Authorship：Lead author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier {BV}  

Abstract
Introduction: The intercellular network of cell-cell communication among osteocytes is mediated by gap junctions. Gap junctional intercellular communication (GJIC) is thought to play an important role in the integration and synchronization of bone remodeling. To further understand the mechanism of bone development it is important to quantify the difference in the GJIC capacity of young and developmentally mature osteocytes.

Materials and methods: We first established an embryonic chick calvaria growth model to show the growth of the calvaria in embryos at 13 to 21days of age. We then applied a fluorescence recovery after photobleaching (FRAP) technique to compare the difference in the GJIC capacity of young osteocytes with that of developmentally mature osteocytes. Finally, we quantified the dye (Calcein) diffusion from the FRAP data using a mathematic model of simple diffusion which was also used to identify simple diffusion GJIC pattern cells (fitted model) and accelerated diffusion GJIC pattern cells (non-fitted model).

Results: The relationship between the longest medial-lateral length of the calvaria (frontal bone) and the embryonic age fit a logarithmic growth model: length=5.144×ln(day)-11.340. The morphometric data during osteocyte differentiation showed that the cellular body becomes more spindle-shaped and that the cell body volume decreased by approximately 22% with an increase in the length of the processes between the cells. However, there were no significant differences in the cellular body surface area or in the distance between the mass centres of the cells. The dye-displacement rate in young osteocytes was significantly higher than that in developmentally mature osteocytes: dye displacement only occurred in 26.88% of the developmentally mature osteocytes, while it occurred in 64.38% of the young osteocytes. Additionally, in all recovered osteocytes, 36% of the developmentally mature osteocytes comprised non-fitted model cells while 53.19% of the young osteocytes were the non-fitted model, which indicates the active transduction of dye molecules. However, there were no statistically significant differences between the young and developmentally mature osteocytes with regard to the diffusion coefficient, permeability coefficient, or permeance of the osteocyte processes, which were 3.93±3.77 (×10(-8)cm(2)/s), 5.12±4.56 (×10(-5)cm(2)/s) and 2.99±2.47 (×10(-13)cm(2)/s) (mean±SD), respectively.

Conclusions: These experiments comprehensively quantified the GJIC capacity in the embryonic chick calvaria and indicated that the cell-cell communication capacity of the osteocytes in the embryonic chick calvaria was related to their development.

Keywords: Connexins; Fluorescence recovery after photobleaching; Gap-junctional intercellular communication; Mathematic model of simple diffusion; Osteocyte transformation; Osteocytes.

DOI： 10.1016/j.bone.2016.06.016

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Authorship：Corresponding author   Language：English   Publishing type：Research paper (scientific journal)   Publisher：Wiley  

Abstract

To explore the potential mechanisms which O‐linked‐N‐acetylglucosaminylation (O‐GlcNAcylation) regulates osteogenesis, a publicly RNA‐seq dataset was re‐analyzed with literature‐mining and showed the primary targets of O‐GlcNAcylation in osteoblasts are mitochondria/cytoskeleton. Although the O‐GlcNAcylation‐regulated mitochondria/cytoskeleton has been extensively studied, its specific role during osteogenesis remains unclear. To address this, we knocked out Ogt (Ogt‐KO) in MC3T3‐E1 osteoblastic cells. Then, significantly reduced osteoblast differentiation, motility, proliferation, mitochondria–endoplasmic reticulum (Mito–ER) coupling, volume of ER, nuclear tubulins, and oxygen metabolism were observed in Ogt‐KO cells. Through artificial intelligence (AI)‐predicted cellular structures, the time‐lapse live cells imaging with reactive‐oxygen‐species/hypoxia staining showed that lower cell proliferation and altered oxygen metabolism in the Ogt‐KO cells were correlated with the Mito–ER coupling. Bioinformatics analysis, combined with correlated mRNA and protein expression, suggested that Ezh2 and its downstream targets (Opa1, Gsk3a, Wnt3a, Hif1a, and Hspa9) may be involved in O‐GlcNAcylation‐regulated Mito–ER coupling, ultimately impacting osteoblast differentiation. In conclusion, our findings indicate that O‐GlcNAcylation‐regulated osteoblast differentiation is linked to morphological changes in mitochondria, cytoskeleton, and ER, with Ezh2 potentially playing a crucial role.

DOI： 10.1002/biof.2131

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Language：English   Publishing type：Research paper (scientific journal)  

Mesenchymal stem cells (MSCs) and macrophages collaboratively contribute to bone regeneration after injury. However, detailed mechanisms underlying the interaction between MSCs and inflammatory macrophages (M1) remain unclear. A macrophage-depleted tooth extraction model was generated in 5-wk-old female C57BL/6J mice using clodronate liposome (12.5 mg/kg/mouse, intraperitoneally) or saline injection (control) before maxillary first molar extraction. Mice were sacrificed on days 1, 3, 5, 7, and 10 after tooth extraction (n = 4). Regenerated bone volume evaluation of tooth extraction socket (TES) and histochemical analysis of CD80+M1, CD206+M2 (anti-inflammatory macrophages), PDGFRα+MSC, and TNF-α+ cells were performed. In vitro, isolated MSCs with or without TNF-α stimulation (10 ng/mL, 24 h, n = 3) were bulk RNA-sequenced (RNA-Seq) to identify TNF-α stimulation-specific MSC transcriptomes. Day 7 micro-CT and HE staining revealed significantly lower mean bone volume (clodronate vs control: 0.01 mm3 vs 0.02 mm3, p<.0001) and mean percentage of regenerated bone area per total TES in clodronate group (41.97% vs 54.03%, p<.0001). Clodronate group showed significant reduction in mean number of CD80+, TNF-α+, PDGFRα+, and CD80+TNF-α+ cells on day 5 (306.5 vs 558.8, p<.0001; 280.5 vs 543.8, p<.0001; 365.0 vs 633.0, p<.0001, 29.0 vs 42.5, p<.0001), while these cells recovered significantly on day 7 (493.3 vs 396.0, p=.0004; 479.3 vs 384.5, p=.0008; 593.0 vs 473.0, p=.0010, 41.0 vs 32.5, p=.0003). RNA-Seq analysis showed that 15 genes (|log2FC| > 5.0, log2TPM > 5) after TNF-α stimulation were candidates for regulating MSC's immunomodulatory capacity. In vivo, Clec4e and Gbp6 are involved in inflammation and bone formation. Clec4e, Gbp6, and Cxcl10 knockdown increased osteogenic differentiation of MSCs in vitro. Temporal reduction followed by apparent recovery of TNF-α-producing M1 macrophages and MSCs after temporal macrophage depletion suggests that TNF-α activated MSCs during TES healing. In vitro mimicking the effect of TNF-α on MSCs indicated that there are 15 candidate MSC genes for regulation of immunomodulatory capacity.

DOI： 10.1093/jbmrpl/ziae085

PubMed

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MDPI AG  

The pathology of medication-related osteonecrosis of the jaw (MRONJ), often associated with antiresorptive therapy, is still not fully understood. Osteocyte networks are known to play a critical role in maintaining bone homeostasis and repair, but the exact condition of these networks in MRONJ is unknown. On the other hand, the local application of E-coli-derived Recombinant Human Bone Morphogenetic Protein 2/β-Tricalcium phosphate (E-rhBMP-2/β-TCP) has been shown to promote bone regeneration and mitigate osteonecrosis in MRONJ-like mouse models, indicating its potential therapeutic application for the treatment of MRONJ. However, the detailed effect of BMP-2 treatment on restoring bone integrity, including its osteocyte network, in an MRONJ condition remains unclear. Therefore, in the present study, by applying a scanning electron microscope (SEM) analysis and a 3D osteocyte network reconstruction workflow on the alveolar bone surrounding the tooth extraction socket of an MRONJ-like mouse model, we examined the effectiveness of BMP-2/β-TCP therapy on the alleviation of MRONJ-related bone necrosis with a particular focus on the osteocyte network and alveolar bone microstructure (microcrack accumulation). The 3D osteocyte dendritic analysis showed a significant decrease in osteocyte dendritic parameters along with a delay in bone remodeling in the MRONJ group compared to the healthy counterpart. The SEM analysis also revealed a notable increase in the number of microcracks in the alveolar bone surface in the MRONJ group compared to the healthy group. In contrast, all of those parameters were restored in the E-rhBMP-2/β-TCP-treated group to levels that were almost similar to those in the healthy group. In summary, our study reveals that MRONJ induces osteocyte network degradation and microcrack accumulation, while application of E-rhBMP-2/β-TCP can restore a compromised osteocyte network and abrogate microcrack accumulation in MRONJ.

DOI： 10.3390/ijms25126648

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

Abstract
Objective
Mammalian somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) via the forced expression of Yamanaka reprogramming factors. However, only a limited population of the cells that pass through a particular pathway can metamorphose into iPSCs, while the others do not. This study aimed to clarify the pathways that chondrocytes follow during the reprogramming process.

Design
The fate of human articular chondrocytes under reprogramming was investigated through a time-coursed single-cell transcriptomic analysis, which we termed an inverse genetic approach. The iPS interference technique was also employed to verify that chondrocytes inversely return to pluripotency following the proper differentiation pathway.

Results
We confirmed that human chondrocytes could be converted into cells with an iPSC phenotype. Moreover, it was clarified that a limited population that underwent the silencing of SOX9, a master gene for chondrogenesis, at a specific point during the proper transcriptome transition pathway, could eventually become iPSCs. Interestingly, the other cells, which failed to be reprogrammed, followed a distinct pathway toward cells with a surface zone chondrocyte phenotype. The critical involvement of cellular communication network factors (CCNs) in this process was indicated. The idea that chondrocytes, when reprogrammed into iPSCs, follow the differentiation pathway backward was supported by the successful iPS interference using SOX9.

Conclusions
This inverse genetic strategy may be useful for seeking candidates for the master genes for the differentiation of various somatic cells. The utility of CCNs in articular cartilage regeneration is also supported.

Keywords
cartilagechondrocytedifferentiationreprogrammingCCN family

DOI： 10.1016/j.joca.2024.06.009

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MDPI AG  

The oral mucosa functions as a physico-chemical and immune barrier to external stimuli, and an adequate width of the keratinized mucosa around the teeth or implants is crucial to maintaining them in a healthy and stable condition. In this study, for the first time, bulk RNA-seq analysis was performed to explore the gene expression of laser microdissected epithelium and lamina propria from mice, aiming to investigate the differences between keratinized and non-keratinized oral mucosa. Based on the differentially expressed genes (DEGs) and Gene Ontology (GO) Enrichment Analysis, bone morphogenetic protein 2 (BMP-2) was identified to be a potential regulator of oral mucosal keratinization. Monoculture and epithelial–mesenchymal cell co-culture models in the air–liquid interface (ALI) indicated that BMP-2 has direct and positive effects on epithelial keratinization and proliferation. We further performed bulk RNA-seq of the ALI monoculture stimulated with BMP-2 in an attempt to identify the downstream factors promoting epithelial keratinization and proliferation. Analysis of the DEGs identified, among others, IGF2, ID1, LTBP1, LOX, SERPINE1, IL24, and MMP1 as key factors. In summary, these results revealed the involvement of a well-known growth factor responsible for bone development, BMP-2, in the mechanism of oral mucosal keratinization and proliferation, and pointed out the possible downstream genes involved in this mechanism.

DOI： 10.3390/cells13100807

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Oxford University Press (OUP)  

Cherubism (OMIM 118400) is a rare craniofacial disorder in children characterized by destructive jawbone expansion due to the growth of inflammatory fibrous lesions. Our previous studies have shown that gain-of-function mutations in SH3 domain-binding protein 2 (SH3BP2) are responsible for cherubism and that a knock-in mouse model for cherubism recapitulates the features of cherubism, such as increased osteoclast formation and jawbone destruction. To date, SH3BP2 is the only gene identified to be responsible for cherubism. Since not all patients clinically diagnosed with cherubism had mutations in SH3BP2, we hypothesized that there may be novel cherubism genes and that these genes may play a role in jawbone homeostasis. Here, using whole exome sequencing, we identified homozygous loss-of-function variants in the opioid growth factor receptor like 1 (OGFRL1) gene in two independent autosomal recessive cherubism families from Syria and India. The newly identified pathogenic homozygous variants were not reported in any variant databases, suggesting that OGFRL1 is a novel gene responsible for cherubism. Single cell analysis of mouse jawbone tissue revealed that Ogfrl1 is highly expressed in myeloid lineage cells. We generated OGFRL1 knockout mice and mice carrying the Syrian frameshift mutation to understand the in vivo role of OGFRL1. However, neither mouse model recapitulated human cherubism or the phenotypes exhibited by SH3BP2 cherubism mice under physiological and periodontitis conditions. Unlike bone marrow-derived M-CSF-dependent macrophages (BMMs) carrying the SH3BP2 cherubism mutation, BMMs lacking OGFRL1 or carrying the Syrian mutation showed no difference in TNF-ɑ mRNA induction by LPS or TNF-ɑ compared to wild-type BMMs. Osteoclast formation induced by receptor activator of NF-κB ligand (RANKL) was also comparable. These results suggest that the loss-of-function effects of OGFRL1 in humans differ from those in mice and highlight the fact that mice are not always an ideal model for studying rare craniofacial bone disorders.

DOI： 10.1093/jbmrpl/ziae050

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Wiley  

Abstract

T‐cell acute leukemia and lymphoma have a poor prognosis. Although new therapeutic agents have been developed, their therapeutic effects are suboptimal. α‐Pinene, a monoterpene compound, has an antitumor effect on solid tumors; however, few comprehensive investigations have been conducted on its impact on hematologic malignancies. This report provides a comprehensive analysis of the potential benefits of using α‐pinene as an antitumor agent for the treatment of T‐cell tumors. We found that α‐pinene inhibited the proliferation of hematologic malignancies, especially in T‐cell tumor cell lines EL‐4 and Molt‐4, induced mitochondrial dysfunction and reactive oxygen species accumulation, and inhibited NF‐κB p65 translocation into the nucleus, leading to robust apoptosis in EL‐4 cells. Collectively, these findings suggest that α‐pinene has potential as a therapeutic agent for T‐cell malignancies, and further investigation is warranted.

DOI： 10.1111/cas.16086

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.archoralbio.2023.105797

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1016/j.stem.2023.08.004

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/ijms24087394

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Wiley  

Abstract

Objective

The temporomandibular joint (TMJ) is anatomically comprised of the mandibular condylar cartilage (CC) lined with fibrocartilaginous superficial zone and is crucial for eating and dental occlusion. TMJ osteoarthritis (OA) leads to pain, joint dysfunction and permanent loss of cartilage tissue. However, there are no drugs clinically available that ameliorate OA and little is known about global profiles of genes that contribute to TMJ OA. Furthermore, animal models that recapitulate the complexity of signalling pathways contributing to OA pathogenesis are crucial for designing novel biologics that thwart OA progression. We have previously developed a New Zealand white rabbit TMJ injury model that demonstrates CC degeneration. Here, we performed genome‐wide profiling to identify new signalling pathways critical for cellular functions during OA pathology.

Materials and Methods

Temporomandibular joint OA was surgically induced in New Zealand white rabbits. Three months following injury, we performed global gene expression profiling of the TMJ condyle. RNA samples from TMJ condyles were subjected to sequencing. After raw RNA‐seq data were mapped to relevant genomes, differential expression was analysed with DESeq2. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway analysis were conducted.

Results/Conclusions

Our study revealed multiple pathways altered during TMJ OA induction including the Wnt, Notch and PI3K‐Akt signalling pathways. We demonstrate an animal model that recapitulates the complexity of the cues and signals underlying TMJ OA pathogenesis, which is essential for developing and testing novel pharmacologic agents to treat OA.

DOI： 10.1111/ocr.12649

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：MDPI AG  

Tumor angiogenesis is one of the hallmarks of solid tumor development. The progressive tumor cells produce the angiogenic factors and promote tumor angiogenesis. However, how the tumor stromal cells influence tumor vascularization is still unclear. In the present study, we evaluated the effects of oral squamous cell carcinoma (OSCC) stromal cells on tumor vascularization. The tumor stromal cells were isolated from two OSCC patients with different subtypes: low invasive verrucous squamous carcinoma (VSCC) and highly invasive squamous cell carcinoma (SCC) and co-xenografted with the human OSCC cell line (HSC-2) on nude mice. In comparison, the CD34+ vessels in HSC-2+VSCC were larger than in HSC-2+SCC. Interestingly, the vessels in the HSC-2+VSCC expressed vascular endothelial cadherin (VE-cadherin), indicating well-formed vascularization. Our microarray data revealed that the expression of extracellular superoxide dismutase, SOD3 mRNA is higher in VSCC stromal cells than in SCC stromal cells. Moreover, we observed that SOD3 colocalized with VE-cadherin on endothelial cells of low invasive stroma xenograft. These data suggested that SOD3 expression in stromal cells may potentially regulate tumor vascularization in OSCC. Thus, our study suggests the potential interest in SOD3-related vascular integrity for a better OSCC therapeutic strategy.

DOI： 10.3390/biomedicines10112729

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.3390/cells11192970

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1007/s00774-022-01321-x

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media {LLC}  

<jats:title>Abstract</jats:title><jats:p>Bone loss due to smoking represents a major risk factor for fractures and bone osteoporosis. Signaling through the aryl hydrocarbon receptor (AhR) and its ligands contributes to both bone homeostasis and inflammatory diseases. It remains unclear whether the same AhR signaling axis affects the temporomandibular joint (TMJ). The aim of this study was to investigate possible mechanisms which mediate bone loss in the TMJ due to smoking. In particular, whether benzo[<jats:italic>a</jats:italic>]pyrene (B[<jats:italic>a</jats:italic>]P), a carcinogen of tobacco smoke, induces expression of the AhR target gene, Cyp1a1, in mandibular condyles. Possible functions of an endogenous ligand of FICZ, were also investigated in a TMJ-osteoarthritis (OA) mouse model. B[<jats:italic>a</jats:italic>]P was administered orally to wild-type and <jats:italic>AhR</jats:italic><jats:sup>−/−</jats:sup> mice and bone metabolism was subsequently examined. TMJ-OA was induced in wild-type mice with forceful opening of the mouth. Therapeutic functions of FICZ were detected with μCT and histology. Exposure to B[<jats:italic>a</jats:italic>]P accelerated bone loss in the mandibular subchondral bone. This bone loss manifested with osteoclastic bone resorption and upregulated expression of Cyp1a1 in an <jats:italic>AhR</jats:italic>-dependent manner. In a mouse model of TMJ-OA, FICZ exhibited a dose-dependent rescue of mandibular subchondral bone loss by repressing osteoclast activity. Meanwhile, in vitro, pre-treatment with FICZ reduced RANKL-mediated osteoclastogenesis. B[<jats:italic>a</jats:italic>]P regulates mandibular subchondral bone metabolism via the Cyp1a1. The AhR ligand, FICZ, can prevent TMJ-OA by regulating osteoclast differentiation.</jats:p>

DOI： 10.1038/s41598-021-94470-4

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Springer Science and Business Media {LLC}  

Abstract:
Bone is a highly dynamic organ that undergoes remodeling equally regulated by osteoblast-mediated bone formation and osteoclast-mediated bone resorption. To clarify the regulation of osteoblastogenesis, primary murine osteoblasts are required for an in vitro study. Primary osteoblasts are isolated from neonatal calvariae through digestion with collagenase. However, the number of cells collected from one pup is not sufficient for further in vitro experiments, leading to an increase in the use of euthanized pups. We hypothesized that the viscosity of digested calvariae and digestion solution supplemented with collagenase results in cell clumping and reduction of isolated cells from bones. We simply added Benzonase, a genetically engineered endonuclease that shears all forms of DNAs/RNAs, in order to reduce nucleic acid-mediated viscosity. We found that addition of Benzonase increased the number of collected osteoblasts by three fold compared to that without Benzonase through reduction of viscosity. Additionally, Benzonase has no effect on cellular identity and function. The new osteoblast isolation protocol with Benzonase minimizes the number of neonatal pups required for an in vitro study and expands the concept that isolation of other populations of cells including osteocytes that are difficult to be purified could be modified by Benzonase.

DOI： 10.1038/s41598-021-87716-8

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier {BV}  

DOI： 10.1016/j.bbadis.2021.166236

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Wiley  

Abstract

Diamond‐Blackfan anemia (DBA) is a genetic disorder caused by mutations in genes encoding ribosomal proteins and characterized by erythroid aplasia and various physical abnormalities. Although accumulating evidence suggests that defective ribosome biogenesis leads to p53‐mediated apoptosis in erythroid progenitor cells, little is known regarding the underlying causes of the physical abnormalities. In this study, we established induced pluripotent stem cells from a DBA patient with RPL5 haploinsufficiency. These cells retained the ability to differentiate into osteoblasts and chondrocytes. However, RPL5 haploinsufficiency impaired the production of mucins and increased apoptosis in differentiated chondrocytes. Increased expression of the pro‐apoptotic genes BAX and CASP9 further indicated that RPL5 haploinsufficiency triggered p53‐mediated apoptosis in chondrocytes. Murine double minute 2 (MDM2), the primary negative regulator of p53, plays a crucial role in erythroid aplasia in DBA patient. We found the phosphorylation level of MDM2 was significantly decreased in RPL5 haploinsufficient chondrocytes. In stark contrast, we found no evidence that RPL5 haploinsufficiency impaired osteogenesis. Collectively, our data support a model in which RPL5 haploinsufficiency specifically induces p53‐mediated apoptosis in chondrocytes through MDM2 inhibition, which leads to physical abnormalities in DBA patients.

DOI： 10.1111/pin.13168

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Other Link： https://onlinelibrary.wiley.com/doi/full-xml/10.1111/pin.13168

Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1002/biof.1774

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

Abstract
Introduction: The Wnt signaling pathway acts as a key regulator of skeletal development and its homeostasis. However, the potential role of Wnt1 in the mechanotransduction machinery of orthodontic tooth movement-initiated bone remodeling is still unclear. Hence, this study focused on the regulatory dynamics of the Wnt1 expression in both the periodontal ligament (PDL) and osteocytes in vivo and in vitro.

Methods: The Wnt1 expression in the orthodontically moved maxillary first molar in mice was assessed at 0, 1, and 5 days, on both the compression and tension sides. Primary isolated human PDL (hPDL) fibroblasts, as well as murine long-bone osteocyte-Y4 (MLO-Y4) cells, were exposed to continuous compressive force and static tensile force.

Results: The relative quantification of immunodetection showed that orthodontic tooth movement significantly stimulated the Wnt1 expression in both the PDL and alveolar osteocytes on the tension side on day 5, whereas the expression on the compression side did not change. This increase in the Wnt1 expression, shown in vivo, was also noted after the application of 12% static tensile force in isolated hPDL fibroblasts and 20% in MLO-Y4 cells. In contrast, a compressive force led to the attenuation of the Wnt1 gene expression in both hPDL fibroblasts and MLO-Y4 cells in a force-dependent manner. In the osteocyte-PDL coculture system, recombinant sclerostin attenuated Wnt1 in PDL, whereas the antisclerostin antibody upregulated its gene expression, indicating that mechanically-driven Wnt1 signaling in PDL might be regulated by osteocytic sclerostin.

Conclusions: Our findings provide that Wnt1 signaling plays a vital role in tooth movement-initiated bone remodeling via innovative mechanotransduction approaches.

DOI： 10.1016/j.ajodo.2020.08.006

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

Abstract
Pseudomonas aeruginosa (P. aeruginosa) is associated with periapical periodontitis. The lesions are characterized by a disorder in osteoblast metabolism. Quorum sensing molecular N-(3-oxododecanoyl)-homoserine lactone (AHL) is secreted by P. aeruginosa and governs the expression of numerous virulence factors. AHL can trigger intracellular calcium ([Ca2+]i) fluctuations in many host cells. However, it is unclear whether AHL can regulate osteoblast metabolism by affecting [Ca2+]i changes or its spatial correlation. We explored AHL-induced apoptosis and differentiation in pre-osteoblastic MC3T3-E1 cells and evaluated [Ca2+]i mobilization using several extraction methods. The spatial distribution pattern of [Ca2+]i among cells was investigated by Moran's I, an index of spatial autocorrelation. We found that 30 μM and 50 μM AHL triggered opposing osteoblast fates. At 50 μM, AHL inhibited osteoblast differentiation by promoting mitochondrial-dependent apoptosis and negatively regulating osteogenic marker genes, including Runx2, Osterix, bone sialoprotein (Bsp), and osteocalcin (OCN). In contrast, prolonged treatment with 30 μM AHL promoted osteoblast differentiation concomitantly with cell apoptosis. The elevation of [Ca2+]i levels in osteoblasts treated with 50 μM AHL was spatially autocorrelated, while no such phenomenon was observed in 30 μM AHL-treated osteoblasts. The blocking of cell-to-cell spatial autocorrelation in the osteoblasts provoked by 50 μM AHL significantly inhibited apoptosis and partially restored differentiation. Our observations suggest that AHL affects the fate of osteoblasts (apoptosis and differentiation) by affecting the spatial correlation of [Ca2+]i changes. Thus, AHL acts as a double-edged sword for osteoblast function.

Keywords: Apoptosis; Calcium; Differentiation; N-(3-oxododecanoyl)-homoserine lactone; Osteoblast metabolism; Spatial autocorrelation.

DOI： 10.1016/j.cellsig.2020.109740

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Elsevier BV  

Abstract
Objective
Porphyromonas gingivalis (P. gingivalis) is a major bacterium responsible for the progression of periodontitis. P. gingivalis produces small vesicles called outer membrane vesicles (OMVs) containing virulence factors. Increasing evidence suggests a close relationship between periodontitis and respiratory system diseases, such as aspiration pneumonia. However, little is known about whether P. gingivalis OMVs give rise to the impediment of lung epithelial cells. We investigated the effect of the OMVs on cell viability and tight junctions of lung epithelial cells.

Design
Human lung epithelial A549 cells were treated with P. gingivalis OMVs. Cell viability was evaluated, and cell morphology was examined using scanning electron and phase contrast microscopies. To detect apoptosis induced by P. gingivalis OMVs, activation of caspase-3 and poly ADP-ribose polymerase (PARP) cleavage was examined by using Western blotting. Immunocytochemistry was performed to stain tight junction proteins.

Results
P. gingivalis OMVs decreased cell viability in A549 cells in a dose- and time-dependent manner. Microscopic analysis revealed that the OMVs induced morphological changes leading to irregular cell membrane structures. The OMVs caused cell shrinkage, membrane blebbing, and cytoplasmic expulsion in a dose-dependent manner. Western blot analysis showed the OMVs induced caspase-3 activation and PARP cleavage. Treatment with the OMVs disrupted the intact distributions of tight junction proteins.

Conclusions
These results indicate that P. gingivalis OMVs induced cell death by destroying the barrier system in lung epithelial cells. Our present study raises the possibility that P. gingivalis OMVs is an important factor in the engagement of periodontitis with respiratory system diseases.

DOI： 10.1016/j.archoralbio.2020.104841

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Authorship：Lead author   Language：English   Publishing type：Part of collection (book)   Publisher：Springer US  

DOI： 10.1007/7651_2020_322

Web of Science

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Authorship：Lead author,　Last author   Language：English   Publishing type：Doctoral thesis  

DOI： 10.18926/60928

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Language：English   Publishing type：Research paper (scientific journal)   Publisher：Wiley  

Abstract
Human periodontal ligament (hPDL) fibroblasts are thought to receive mechanical stress (MS) produced by orthodontic tooth movement, thereby regulating alveolar bone remodeling. However, the role of intracellular calcium ([Ca2+]i)-based mechanotransduction is not fully understood. We explored the MS-induced [Ca2+]i responses both in isolated hPDL fibroblasts and in intact hPDL tissue and investigated its possible role in alveolar bone remodeling. hPDL fibroblasts were obtained from healthy donors' premolars that had been extracted for orthodontic reasons. The oscillatory [Ca2+]i activity induced by static compressive force was measured by a live-cell Ca2+ imaging system and evaluated by several feature extraction method. The spatial pattern of cell-cell communication was investigated by Moran's I, an index of spatial autocorrelation and the gap junction (GJ) inhibitor. The Ca2+-transporting ionophore A23187 was used to further investigate the role of [Ca2+]i up-regulation in hPDL cell behavior. hPDL fibroblasts displayed autonomous [Ca2+]i responses. Compressive MS activated this autonomous responsive behavior with an increased percentage of responsive cells both in vitro and ex vivo. The integration, variance, maximum amplitude, waveform length, and index J in the [Ca2+]i responses were also significantly increased, whereas the mean power frequency was attenuated in response to MS. The increased Moran's I after MS indicated that MS might affect the pattern of cell-cell communication via GJs. Similar to the findings of MS-mediated regulation, the A23187-mediated [Ca2+]i uptake resulted in the up-regulation of receptor activator of NF-κB ligand (Rankl) and Sost along with increased sclerostin immunoreactivity, suggesting that [Ca2+]i signaling networks may be involved in bone remodeling. In addition, A23187-treated hPDL fibroblasts also showed the suppression of osteogenic differentiation and mineralization. Our findings suggest that augmented MS-mediated [Ca2+]i oscillations in hPDL fibroblasts enhance the production and release of bone regulatory signals via Rankl/Osteoprotegerin and the canonical Wnt/β-catenin pathway as an early process in tooth movement-initiated alveolar bone remodeling.

DOI： 10.1096/fj.201900484r

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Other Link： https://onlinelibrary.wiley.com/doi/full-xml/10.1096/fj.201900484R

Language：Chinese   Publishing type：Research paper (scientific journal)  

DOI： 10.13701/j.cnki.kqyxyj.2018.08.023

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Language：English   Publishing type：Research paper (scientific journal)  

DOI： 10.1177/0022034518771331

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Language：Chinese   Publishing type：Research paper (scientific journal)  

DOI： 10.13241/j.cnki.pmb.2017.02.033

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Language：Chinese   Publishing type：Research paper (scientific journal)  

DOI： 10.13241/j.cnki.pmb.2017.07.026

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Language：Chinese   Publishing type：Research paper (scientific journal)  

DOI： 10.13929/j.1003-3289.201606089

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Authorship：Lead author,　Last author   Language：Chinese   Publishing type：Master’s thesis  

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Language：English   Publishing type：Research paper, summary (international conference)   Publisher：International Association for Dental Research (IADR)  

Poster Presentation on 102nd General Session & Exhibition of the IADR (2024).
Meeting: 2024 IADR/AADOCR/CADR General Session (New Orleans, Louisiana)
Location: New Orleans, Louisiana
Year: 2024
Final Presentation ID: 0964
Abstract Category|Abstract Category(s): Dental Materials 5: Biocompatibility, Bioengineering and Biologic Effects of Materials

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Other Link： https://www.iadr.org/2024iags

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：American Society of Hematology (ASH)  

Poster Presentation on 2017 ASBMR Annual Metting.
Available at https://ashpublications.org/blood/article/142/Supplement%201/1441/503011/Alpha-Pinene-As-a-Novel-Therapeutic-Agent-for-T

DOI： 10.1182/blood-2023-181447

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Other Link： https://academy.hematology.org/local/catalog/view/product.php?productid=1641&productcourse=&globalid=&bundleid=1641&_gl=1*1x0jmyj*_ga*MTg0NjU2ODYzNy4xNzEyNjIzMzA3*_ga_6P2YRXBKYX*MTcxMjYyMzMwNi4xLjEuMTcxMjYyMzY3NS4wLjAuMA..&_ga=2.76891094.2128980305.1712623307-1846568637.1712623307

Language：Japanese   Publishing type：Research paper, summary (national, other academic conference)   Publisher：Japanese Orthodontic Society  

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Language：Japanese   Publishing type：Research paper, summary (national, other academic conference)   Publisher：Japanese Orthodontic Society  

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Language：English   Publishing type：Research paper, summary (national, other academic conference)   Publisher：Japanese Pharmacological Society  

DOI： 10.1254/jpssuppl.95.0_2-yia-43

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Other Link： https://www.jstage.jst.go.jp/article/jpssuppl/95/0/95_2-YIA-43/_pdf

Authorship：Lead author   Language：English   Publisher：Springer Science and Business Media LLC  

DOI： 10.1007/s00774-020-01155-5

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Other Link： https://link.springer.com/article/10.1007/s00774-020-01155-5/fulltext.html

Authorship：Lead author   Language：English   Publishing type：Research paper, summary (international conference)   Publisher：The American Society for Bone and Mineral Research (ASBMR)  

Poster Presentation on 2020 ASBMR Annual Meeting.
Available at https://www.asbmr.org/meetings/annualmeeting/AbstractDetail?aid=d05390e3-a2f9-4f89-a014-c0391ed39715

Poster Sessions, Presentation Number: P-319
Session: Poster Presentations
Friday, September 11, 2020 10:00 AM - 4:00 PM

DOI： 10.1002/jbmr.4206

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Other Link： https://www.asbmr.org/2020-abstracts

Authorship：Lead author   Language：English   Publishing type：Research paper, summary (national, other academic conference)   Publisher：Japanese Orthodontic Society  

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Language：English   Publishing type：Research paper, summary (national, other academic conference)   Publisher：Japanese Society for Bone and Mineral Research  

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Authorship：Lead author   Language：English   Publishing type：Research paper, summary (international conference)   Publisher：The American Society for Bone and Mineral Research (ASBMR)  

Poster Presentation on 2019 ASBMR Annual Meeting.
Available at https://www.asbmr.org/meetings/annualmeeting/AbstractDetail?aid=65f25eb6-4dd7-42de-adaf-04ba34206836

Poster Sessions, Presentation Number: SAT-562
Session: Poster Session I
Saturday, September 21, 2019 12:30 PM - 2:30 PM, Orange County Convention Center , West Hall C

DOI： 10.1002/jbmr.3936

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Other Link： https://www.asbmr.org/meetings/2019-abstracts

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：The American Society for Bone and Mineral Research (ASBMR)  

Poster Presentation on 2019 ASBMR Annual Meeting.
Available at https://www.asbmr.org/meetings/annualmeeting/AbstractDetail?aid=32f940ac-dc30-4f68-a544-b807652485ba

Poster Sessions, Presentation Number: LB SAT-1007
Session: Late-Breaking Posters I
Saturday, September 21, 2019 12:30 PM - 2:30 PM, Orange County Convention Center , West Hall C

DOI： 10.1002/jbmr.3936

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Other Link： https://www.asbmr.org/meetings/2019-abstracts

Authorship：Lead author   Language：English   Publishing type：Research paper, summary (international conference)   Publisher：The American Society for Bone and Mineral Research (ASBMR)  

Poster Presentation on 2019 ASBMR Annual Meeting.
Available at https://www.asbmr.org/meetings/annualmeeting/AbstractDetail?aid=12917fbd-723a-482e-8a36-9efe018e9f9e

Poster Sessions, Presentation Number: SUN-577
Session: Poster Session II
Sunday, September 22, 2019 12:30 PM - 2:30 PM, Orange County Convention Center , West Hall C

DOI： 10.1002/jbmr.3936

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Other Link： https://www.asbmr.org/meetings/2019-abstracts

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：The American Society for Bone and Mineral Research (ASBMR)  

Poster Presentation on 2019 ASBMR Annual Meeting.
Available at https://www.asbmr.org/meetings/annualmeeting/AbstractDetail?aid=caf69d62-b8d2-4e67-a1d5-ab90a5fc32d6

Poster Sessions, Presentation Number: LB SAT-1033
Session: Late-Breaking Posters I
Saturday, September 21, 2019 12:30 PM - 2:30 PM, Orange County Convention Center , West Hall C

DOI： 10.1002/jbmr.3936

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Other Link： https://www.asbmr.org/meetings/2019-abstracts

Language：English   Publishing type：Research paper, summary (international conference)  

Poster Presentation on 2017 ASBMR Annual Meeting.
Available at https://www.asbmr.org/meetings/annualmeeting/AbstractDetail?aid=7a11a0e6-5e92-4c4b-a2a9-77205d532d05

Poster Sessions, Presentation Number: SAT-0398
Session: Poster Session I and Poster Tours
Saturday, September 29, 2018 12:30 PM - 2:30 PM, Palais des congrès de Montréal, ASBMR Discovery Hall - Exhibit Hall 220 B-E

DOI： 10.1002/jbmr.3621

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Other Link： https://www.asbmr.org/meetings/2018-abstracts

Language：Japanese   Publishing type：Research paper, summary (national, other academic conference)   Publisher：Japanese Society for Bone and Mineral Research  

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Authorship：Lead author   Language：English   Publishing type：Research paper, summary (international conference)   Publisher：The American Society for Bone and Mineral Research (ASBMR)  

Poster Presentation on 2017 ASBMR Annual Meeting.
Available at https://www.asbmr.org/meetings/annualmeeting/AbstractDetail?aid=5db6e683-b053-469e-ab84-b26b8cd6bca3

Poster Sessions, Presentation Number: SA0116
Session: Poster Session I & Poster Tours
Saturday, September 9, 2017 12:30 PM - 2:30 PM, Colorado Convention Center, ASBMR Discovery Hall - Exhibit Hall A & B1

DOI： 10.1002/jbmr.3363

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Other Link： https://www.asbmr.org/meetings/2017-abstracts

Language：English   Publishing type：Research paper, summary (international conference)   Publisher：The American Society for Bone and Mineral Research (ASBMR)  

Poster Presentation on 2017 ASBMR Annual Meeting.
Available at https://www.asbmr.org/meetings/annualmeeting/AbstractDetail?aid=633f6ff6-620a-4750-9839-6e1331d7a78c

Poster Sessions, Presentation Number: SU0109
Session: Poster Session II & Poster Tours
Sunday, September 10, 2017 12:30 PM - 2:30 PM, Colorado Convention Center, ASBMR Discovery Hall - Exhibit Hall A & B1

DOI： 10.1002/jbmr.3363

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Other Link： https://www.asbmr.org/meetings/2017-abstracts

Language：Japanese   Publishing type：Research paper, summary (national, other academic conference)   Publisher：Japanese Society for Bone and Mineral Research  

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Language：Japanese   Publishing type：Research paper, summary (national, other academic conference)   Publisher：Japanese Society for Bone and Mineral Research  

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Authorship：Lead author   Language：English   Publishing type：Research paper, summary (international conference)   Publisher：The American Society for Bone and Mineral Research (ASBMR)  

Poster Presentation on 2016 ASBMR Annual Meeting.
Available at https://www.asbmr.org/education/AbstractDetail?aid=3aa4ea61-d454-4d7c-8cd5-1ef4e5ef4329

Poster Sessions, Presentation Number: SU0179
Session: Poster Session II & Poster Tours
Sunday, September 18, 2016 12:30 PM - 2:30 PM, Georgia World Congress Center, ASBMR Discovery Hall - Expo Hall A1

DOI： 10.1002/jbmr.3107

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Other Link： https://www.asbmr.org/meetings/2016-abstracts

Authorship：Lead author   Language：English   Publishing type：Research paper, summary (national, other academic conference)   Publisher：Japanese Society for Bone and Mineral Research  

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Event date： 2018.6.21 - 2018.6.23

Language：English   Presentation type：Oral presentation (general)  

Venue：Osaka International House Foundation（i-house）  

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Other Link： https://www.jsbm.biz/

Event date： 2017.9.16 - 2017.9.18

Language：English   Presentation type：Oral presentation (general)  

Venue：Matsumoto Dental University Campus  

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Other Link： https://www.jaob.jp/publication/meet_abstract_59.html

Language：English   Presentation type：Oral presentation (general)  

Venue：Basic medical lecture building 1st floor multipurpose room  

Okayama University has been actively engaged in exchanges with foreign universities, including the Okayama University-Northeastern China Graduate Student Exchange (O-NECUS Program) and the JICA Six-University Myanmar Medical Education Enhancement Project. In the Shikata area, more than 80 international students (regular students, special auditing students, special research students, visiting foreign researchers, etc.) from China, Myanmar, Indonesia, and other countries are enrolled, and the number continues to increase. In the laboratory, graduate students and faculty members are actively conducting research in a global environment.

In order to promote the “Advanced Human Resource Development Program to Promote Sustainable Healthcare Models in ASEAN Core City Regions ” and ” A special PhD course for organ regeneration, tissue reconstruction and integrative biology to collaborate with ASEAN leading medical universities,” as well as to promote exchange and information sharing among international students, faculty members, and staff, the 7th International Student Research Presentation was held.

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Other Link： https://www.mdps.okayama-u.ac.jp/en/news_events/7th-international-student-research-presentation-notice/

Language：English   Presentation type：Symposium, workshop panel (nominated)  

Venue：Online  

This Bilateral Joint Research Project, supported by STINT and JSPS, aims to promote international and interdisciplinary collaborations between Linkoping and Okayama Universities in Bone Biology and Material Science and Engineering.

This kick-off meeting will gather multidisciplinary speakers on topics ranging from understanding the theoretical basis of bone development and metabolism to developing novel materials for application in bone tissue engineering.

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Language：English   Presentation type：Oral presentation (general)  

Venue：Lecture Room 1, Dental School of Okayama University  

The "Okayama Dental Association," composed of faculty and students from the Okayama University Faculty of Dentistry, held the "4th Okayama Medical Education International Symposium" in the first lecture hall of our Faculty of Dentistry on December 14th. This symposium was held with the aim of building international collaborations among dental universities in the fields of education and research. Deans or hospital directors from two universities in Indonesia, two in Vietnam, and one each from the Netherlands, the United States, and China were invited to share information about their unique education and research systems. More than 200 dentistry faculty members and students participated, and there were lectures (two sessions) by invited speakers, as well as presentations and poster sessions by ODAPUS students, O-NECUS students, graduate students, and young researchers from abroad. The symposium began with a lecture by Associate Professor Muhammad Ruslin from Hasanuddin University, and included research presentations and introductions of universities by ten researchers and seven undergraduate and graduate students, including ODAPUS and O-NECUS exchange students. There was a Q&A session for the 24 posters presented on the floor, and lively discussions were held in all sessions. Eman A. Taha (in the field of dental pharmacology), Ha Thi Thu Nguyen (in the field of implant regenerative prosthodontics), Islam Md Monirui (in the field of preventive dentistry), and Assistant Professor Hokuto Kawai (in the field of oral pathology) were awarded the Excellent Poster Award for their outstanding poster presentations. The symposium was actively participated in not only by faculty members but also by undergraduate and graduate students involved in various exchange programs. It was a good opportunity for the next generation of dental students and young researchers, who will lead the future development of dentistry and dental care, to gain the knowledge and experience necessary for international academic exchange.

ODAPUS: Okayama University Dental School Short-term Study Abroad Program for Undergraduate Students
O-NECUS: Okayama University-North East China Universities platform, 'Graduate' Student Exchange Program

The symposium was conducted with support from Astellas Pharma Educational Support and Alpha Bio Co., Ltd.

Presenters and Contents:

Associate Professor Muhammad Ruslin (Hasanuddin University): Current state and future of dental care in Indonesia
Associate Professor Tong Minh Son (Hanoi Medical University): Oral health status of the elderly in Vietnam
Professor Vu Quang Hung (Haiphong Medical University): Introduction to Haiphong Medical University
Associate Professor Tianna Wahyu Utami (Gadjah Mada University): Efficacy of oil from Scorodocarpus borneensis leaves
Professor Hongchen Sun (China Medical University): Role of Acvr1 in dentin formation
Professor Albert Feilzer (Academic Center for Dentistry Amsterdam): Side effects of dental materials
Professor Igor Spigelman (University of California, Los Angeles (UCLA)): About novel non-psychoactive cannabinoids for chronic pain treatment
Yuji Komiyama (5th year, Faculty of Dentistry, our university), Kazuho Natsume (3rd year, same), Dr. Nur Mohammad Hassan (Charles Sturt University): Lectures related to the ODAPUS program
Yuhan He (China Medical University, O-NECUS student in 2019), Qianqian Lu (Jilin University, O-NECUS student in 2019): Introduction to their home universities
Akhter Mst Nahid (Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, our university, in the field of biomaterials), May Wathone Oo (in the field of oral pathology), Ei Ei Hsu Hlaing (in the field of orthodontics): Research presentations
Associate Professor Heni Susilowati (Gadjah Mada University), Dr. Pham Thanh Hai (Haiphong Medical University): Lectures on research content
[Contact for inquiries]
Department of Oral Morphology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University
Mika Ikegame, Hirohiko Okamura
TEL: 086-235-6632
E-mail: ikegame@md.okayama-u.ac.jp; hiro-okamura@okayama-u.ac.jp

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Work type：Database science   Location：Mendeley Data  

All raw data, processed results, and analysis procedures of the below research article:
Wang, Ziyi, Yao Weng, Yoshihito Ishihara, Naoya Odagaki, Ei Ei Hsu Hlaing, Takashi Izawa, Hirohiko Okamura, and Hiroshi Kamioka. "Loading history changes the morphology and compressive force-induced expression of receptor activator of nuclear factor kappa B ligand/osteoprotegerin in MLO-Y4 osteocytes." PeerJ 8 (2020): e10244.

DOI： 10.17632/2yfd2w8jfp.1

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Work type：Database science   Location：Mendeley Data  

Raw time-lapse images of calcium changes in MC3T3 cells for the below research article:
All raw data, processed results, and analysis procedures of the below research article:
Guo, Jiajie, Ziyi Wang, Yao Weng, Haoze Yuan, Kaya Yoshida, Mika Ikegame, Kenta Uchibe et al. "N-(3-oxododecanoyl)-homoserine lactone regulates osteoblast apoptosis and differentiation by mediating intracellular calcium." Cellular Signalling 75 (2020): 109740.

DOI： 10.17632/2hxv9cr73m.1

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