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

BACKGROUND: Diabetes treatments by the control of sodium-glucose cotransporter 2 (SGLT2) is commonly conducted while there are still uncertainties about the mechanisms for the SGLT2 overexpression in kidneys with diabetes. Previously, we have reported that glomeruli and proximal tubules with diabetic nephropathy express toll-like receptor TLR2/4, and that the TLR ligand lipopolysaccharide (LPS) of periodontal pathogens have caused nephropathy in diabetic model mice. Recently, many researchers suggested that the periodontal pathogenic bacteria Fusobacterium (F.) nucleatum has the TLR4-associated strong activator of the colorectal inflammation and cancer. The present study aimed to investigate the possibility of F. nucleatum as an exacerbation factor of diabetes through the renal SGLT2 induction. METHODS: The induction of the SGLT2 by F. nucleatum LPS (Fn-LPS) were investigated in the streptozotocin-induced diabetic mouse renal tissue and cultured renal proximal epithelial cells. The changes of blood glucose levels and survival curves in diabetic mice with Fn-LPS were analyzed. The Fn-LPS-induced SGLT2 production in the diabetic mouse renal tissue and in the cultured proximal epithelial cells was examined by ELISA, quantitative RT-PCR, and immunohistochemical analysis. RESULTS: The SGLT2 expression in the cultured mouse tubular epithelial cells was significantly increased by TNF- or co-culture with Fn-LPS-supplemented J774.1 cells. The period to reach diabetic condition was significantly shorter in Fn-LPS-administered diabetic mice than in diabetic mice. All Fn-LPS-administered-diabetic mice reached humane endpoints during the healthy period of all of the mice administered Fn-LPS only. The promotion of the SGLT2 expression at the inner lumen of proximal tubules were stronger in the Fn-LPS-administered-diabetic mice than in diabetic mice. The renal tissue SGLT2 mRNA amounts and the number of renal proximal tubules with overexpressed SGLT2 in the lumen were more in the Fn-LPS-administered-diabetic mice than in diabetic mice. CONCLUSIONS: This study suggests that F. nucleatum causes the promotion of diabetes through the overexpression of SGLT2 in proximal tubules under the diabetic condition. Periodontitis with F. nucleatum may be a diabetic exacerbating factor.

DOI： 10.1186/s12882-025-03965-z

The Abnormal Expression of Tubular SGLT2 and GULT2 in Diabetes Model Mice with Malocclusion-Induced Hyperglycemia. 査読国際誌

Koichiro Kajiwara, Sachio Tamaoki, Yoshihiko Sawa

Biomedicines 13 ( 2 ) 2025年1月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Background: A relationship between malocclusion and the promotion of diabetes has been suggested. In hyperglycemia, the expression of sodium-glucose cotransporter 2 (SGLT2) and the facilitative glucose transporter 2 (GLUT2) is upregulated in proximal tubular cells, leading to an increase in renal glucose reabsorption. The present study aimed to investigate whether malocclusion contributes to diabetic exacerbation. Methods: Streptozotocin (STZ)-induced diabetic mice with malocclusion due to cutting molars were investigated based on increased blood glucose levels. PCR and immunohistochemical analyses were performed on diabetic mice kidneys to investigate the expression of SGLT2 and GLUT2. Results: Animal experiments were performed using 32 mice for 21 days. The time to reach a diabetic condition in STZ-administered mice was shorter with malocclusion than without malocclusion. The increase and mean blood glucose levels in STZ-administered mice were steeper and higher with malocclusion than without malocclusion. Urea albumin, BUN, and CRE levels were higher in diabetic mice with malocclusion than in diabetic mice without. Immunoreaction with anti-SGLT2 and anti-GLUT2 in the renal tissue of STZ-administered mice was stronger with malocclusion than without malocclusion. The amounts of SGLT2 and GLUT2 mRNA in the renal tissue in STZ-administered mice were higher with malocclusion than without malocclusion. The amounts of TNF-a and IL-6 mRNA in the large intestinal tissue in STZ-administered mice were higher with malocclusion than without malocclusion. Conclusions: Our results indicate that malocclusion accelerates the tubular expression of SGLT2 and GLUT2 under hyperglycemia. Malocclusion may be a diabetes-exacerbating factor with increased poor glycemic control due to shortened occlusion time resulting from swallowing food without chewing.

DOI： 10.3390/biomedicines13020267

ポドプラニンノックアウトマウスにおける下顎前歯歯槽骨の骨リモデリング機構について

梶原弘一郎, 沢禎彦, 玉置幸雄

日本矯正歯科学会大会プログラム・抄録集 83回 187 - 187 2024年10月

Morphological Study for the Osteocytes in Podoplanin-Conditional Knockout Mice 査読

Kyoko Osawa, Takenori Kanai, Natsumi Ushijima, Koichiro Kajiwara, Yoshihiko Sawa, Yoshiaki Sato

Journal of Hard Tissue Biology 32 ( 4 ) 213 - 222 2023年10月

糖尿病マウス腎におけるSGLT2 and VCAM-1発現について

梶原弘一郎, 沢禎彦

Journal of Oral Biosciences Supplement 2023 [P3 - 05 2023年9月

P.gingivalis LPS誘導性糖尿病モデルマウス腎でのSGLT2過剰発現について(Overexpression of SGLT2 in the kidney of a P.gingivalis LPS-induced diabetic nephropathy mouse model) 査読

梶原弘一郎, 沢禎彦

Journal of Oral Biosciences Supplement 2022 370 - 370 2022年9月

Maxillofacial morphological characteristics in growing orthodontic patients with non‐syndromic oligodontia 査読

Taiki Suyama, Hiroyuki Ishikawa, Sachio Tamaoki, Remi Higa, Shunsuke Takata, Yoshihiko Sawa

Orthodontics & Craniofacial Research 2021年12月

Overexpression of SGLT2 in the kidney of a P. gingivalis LPS-induced diabetic nephropathy mouse model. 査読国際誌

Koichiro Kajiwara, Yoshihiko Sawa

BMC Nephrology 22 ( 1 ) 287 - 287 2021年8月

担当区分：最終著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

BACKGROUND: The overexpression of sodium-glucose cotransporter 2 (SGLT2) in diabetic kidneys has been reported. It has also been established that the diabetic glomerular endothelium expresses the toll-like receptors TLR2 and TLR4. The present study aims to examine the renal SGLT2 induction by the TLR2/4 ligand Porphyromonas (P.) gingivalis lipopolysaccharide (Pg-LPS) in mouse diabetic nephropathy. METHODS: Immunohistochemical study and tissue RT-PCR analyses were performed on mouse kidneys in streptozotocin (STZ)-induced diabetic ICR mice (STZ-ICR), in healthy ICR mice administered Pg-LPS (LPS-ICR), and in diabetic ICR mouse kidneys with Pg-LPS-induced nephropathy (LPS-STZ). RESULTS: In the quantitative analysis of blood sugar levels, the mean time to reach 600 mg/dl was shorter in the LPS-STZ than in the STZ-ICR kidneys. The rise in blood glucose levels was significantly steeper in the LPS-STZ than in the STZ-ICR kidneys. According to these data the LPS-STZ model suggests a marked glucose intolerance. The expression of SGLT2 was significantly stronger in the whole of the renal parenchyma of the LPS-STZ than in the LPS-ICR or in the STZ-ICR. The expression of SGLT2 was observed both in the renal tubules and around the renal tubules, and in the glomeruli of the LPS-STZ kidneys. In the analysis by tissue real-time PCR and cell ELISA, the expression of the SGLT2 gene and protein was significantly stronger in the LPS-STZ than in the LPS-ICR or in the STZ-ICR. There were no differences in the renal SGLT2 production in the LPS-ICR and the STZ-ICR kidneys. CONCLUSIONS: Abnormally high renal expression of SGLT2 occurs in diabetic kidneys with P. gingivalis LPS. Periodontitis may be an exacerbating factor in diabetic nephropathy as well as in diabetes.

DOI： 10.1186/s12882-021-02506-8

Expression of SARS-CoV-2 entry factors in human oral tissue. 査読国際誌

Yoshihiko Sawa, Soichiro Ibaragi, Tatsuo Okui, Junro Yamashita, Tetsuro Ikebe, Hiroyuki Harada

Journal of Anatomy 238 ( 6 ) 1341 - 1354 2021年6月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

The distribution of cells expressing SARS-CoV-2 entry factor angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) in human oral tissues were tested. The investigation was conducted with normal flesh tissue and paraffin-embedded specimens. The ACE2 and TMPRSS2 expression was detected with all subjects in the normal mucosa of the keratinized stratified squamous epithelia of the tongue and non-keratinized stratified squamous epithelia of the lip and cheek. It was found that ACE2 is expressed in the cytoplasm and on the cell membrane mainly in the stratum granulosum of the epithelia while the TMPRSS2 is strongly expressed on the cell membrane mainly in the stratum granulosum and stratum spinosum, but not in the stratum basale. Antibodies' reactions for ACE2 and TMPRSS2 were not observed in the nuclei or keratin layer. The expression of ACE2 and TMPRSS2 in the oral epithelia appears to be general, and the expression was also observed in the mucous and serous acini of the labial glands. The SARS-CoV-2 may transiently attach to the oral mucosa and the minor salivary glands which are present under all of the oral mucosa. The oral cavity can be considered an important organ for SARS-CoV-2 attachment and may provide a preventive medical avenue to guard against COVID-19 by preventing saliva from scattering.

DOI： 10.1111/joa.13391

Immunohistochemical study for the expression of leukocyte adhesion molecules, and FGF23 and ACE2 in P. gingivalis LPS-induced diabetic nephropathy. 査読国際誌

Koichiro Kajiwara, Yoshihiko Sawa, Takahiro Fujita, Sachio Tamaoki

BMC Nephrology 22 ( 1 ) 3 - 3 2021年1月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

OBJECTIVE: The present study aims to examine the expression of leukocyte adhesion molecules and renal metabolic factors in diabetic mouse kidneys with periodontal pathogen Pg-LPS-induced nephropathy. BACKGROUND: We recently reported that the glomerular endothelium expresses toll-like receptor (TLR)2 and TLR4 in diabetic environments and TLR2/4 ligand Porphyromonas (P.) gingivalis lipopolysaccharides (Pg-LPS) induce nephropathy in diabetic mice. It is thought that Pg-LPS promotes the chronic inflammation with the overexpression of leukocyte adhesion molecules and renal-specific metabolic enzymes by the recognition of Pg-LPS via TLR in the diabetic kidneys. There have been no reports of the effects of periodontopathic bacteria on the expression of leukocyte adhesion molecules and the accumulation of physiologically active substances in the kidney. METHODS: The immunohistochemical investigation was performed on diabetic mouse kidney with Pg-LPS-induced nephropathy with glomerulosclerosis in glomeruli. RESULTS: There were no vessels which expressed vascular cell adhesion molecule-1 (VCAM-1), E-selectin, or fibroblast growth factor (FGF) 23 in streptozotocin (STZ)-induced diabetic ICR mice (STZ-ICR), or in healthy ICR mice administered Pg-LPS (LPS-ICR). However, in diabetic ICR mouse kidneys with Pg-LPS-induced nephropathy (LPS-STZ) the expression of VCAM-1 and the accumulation of FGF23 were observed in renal tubules and glomeruli, and the expression of E-selectin was observed in renal parenchyma and glomeruli. The angiotensin-converting enzyme 2 (ACE2) was detected in the proximal tubules but not in other regions of ICR, STZ-ICR, or LPS-ICR. In LPS-STZ ACE2 was detected both in renal tubules as well as in glomeruli. The Mac-1 and podoplanin-positive cells increased in the renal parenchyma with diabetic condition and there was the distribution of a large number of Mac-1-positive cells in LPS-STZ. CONCLUSIONS: The Pg-LPS may induce diabetic renal inflammation such as glomerulosclerosis and tubulitis with infiltration of Mac-1/podoplanin positive macrophages via glomerular overexpression of VCAM-1 and E-selectin, resulting in accumulation of both ACE2 and FGF23 which were unmetabolized with the inflammation-induced kidney damage under the diabetic condition. Periodontitis may be a critical factor in the progress of nephropathy in diabetic patients.

DOI： 10.1186/s12882-020-02203-y

Cancellation of the Calcification in Cultured Osteoblasts by CLEC-2

Takenori Kanai, Yoshihiko Sawa, Yoshiaki Sato

Journal of Hard Tissue Biology 30 ( 1 ) 53 - 62 2021年

Effect of bisphosphonates on healing of tooth extraction wounds in infectious osteomyelitis of the jaw 査読国際誌

Junro Yamashita, Naruhiko Sawa, Yoshihiko Sawa, Shoji Miyazono

Bone 143 115611 - 115611 2020年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier BV

OBJECTIVES: Antiresorptive agent-related osteonecrosis of the jaw (ARONJ) and infectious osteomyelitis of the jaw (OMJ) in antiresorptive-naïve patients are different disease entities. Although osteoclast inhibition is at the center of the pathogenesis of ARONJ, the role of osteoclast inhibition in infectious OMJ is unknown. The objective of this study was to determine the effect of bisphosphonate osteoclast inhibition in infectious OMJ. METHODS: Osteomyelitis was induced in mice by S. aureus inoculation. The establishment of OMJ was verified by the culture of bone marrow samples obtained from the mandible. Infected animals received either zoledronic acid (ZA) or saline starting at week-2. Treated animals along with non-infected animals underwent tooth extractions at week-4 post-infection. Healing was assessed every week using in vivo micro-computed tomography and intraoral photos. Animals were euthanized at week-8 and cervical lymph nodes were assessed for lymphatic and blood vessels. RESULTS: Tooth extraction wounds did not heal in animals with OMJ. These wounds were characterized by incomplete soft tissue coverage, sporadic bone fill in the sockets, and inflammatory cell accumulation in the connective tissue at 4 weeks after tooth extractions. Conversely, the majority of tooth extraction wounds in the infected animals treated with ZA had improved healing with better bone fill than even non-infected control animals. Consistently, atrophic lymphatic vessels were noted in the draining lymph nodes in animals with OMJ. However, infected animals treated with ZA had lymphatic vessels that were unaltered and showed a similar appearance to those in non-infected control animals. CONCLUSION: ZA treatment promoted wound healing in the jaw with infectious osteomyelitis. Clearly, antiresorptive therapy is contraindicated in patients with ARONJ. However, our finding suggests that osteoclast inhibition is potentially an effectual remedy for infectious OMJ in antiresorptive-naïve patients.

DOI： 10.1016/j.bone.2020.115611

Alternating Differentiation and Dedifferentiation between Mature Osteoblasts and Osteocytes. 国際誌

Naruhiko Sawa, Hiroki Fujimoto, Yoshihiko Sawa, Junro Yamashita

Scientific reports 9 ( 1 ) 13842 - 13842 2019年9月

記述言語：英語掲載種別：研究論文（学術雑誌）

Osteocytes are terminally differentiated osteoblasts embedded in the bone matrix. Evidence indicates that cells in the mesenchymal lineage possess plasticity. However, whether or not osteocytes have the capacity to dedifferentiate back into osteoblasts is unclear. This study aimed to clarify the dedifferentiation potential of osteocytes. Mouse calvarial osteoblasts were isolated and maintained in normal two-dimensional (2D) or collagen gel three-dimensional (3D) cultures. In 2D cultures, osteoblasts exhibited a typical fibroblast-like shape with high Alpl and minimal Sost, Fgf23, and Dmp1 expression and osteoblasts formed mineralised nodules. When these osteoblasts were transferred into 3D cultures, they showed a stellate shape with diminished cytoplasm and numerous long processes and expression of Alpl decreased while Sost, Fgf23, and Dmp1 were significantly increased. These cells were in cell cycle arrest and showed suppressed mineralisation, indicating that they were osteocytes. When these osteocytes were recovered from 3D cultures and cultured two-dimensionally again, they regained adequate cytoplasm and lost the long processes, resulting in a fibroblast-like shape. These cells showed high Alpl and low Sost, Fgf23, and Dmp1 expression with a high mineralisation capability, indicating that they were osteoblasts. This report shows that osteocytes possess the capacity to dedifferentiate back into mature osteoblasts without gene manipulation.

DOI： 10.1038/s41598-019-50236-7

Alternating Differentiation and Dedifferentiation between Mature Osteoblasts and Osteocytes 査読国際誌

Sawa N, Fujimoto H, Sawa Y, Yamashita J

Scientific Reports 25;9 ( 1 ) 13842 - 21788 2019年9月

Maxillofacial morphological factors related to acceleration of maxillary growth attributed to facial mask treatment: a structural superimposition study. 査読国際誌

Takashi S Kajii, Yui Sakaguchi, Yoshihiko Sawa, Sachio Tamaoki

Progress in orthodontics 20 ( 1 ) 2 - 2 2019年1月

記述言語：英語掲載種別：研究論文（学術雑誌）

BACKGROUND: Anatomical textbooks mention that the contact between the pterygoid process and the palatine's pyramidal process is not a "suture" but "conjugation.".The aim was to evaluate the maxillofacial morphological factor responding most to the orthopedic force of facial mask treatment, using the structural superimposition analysis. METHODS: Thirty-one girls with Angle Class III malocclusion treated using a facial mask (FM group) and 11 girls with pseudo-Class III malocclusion (pseudo-III group) were examined. Lateral cephalograms at pre- and posttreatment were analyzed to evaluate maxillofacial changes. Cephalometric structural superimposition analysis originating with Björk and Skieller was also performed. RESULTS: In the FM group, a multiple linear regression model showed that maxillary sutural growth was significantly associated with counter-clockwise rotation of the maxilla and treatment changes in the anteroposterior distance from the pterygomaxillary fissure to the maxillary anterior alveolus, not changes in the distance from the nasion to the maxillary anterior alveolus. CONCLUSIONS: Structural superimposition analysis showed that counter-clockwise rotation of the maxilla and changes in the distance from the pterygomaxillary fissure to the maxillary anterior alveolus responded most to the orthopedic force of facial mask treatment. The analysis implicated that the pterygoid fissure-palatine's pyramidal process conjugation responds most to facial mask treatment among maxillofacial sutures and conjugation, and that the difference in the response induces maxillary counter-clockwise rotation.

DOI： 10.1186/s40510-018-0254-9

LpMab-23-recognizing cancer-type podoplanin is a novel predictor for a poor prognosis of early stage tongue cancer 査読国際誌

Akihiro Miyazaki, Hiromi Nakai, Tomoko Sonoda, Yoshihiko Hirohashi, Mika K. Kaneko, Yukinari Kato, Yoshihiko Sawa, Hiroyoshi Hiratsuka

Oncotarget 9 ( 30 ) 21156 - 21165 2018年4月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Impact Journals LLC

Purpose: We report that the reactivity of a novel monoclonal antibody LpMab-23 for human cancer-type podoplanin (PDPN) is a predictor for a poor prognosis of tongue cancer. Patients and Methods: The association between LpMab-23-recognizing cancertype PDPN expression and clinical/pathological features were analyzed on 60 patients with stage I and II tongue cancer treated with transoral resection of the primary tumor. Results: In the mode of invasion, the LpMab-23-dull/negative cases were significantly larger in cases with low-grade malignancies and without late cervical lymph node metastasis, than in cases with high-grade malignancies and the metastasis. In the high-grade malignant cases, LpMab-23-positive cases were significantly larger than LpMab-23-dull/negative cases. The Kaplan-Meier curves of the five-year metastasis-free survival rate (MFS) were significantly lower in the LpMab-23 positive patients than in LpMab-23 dull/negative patients. The LpMab-23- dull/negative cases showed the highest MFS in all of the clinical/pathological features and particularly, the MFS of the LpMab-23 positive cases decreased to less than 60% in the first year. In the Cox proportional hazard regression models a comparison of the numbers of LpMab-23 dull/negative with positive cases showed the highest hazard ratio with statistical significance in all of the clinical/pathological features. Conclusions: LpMab-23 positive cases may be considered to present a useful predictor of poor prognosis for early stage tongue cancer.

DOI： 10.18632/oncotarget.24986

Expression and dynamics of podoplanin in cultured osteoblasts with mechanostress and mineralization stimulus 査読

Tomohiro Takenawa, Takenori Kanai, Tetsuya Kitamura, Yoshitaka Yoshimura, Yoshihiko Sawa, Junichiro Iida

Acta Histochemica et Cytochemica 51 ( 1 ) 41 - 52 2018年

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Japan Society of Histochemistry and Cytochemistry

This study investigates the significance of the expression and dynamics of podoplanin in mechanostress and mineralization in cultured murine osteoblasts. Podoplanin increased in osteoblasts subjected to straining in non-mineralization medium, suggesting that the mechanostress alone is a podoplanin induction factor. In osteoblasts subjected to vertical elongation straining in the mineralization medium, the mRNA amounts of podoplanin, osteopontin, and osteocalcin were significantly larger than those in cells not subjected to straining, suggesting that mechanostress is the cause of a synergistic effect in the expression of these proteins. In osteoblasts in the mineralization medium, significant increases in osteocalcin mRNA occurred earlier in cells subjected to straining than in the cells not subjected to straining, suggesting that the mechanostress is a critical factor to enhance the expression of osteocalcin. Western blot and ELISA analysis showed increased podoplanin production in osteoblasts with longer durations of straining. There was significantly less mineralization product in osteoblasts with antibodies for podoplanin, osteopontin, and osteocalcin. There was also less osteopontin and osteocalcin produced in osteoblasts with anti-podoplanin. These findings suggest that mechanostress induces the production of podoplanin in osteoblasts and that podoplanin may play a role in mineralization in cooperation with bone-associated proteins.

DOI： 10.1267/ahc.17031

The promotion of nephropathy by Porphyromonas gingivalis lipopolysaccharide via toll-like receptors 査読国際誌

Koichiro Kajiwara, Shunsuke Takata, Thao T. To, Kenyo Takara, Yuji Hatakeyama, Sachio Tamaoki, Richard Peters Darveau, Hiroyuki Ishikawa, Yoshihiko Sawa

Diabetology and Metabolic Syndrome 9 ( 1 ) 73 - 73 2017年9月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：BioMed Central Ltd.

Background: Recently, we reported that toll-like receptor (TLR)2 and TLR4 localized on the glomerular endothelium in the glomeruli of streptozotocin (STZ)-induced type 1 diabetic mice and high fat diet feed-induced type 2 diabetic mice, and that periodontal pathogen Porphyromonas gingivalis LPS (Pg-LPS) administration lowered the survival rate of diabetic mice. The present study aims to examine the effect of TLR4 blocking on the suppression of Pg-LPS-induced diabetic nephropathy. Methods: The survival rate and morphological/biochemical features for streptozotocin-induced diabetic mice with Pg-LPS and TLR4 blocker eritoran administration were investigated by reporter gene assay, urine and blood analysis, immunohistochemistry, and real time-PCR. Results and Conclusions: All of the diabetic mice administered Pg-LPS were euthanized within the survival period of almost all of the diabetic mice. The blood urea nitrogen and creatinine, expression of TLR2 and TGF-b, and type 1 collagen accumulation, in the diabetic mice increased significantly with the Pg-LPS administration. In spite of the limited TLR4 activation with Pg-LPS, the TLR4 blocker eritoran decreased blood urea nitrogen and creatinine, and raised the survival rate of the Pg-LPS-administered diabetic mice slightly. The high expression levels of TLR2, TGF-b, and type 1 collagen in Pg-LPS-administered diabetic mice decreased with eritoran. Nuclear STAT3 which enhances TLR2 expression was detected in the TLR2-expressing glomeruli of diabetic mice. The TLR2 and STAT3 gene expression increased by the Pg-LPS administration but decreased with eritoran. These may suggest that Pg-LPS-induced diabetic nephropathy is mainly dependent on TLR2 signaling on glomerular endothelial cells, and that TLR4 blocker eritoran may play a role to slow the progress of diabetic nephropathy.

DOI： 10.1186/s13098-017-0271-8

Distinct Role of Transforming Growth Factor-Beta 1 and Fibroblast Growth Factors in Human Amelobastoma Epithelial Cell Proliferation

Yuko Matsuda, Naoko Kamogashira, Yuji Hatakeyama, Toshinari Mikami, Kazuki Nakashima, Junko Hatakeyama, Sachio Tamaoki, Yoshihiko Sawa, Hiroyuki Ishikawa

Biochemistry and Molecular Biology 2 ( 1 ) 1 - 5 2017年9月

Morphological study of tooth development in podoplanin-deficient mice 査読国際誌

Kenyo Takara, Naoki Maruo, Kyoko Oka, Chiaki Kaji, Yuji Hatakeyama, Naruhiko Sawa, Yukinari Kato, Junro Yamashita, Hiroshi Kojima, Yoshihiko Sawa

PLOS ONE 12 ( 2 ) e0171912 2017年2月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PUBLIC LIBRARY SCIENCE

Podoplanin is a mucin-type highly O-glycosylated glycoprotein identified in several somatyic cells: podocytes, alveolar epithelial cells, lymphatic endothelial cells, lymph node stromal fibroblastic reticular cells, osteocytes, odontoblasts, mesothelial cells, glia cells, and others. It has been reported that podoplanin-RhoA interaction induces cytoskeleton relaxation and cell process stretching in fibroblastic cells and osteocytes, and that podoplanin plays a critical role in type I alveolar cell differentiation. It appears that podoplanin plays a number of different roles in contributing to cell functioning and growth by signaling. However, little is known about the functions of podoplanin in the somatic cells of the adult organism because an absence of podoplanin is lethal at birth by the respiratory failure. In this report, we investigated the tooth germ development in podoplanin-knockout mice, and the dentin formation in podoplanin-conditional knockout mice having neural crest-derived cells with deficiency in podoplanin by the Wnt1 promoter and enhancer-driven Cre recombinase: Wnt1-Cre; Pdpn(Delta/Delta) mice. In the Wnt1-Cre; Pdpn(Delta/Delta)mice, the tooth and alveolar bone showed no morphological abnormalities and grow normally, indicating that podoplanin is not critical in the development of the tooth and bone.

DOI： 10.1371/journal.pone.0171912

Effects of a chemically synthesized leucine-rich amelogenin peptide (csLRAP) on chondrogenic and osteogenic cells 査読

Yuko Matsuda, Yuji Hatakeyama, Kazuki Nakashima, Naoko Kamogashira, Junko Hatakeyama, Sachio Tamaoki, Yoshihiko Sawa, Hiroyuki Ishikawa

Journal of Hard Tissue Biology 26 ( 1 ) 51 - 60 2017年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Society of Hard Tissue Regenerative Biology

Amelogenin is one of the enamel matrices secreted by ameloblasts and is known to have at least 15 alternative mRNA splicing isoforms. The leucine rich amelogenin peptide (LRAP) is one of most abundant amelogenin isoforms and numerous studies have shown that amelogenin peptides including LRAP, are expressed in cartilage and bone. Lysosome-associated membrane protein-1 (LAMP-1) has been identified as amelogenin binding partner, however, the mechanism of action for LRAP in chondrogenesis or osteogenesis is still unclear. This study aimed to assay the effect of chemically synthesized LRAP (csLRAP) on chondrogenic or osteogenic differentiation with the involvement of LAMP-1. The csLRAP used in this study was generated by F-moc solid phase chemistry based on the mouse LRAP amino acid sequence and was added to the culture medium of the chondrogenic cells, ATDC5 and the osteoblast cells, MC3T3-E1. Both chondrogenic and osteoblast cells, in which an immunopositive reaction to LAMP-1 antibody was observed by immunocytochemistry, showed significant suppression in cell number in the presence of csLRAP at 10 μg/ml compared to that in control, but this effect was rescued in the presence of LAMP-1 antibody. On the other hand, the intensity of alcian blue staining in chondrogenic cells and alizarin red staining in osteoblast cells was significantly increased in the presence of csLRAP at a dose of 10 μg/ml after 4 weeks culture, and this effect was suppressed in the presence LAMP-1 antibody. Moreover, the chondrogenic or osteogenic differentiation marker genes including Sox9, Type II Collagen, Type X Collagen, Runx2, Alkaline Phosphatase, and Type I Collagen, were upregulated in the presence of csLRAP after one week of culture and were suppressed in the presence of LAMP-1 antibody. These results suggest that csLRAP could promote osteogenic and chondrogenic differentiation in vitro, and that LAMP-1 may be involved in the differentiation and proliferation of these cells.

DOI： 10.2485/jhtb.26.51

レクチンを用いた歯根膜におけるオキシタラン線維の検出の検討

鴨頭奈央子, 藤田隆寛, 中島一記, 松田裕子, 畠山雄次, 沢禎彦, 石川博之

福岡歯科大学学会雑誌 42 ( 増補 ) 38 - 38 2016年12月

化学合成により作製されたLRAPの骨芽細胞分化に与える影響

松田裕子, 畠山雄次, 沢禎彦, 石川博之

福岡歯科大学学会雑誌 42 ( 増補 ) 35 - 35 2016年12月

Novel Monoclonal Antibody LpMab-17 Developed by CasMab Technology Distinguishes Human Podoplanin from Monkey Podoplanin. 査読国際誌

Yukinari Kato, Satoshi Ogasawara, Hiroharu Oki, Ryusuke Honma, Michiaki Takagi, Yuki Fujii, Takuro Nakamura, Noriko Saidoh, Hazuki Kanno, Mitsuo Umetsu, Satoshi Kamata, Hiroshi Kubo, Mitsuhiro Yamada, Yoshihiko Sawa, Kei-Ichi Morita, Hiroyuki Harada, Hiroyoshi Suzuki, Mika Kato Kaneko

Monoclonal antibodies in immunodiagnosis and immunotherapy 35 ( 2 ) 109 - 16 2016年4月

記述言語：英語掲載種別：研究論文（学術雑誌）

Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein, which possesses a platelet aggregation-stimulating (PLAG) domain in its N-terminus. Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for the binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. Although many anti-PDPN monoclonal antibodies (mAbs) have been established, almost all mAbs bind to PLAG domains. We recently established CasMab technology to produce mAbs against membranous proteins. Using CasMab technology, we produced a novel anti-PDPN mAb, LpMab-17, which binds to non-PLAG domains. LpMab-17 clearly detected endogenous PDPN of cancer cells and normal cells in Western-blot, flow cytometry, and immunohistochemistry. LpMab-17 recognized glycan-deficient PDPN in flow cytometry, indicating that the interaction between LpMab-17 and PDPN is independent of its glycosylation. The minimum epitope of LpMab-17 was identified as Gly77-Asp82 of PDPN using enzyme-linked immunosorbent assay. Of interest, LpMab-17 did not bind to monkey PDPN, whereas the homology is 94% between human PDPN and monkey PDPN, indicating that the epitope of LpMab-17 is unique compared with the other anti-PDPN mAbs. The combination of different epitope-possessing mAbs could be advantageous for the PDPN-targeting diagnosis or therapy.

DOI： 10.1089/mab.2015.0077

Differentiation of Apical Bud Cells in a Newly Developed Apical Bud Transplantation Model Using GFP Transgenic Mice as Donor. 査読国際誌

Naoki Maruo, Ryuji Sakagami, Yasunori Yoshinaga, Kazuhiko Okamura, Yoshihiko Sawa

PLoS one 11 ( 3 ) e0150766 2016年

記述言語：英語掲載種別：研究論文（学術雑誌）

Rodent mandibular incisors have a unique anatomical structure that allows teeth to grow throughout the lifetime of the rodent. This report presents a novel transplantation technique for studying the apical bud differentiation of rodent mandibular incisors. Incisal apical end tissue with green fluorescent protein from transgenic mouse was transplanted to wild type mice, and the development of the transplanted cells were immunohistologically observed for 12 weeks after the transplantation. Results indicate that the green fluorescent apical end tissue replaced the original tissue, and cells from the apical bud differentiated and extended toward the incisal edge direction. The immunostaining with podoplanin also showed that the characteristics of the green fluorescent tissue were identical to those of the original. The green fluorescent cells were only found in the labial side of the incisor up to 4 weeks. After 12 weeks, however, they were also found in the lingual side. Here the green fluorescent cementocyte-like cells were only present in the cementum close to the dentin surface. This study suggests that some of the cells that form the cellular cementum come from the apical tissue including the apical bud in rodent incisors.

DOI： 10.1371/journal.pone.0150766

Intracellular Signaling Pathway Activation via TGF-β Differs in the Anterior and Posterior Axis During Palatal Development

Higa Arisa, Oka Kyoko, Kira-Tatsuoka Michiko, Tamura Shougo, Itaya Satoshi, Toda Masako, Ozaki Masao, Sawa Yoshihiko

Journal of Hard Tissue Biology 25 ( 2 ) 195 - 204 2016年

記述言語：英語出版者・発行元：硬組織再生生物学会

It is important to understand the different mechanisms involved in anterior hard and posterior soft palate development to prevent and treat patients with cleft palate. Genetic analyses of humans and gene-mutated mice with cleft palate have shown that TGF-β signaling has a critical role in palatogenesis. However, the intracellular signaling pathway of TGF-β during palatogenesis in the anterior-posterior axis has not yet been fully understood. In the present study, the expression patterns of intracellular molecules Smad2/3 and phospho-p38 (Pp38) were examined at embryonic days 13.5, 14.0, and 14.5 (E13.5, E14.0, and E14.5) in mice. It was found that Smad3 was activated in anterior palatal mesenchyme and in the medial edge epithelium (MEE) region, with TGF-β3 expressed at E13.5. On the other hand, Pp38 was more expressed in posterior palatal mesenchyme and strongly expressed in the entire palatal epithelium at E13.5. These opposing expression patterns between Smad3 and Pp38 in palatal mesenchyme were also observed at E14.0. Interestingly, Pp38 expression was inhibited in MEE from E14.0. Generally, from E14.5, the tissue specificities of hard and soft palate started showing their characteristics following the activation of cell differentiation in palatal mesenchyme, and the medial edge seam (MES) of the palatal epithelium started to disappear for fusion to occur. At this stage, Smad3 was also more expressed in anterior palatal mesenchyme, while expression of Pp38 was activated in posterior palatal mesenchyme. Pp38 expression was inhibited, but Smad3 was activated in the MES. These results suggest that TGF-β signaling plays various roles, such as in cell proliferation and differentiation of palatal mesenchyme and in the disappearance of the MES, through different intracellular signaling pathways in anterior-posterior palatal mesenchyme and epithelium.

DOI： 10.2485/jhtb.25.195

その他リンク： https://jlc.jst.go.jp/DN/JLC/20022363242?from=CiNii

Immunohistochemical expression of fibrillin-1 and fibrillin-2 during tooth development 査読

M. Kira-Tatsuoka, K. Oka, E. Tsuruga, M. Ozaki, Y. Sawa

Journal of Periodontal Research 50 ( 6 ) 714 - 720 2015年12月

Intermittent Administration of Parathyroid Hormone Ameliorates Periapical Lesions in Mice 査読国際誌

Masato Otawa, Ryuichiro Tanoue, Hirofumi Kido, Yoshihiko Sawa, Junro Yamashita

JOURNAL OF ENDODONTICS 41 ( 5 ) 646 - 651 2015年5月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ELSEVIER SCIENCE INC

Introduction: Intermittent administration of parathy raid hormone (PTH) promotes oral osseous wound healing and protects against ligature-induced alveolar bone loss. However, its therapeutic value on periapical periodontitis is unknown. The goal of this study was to determine the effect of intermittent PTH administration on the progression of periapical periodontitis. Methods: Seven lymphotoxin alpha deficient mice received pulp exposures of mandibular first and second molars. Exposed pulp in the right mandible was covered with plaque-contaminated fibrin, whereas exposed pulp in the left mandible was left open. After 4 weeks, the perk apical tissues were examined to determine the effect of plaque-contaminated fibrin to induce periapical lesions. Fourteen mice received pulp exposure covered with plaque-contaminated fibrin. PTH (40 Ag/kg/d) was administered intermittently to half of the mice for 3 weeks beginning 1 week after pulp exposure. The remaining half received saline injections as the vehicle control. At sacrifice, mandibles and tibiae were harvested and processed for histologic examination. Evaluation of neutrophils and blood vessels was performed after staining with immunofluorescence, and periradicular bone was histomorphometrically analyzed. Results: The exposed pulp covered with plaque-contaminated fibrin resulted in significantly larger periapical lesions compared with the control. Intermittent PTH administration reduced the size of periapical lesions significantly. Significantly less neutrophil infiltration around the root apex was found in PTH-treated animals compared with the control. Conclusions: PTH treatment suppressed periapical inflammation by reducing neutrophil infiltration and protected against tissue destruction by periapical periodontitis.

DOI： 10.1016/j.joen.2014.12.008

歯周病原菌由来LPSによるTLRを介した糖尿病性腎症の促進について

高田俊輔, 梶原弘一郎, 石川博之, 沢禎彦

福岡歯科大学学会雑誌 40 ( 増補 ) 31 - 31 2014年12月

ヒト無色素毛様体上皮細胞およびヒト歯根膜線維芽細胞におけるオキシタラン線維の分解過程

川越慈, 敦賀英知, 石川博之, 沢禎彦

福岡歯科大学学会雑誌 40 ( 増補 ) 36 - 36 2014年12月

マウス下顎切歯歯胚の幹細胞におけるSox2およびOct4の発現査読

丸尾直樹, 岡村和彦, 沢禎彦, 坂上竜資

Journal of Oral Biosciences Supplement 2014 160 - 160 2014年9月

Immunohistochemical Study of Amelogenin and Lysosome-Associate Membrane Proteins (LAMPs) in Cartilage 査読

Yuji Hatakeyama, Junko Hatakeyama, Kyoko Oka, Eichi Tsuruga, Tetsuichiro Inai, Hisashi Anan, Yoshihiko Sawa

INTERNATIONAL JOURNAL OF MORPHOLOGY 32 ( 2 ) 618 - 626 2014年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：SOC CHILENA ANATOMIA

Amelogenin is one of the enamel matrix proteins secreted by ameloblasts during enamel formation in tooth development. Recent studies showed that the amelogenin is expressed in chondrocyte. Lysosome-associated membrane proteins (LAMPs) have been identified as binding partner proteins to amelogenin and it has been suggested they act as signaling receptors of amelogenin. The purpose of this study is to clarify the localization of amelogenin and LAMPs in growth plate cartilage and cartilaginous nodules in micromass culture. Mouse knee joints including tibia growth plate at 4 weeks old and micromass cultures of limb bud mesenchymal cells after 2 weeks were fixed in paraformaldehyde, routinely processed, sections were cut and immunostained with amelogenin, collagen type II and type X, LAMP-1 and -3. The positive immunoreaction of amelogenin was observed both in proliferation and hypertrophic zone cartilage of growth plate after enzymatic pretreatment in immunostaining. Furthermore, cartilaginous nodules in micromass culture were immunopositive to amelogenin. The chondrocytes in the proliferation zone of the growth plate were immunopositive to LAMP-1 but weakly stained in the chondrocytes of hypertrophic zone. These observations indicate that amelogenin may be present in cartilage matrix produced in vivo and in vitro and amelogenin may involve cartilage formation through the LAMP-1 signaling pathway.

Expression of Toll-Like Receptor 2 in Glomerular Endothelial Cells and Promotion of Diabetic Nephropathy by Porphyromonas gingivalis Lipopolysaccharide 査読国際誌

Yoshihiko Sawa, Shunsuke Takata, Yuji Hatakeyama, Hiroyuki Ishikawa, Eichi Tsuruga

PLOS ONE 9 ( 5 ) e97165 2014年5月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：PUBLIC LIBRARY SCIENCE

The toll-like receptor (TLR) has been suggested as a candidate cause for diabetic nephropathy. Recently, we have reported the TLR4 expression in diabetic mouse glomerular endothelium. The study here investigates the effects of the periodontal pathogen Porphyromonas gingivalis lipopolysaccharide (LPS) which is a ligand for TLR2 and TLR4 in diabetic nephropathy. In laser-scanning microscopy of glomeruli of streptozotocin-and a high fat diet feed-induced type I and type II diabetic mice, TLR2 localized on the glomerular endothelium and proximal tubule epithelium. The TLR2 mRNA was detected in diabetic mouse glomeruli by in situ hybridization and in real-time PCR of the renal cortex, the TLR2 mRNA amounts were larger in diabetic mice than in non-diabetic mice. All diabetic mice subjected to repeated LPS administrations died within the survival period of all of the diabetic mice not administered LPS and of all of the non-diabetic LPS-administered mice. The LPS administration promoted the production of urinary protein, the accumulation of type I collagen in the glomeruli, and the increases in IL-6, TNF-alpha, and TGF-beta in the renal cortex of the glomeruli of the diabetic mice. It is thought that blood TLR ligands like Porphyromonas gingivalis LPS induce the glomerular endothelium to produce cytokines which aid glomerulosclerosis. Periodontitis may promote diabetic nephropathy.

DOI： 10.1371/journal.pone.0097165

Microfibril-associated glycoprotein-1 controls human ciliary zonule development in vitro. 査読

Takahiro Fujita, Eichi Tsuruga, Kaori Yamanouchi, Yoshihiko Sawa, Hiroyuki Ishikawa

Acta histochemica et cytochemica 47 ( 1 ) 11 - 7 2014年2月

記述言語：英語掲載種別：研究論文（学術雑誌）

The ciliary zonule in the eye, also known as Zinn's zonule, is composed of oxytalan fibers, which are bundles of microfibrils consisting mainly of fibrillin-1. However, it is still unclear which of the microfibril-associated molecules present in the ciliary zonule controls oxytalan fibers. Microfibril-associated glycoprotein-1 (MAGP-1) is the only microfibril-associated molecule identified in the human ciliary zonule. In the present study, we used siRNA against MAGP-1 in cultures of human non-pigmented ciliary epithelial cells to examine the extracellular deposition and appearance of fibrillin-1 employing Western blotting and immunofluorescence. MAGP-1 suppression led to a reduction of fibrillin-1 deposition. Immunofluorescence also confirmed that RNAi-mediated down-regulation of MAGP-1 led to suppression of fiber development. These results suggest that MAGP-1 plays a crucial role in the extracellular deposition of fibrillin-1 during formation of the human ciliary zonule.

DOI： 10.1267/ahc.13038

Distribution of podoplanin-expressing cells in the mouse nervous systems. 査読

Miwa Tomooka, Chiaki Kaji, Hiroshi Kojima, Yoshihiko Sawa

Acta histochemica et cytochemica 46 ( 6 ) 171 - 7 2013年12月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Podoplanin is a mucin-type glycoprotein which was first identified in podocytes. Recently, podoplanin has been successively reported as a marker for brain and peripheral nerve tumors, however, the distribution of podoplanin-expressing cells in normal nerves has not been fully investigated. This study aims to examine the podoplanin-expressing cell distribution in the mouse head and nervous systems. An immunohistochemical study showed that the podoplanin-positive areas in the mouse peripheral nerve and spinal cord are perineurial fibroblasts, satellite cells in the dorsal root ganglion, glia cells in the ventral and dorsal horns, and schwann cells in the ventral and dorsal roots; in the cranial meninges the dura mater, arachnoid, and pia mater; in the eye the optic nerve, retinal pigment epithelium, chorioidea, sclera, iris, lens epithelium, corneal epithelium, and conjunctival epithelium. In the mouse brain choroid plexus and ependyma were podoplanin-positive, and there were podoplanin-expressing brain parenchymal cells in the nuclei and cortex. The podoplanin-expressing cells were astrocyte marker GFAP-positive and there were no differences in the double positive cell distribution of several portions in the brain parenchyma except for the fornix. The results suggest that podoplanin may play a common role in nervous system support cells and eye constituents.

DOI： 10.1267/ahc.13035

Leucine Rich Amelogenin Peptideによる軟骨形成の誘導

畠山純子, 畠山雄次, 松本典祥, 春名千英子, 諸冨孝彦, 泉利雄, 沢禎彦, 笹野泰之, 阿南壽

日本歯科保存学雑誌 56 ( 6 ) 560 - 569 2013年12月

記述言語：日本語出版者・発行元：(NPO)日本歯科保存学会

目的:アメロジェニンスプライシングバリアントの一つであるleucine rich amelogenin peptide(LRAP)は,破骨細胞と骨芽細胞の成熟,歯根膜細胞の遊走および増殖を促進するシグナルペプチドとして示唆されている.近年,軟骨にアメロジェニンの発現が報告されている.しかし,LRAPが軟骨形成に関与するかについての詳細は不明である.本研究の目的は,軟骨形成におけるLRAPの機能を明らかにすることである.材料と方法:C57BL6/CrSlcマウスの胎生10日齢の肢芽を摘出後,トリプシン/コラゲナーゼ処理により細胞を分離し,マイクロマスカルチャーを行った.ブタリコンビナントLRAP(マウスLRAPと〜85%の相同性,Dr.Gibson(ペンシルバニア大学,USA)より供与)と陽性コントロールとしてマウスリコンビナントGrowth Differentiation Factor-5(GDF-5,R & D Systems,USA)を,1,10,100ng/mlずつ培地に添加した.培養開始2,4,6日後にアルシアンブルー染色を行った.24,48時間後にBrdU assayを行い,細胞の増殖を確認した.12,24,48時間後にRNAを抽出し,軟骨のマーカーであるSox9,II型コラーゲン(Col2a1)およびAggrecanの半定量的RT-PCRを行った.有意差検定にはANOVAとBonferroni's post hoc testを用いた.結果:添加24・48時間後で,LRAP添加0.1,1,10ng/mlのいずれにおいても,濃度依存的にBrdU陽性細胞がコントロール群と比較して有意に増加していた.コントロール群では添加2日後にアルシアンブルー染色陽性結節の出現がみられ,培養期間が長くなると陽性結節の染色性が増加したが,LRAPおよびGDF-5を添加した群では,濃度依存的・培養期間依存的にアルシアンブルーの染色性が強くなった.LRAP添加群では,24・48時間後にSox9の遺伝子発現が,また48時間後にCol2a1およびAggrecanの遺伝子発現上昇がコントロール群と比較して有意に上昇した.結論:これらの知見は,アメロジェニンスプライシングバリアントの一つであるLRAPが,軟骨形成を促進していることを示唆している.(著者抄録)

その他リンク： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2013&ichushi_jid=J01092&link_issn=&doc_id=20140116190010&doc_link_id=%2Fdo1conde%2F2013%2F005606%2F010%2F0560-0569%26dl%3D0&url=https%3A%2F%2Fwww.medicalonline.jp%2Fjamas.php%3FGoodsID%3D%2Fdo1conde%2F2013%2F005606%2F010%2F0560-0569%26dl%3D0&type=MedicalOnline&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00004_2.gif

歯の発生におけるLecin rich Amelogenin peptideの発現解析

畠山純子, 畠山雄次, 松本典祥, 春名千英子, 諸富孝彦, 泉利雄, 沢禎彦, 阿南壽

福岡歯科大学学会雑誌 39 ( 増補 ) 36 - 36 2013年12月

I型とII型糖尿病モデルマウスにおける腎リンパ管新生について

内山貴誠, 高田俊輔, 石川博之, 沢禎彦

福岡歯科大学学会雑誌 39 ( 増補 ) 28 - 28 2013年12月

糖尿病モデルマウス腎糸球体におけるTLR2とTLR4の発現

高田俊輔, 内山貴誠, 石川博之, 沢禎彦

福岡歯科大学学会雑誌 39 ( 増補 ) 36 - 36 2013年12月

Quantitative evaluation of myofibroblast apoptosis during wound healing in rat palate after post-operative administration of basic fibroblast growth factor (bFGF). 国際誌

Yuichiro Hata, Hiroyuki Ishikawa, Takeshi Ueki, Takashi S Kajii, Sachio Tamaoki, Eichi Tsuruga, Yoshihiko Sawa, Kunihisa Taniguchi

Acta odontologica Scandinavica 71 ( 6 ) 1501 - 7 2013年11月

記述言語：英語掲載種別：研究論文（学術雑誌）

OBJECTIVE: Excessive wound contraction apparently inhibits maxillary growth; thus, myofibroblast apoptosis needs to be accelerated in mucoperiosteal denudation after palatoplasty. The aim of this study was to evaluate myofibroblast apoptosis during wound healing in mucoperiosteal denudation of rat palates immediately after post-operative administration of basic fibroblast growth factor (bFGF). MATERIALS AND METHODS: A total of 100 male Wistar rats aged 20 days were divided into control, scar, sham and bFGF groups (n = 25 each). In the scar, sham and bFGF groups, mucoperiosteum was removed from the palate and fibrin glue was applied to the exposed bone surface immediately after surgery. In the bFGF group, 10 μL of 2 μg/μL bFGF solution was injected into the operated area beneath the fibrin glue. At 2, 5, 7, 14 and 28 days post-operatively, myofibroblast apoptosis during the wound healing process was investigated by double immunofluorescence staining. The apoptotic area of myofibroblasts was measured using image software. RESULTS: In the bFGF group, at 2 days, apoptosis of myofibroblasts in the lamina propria and submucosa was marked, as compared with the other three groups and apoptosis of myofibroblasts was scarcely seen at 5 days. At 5 and 7 days, the apoptotic area of myofibroblasts in the bFGF group was statistically significantly smaller when compared to the scar and sham groups. CONCLUSION: The results confirmed that bFGF injection immediately after surgery accelerated apoptosis of myofibroblasts in mucoperiosteal denudation of rats. This may reduce maxillary growth retardation due to excessive wound contraction.

DOI： 10.3109/00016357.2013.773370

Altered dynamics in the renal lymphatic circulation of type 1 and type 2 diabetic mice.

Takanobu Uchiyama, Shunsuke Takata, Hiroyuki Ishikawa, Yoshihiko Sawa

Acta histochemica et cytochemica 46 ( 2 ) 97 - 104 2013年4月

記述言語：英語掲載種別：研究論文（学術雑誌）

The dynamics of the renal lymphatic circulation in diabetic nephropathy is not fully elucidated. The present study evaluated the effect of diabetic nephropathy on the renal lymphatic circulation in streptozotocin (STZ)-induced type 1 diabetic mice (ICR-STZ) and in type 2 diabetic KK/Ta mice which were fed a high fat diet (KK/Ta-HF). The diabetic mouse kidneys developed edema because of the nephropathy. In control mice renal lymphatic vessels distributed in the cortex but rarely in the medulla while in ICR-STZ and KK/Ta-HF mice, there were many lymphatic vessels with small lumen in both cortex and medulla. Total numbers and areas of renal blood vessels in the diabetic mice were similar to those in the controls while the total numbers and areas of renal lymphatic vessels were larger in diabetic mice than in the controls. There were statistically significant differences in the numbers of lymphatic vessels with diameters of 50-100 µm between the ICR-STZ and the control ICR mice, and in the numbers of lymphatic capillaries with diameters smaller than 50 µm between the KK/Ta-HF and the control KK/Ta mice. The diabetic nephropathy may induce the lymphangiogenesis or result in at least the renal lymphatic vessel expansion.

DOI： 10.1267/ahc.13006

Immunohistochemical Study of Lysosome-Associated Membrane Proteins During Periodontal Ligament Development 査読

Yuji Hatakeyama, Junko Hatakeyama, Kyoko Oka, Eichi Tsuruga, Tetsuichiro Inai, Yoshihiko Sawa

JOURNAL OF HARD TISSUE BIOLOGY 22 ( 2 ) 233 - 239 2013年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：JOURNAL HARD TISSUE BIOLOGY

Amelogenin is an enamel matrix protein also expressed in periodontal tissues. Recent studies suggest that lysosome-associated membrane proteins (LAMPs) can bind cell surface amelogenin and act as signaling receptors for amelogenin in periodontal ligament (PDL) cells and cementoblasts in vitro. However, whether amelogenin and LAMPs are involved in the development of periodontal tissues is unclear. To address this issue, we used immunohistochemistry to profile LAMPs and amelogenin expression during periodontal tissue development. Paraffin sections from mice at embryonic day 15 (E15) and 1, 2, and 4 weeks were made, and serial sections at each age were stained with hematoxylin-eosin, followed by immunostaining with anti-amelogenin, LAMP-1, and LAMP-3. At El 5, dental follicle cells, which are the source of periodontal tissues, were amelogenin-positive and weakly positive for LAMP-1 and LAMP-3. At 1 week, dental follicle cells were immunopositive for amelogenin and LAMP-1 but weakly positive for LAMP-3. At 2 weeks, PDL cells were positive for amelogenin and LAMP-1 but negative for LAMP-3. At 4 weeks, PDL cells were immunopositive for both amelogenin and LAMP-1. These results suggest that LAMP-1 is expressed in both the dental follicle and PDL, and that LAMP-1 and LAMP-3 have temporally distinct roles as signaling receptors for amelogenin.

Case of autoimmune hepatitis with markedly enlarged hepatoduodenal ligament lymph nodes. 国際誌

Hideki Fujii, Naoki Ohnishi, Kazuho Shimura, Masafumi Sakamoto, Tohru Ohkawara, Yoshihiko Sawa, Koichi Nishida, Yasuo Ohkawara, Tatsuro Kobata, Kanji Yamaguchi, Yoshito Itoh

World journal of gastroenterology 19 ( 11 ) 1834 - 40 2013年3月

記述言語：英語掲載種別：研究論文（学術雑誌）

Autoimmune hepatitis (AIH) is a necroinflammatory liver disease of unknown etiology. The disease is characterized histologically by interface hepatitis, biochemically by increased aspartate aminotransferase and alanine aminotransferase levels, and serologically by increased autoantibodies and immunoglobulin G levels. Here we discuss AIH in a previously healthy 37-year-old male with highly elevated serum levels of soluble interleukin-2 receptor and markedly enlarged hepatoduodenal ligament lymph nodes (HLLNs, diameter, 50 mm). Based on these observations, the differential diagnoses were AIH, lymphoma, or Castleman's disease. Liver biopsy revealed the features of interface hepatitis without bridging fibrosis along with plasma cell infiltration which is the typical characteristics of acute AIH. Lymph node biopsy revealed lymphoid follicles with inflammatory lymphocytic infiltration; immunohistochemical examination excluded the presence of lymphoma cells. Thereafter, he was administered corticosteroid therapy: after 2 mo, the enlarged liver reached an almost normal size and the enlarged HLLNs reduced in size. We could not find AIH cases with such enlarged lymph nodes (diameter, 50 mm) in our literature review. Hence, we speculate that markedly enlarged lymph nodes observed in our patient may be caused by a highly activated, humoral immune response in AIH.

DOI： 10.3748/wjg.v19.i11.1834

Expression of Toll-Like Receptor 4 in Glomerular Endothelial Cells under Diabetic Conditions.

Shunsuke Takata, Yoshihiko Sawa, Takanobu Uchiyama, Hiroyuki Ishikawa

Acta histochemica et cytochemica 46 ( 1 ) 35 - 42 2013年2月

記述言語：英語掲載種別：研究論文（学術雑誌）

Diabetic conditions promote glomerulosclerosis by mesangial cells but the mechanisms are not fully elucidated. The present study evaluated the expression of toll-like receptor 4 in glomerular endothelial cells in the streptozotocin (STZ)-induced type 1 diabetic mouse (ICR-STZ) and the type 2 diabetic KK/TaJcl mouse which were fed a high fat diet feed (KK/Ta-HF). In the ICR-STZ and KK/Ta-HF almost glomeruli were immunostained with anti-TLR4 but there was no glomerulus immunostained by ani-TLR4 in the control ICR and KK/Ta. Laser-scanning confocal microscopy showed that the TLR4-positive region did not coincide with the podoplanin-positive region but coincide with the PECAM-1- and VE-cadherin-positive regions in the glomeruli of the ICR-STZ and KK/Ta-HF. The in situ hybridization showed that almost signals for TLR4 mRNA were present in the glomerulus of the ICR-STZ and KK/Ta-HF to a stronger extent than in the control ICR and KK/Ta. These suggest that glomerular endothelial cells usually express the TLR4 gene and hyperglycemia in the diabetic condition induces the TLR4 protein expression in the glomerular capillary endothelial cells. Cytokine productions through the TLR signaling pathway in glomerular endothelial cells may allow mesangial cells to produce extracellular matrix proteins in the diabetic milieu.

DOI： 10.1267/ahc.13002

Matrix Metalloproteinase-2 Degrades Fibrillin-1 and Fibrillin-2 of Oxytalan Fibers in the Human Eye and Periodontal Ligaments In Vitro 査読

Megumi Kawagoe, Eichi Tsuruga, Kyoko Oka, Yoshihiko Sawa, Hiroyuki Ishikawa

ACTA HISTOCHEMICA ET CYTOCHEMICA 46 ( 5 ) 153 - 159 2013年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：JAPAN SOC HISTOCHEMISTRY & CYTOCHEMISTRY

Oxytalan fibers are distributed in the eye and periodontal ligaments (PDL). The ciliary zonule, known as Zinn's zonule, in the eye is composed of oxytalan fibers, which are bundles of microfibrils consisting mainly of fibrillin-1 and fibrillin-2. As turnover of oxytalan fibers is slow during life, their degradation mechanism remains unclarified. This study was performed to examine degradation pattern of fibrillin-1 and fibrillin-2 by experimental MMP activation. We cultured human non-pigmented ciliary epithelial cells (HNPCEC) and PDL fibroblasts for 7 days, then treated them with concanavalin A to activate matrix metalloproteinase (MMP)-2, and examined the degradation of fibrillin-1 and fibrillin-2 for 72 hr using immunofluorescence. At 7 days of HNPCEC culture, fibrillin-1-positive fibers were observed, some of which merged with fibrillin-2. After MMP-2 activation, fibrillin-1-positive fibers became thin and disappeared by 72 hr, while fibrillin-2-positive fibers disappeared almost completely within 24 hr. At 7 days of PDL fibroblast culture, fibrillin-1-positive fibers were mostly merged with fibrillin-2. After MMP-2 activation, fibrillin-1-positive fibers became thin by 24 hr and had almost disappeared by 48 hr, while fibrillin-2-positive fibers decreased constantly after 24 hr. A MMP-2 inhibitor completely suppressed these degradations. These results suggest that the patterns of fibrillin-1 and fibrillin-2 degradation differ between the eye and the PDL, possibly reflecting the sensitivity of fibrillin-1 and fibrillin-2 of each type of oxytalan fiber against MMP-2.

DOI： 10.1267/ahc.13024

Cellular turnover in epithelial rests of Malassez in the periodontal ligament of the mouse molar 査読国際誌

Kyoko Oka, Masakazu Morokuma, Kyoko Imanaka-Yoshida, Yoshihiko Sawa, Keitaro Isokawa, Masaki J Honda

European Journal of Oral Sciences 120 ( 6 ) 484 - 494 2012年12月

記述言語：英語掲載種別：研究論文（学術雑誌）

Fragments of Hertwig's epithelial root sheath persist in the periodontal ligament (PDL) in small clusters known as epithelial rests of Malassez (ERM). It is generally agreed that ERM are maintained as a quiescent and exclusively dental epithelial cluster in PDL. However, we speculate that homeostasis and cellular turnover underlies cluster maintenance. We also hypothesize that the fate of ERM clusters - diminishing or remaining - might be regulated via the presence or absence of epithelial stem cells therein. Histological analysis of aging mouse molar PDL showed that ERM clusters gradually increase in size with increasing age. Immunocytochemistry and cell culture revealed that ERM clusters contained Ki67-positive cells and were able to expand when brought in culture. The TdT-mediated biotin-dUTP nick-end labeling (TUNEL) procedure also detected signs of apoptosis. Finally, we identified putative epithelial stem cells in the clusters by 5-bromo-2′-deoxyuridine (BrdU) pulse-chase experiments and immunohistochemistry, using the stem-cell marker leucine-rich repeat-containing G-protein-coupled receptor 5 (Lgr5). The results suggest that ERM clusters are maintained in the PDL, via cellular turnover, throughout life. © 2012 Eur J Oral Sci.

DOI： 10.1111/eos.12003

Latent transforming growth factor-β binding protein 2 negatively regulates coalescence of oxytalan fibers induced by stretching stress 査読国際誌

Eichi Tsuruga, Kyoko Oka, Yuji Hatakeyama, Keitaro Isokawa, Yoshihiko Sawa

Connective Tissue Research 53 ( 6 ) 521 - 527 2012年12月

記述言語：英語掲載種別：研究論文（学術雑誌）

Oxytalan fibers are extracellular matrix components consisting of pure microfibrils. However, the mechanism whereby oxytalan fibers develop is not fully understood. We have previously reported that in human periodontal ligament (PDL) fibroblasts subjected to stretching stress, bundles of oxytalan fibers coalesce under the control of fibulin-5. Latent transforming growth factor-β binding protein 2 (LTBP-2) is known to bind to fibulin-5. The purpose of this study was to clarify the role of LTBP-2 in the coalescence of oxytalan fibers. We subjected PDL fibroblasts to stretching in order to examine the effects of LTBP-2 on the coalescence of oxytalan fibers in cell/matrix layers. Interaction of LTBP-2 with fibulin-5 was examined by immunoprecipitation assay, and changes in LTBP-2 deposition upon stretching were investigated by Western blotting and immunofluorescence assays. We used small interfering RNA against LTBP-2 in PDL cell culture and examined the appearance of oxytalan fibers on the basis of immunofluorescence. Stretching induced coalescence of oxytalan fibers, but did not affect LTBP-2 expression. The amount of extracellularly deposited LTBP-2 was decreased by about 70% as a result of stretching, compared with the control. LTBP-2 interacted with fibulin-5 on the fibers, and stretching decreased the amount of the LTBP-2 interacted with fibulin-5 by about 60%. Oxytalan fiber coalescence did not occur when LTBP-2 was suppressed by about 95%, whereas it occurred when LTBP-2 was suppressed by about 40%, fibulin-5 being colocalized with oxytalan fibers. These results suggest that LTBP-2, in response to tension stress, may negatively control the function of fibulin-5, thereby modulating the mechanism of oxytalan fiber coalescence. Copyright © Informa Healthcare USA, Inc.

DOI： 10.3109/03008207.2012.702816

Chimeric anti-podoplanin antibody suppresses tumor metastasis through neutralization and antibody-dependent cellular cytotoxicity 査読国際誌

Mika Kato Kaneko, Akiko Kunita, Shinji Abe, Yuta Tsujimoto, Masashi Fukayama, Kaoru Goto, Yoshihiko Sawa, Yasuhiko Nishioka, Yukinari Kato

Cancer Science 103 ( 11 ) 1913 - 1919 2012年11月

記述言語：英語掲載種別：研究論文（学術雑誌）

Podoplanin is a platelet aggregation-inducing factor associated with tumor metastasis, malignant progression, and cancer stem cells. We produced a rat-human chimeric anti-podoplanin mAb, NZ-8, from rat anti-podoplanin mAb (NZ-1). Although both NZ-1 and NZ-8 possess high binding affinities and high neutralizing activities of platelet aggregation, the antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity of NZ-8 were much higher than NZ-1. Furthermore, both NZ-1 and NZ-8 inhibited the growth of podoplanin-expressing tumors in vivo. Both NZ-1 and NZ-8 also suppressed hematogenous metastasis of podoplanin-expressing tumors. These results suggest that anti-podoplanin mAbs suppressed hematogenous metastasis by both neutralization and antibody-dependent cellular cytotoxicity/complement-dependent cytotoxicity activities. Targeting therapy to podoplanin-expressing tumors should be useful as a novel immunotherapy. © 2012 Japanese Cancer Association.

DOI： 10.1111/j.1349-7006.2012.02385.x

歯周組織においてFibulin-4はLOXL2/エラスチン複合体形成を調節する

山内由宣, 中冨佑香, 中島一記, 敦賀英知, 沢禎彦, 石川博之

福岡歯科大学学会雑誌 38 ( 増補 ) 37 - 37 2012年11月

小児癌の抗癌剤治療が引き起こす永久歯の形成障害について

西村紗和, 沢禎彦, 石川博之

福岡歯科大学学会雑誌 38 ( 増補 ) 29 - 29 2012年11月

Intracellular interaction of EMILIN-1 with fibrillin-1 in human periodontal ligament cells 査読

Yuka Nakatomi, Eichi Tsuruga, Yoshinori Yamauchi, Megumi Kawagoe, Kaori Yamanouchi, Kazuki Nakashima, Yoshihiko Sawa, Hiroyuki Ishikawa

Orthodontic Waves 71 ( 2 ) 66 - 69 2012年6月

記述言語：英語掲載種別：研究論文（学術雑誌）

The elastic system fibers comprise oxytalan, elaunin and elastic fibers, differing in their relative microfibril and elastin contents. Human periodontal ligaments (PDLs) contain oxytalan fibers (pure microfibrils), which are composed mainly of fibrillin-1, the major component of microfibrils. We recently demonstrated that EMILIN-1, located at the interface between elastin and microfibrils, controls the amount of fibrillin-1 assembly in PDL fibroblast cell/matrix layers [8], although the mechanism involved was unclear. We subjected cultured PDL fibroblasts to immunofluorescence and immunoprecipitation assays in order to examine the intracellular interaction of EMILIN-1 with fibrillin-1. Immunofluorescence showed that EMILIN-1 was colocalized with fibrillin-1, both inside and outside the cells. Additionally, EMILIN-1 formed a complex with fibrillin-1 in the intracellular fraction. These results suggest that EMILIN-1 may form complexes with fibrillin-1 in cellular vesicles, thus contributing effectively to the initial assembly of pericellular fibrillin-1 during the process of oxytalan fiber formation. © 2012 Elsevier Ltd and the Japanese Orthodontic Society.

DOI： 10.1016/j.odw.2012.01.002

The expression of podoplanin and classic cadherins in the mouse brain 査読国際誌

Chiaki Kaji, Miwa Tomooka, Yukinari Kato, Hiroshi Kojima, Yoshihiko Sawa

Journal of Anatomy 220 ( 5 ) 435 - 446 2012年5月

記述言語：英語掲載種別：研究論文（学術雑誌）

Podoplanin is a transmembrane glycoprotein indirectly linked to classic cadherins through ezrin-actin networks. Recently, the overexpression of podoplanin in high-grade malignancy brain tumors has been reported. The aim of this study was to investigate the expression of podoplanin and classic cadherins in the mouse brain. Immunohistochemistry showed that podoplanin was expressed on ependymal cells and choroid plexus epithelial cells at the ventricle side of the cell surface and at the cell-cell junctions, and on retinal pigment epithelial cells and in the pia mater
P-cadherin between choroid plexus epithelial cells and endothelial cells at the basement membrane side of cell surface, and between retinal pigment epithelial cells
VE-cadherin on the PECAM-1 positive-choroid plexus endothelial cells of the fibrovascular core
and N-cadherin on the cell surface and at the cell-cell junctions of ependymal cells, and in the pia mater. The regions expressing podoplanin, P-cadherin, and VE-cadherin did not coincide. In real-time PCR analysis, the amounts of podoplanin and P- and N-cadherin mRNA were larger in the ventricular wall with choroid plexus than in the abdominal aorta and cerebrum. In the RT-PCR analysis, the intensities of amplicon for VE-cadherin mRNA were the same for the abdominal aorta, cerebrum, and ventricular wall with the choroid plexus, suggesting that mouse ependymal cells, choroid plexus epithelial cells, and glial cells under the pia mater have the ability to express podoplanin and P- and N-cadherins. Glial cells and retinal pigment epithelial cells may create barriers by podoplanin and classic cadherins as a rate-determining step for transmission of blood components. © 2012 The Authors. Journal of Anatomy © 2012 Anatomical Society.

DOI： 10.1111/j.1469-7580.2012.01484.x

Factors associated with the overall survival of elderly patients with hepatocellular carcinoma. 国際誌

Hideki Fujii, Yoshito Itoh, Naoki Ohnishi, Masafumi Sakamoto, Tohru Ohkawara, Yoshihiko Sawa, Koichi Nishida, Yasuo Ohkawara, Kanji Yamaguchi, Masahito Minami, Takeshi Okanoue

World journal of gastroenterology 18 ( 16 ) 1926 - 32 2012年4月

記述言語：英語掲載種別：研究論文（学術雑誌）

AIM: To identify the factors associated with overall survival of elderly patients with hepatocellular carcinoma (HCC). METHODS: A total of 286 patients with HCC (male/female: 178/108, age: 46-100 years), who were diagnosed and treated by appropriate therapeutic procedures between January 2000 and December 2010, were enrolled in this study. Patients were stratified into two groups on the basis of age: Elderly (≥ 75 years old) and non-elderly (< 75 years old). Baseline clinical characteristics as well as cumulative survival rates were then compared between the two groups. Univariate and multivariate analyses were used to identify the factors associated with prolonged overall survival of patients in each group. Cumulative survival rates in the two groups were calculated separately for each modified Japan Integrated Stage score (mJIS score) category by the Kaplan-Meier method. In addition, we compared the cumulative survival rates of elderly and non-elderly patients with good hepatic reserve capacity (≤ 2 points as per mJIS). RESULTS: In the elderly group, the proportion of female patients, patients with absence of hepatitis B or hepatitis C viral infection, and patients with coexisting extrahepatic comorbid illness was higher (56.8% vs 31.1%, P < 0.001; 27.0% vs 16.0%, P = 0.038; 33.8% vs 22.2%, P = 0.047; respectively) than that in the non-elderly group. In the non-elderly group, the proportion of hepatitis B virus (HBV)-infected patients was higher than that in the elderly group (9.4% vs 0%, P = 0.006). The cumulative survival rates in the elderly group were 53.7% at 3 years and 32.9% at 5 years, which were equivalent to those in the non-elderly group (55.9% and 39.4%, respectively), as shown by a log-rank test (P = 0.601). In multivariate analysis, prolonged survival was significantly associated with the extent of liver damage and stage (P < 0.001 and P < 0.001, respectively), but was not associated with patient age. However, on individual evaluation of factors in both groups, stage was significantly (P < 0.001) associated with prolonged survival. Regarding mJIS scores of ≤ 2, the rate of female patients with this score was higher in the elderly group when compared to that in the non-elderly group (P = 0.012) and patients ≥ 80 years of age tended to demonstrate shortened survival. CONCLUSION: Survival of elderly HCC patients was associated with liver damage and stage, but not age, except for patients ≥ 80 years with mJIS score ≤ 2.

DOI： 10.3748/wjg.v18.i16.1926

Roles of Collagen and Periostin Expression by Cranial Neural Crest Cells during Soft Palate Development 査読国際誌

Kyoko Oka, Masaki J. Honda, Eichi Tsuruga, Yuji Hatakeyama, Keitaro Isokawa, Yoshihiko Sawa

Journal of Histochemistry and Cytochemistry 60 ( 1 ) 57 - 68 2012年1月

記述言語：英語掲載種別：研究論文（学術雑誌）

The tissue in the palatal region can be divided into the hard and the soft palates, each having a specialized function such as occlusion, speech, or swallowing. Therefore, an understanding of the mechanism of palatogenesis in relation to the function of each region is important. However, in comparison with the hard palate, there is still a lack of information about the mechanisms of soft palate development. In this study, the authors investigated the contribution of cranial neural crest (CNC) cells to development of both hard and soft palates. They also demonstrated a unique pattern of periostin expression during soft palate development, which was closely related to that of collagen type I (Col I) in palatine aponeurosis. Furthermore, organ culture analysis showed that exogenous transforming growth factor-β (TGF-β) induced the expression of both periostin and Col I. These novel patterns of expression in the extracellular matrix (ECM) induced by CNC cells suggest that these cells may help to determine the character of both the hard and soft palates through ECM induction. TGF-β signaling appears to be one of the mediators of Col I and periostin expression in the formation of functional structures during soft palate development. © The Histochemical Society 2012.

DOI： 10.1369/0022155411427059

Immunohistochemical examination of novel rat monoclonal antibodies against mouse and human podoplanin 査読

Chiaki Kaji, Yuta Tsujimoto, Mika Kato Kaneko, Yukinari Kato, Yoshihiko Sawa

Acta Histochemica et Cytochemica 45 ( 4 ) 227 - 237 2012年

記述言語：英語掲載種別：研究論文（学術雑誌）

This study aims to develop new monoclonal antibodies (mAbs) against mouse and human podoplanin. Rats were immunized with synthetic peptides, corresponding to amino acids 38-51 of mouse podoplanin or human podoplanin which is 100% homologous to the same site of monkey podoplanin
anti-mouse podoplanin mAb PMab-1 (IgG2a) and anti-human mAb NZ-1.2 (IgG2a) were established. In immunocytochemistry, the mouse melanoma B16-F10 and mouse podoplanin (mPDPN)-expressed CHO transfectant were stained by PMab-1
human lymphatic endothelial cells (LEC) and human podoplanin (hPDPN)-expressed squamous cell carcinoma HSC3 transfectant, were stained by NZ-1.2. Western-blot analysis detected an about 40-kDa protein in CHO-mPDPN and B16-F10 by PMab-1, and in HSC3-hPDPN and LEC by NZ-1.2. In frozen sections, PMab-1 reacted with mouse kidney, pulmonary alveoli, pulmonary pleura, and salivary gland myoepithelial cells while NZ-1.2 reacted to the human salivary gland myoepithelial cells. The immunostaining of paraffin-embedded sections also showed the reaction of PMab-1 or NZ-1.2 to the mouse or monkey kidney glomerulus, pulmonary alveoli, and lung lymphatic vessels. These results indicate that the two novel rat mAbs to the mouse and human/monkey podoplanin are useful for Western-blot and immunostaining of somatic tissues on paraffin-embedded sections as well as frozen sections. © 2012 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.12008

Fibrillin-1 and Fibrillin-2 Are Essential for Formation of Thick Oxytalan Fibers in Human Nonpigmented Ciliary Epithelial Cells In Vitro 査読国際誌

Kaori Yamanouchi, Eichi Tsuruga, Kyoko Oka, Yoshihiko Sawa, Hiroyuki Ishikawa

CONNECTIVE TISSUE RESEARCH 53 ( 1 ) 14 - 20 2012年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：INFORMA HEALTHCARE

The ciliary zonule, also known as Zinn's zonule, is composed of oxytalan fibers. However, the mechanism by which epithelial cells in the ciliary body form these fibers in not fully understood. We examined human nonpigmented ciliary epithelial cells to determine the appearance and amount of oxytalan fibers in terms of positivity for their major components, fibrillin-1 and fibrillin-2. Examination of fibrillin-1 and fibrillin-2 expression by immunofluorescence revealed that thin fibers positive for fibrillin-1 on Day 2 changed to thick fibers by Day 8. The fibers positive for fibrillin-2 appeared on the thick fibrillin-1-positive fibers after Day 4. Northern blot analysis revealed that the level of fibrillin-1 did not change markedly, while induction of fibrillin-2 gene was evident on Day 5. Western blot analysis showed that fibrillin-1 deposition increased gradually, while that of fibrillin-2 increased markedly from Day 5 to Day 8. Fibrillin-1 suppression did not lead to the formation of fibrillin-2-positive thick fibers, whereas fibrillin-2 suppression led to the formation of fibrillin-1-positive thin fibers, but not thick fibers. These results suggest that both fibrillin-1 and fibrillin-2 are essential for the formation of thick oxytalan fibers in the ciliary zonule and are informative for clarifying the mechanism of homeostasis of the ocular matrix.

DOI： 10.3109/03008207.2011.602767

Immunohistochemical and immunocytochemical localization of amylase in rat parotid glands and von Ebner's glands by ion etching-immunoscanning electron microscopy 査読

Junko Yahiro, Tetsuichiro Inai, Akihito Tsutsui, Atsuko Sato, Toshikazu Nagato, Kunihisa Taniguchi, Eichi Tsuruga, Yoshihiko Sawa

Acta Histochemica et Cytochemica 44 ( 5 ) 201 - 212 2011年8月

記述言語：英語掲載種別：研究論文（学術雑誌）

The distribution of amylase in rat parotid glands and von Ebner's glands was examined using ion etching-immunoscanning electron microscopy, which enables both light and electron microscopic observations of identical semi-thin resin sections immunolabeled with anti-α- amylase and immunogold in association with silver enhancement. At the light microscopic level, most acinar secretory granules (SG) and striated duct secretions of parotid glands were strongly stained dark brown. In von Ebner's glands, acinar SG and duct secretions were weakly to strongly stained light to dark brown. At the electron microscopic level, labeling was observed as bright gold-silver particles. The labeling intensity of acinar SG of parotid glands was higher than that of von Ebner's glands. In parotid glands, weak labeling of SG in transitional cells between acini and intercalated ducts, very weak labeling of SG in intercalated ducts, and strong labeling of striated duct secretions were observed. In von Ebner's glands, the secretions and some SG of interlobular ducts were strongly labeled compared to those of intralobular ducts and SG of acini. Less amylase was synthesized in von Ebner's acini compared to parotid acini, whereas von Ebner's ducts may secrete significantly more amylase to modify saliva than parotid ducts. © 2011 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.10039

Establishment of a novel monoclonal antibody SMab-1 specific for IDH1-R132S mutation 査読国際誌

Mika Kato Kaneko, Wei Tian, Shingo Takano, Hiroyuki Suzuki, Yoshihiko Sawa, Yasukazu Hozumi, Kaoru Goto, Kentaro Yamazaki, Chifumi Kitanaka, Yukinari Kato

Biochemical and Biophysical Research Communications 406 ( 4 ) 608 - 613 2011年3月

記述言語：英語掲載種別：研究論文（学術雑誌）

Isocitrate dehydrogenase 1 (IDH1) mutations, which are early and frequent genetic alterations in gliomas, are specific to a single codon in the conserved and functionally important Arginine 132 (R132) in IDH1. We earlier established a monoclonal antibody (mAb), IMab-1, which is specific for R132H-containing IDH1 (IDH1-R132H), the most frequent IDH1 mutation in gliomas. To establish IDH1-R132S-specific mAb, we immunized mice with R132S-containing IDH1 (IDH1-R132S) peptide. After cell fusion using Sendai virus envelope, IDH1-R132S-specific mAbs were screened in ELISA. One mAb, SMab-1, reacted with the IDH1-R132S peptide, but not with other IDH1 mutants. Western-blot analysis showed that SMab-1 reacted only with the IDH1-R132S protein, not with IDH1-WT protein or IDH1 mutants, indicating that SMab-1 is IDH1-R132S-specific. Furthermore, SMab-1 specifically stained the IDH1-R132S-expressing glioblastoma cells in immunocytochemistry and immunohistochemistry, but did not react with IDH1-WT or IDH1-R132H-containing glioblastoma cells. We newly established an anti-IDH1-R132S-specific mAb SMab-1 for use in diagnosis of mutation-bearing gliomas. © 2011 Elsevier Inc.

DOI： 10.1016/j.bbrc.2011.02.102

Stretch stimuli increase fibulin-5/EMILIN-1 complex on oxytalan fibers in human periodontal ligament cells 査読

Kazuki Nakashima, Eichi Tsuruga, Yuka Nakatomi, Yoshinori Yamauchi, Yuichiro Hata, Sachio Tamaoki, Yoshihiko Sawa, Hiroyuki Ishikawa

Orthodontic Waves 70 ( 1 ) 15 - 20 2011年3月

記述言語：英語掲載種別：研究論文（学術雑誌）

Periodontal ligaments (PDLs) are continuously exposed to various functional forces, such as tooth movement and occlusal loading. Human PDLs comprise elastic system fibers as well as collagen fibers. The elastic system fibers in turn comprise oxytalan, elaunin and elastic fibers, which differ in their relative microfibril and elastin contents. Human PDLs contain oxytalan fibers (pure microfibrils), which are composed mainly of fibrillin-1 (the major component of microfibrils). We have previously reported that bundles of oxytalan fibers in cultured PDL cells coalesce in response to mechanical strain, and that this coalescence is controlled by fibulin-5. However, the relationship between fibulin-5 and other fibrillin-1-binding molecules is unclear. In the present study we investigated whether fibulin-5 and EMILIN-1 (both of which are fibrillin-1-binding molecules) contribute to the formation of oxytalan fibers upon exposure to mechanical strain. We subjected PDL cells to stretching in order to examine the role of fibulin-5 and EMILIN-1 in the formation of oxytalan fibers in cell/matrix layers. We examined the relationship between fibulin-5 and EMILIN-1 in PDL cell cultures using immunofluorescence and immunoprecipitation assay. Immunofluorescence showed that fibulin-5 and EMILIN-1 were colocalized on fibrillin-1-positive oxytalan fibers. Fibulin-5 formed a complex with EMILIN-1, and stretching increased the amount of this complex relative to cells that were not subjected to stretching. These results suggest that the expression of the fibulin-5/EMILIN-1 complex is upregulated in response to tension strain, and may control the formation of oxytalan fibers in PDLs. © 2010 Japanese Orthodontic Society.

DOI： 10.1016/j.odw.2010.09.001

EMILIN-1 regulates the amount of oxytalan fiber formation in periodontal ligaments in vitro 査読国際誌

Yuka Nakatomi, Eichi Tsuruga, Kazuki Nakashima, Yoshihiko Sawa, Hiroyuki Ishikawa

Connective Tissue Research 52 ( 1 ) 30 - 35 2011年2月

記述言語：英語掲載種別：研究論文（学術雑誌）

The elastic system fibers comprise oxytalan, elaunin, and elastic fibers, differing in their relative microfibril and elastin contents. Among them, human periodontal ligament (PDL) contains only oxytalan fibers (pure microfibrils). Elastin microfibril interface-located protein-1 (EMILIN-1) is localized at the interface between microfibrils and elastin. We hypothesized that EMILIN-1 may contribute to the formation of oxytalan fibers. We used a small interfering RNA (siRNA) for EMILIN-1 in PDL cell culture to examine the extracellular deposition of fibrillin-1 (the major component of microfibrils). EMILIN-1 was labeled on microfibrils positive for fibrillin-1 and was colocalized with fibrillin-1 upon immunoprecipitation assay. EMILIN-1 suppression reduced the level of fibrillin-1 deposition to 23 of the control, and this was responsible for the diminution of fibrillin-1 deposition revealed by immunofluorescence. These results suggest that EMILIN-1 may regulate the formation of oxytalan fibers and play a role in their homeostasis. © 2011 Informa Healthcare USA, Inc.

DOI： 10.3109/03008207.2010.502982

Expression of podoplanin and classical cadherins in salivary gland epithelial cells of klotho-deficient mice 査読

Ikuko Amano, Yuri Imaizumi, Chiaki Kaji, Hiroshi Kojima, Yoshihiko Sawa

Acta Histochemica et Cytochemica 44 ( 6 ) 267 - 276 2011年

記述言語：英語掲載種別：研究論文（学術雑誌）

We have recently shown that salivary gland myoepithelial cells express podoplanin. Podoplanin indirectly binds the actin filament network which links classical cadherins. The study here is aimed to investigate the expression of podoplanin and cadherins on salivary gland myoepithelial cells and the changes in the aging cells using klotho-deficient (kl/kl) mice. The submandibular glands of kl/kl mouse lack granular ducts which express klotho in wild type mice, suggesting that klotho may be a gene responsible for granular duct development. Although aging resulted in growth suppression of myoepithelial cells because of the sparse distribution of the cells in kl/kl mouse salivary glands, the expression of podoplanin and Ecadherin was shown in aging myoepithelial cells. It is thought that podoplanin participates in the actin-E-cadherin networks which are maintained in aging myoepithelial cells. It was also shown that granular ducts were filled with P-cadherin, and that the P-cadherin amount was larger in the wild type mouse submandibular glands than in the sublingual and parotid glands of wild type mouse, and in the submandibular glands of kl/kl mouse. These findings suggest that the granular duct is an organ secreting soluble P-cadherin into the saliva. © 2011 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.11037

The effect of valproic Acid on mesenchymal pluripotent cell proliferation and differentiation in extracellular matrices. 国際誌

Yuji Hatakeyama, Junko Hatakeyama, Atsushi Takahashi, Kyoko Oka, Eichi Tsuruga, Tetsuichiro Inai, Yoshihiko Sawa

Drug target insights 5 1 - 9 2011年

記述言語：英語掲載種別：研究論文（学術雑誌）

Valproic acid (2-n-propylpentanoic acid, VPA) is a widely used antiepileptic and anticonvulsant drug. Previous studies have reported that VPA effects osteogenesis in vivo and in vitro, yet it remains unclear whether VPA promotes cell differentiation of osteoblasts derived from mesenchymal cells. The purpose of this study was to clarify the effect of VPA on undifferentiated pluripotent mesenchymal cell proliferation and differentiation into osteoblasts while analyzing the impact of the absence or presence of extracellular matrices (ECMs). Mouse mesenchymal cells were cultured on non-coated plastic, type I collagen-coated, and fibronectin-coated plates in the absence or presence of VPA. A cell proliferation assay was performed in which modified formazan dye content was analyzed and proliferation nuclear antigen (PCNA)-positive cells were counted at various concentrations of VPA. A high concentration of VPA did not clearly alter cell morphology, but large numbers of stress fibers were observed in these cells and the cell proliferation ratio was decreased with positive PCNA counts. In the presence of matrices, the cell proliferation ratio decreased at low VPA concentrations compared with the ratio obtained in the absence of these ECMs. On the other hand, VPA promoted osteoblastic differentiation in the presence of type I collagen. These findings indicate that for undifferentiated mesenchymal cells, VPA promotes a decrease in the cell proliferation rate in the presence of ECMs and promotes osteoblastic differentiation, both of which could provide insight into additional mechanisms of osteoblastic cell differentiation caused by VPA.

DOI： 10.4137/DTI.S6534

歯根膜オキシタラン線維形成過程におけるEMILIN-1の機能

中冨佑香, 中島一記, 久永豊, 敦賀英知, 沢禎彦, 石川博之

日本矯正歯科学会大会プログラム・抄録集 69回 184 - 184 2010年9月

Immunoelectron microscopic study of podoplanin localization in mouse salivary gland myoepithelium 査読

Minoru Hata, Ikuko Amano, Eichi Tsuruga, Hiroshi Kojima, Yoshihiko Sawa

Acta Histochemica et Cytochemica 43 ( 2 ) 77 - 82 2010年

記述言語：英語掲載種別：研究論文（学術雑誌）

We have recently reported that salivary gland cells express the lymphatic endothelial cell marker podoplanin. The present study was aimed to immunohistochemically investigate the expression of the myoepithelial cell marker α-smooth muscle actin (SMA) on podoplaninpositive cells in mouse parotid and sublingual glands, and to elucidate podoplanin localization in salivary gland myoepithelial cells by immunoelectron microscopic study. The distribution of myoepithelial cells expressing podoplanin and α-SMA was examined by immunofluorescent staining, and the localization of reaction products of anti-podoplanin antibody was investigated by pre-embedded immunoelectron microscopic method. In immunohistochemistry, the surfaces of both the mucous acini terminal portion and ducts were covered by a number of extensive myoepithelial cellular processes expressing podoplanin, and the immunostaining level with anti-podoplanin antibody to myoepithelial cells completely coincided with the immunostaining level with anti-α-SMA antibody. These findings suggest that podoplanin is a salivary gland myoepithelial cell antigen, and that the detection level directly reflects the myoepithelial cell distribution. In immunoelectron microscopic study, a number of reaction products with anti-podoplanin antibody were found at the Golgi apparatus binding to the endoplasmic reticulum in the cytoplasm of myoepithelial cells between sublingual gland acinar cells, and were also found at the myoepithelial cell membrane. These findings suggest that salivary gland myoepithelial cells constantly produce podoplanin and glycosylate at the Golgi apparatus, and transport them to the cell membrane. Podoplanin may be involved in maintaining the homeostasis of myoepithelial cells through its characteristic as a mucin-type transmembrane glycoprotein. © 2010 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.10011

Fibulin-4 and -5, but not Fibulin-2, are Associated with Tropoelastin Deposition in Elastin-Producing Cell Culture 査読

Yoshinori Yamauchi, Eichi Tsuruga, Kazuki Nakashima, Yoshihiko Sawa, Hiroyuki Ishikawa

Acta Histochemica et Cytochemica 43 ( 6 ) 131 - 138 2010年

記述言語：英語掲載種別：研究論文（学術雑誌）

201E0l aTshteic J sapyasnte Smo cfiibeteyr so fc oHnisstioscth oefm misitcrryo afinbdri lCs ya-nd tropoelastin. During development, microfibrils act as a template on which tropoelastin is deposited. Fibrillin-1 is the major component of microfibrils. It is not clear whether elastic fiber-associated molecules, such as fibulins, contribute to tropoelastin deposition. Among the fibulin family, fibulin-2, -4 and -5 are capable of binding to tropoelastin and fibrillin-1. In the present study, we used the RNA interference (RNAi) technique to establish individual gene-specific knockdown of fibulin-2, -4 and -5 in elastin-producing cells (human gingival fibroblasts
HGF). We then examined the extracellular deposition of tropoelastin using immunofluorescence. RNAi-mediated downregulation of fibulin-4 and -5 was responsible for the diminution of tropoelastin deposition. Suppression of fibulin-5 appeared to inhibit the formation of fibrillin-1 microfibrils, while that of fibulin-4 did not. Similar results to those for HGF were obtained with human dermal fibroblasts. These results suggest that fibulin-4 and -5 may be associated in different ways with the extracellular deposition of tropoelastin during elastic fiber formation in elastinproducing cells in culture. © 2010 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.10026

Immunohistochemical examination on the distribution of cells expressed lymphatic endothelial marker podoplanin and LYVE-1 in the mouse tongue tissue 査読

Yuya Noda, Ikuko Amano, Minoru Hata, Hiroshi Kojima, Yoshihiko Sawa

Acta Histochemica et Cytochemica 43 ( 2 ) 61 - 68 2010年

記述言語：英語掲載種別：研究論文（学術雑誌）

The clinical study for lingual disease requires the detailed investigation of the lingual lymphatic network and lymphatic marker-positive cells. Recently, it has been reported that several tissue cells and leukocytes express lymphatic markers, LYVE-1 and podoplanin. This study was aimed to clarify the lingual distribution of cells expressing LYVE-1 and podoplanin. In the mouse tongue, podoplanin is expressed in nerve sheaths, lingual gland myoepithelial cells, and lymphatic vessels. LYVE-1 is expressed in the macrophage marker Mac-1-positive cells as well as lymphatic vessels, while factor-VIII was detected in only blood endothelial cells. α-SMA was detected in vascular smooth muscle and myoepithelial cells. Therefore, identification of lymphatic vessels in lingual glands, the combination of LYVE-1 and factor-VIII, or LYVE-1 and Mac-1 is useful because myoepithelial cells express podoplanin and α-SMA. The immunostaining of factor-VIII on lymphatic vessels was masked by the immunostaining to LYVE-1 or podoplanin because lymphatic vessels express factor-VIII to a far lesser extent than blood vessels. Therefore, except for the salivary glands, the combination of podoplanin and α-SMA, or factor-VIII is useful to identify lymphatic vessels and blood vessels with smooth muscle, or blood capillaries. © 2010 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.10008

Immunohistochemical examination for the distribution of podoplanin-expressing cells in developing mouse molar tooth germs 査読

Yuri Imaizumi, Ikuko Amano, Eichi Tsuruga, Hiroshi Kojima, Yoshihiko Sawa

Acta Histochemica et Cytochemica 43 ( 5 ) 115 - 121 2010年

記述言語：英語掲載種別：研究論文（学術雑誌）

We recently reported the expression of podoplanin in the apical bud of adult mouse incisal tooth. This study was aimed to investigate the distribution of podoplanin-expressing cells in mouse tooth germs at several developing stages. At the bud stage podoplanin was expressed in oral mucous epithelia and in a tooth bud. At the cap stage podoplanin was expressed on inner and outer enamel epithelia but not in mesenchymal cells expressing the neural crest stem cell marker nestin. At the early bell stage nestin and podoplanin were expressed in cervical loop and odontoblasts. At the root formation stage both nestin and podoplanin were weakly expressed in odontoblasts generating radicular dentin. Podoplanin expression was also found in the Hertwig epithelial sheath. These results suggest that epithelial cells of developing tooth germ acquire the ability to express nestin, and that tooth germ epithelial cells maintain the ability to express podoplanin in oral mucous epithelia. The expression of podoplanin in odontoblasts was induced as tooth germ development advanced, but was suppressed with the completion of the primary dentin, suggesting that podoplanin may be involved in the cell growth of odontoblasts. Nestin may function as an intermediate filament that binds podoplanin in odontoblasts. © 2010 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.10023

Fibulin-5 contributes to microfibril assembly in human periodontal ligament cells 査読

Yutaka Hisanaga, Kazuki Nakashima, Eichi Tsuruga, Yuka Nakatomi, Yuji Hatakeyama, Hiroyuki Ishikawa, Yoshihiko Sawa

Acta Histochemica et Cytochemica 42 ( 5 ) 151 - 157 2009年10月

記述言語：英語掲載種別：研究論文（学術雑誌）

The elastic system fibers comprise oxytalan, elaunin and elastic fibers, which differ in their relative microfibril and elastin content. Human periodontal ligaments (PDL) contain only oxytalan fibers (pure microfibrils) among them. Since fibulin-5 regulates the organization of elastic fibers to link the fibers to cells, we hypothesized that fibulin-5 may contribute to the formation of oxytalan fibers. We used siRNA for fibulin-5 in PDL cell culture to examine the extracellular deposition of fibrillin-1 and -2, which are the major components of microfibrils. Fibulin-5 was labeled on microfibrils positive for fibrillin-1 and -2. Fibulin-5 suppression reduced the level of fibrillin-1 and -2 deposition to 60% of the control level. These results suggest that fibulin-5 may control the formation of oxytalan fibers, and play a role in the homeostasis of oxytalan fibers. © 2009 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.09021

Stretching stimulates fibulin-5 expression and controls microfibril bundles in human periodontal ligament cells 査読

K. Nakashima, E. Tsuruga, Y. Hisanaga, H. Ishikawa, Y. Sawa

Journal of Periodontal Research 44 ( 5 ) 622 - 627 2009年10月

記述言語：英語掲載種別：研究論文（学術雑誌）

Background and Objective: The elastic fiber system comprises oxytalan, elaunin and elastic fibers, differing in their relative microfibril and elastin contents. Human periodontal ligaments contain oxytalan fibers (pure microfibrils). Periodontal ligaments are continuously exposed to various functional forces, such as tooth movement and occlusal loading. We have reported that bundles of microfibrils coalesce in response to mechanical strain in cultured periodontal ligament fibroblasts, as assessed in terms of their positivity for fibrillin-1 (the major component of microfibrils). However, the mechanism of microfibril coalescence is unclear. We hypothesized that the fibrillin-1-binding molecule, fibulin-5, contributes to oxytalan fiber formation under mechanical stra Material and Methods: We subjected periodontal ligament fibroblasts to stretching in order to examine the effects of fibulin-5 on the formation of oxytalan fibers in cell/matrix layers. We transfected periodontal ligament cells with small interference RNA for fibulin-5, then examined oxytalan fibers using immunofluorescence and electron microscopy. Results: Immunofluorescence showed that fibrillin-1-positive microfibrils coalesced as a result of stretching, compared with cells that were not subjected to stretching. Fibulin-5 colocalized on fibrillin-1-positive microfibrils. Stretching increased fibulin-5 gene expression and protein deposition. Immunofluorescence and immunogold electron microscopy analysis revealed that fibulin-5 suppression inhibited the coalescence of microfibrils under stretching conditions. Conclusion: These results suggest that fibulin-5 up-regulated in response to tension strain may control the formation of microfibril bundles in periodontal ligament. © 2008 Blackwell Munksgaard.

DOI： 10.1111/j.1600-0765.2008.01170.x

Integrin αvβ3 regulates microfibril assembly in human periodontal ligament cells 査読

E. Tsuruga, A. Sato, T. Ueki, K. Nakashima, Y. Nakatomi, H. Ishikawa, T. Yajima, Y. Sawa

Tissue and Cell 41 ( 2 ) 85 - 89 2009年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier Ltd

Fibrillin-1 is the major structural component of extracellular microfibrils. However, the mechanism by which extracellular fibrillin-1 assembles into microfibrils is not fully understood. Fibrillin-1 contains the Arg-Gly-Asp (RGD) motif, which may allow binding to RGD-recognizing integrins. We hypothesized that integrin αvβ3 on the cell surface of human periodontal ligament (PDL) fibroblasts may influence fibrillin-1 assembly into cell/matrix layers. We treated PDL fibroblasts with an integrin αvβ3-specific antagonist to examine fibrillin-1 assembly. Western blotting and immunofluorescence analysis showed that treatment with the integrin αvβ3 antagonist at 5 μM clearly abolished fibrillin-1 deposition. These results provide for the first time evidence that integrin αvβ3 regulates extracellular assembly of fibrillin-1, thereby modulating cell-mediated homeostasis of microfibrils. © 2008 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.tice.2008.07.005

Expression of podoplanin in the mouse tooth germ and apical bud cells 査読

Yoshihiko Sawa, Kana Iwasawa, Hiroyuki Ishikawa

Acta Histochemica et Cytochemica 41 ( 5 ) 121 - 126 2008年10月

記述言語：英語掲載種別：研究論文（学術雑誌）

This study was designed to investigate the distribution of cells expressing podoplanin in the mouse tooth bud. Podoplanin expression was detected in enamel epithelia of the cervical loop at cell-cell contacts strongly, and weakly on the loosely aggregated stellate reticulum in the center and the neighboring stratum intermedium. Odontoblasts exhibited intense podoplanin expression at the junction with predentin while no expression was detected in the enamel organ containing ameloblasts. These results suggest that proliferating inner and outer enamel epithelia express podoplanin but that the expression is suppressed in the differentiated epithelia containing ameloblasts. On the other hand the podoplanin expression occurs in the differentiating odontoblasts and the expression is sustained in differentiated odontoblasts, indicating that odontoblasts have the strong ability to express podoplanin. In cultured apical bud cells podoplanin was detected at cell-cell contacts. In real-time PCR analysis the amount of podoplanin mRNA of the apical buds was 2-fold compared with the amount of kidney used as a positive control. These findings indicate that apical bud cells have the strong ability to express the podoplanin gene. Podoplanin is a mucin-type glycoprotein negatively charged by extensive O-glycosylation and a high content of sialic acid, which expresses the adhesive property. The podoplanin may contribute to form odontoblastic fiber or function as the anchorage to the tooth development and in proliferating epithelial cells of cervical loop and apical bud. © 2008 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.08019

Induction of ICAM-1 and VCAM-1 on the mouse lingual lymphatic endothelium with TNF-α 査読

Kana Iwasawa, Takeshi Kameyama, Hiroyuki Ishikawa, Yoshihiko Sawa

Acta Histochemica et Cytochemica 41 ( 5 ) 115 - 120 2008年10月

記述言語：英語掲載種別：研究論文（学術雑誌）

This study investigated the TNF-α-induced ICAM-1 and VCAM-1 expression on mouse lingual lymphatic vessels. All podoplanin-positive lymphatic vessels expressed PECAM-1. In the lamina propria mucosae of TNF-α-treated tongue, almost all initial lymphatics expressed ICAM-1. There were initial lymphatics with the VCAM-1 expression and also the vessels without the expression. In the tunica muscularis of TNF-α-treated tongue, collecting lymphatic vessels expressed ICAM-1, but rarely expressed VCAM-1 whereas blood vessels simultaneously expressed ICAM-1 and VCAM-1. The ICAM-1-positive rate increased with TNF-α to 75% from 10% on initial lymphatics, and to 40% from 0% on collecting lymphatic vessels while it increased to 90% from 45% on blood vessels. The VCAM-1-positive rate increased with TNF-α to 30% from 0% on initial lymphatics, and to 5% from 0% on collecting lymphatic vessels while it increased to 75% from 5% on blood vessels. These findings suggest that the lingual lymphatic endothelium has the ability to express ICAM-1, and VCAM-1 to a lesser extent than the ICAM-1 induction with TNF-α, and that the ICAM-1 and VCAM-1 induction predominantly occurs in the initial lymphatics compared with collecting lymphatic vessels. © 2008 The Japan Society of Histochemistry and Cytochemistry.

DOI： 10.1267/ahc.08017

Expression of podoplanin in the mouse salivary glands 査読国際誌

Minoru Hata, Takeshi Ueki, Atsuko Sato, Hiroshi Kojima, Yoshihiko Sawa

Archives of Oral Biology 53 ( 9 ) 835 - 841 2008年9月

記述言語：英語掲載種別：研究論文（学術雑誌）

Objective: Podoplanin is one of the most highly expressed lymphatic-specific genes. Here, we report the distribution of cells expressing podoplanin in mouse salivary glands. Design: We immunohistochemically investigated the distribution of cells expressing podoplanin in mouse major salivary glands by laser-scanning microscopy. The expression of endothelial cell marker PECAM-1 was tested to discriminate lymphatic endothelium from salivary gland cells, and myoepithelial cells were identified by an antibody for P-cadherin. Results: The podoplanin expression was rarely found in acini of the parotid gland but clearly found at the basal portion of acini in the submandibular and sublingual glands. The number of portion reacted with anti-podoplanin is greater in the sublingual gland than in the submandibular gland. The expression was also found at the basal portion of ducts in all major salivary glands. The P-cadherin expression was rarely found in acini of the parotid gland but found in acini of the sublingual gland and on ducts in parotid and sublingual glands, corresponding to the area of podoplanin expression. Conclusions: It was suggested that the acinar and myoepithelial cells in the salivary glands have the ability to express podoplanin, and that the expression may be concerned with the mucous saliva excretion. © 2008 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.archoralbio.2008.02.006

Leukocyte adhesion molecule and chemokine production through lipoteichoic acid recognition by toll-like receptor 2 in cultured human lymphatic endothelium 査読国際誌

Yoshihiko Sawa, Eichi Tsuruga, Kana Iwasawa, Hiroyuki Ishikawa, Shigemitsu Yoshida

Cell and Tissue Research 333 ( 2 ) 237 - 252 2008年8月

記述言語：英語掲載種別：研究論文（学術雑誌）

We have recently reported that the human lymphatic endothelium has toll-like receptor 4 (TLR4)-mediated lipopolysaccharide recognition mechanisms that induce the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Although ligand engagement with TLR2 enables activation of the MyD88-dependent pathway similarly to TLR4, whether TLR2 ligands such as lipoteichoic acid (LTA) trigger the activation of lymphatic endothelium remains unclear. This study has been designed to investigate the expression dynamics of LTA-induced leukocyte adhesion molecules and chemokines in cultured human lymphatic endothelium (LEC). Reverse transcription/polymerase chain reaction (RT-PCR) and real-time quantitative PCR analyses have shown that LEC usually expresses TLR2 and increases TLR2 gene expression on LTA treatment. Indeed, LTA-treated LEC increases the expression of E-selectin, ICAM-1, and VCAM-1 but does not alter the gene expression of ICAM-2, ICAM-3, junctional adhesion molecule-1 (JAM-1), JAM-3, or platelet endothelial cell adhesion molecule-1 (PECAM-1). The expression of LTA-induced E-selectin, ICAM-1, and VCAM-1 in LEC is suppressed by anti-TLR2 but not by anti-TLR4 and is also suppressed by TLR2-specific short interfering RNA (siRNA) but not by siRNA for TLR4. The expression of CCL2, CCL5, and CCL20 (Cys-Cys motif chemokines) and of CXCL1, CXCL3, CXCL5, CXCL6, and CXCL8 (Cys-X-Cys motif chemokines) was induced in LEC with LTA. These data suggest that the human lymphatic endothelial phenotype has TLR2-mediated LTA-recognition mechanisms, resulting in increased expression of inflammatory leukocyte adhesion molecules and phagocyte-attractive chemokines. The human lymphatic endothelium may thus function to collect leukocytes from tissues into lymphatic vessels by means of immunologically functional molecules. © 2008 Springer-Verlag.

DOI： 10.1007/s00441-008-0625-5

The expression of E-selectin and chemokines in the cultured human lymphatic endothelium with lipopolysaccharides 査読国際誌

Yoshihiko Sawa, Eichi Tsuruga

Journal of Anatomy 212 ( 5 ) 654 - 663 2008年5月

記述言語：英語掲載種別：研究論文（学術雑誌）

This study investigated the expression of selectins and chemokines in cultured human lymphatic endothelial cells stimulated with lipopolysaccharides. In microarray, vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 gene expressions in the lymphatic endothelium with lipopolysaccharides did not change at 0.5 h but increased two- to three-fold at 12 h, whereas E-selectin increased 10-fold at 0.5 h and 68-fold at 12 h compared with untreated cells. The E-selectin mRNA and protein increased in the lymphatic endothelial cells with lipopolysaccharides at more than two-fold levels compared with human umbilical vein endothelial cells. Induction of Cys-Cys chemokine ligand 2, 3, 5, 7, 8 and 20 mRNAs in the lymphatic endothelial cells with lipopolysaccharides was detected in microarray and real-time PCR. The Cys-Cys chemokine ligand 2, 5 and 20 mRNA amounts in cells with high concentration lipopolysaccharides were larger in the lymphatic endothelial cells than in human umbilical vein endothelial cells. The Cys-Cys chemokine ligand 3 and 8 mRNAs were not detected in human umbilical vein endothelial cells. Induction of Cys-X-Cys chemokine ligand 1, 3, 5, 6 and 8 mRNAs was detected in the lymphatic endothelial cells with lipopolysaccharides. The Cys-X-Cys chemokine ligand 3, 5 and 8 mRNA amounts in cells with high concentration lipopolysaccharides were larger in the lymphatic endothelial cells than in human umbilical vein endothelial cells. In conclusion, it was demonstrated that the cultured human lymphatic endothelial cells express E-selectin and phagocyte-attractive chemokine genes. © 2008 The Authors Journal compilation © 2008 Anatomical Society of Great Britain and Ireland.

DOI： 10.1111/j.1469-7580.2008.00892.x

Expression of junctional adhesion molecules on the human lymphatic endothelium 査読国際誌

Takeshi Ueki, Kana Iwasawa, Hiroyuki Ishikawa, Yoshihiko Sawa

MICROVASCULAR RESEARCH 75 ( 2 ) 269 - 278 2008年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE

Human lymphatic vessels express several leukocyte adhesion molecules. The study here investigated the expression of three junctional adhesion molecules (JAM) which are a newly reported glycoprotein family of adhesion molecules on human lymphatic endothelium. In this study, JAM-1 and JAM-3 but not JAM-2 were detected in cultured human neonatal dermal lymphatic endothelial cells (LEC) at the gene and protein levels by microarray, RT-PCR, real-time PCR, and immunohistochemical analysis. The JAM-I and JAM-3 expression was not altered in the TNF-alpha-treated LEC or in the untreated cells. In human tissue, the expression of JAM-1, and the expression of JAM-1, JAM-2, and JAM-3 were observed in collecting lymphatic vessels of uninflamed small intestine, and in initial lymphatics of inflamed tongue and uninflamed gingival tissue. It is thought that JAM-2 mRNA could be produced in mature vascular endothelium but not in cultured cells, and that human intestinal and oral lymphatic vessels usually express JAM-1, JAM-2, and JAM-3. There were initial lymphatics simultaneously expressing JAM-1, JAM-2, and JAM-3 in the mucosal connective tissue papillae of gingival tissue. The three JAM expressions on the lymphatic endothelium may contribute to both seal the cell-cell contact at interendothelial junctions and also allow lymphocytes to transmigrate into lymphatic vessels from tissue, independent of inflammatory cytokines. (c) 2007 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.mvr.2007.07.005

LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 expression in human lymphatic endothelium 査読国際誌

Yoshihiko Sawa, Takeshi Ueki, Minoru Hata, Kana Iwasawa, Eichi Tsuruga, Hiroshi Kojima, Hiroyuki Ishikawa, Shigemitsu Yoshida

Journal of Histochemistry and Cytochemistry 56 ( 2 ) 97 - 109 2008年2月

記述言語：英語掲載種別：研究論文（学術雑誌）

We have previously reported the TLR4 expression in human intestinal lymphatic vessels. In the study here, microarray analysis showed the expression of the TLR4, MD-2, CD14, MyD88, TIRAP, TRAM, IRAK1, and TRAF6 genes in cultured human neonatal dermal lymphatic microvascular endothelial cells (LEC). The microarray analysis also showed that LEC expressed genes of IL-6, IL-8, VCAM-1, and ICAM-1, and the real-time quantitative PCR analysis showed that mRNA production was increased by lipopolysaccharide (LPS). The LPS-induced IL-6, IL-8, VCAM-1, and ICAM-1 production in LEC was suppressed by the introduction of TLR4-specific small interfering RNA, and also by anti-TLR4, nobiletin, and CAPE pretreatment. These findings suggest that LEC has TLR4-mediated LPS recognition mechanisms that involve at least activation of NF-κB, resulting in increased expression of IL-6, IL-8, VCAM-1, and ICAM-1. Both the LPS effect on the gene expression and also the suppression by nobiletin and CAPE pre-treatment on the protein production were larger in IL-6 and in VCAM-1 than in IL-8 and in ICAM-1 in LEC. The signal transduction of NF-κB and AP-1-dependent pathway may be more critical for the expression of IL-6 and VCAM-1 than that of IL-8 and ICAM-1 in LEC. © The Histochemical Society, Inc.

DOI： 10.1369/jhc.7A7299.2007

Effects of bFGF on suppression of collagen type I accumulation and scar tissue formation during wound healing after mucoperiosteal denudation of rat palate 査読国際誌

Wookjin Choi, Hitoshi Kawanabe, Yoshihiko Sawa, Kunihisa Taniguchi, Hiroyuki Ishikawa

ACTA ODONTOLOGICA SCANDINAVICA 66 ( 1 ) 31 - 37 2008年

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：TAYLOR & FRANCIS AS

Objective. The purpose of this study was to evaluate the effect of basic fibroblast growth factor (bFGF) on collagen changes after mucoperiosteal denudation of rat palate. Material and methods. A total of 36 male Wistar rats were divided into control, scar, sham, and bFGF groups. In the scar, sham, and bFGF groups, lateral palatal mucoperiosteum was excised to form scar tissue on the palate. In the bFGF group, bFGF solution was injected into the operated area 1 week postoperatively. At 6 weeks postoperatively, the distribution of collagen type I and the 3-dimensional structure of collagen fibers were investigated under immunofluorescent and scanning electron microscopy. Result. In the bFGF group, weakly immunostained submucosa was clearly distinguishable from the strongly immunostained cervical periodontal ligament and gingiva. Collagen fibers running from submucosal tissue into the surface of underlying palatal bone comprised loosely arranged collagen fibrils. Lumen structures in collagen fibers resembled those in the control group. Conclusion. Administration of bFGF for suppression of collagen type I generation could suppress scar tissue formation and reduce connective strength with adjacent teeth and palatal bone.

DOI： 10.1080/00016350701884612

Effects of TNF-alpha on leukocyte adhesion molecule expressions in cultured human lymphatic endothelium 査読国際誌

Yoshihiko Sawa, Yukitaka Sugimoto, Takeshi Ueki, Hiroyuki Ishikawa, Atuko Sato, Toshikazu Nagato, Shigemitsu Yoshida

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY 55 ( 7 ) 721 - 733 2007年7月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：HISTOCHEMICAL SOC INC

TNF-alpha alters leukocyte adhesion molecule expression of cultured endothelial cells like human umbilical vein endothelial cells (HUVEC). This study was designed to investigate the changes in vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), and platelet endothelial cell adhesion molecule-1 (PECAM-1) expression with TNF-alpha stimulation in cultured human neonatal dermal lymphatic endothelial cells (HNDLEC). The real-time quantitative PCR analysis on HNDLEC showed that TNF-alpha. treatment leads to increases of VCAM-1 and ICAM-1 mRNAs to the 10.8- and 48.2-fold levels of untreated cells and leads to a reduction of PECAM-1 mRNA to the 0.42-fold level of untreated cells. Western blot and immunohistochemical analysis showed that TNF-a leads to VCAM-1 and ICAM-1 expressions that were inhibited by antiserum to human TNF receptor or by AP-1 inhibitor nobiletin. In flow cytometry analysis, the number of VCAM-1- and ICAM-1-positive cells increased, and PECAM-1-positive cells decreased with TNF-alpha treatment. Regarding protein amounts produced in cells and amounts expressed on the cell surface, VCAM-1 and ICAM-1 increased in HNDLEC and HUVEC, and PECAM-1 decreased in HNDLEC in a TNF-a concentration-dependent manner. VCAM-1, ICAM-1, and PECAM-1 protein amounts in TNF-alpha-stimulated cells were lower in HNDLEC than in HUVEC. This suggests that the lymphatic endothelium has the TNF-alpha-induced signaling pathway, resulting in increased VCAM-1 and ICAM-1 expression to a weaker extent than blood endothelium and PECAM-1 reduction to a stronger extent than blood endothelium.

DOI： 10.1369/jhc.6A7171.2007

[A case of recrudescent groove pancreatitis, which was in remission by proximal gastrectomy with vagotomy for gastric cancer].

Hideki Aragane, Hideki Fujii, Takaharu You, Atuhiro Morita, Morimichi Miyazaki, Kiyoshi Morita, Tohru Ohkawara, Shinji Fukumitu, Yoshihiko Sawa, Yasuo Ohkawara

Nihon Shokakibyo Gakkai zasshi = The Japanese journal of gastro-enterology 103 ( 5 ) 537 - 42 2006年5月

記述言語：日本語掲載種別：研究論文（学術雑誌）

This report describes our experience with a 60 year old male who suffered from a recrudescence of groove pancreatitis. He had been treated by conservative medication therapy by proton pump inhibitor used for therapy of duodenal ulcer, and was in remission. During a follow-up one year later, endoscopy revealed gastric cancer, for which a proximal gastrectomy and vagotomy were performed. The patient continues to remain in remission for the groove pancreatitis. Our experience with the clinical course of this disease, in which treatment for duodenal ulcer was used effectively, offers new insights into the progression and therapy of groove pancreatitis.

Intracellular distribution of desmoplakin in human odontoblasts 査読国際誌

Y Sawa, S Kuroshima, Y Yamaoka, S Yoshida

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY 53 ( 9 ) 1099 - 1108 2005年9月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：HISTOCHEMICAL SOC INC

Coexpression of desmosomal proteins and vimentin has been reported in a specific mesenchymal phenotype. This study investigated the expression of vimentin-binding desmosomal proteins in human dental pulp fibroblasts (DPF) and odontoblasts. The dental pulp has no cells expressing desmocollin (DSC) 1-3, desmoglein (DSG) 1-3, junction plakoglobin (JUP), or desmoplakin (DPK) 1 and 2 except for odontoblasts expressing DPK. A confocal image by laser-scanning microscopy demonstrated the diffuse distribution of DPK in the cytoplasm throughout the odontoblast processes. In culture, the mRNA expression of JUP and DPK1, but not DSC1-3 and DSG1-3, was detected in all DPF clones tested and also in odontoblast-like cells (OB) expressing osteocalcin and dentin sialophosphoprotein mRNAs established in the differentiation medium. The DPF having the potential to differentiate into OB expressed vimentin, but not DPK before culturing in the differentiation medium, whereas OB expressed vimentin-binding DPK1. These results suggest that DPF usually expresses DPK1 mRNA, and that the DPK1 production and the bonding of vimentin to DPK1 occur in DPF with the differentiation into odontoblasts.

DOI： 10.1369/jhc.4A6525.2005

Expression of cys-cys chemokine ligand 21 on human gingival lymphatic vessels 査読国際誌

S Kuroshima, Y Sawa, Y Yamaoka, K Notani, S Yoshida, N Inoue

TISSUE & CELL 36 ( 2 ) 121 - 127 2004年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：CHURCHILL LIVINGSTONE

The cys-cys (C-C) chemokine ligand 21 is a member of the C-C chemokines that constitute a group of heparin-binding cytokines with a pattern of four or six conserved cysteines. The CCL21 is known to be expressed in secondary lymphoid tissues, however it has rarely been reported for the expression on peripheral lymphatic vessels in somatic tissue. Here we investigated the expression of CCL21 on lymphatic vessels identified by anti-desmoplakin in uninflamed and inflamed human gingiva. In uninflamed tissue the expression of CCL21 was detected on lymphatic vessels in gingiva. In uninflamed gingiva the expression of CCL21 was detected on all lymphatic capillaries of the mucosal connective tissue papillae. There were two types of collecting lymphatic vessels in the lamina propria mucosae expressing CCL21 strongly or very weakly. In inflamed gingiva no expression of CCL21 was detected on lymphatic vessels. In all tissue sections no blood vessels expressing CCL21 were observed. These results may suggest that the expression of CCL21 is predominantly induced in the peripheral lymphatic endothelium of the uninflamed mucosal microcirculation, and that under inflamed conditions a reduction of CCL21 occurs in lymphatic endothelium. (C) 2003 Elsevier Ltd. All rights reserved.

DOI： 10.1016/j.tice.2003.10.004

Reduction of alkaline phosphatase activity in aged human osteogenic periodontal ligament fibroblasts exhibiting short telomeres 査読国際誌

Y Sawa, Y Yamaoka, S Kuroshima, S Yoshida

CELL AND TISSUE RESEARCH 315 ( 3 ) 331 - 337 2004年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：SPRINGER-VERLAG

The osteogenic cell type of human periodontal ligament fibroblasts (PDLF) undergoes senescence at finite population doubling numbers unrelated to donor ages. This study investigated telomere lengths of osteogenic PDLF from differently aged donors and alterations of the osteoblast-like properties in the aged PDLF with short telomeres. Telomere lengths of osteogenic PDLF were biased towards long or short among all 15- to 51-year-old individuals, and did not show a normal distribution by Pearson's test or a correlation to donor age by simple regression analysis. In osteogenic PDLF, senescence-associated beta-galactosidase was expressed in 78.5% of cells in the clones with short telomeres (mean 3.02 kbp), and in 9.4% of cells in the clones with long telomeres (mean 13.06 kbp). These results suggest that human periodontium comprises aged osteogenic PDLF without correlation to age. Osteogenic PDLF with long telomeres strongly expressed alkaline phosphatase (ALPase) activity whereas cells with short telomeres expressed ALPase activity to a weaker extent. Total activity of ALPase in the clones of osteogenic PDLF with long telomeres was significantly higher than that in the clones with short telomeres. The produced amounts of both osteopontin and osteocalcin in the clones of osteogenic PDLF with long telomeres were slightly but statistically significantly smaller than those in the clones with short telomeres. These findings suggest that aged osteogenic PDLF reduce the expression of ALPase activity but that there is not a critical alteration in bone-associated protein production. Aged osteogenic PDLF may impair the ability to induce ALPase-dependent calcification.

DOI： 10.1007/s00441-003-0837-7

Expression of Toll-like receptors 2 and 4 on human intestinal lymphatic vessels 査読国際誌

Microvascular Research 67 ( 1 ) 90 - 95 2004年1月

Plastic casts and confocal laser scanning microscopy applied to the observation of enamel tubules in the red Kangaroo (Macropus rufus).

Masatsugu Suzuki, Natsumi Ushijima, Ayako Kohno, Yoshihiko Sawa, Shigemitsu Yoshida, Mitsuo Sekikawa, Noriyuki Ohtaishi

Anatomical science international 78 ( 1 ) 53 - 61 2003年3月

記述言語：英語掲載種別：研究論文（学術雑誌）

Scanning electron microscopy for plastic casts and confocal laser scanning microscopy for Villanueva bone-stained ground sections were used together to observe enamel tubules in red kangaroo molars. Although the tubular structures such as terminals, bends, expansions, splits, divergences and rejoinings in this species were within the variations of marsupial species, their morphological characteristics were demonstrated with extremely clear and persuasive images. Thus, the combined observations of plastic casts by scanning electron microscopy and Villanueva bone-stain sections by confocal laser scanning microscopy were found to be of value for the investigation of enamel tubules and tubular structures in other hard tissues.

Production of colony-stimulating factor in human dental pulp fibroblasts 査読

Y Sawa, Y Horie, Y Yamaoka, N Ebata, T Kim, S Yoshida

JOURNAL OF DENTAL RESEARCH 82 ( 2 ) 96 - 100 2003年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：INT AMER ASSOC DENTAL RESEARCHI A D R/A A D R

Class II major histocompatilibity complex (MHC)-expressing cells are usually distributed in dental pulp, and it was postulated that the colony-stimulating factor (CSF) derived from dental pulp fibroblasts contributes to the migration of class II MHC-expressing cells into pulp tissue. This study aimed to investigate the CSF production of human dental pulp fibroblasts. In pulp tissue sections, granulocyte (G)-CSF was detected from normal teeth, while G-CSF, macrophage (M)-CSF, and granulocyte-macrophage (GM)-CSF were detected from teeth with dentinal caries. In cultured dental pulp fibroblasts, G-CSF was detected by immunostaining, immunoprecipitation, and ELISA, and mRNAs of G-CSF, M-CSF, and GM-CSF were detected by RT-PCR. The dental pulp fibroblasts cultured with TNF-alpha were found to increase the G-CSF expression and to produce M-CSF and GM-CSF. These findings suggest that dental pulp fibroblasts usually produce G-CSF. In the presence of TNF-alpha, dental pulp fibroblast express M-CSF and GM-CSF.

Cultured periodontal ligament fibroblasts express diverse connexins 査読国際誌

Y Yamaoka, Y Sawa, N Ebata, N Ibuki, S Yoshida

TISSUE & CELL 34 ( 6 ) 375 - 380 2002年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：CHURCHILL LIVINGSTONE

Recent studies have suggested multiple functions of periodontal ligament fibroblasts (PDLFs) which may relate to the permeability of gap junctions composed of various types of connexins (Cxs). At present, 15 types of Cxs are known to exist, and six of their antibodies, anti-Cx26, Cx32, Cx37, Cx40, Cx43, and Cx45 are commercially available. This study aims to examine which types of Cxs are expressed in cultured PDLFs by an immunohistochemical method, western blotting, and RT-PCR.
The study confirmed the expressions of Cx32, Cx40, Cx43, and Cx45 in PDLFs, while Cx26 and Cx37 were not detected. Considering previous reports, Cx32 may relate to the secretory function, and Cx40 and Cx45 to the contractile function of PDLFs, however, a function for Cx43 has not been specified. In the immunohistochemical examination, different localizations of Cx40/43 and Cx32/45 were established. The former were observed punctately, suggesting that a large part of Cx40/43 may exist in the cell membrane and construct gap junctions. In contrast, the latter were observed uniformly in all the cells, indicating that they are present both in the cell membrane and in the cytoplasm of the cells. (C) 2002 Elsevier Science Ltd. All rights reserved.

DOI： 10.1016/S0040-8166(02)00038-1

Different expressions of connexin 43 and 32 in the fibroblasts of human dental pulp 査読

N Ibuki, Y Yamaoka, Y Sawa, T Kawasaki, S Yoshida

TISSUE & CELL 34 ( 3 ) 170 - 176 2002年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：CHURCHILL LIVINGSTONE

The expression and localization of gap junctional proteins connexin (Cx) 26, 32, and 43 was examined in human dental pulp. Dental pulp tissues were obtained from human third molars immediately after extraction. Some pulp tissues were used for cell culture, and the rest for histological observations. Immunostaining for cultured dental pulp fibroblasts (DPFs) showed that Cx32 and 43 were expressed in human DPFs, and proteins corresponding to 27 (Cx32) and 43 kDa (Cx43) were identified by Western blot analysis. Immunostaining for tissue sections showed that the expression of Cx32 and 43 was observed in the entire region of the pulp and further strong expression of Cx32 was established beneath the cell-rich zone. Considering the close relationship between Cx types and cell functions, the results indicate that DPFs beneath the cell-rich zone may have specific, Cx32-related functions. The cell rich zone is thought to contain progenitor odontoblasts that can be induced to differentiate into mature odontoblasts in response to wounding. Therefore, it may be hypothesized that DPFs just beneath the cell-rich zone produce proteins and induce odontoblast differentiation from the cells in the cell-rich zone. (C) 2002 Elsevier Science Ltd. All rights reserved.

DOI： 10.1016/S0040-8166(02)00028-9

Immunoelectron microscopic study of PECAM-1 expression on lymphatic endothelium of the human tongue 査読

N Ebata, Y Sawa, Y Nodasaka, Y Yamaoka, S Yoshida, Y Totsuka

TISSUE & CELL 33 ( 3 ) 211 - 218 2001年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：CHURCHILL LIVINGSTONE

The expression of platelet-endothelial cell adhesion molecule-1 (PECAM-1) on lymphatic and blood vessels of the human tongue was examined with fluorescence and transmission electron microscopy (TEM), The study used anti-desmoplakins antiserum for light microscopic identification of the lymphatic vessels, plus a pre-embedding immunogold electron microscopic technique for TEM observations. Before making TEM observations, cryostat serial sections were immunostained with anti-desmoplakins or anti-PECAM-l and then embedded. Semithin sections from each cryostat section were photographed under a light microscope and compared in order to identify the lymphatic vessels expressing PECAM-1. In fluorescence microscopy, PECAM-1 expression on lymphatic vessels was weaker than that on blood vessels, TEM observations showed that PECAM-1 expression on the blood vessels was observed only on the luminal surface of the endothelium. In lymphatic vessels, PECAM-1 expression was found both on the luminal and abluminal surfaces of the endothelium. The density of the PECAM-1 reaction products was lower in lymphatic vessels than in blood vessels. The density of PECAM-1 reaction products on the luminal surface of lymphatic vessels was higher than on the abluminal surfaces. The results suggest that blood vessels are more active than lymphatic vessels in leukocyte migration. The expression of PECAM-1 on the abluminal surface of lymphatic endothelium may allow leukocytes to adhere to the endothelium and interact in their migration from tissue into lymphatic vessels. (C) 2001 Harcourt Publishers Ltd.

Desmoplakin as a specific marker of lymphatic vessels 査読

N Ebata, Y Nodasaka, Y Sawa, Y Yamaoka, S Makino, Y Totsuka, S Yoshida

MICROVASCULAR RESEARCH 61 ( 1 ) 40 - 48 2001年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE

The usefulness of immunostaining with anti-desmoplakin antibody for light microscopic identification of lymphatic vessels was examined in cryostat sections of the human tongue. The results were compared with laminin, 5'-nucleotidase (5'-Nase), and factor VIII staining. Immunoelectron microscopic observation was also performed to confirm that the vessels reacting with anti-desmoplakin were lymphatic vessels. Under the immunoelectron microscopic, the vessels reacting with anti-desmoplakin showed ultrastructural features characteristic of lymphatic vessels: thin endothelial walls, no or incomplete basal lamina, open junctions, and overlapping endothelium. In general, lymphatic vessels identified by anti-desmoplakin reacted strongly with 5'-Nase, but showed weak or no reactivity with anti-laminin and anti-factor VIII. Blood vessels showed no reactivity with anti-desmoplakin, but reacted strongly with anti-laminin and anti-factor VIII. However, some blood and lymphatic vessels showed intermediate reactivity with anti-laminin, anti-factor VIII, and 5'-Nase. It was difficult to identify these as blood or lymphatic vessels only by the reactivity differences. The results indicate that anti-desmoplakin antibody specifically distinguishes lymphatic vessels and is useful for studying the fine distribution of lymphatic vessels under light microscopy. (C) 2000 Academic Press.

DOI： 10.1006/mvre.2000.2280

Double expressions of connexin 43 and 32 in human periodontal ligament fibroblasts 査読

Y Yamaoka, Y Sawa, N Ebata, N Ibuki, S Yoshida, T Kawasaki

TISSUE & CELL 32 ( 4 ) 328 - 335 2000年8月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：CHURCHILL LIVINGSTONE

The expressions of connexin 43 and 32 in cultured and intact human periodontal ligament fibroblasts (PDLFs) were examined using immunohistochemical methods, and western blot analysis was conducted with anti-connexin 43 and 32 in cultured PDLFs. The PDLFs both in cultured cells and tissue sections reacted with anti-connexin 43 and 32, and western blot analysis showed bands of approximately 43 kD and 27 kD reacted with anti-connexin 43 and 32 respectively, suggesting the existence of gap junctions in human PDLFs, In cultured PDLFs there were no reaction products of connexin 43 when the cells were not in contact with adjacent cells, but reaction products were increasingly observed with increases in cell-cell contacts. Different from connexin 43, the reaction products of connexin 32 were found in the cytoplasm, regardless of whether the cells were or were not in contact with adjacent cells. Further, the reaction activity of connexin 32 varied among PDLFs; some were strong, some moderate, and some weak. The expressions of connexin 43 and 32 in human PDLFs are suggested to be related to the regulation of two different functions of the PDLFs. (C) 2000 Harcourt Publishers Ltd.

Desmosomal proteins in cultured and intact human periodontal ligament fibroblasts 査読

Y Yamaoka, Y Sawa, N Ebata, S Yoshida, T Kawasaki

TISSUE & CELL 31 ( 6 ) 605 - 609 1999年12月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：CHURCHILL LIVINGSTONE

This study examined the kinds of desmosomal proteins in the human periodontal ligament fibroblasts (PDLFs), The PDLFs obtained from young and older patients were cultured and the amounts of desmosomal proteins were measured by ELISA with antibodies to desmoplakins, desmogleins, and desmocollins. Cultured cells and tissue sections of the human periodontal ligament were immunostained with the same antibodies. Expression of desmosomal proteins in the PDLFs was clearly demonstrated both by ELISA and the immunohistochemical studies, suggesting the existence of desmosome-like junctions in the PDLFs, The junctions are considered to protect gap junctions in the PDLFs against cell transformation caused by cell contraction, which may relate to tooth eruption and repair of periodontal tissue, and/or strong occlusal forces, Statistically significant differences (P<0.0001) in the expression of desmoplakins and desmogleins between younger and older patients were observed in this study, (C) 1999 Harcourt Publishers Ltd.

Transcriptional activation of mRNA of intercellular adhesion molecule 1 and induction of its cell surface expression in normal human gingival fibroblasts by Mycoplasma salivarium and Mycoplasma fermentans 査読

L Dong, KI Shibata, Y Sawa, A Hasebe, Y Yamaoka, S Yoshida, T Watanabe

INFECTION AND IMMUNITY 67 ( 6 ) 3061 - 3065 1999年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：AMER SOC MICROBIOLOGY

Lipoproteins in the cell membranes of both Mycoplasma salivarium and Mycoplasma fermentans were demonstrated to trigger the transcription of intercellular adhesion molecule-1 mRNA in normal fibroblasts isolated from human gingival tissue and to induce its cell surface expression by a mechanism distinct from that of Escherichia call lipopolysaccharide. The lipid moiety of the lipoproteins was suggested to play a key role in the expression of the activity.

Lymphatic endothelium of the human tongue expresses multiple leukocyte adhesion molecules 査読

N Ebata, Y Sawa, Y Ashikaga, Y Yamaoka, M Suzuki, Totsuka, V, S Yoshida

TISSUE & CELL 31 ( 1 ) 34 - 38 1999年2月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：CHURCHILL LIVINGSTONE

The expression of leukocyte adhesion molecules on lymphatic vessels of the human tongue was examined using histochemical and immunohistochemical methods. Three different types of lymphatic vessels were distinguished: type I vessels expressed intercellular adhesion molecule-1 (ICAM-1), platelet-endothelial cell adhesion molecule-1 (PECAM-1), and endothelial cell-selectin (ELAM-1); type II vessels expressed ICAM-1 and PECAM-1; and type III vessels expressed PECAM-1 only, The lymphatic vessels located very close to the oral epithelium (lymphatic capillaries) and the other lymphatic vessels near the oral epithelium were type I. The lymphatic vessels in the submucosal connective tissue (collecting lymphatic vessels) were type II and type III. The results suggest that there may be functional differences in the lymphatic endothelium, where lymphatic capillaries are more active than collecting lymphatic vessels in lymphocyte migration from tissue into the lymphatic vessels.

　前のページ - 次のページ　

1