The Experts below are selected from a list of 2031 Experts worldwide ranked by ideXlab platform
Martha J. Somerman - One of the best experts on this subject based on the ideXlab platform.
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Cementoblasts express pyrophosphate regulators in a time and space restricted manner.
2013Co-Authors: Brian L. Foster, Kanako J. Nagatomo, Francisco H. Nociti, Hanson Fong, Daisy Dunn, Anne B. Tran, Wei Wang, Sonoko Narisawa, Jose Luis Millán, Martha J. SomermanAbstract:TNAP was expressed strongly in all the periodontal tissues in (A) Ank; Enpp1+/+ control as well as both (B, C) Ank and Enpp1−/− models. Loss of ANK or NPP1 did not alter Cementoblast TNAP expression. ANK was localized selectively to Cementoblasts in (D) control, and was increased when (F) Enpp1 was ablated. Like ANK, NPP1 was found at selectively greater concentrations in Cementoblasts in (G) controls, and was increased upon (H) Ank−/−. Specificity of antibody staining was confirmed in null mice in (E) and (I). All panels are mandibular first molar teeth at 26 dpn. Abbreviations: d = dentin; c = acellular cementum; p = periodontal ligament; b = bone. Cervical cementum is indicated by opposing black arrows. Scale bar = 100 µm.
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Pyrophosphate does not affect Cementoblast proliferation or collagen synthesis, in vitro.
2013Co-Authors: Brian L. Foster, Kanako J. Nagatomo, Francisco H. Nociti, Hanson Fong, Daisy Dunn, Anne B. Tran, Wei Wang, Sonoko Narisawa, Jose Luis Millán, Martha J. SomermanAbstract:(A) Cell proliferation was assayed by MTS assay where absorbance at 570 nm is proportional to the number of living cells in culture. No difference in OCCM.30 Cementoblast cell number was found between non-mineralizing (AA) and mineralizing (AA + BGP) treatments at concurrent time points, including with doses of 10 or 100 µM PPi. (B) Picrosirius red dye was used to stain collagen deposited by Cementoblasts at days 3, 5, and 7. (C) Quantification of the collagen-binding assay did not identify any treatment differences for collagen deposition at any of the time points. For both (A) and (C), graphs show mean +/− SD for n = 3 samples, and no intergroup significant differences (at the same time point) were identified by one-way ANOVA and post-hoc Tukey analysis, for α = 0.05.
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bone morphogenetic protein 7 enhances Cementoblast function in vitro
Journal of Periodontology, 2010Co-Authors: Sema S. Hakki, Brian L. Foster, Martha J. Somerman, Kanako J. Nagatomo, Erdogan E. Hakki, Buket S Bozkurt, Rahime M. NohutcuAbstract:Background: Bone morphogenetic protein (BMP)-7 is a potent bone-inducing factor and was shown to promote periodontal regeneration in vivo and in vitro; however, to our knowledge, the specific effect of BMP-7 on Cementoblasts has not been defined. We aimed to investigate the effects of BMP-7 on Cementoblasts, which are cells responsible for tooth root–cementum formation. We hypothesized that BMP-7 would regulate mineralized tissue–associated genes in Cementoblasts and influence the expression profile of genes associated with Cementoblast extracellular matrix (ECM) and cell adhesion molecules (CAMs).Methods: A murine immortalized Cementoblast cell line (OCCM.30) was cultured with and without 50 ng/ml BMP-7. After 72 hours, total RNA was isolated, and mRNA levels for bone/cementum markers, including bone sialoprotein (BSP), osteocalcin (OCN), osteopontin (OPN), and runt-related transcription factor-2 (Runx2), were investigated by real-time quantitative reverse transcription-polymerase chain reaction (Q-PCR)....
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Bone Morphogenetic Protein‐7 Enhances Cementoblast Function In Vitro
Journal of periodontology, 2010Co-Authors: Sema S. Hakki, Brian L. Foster, Martha J. Somerman, Kanako J. Nagatomo, S. Buket Bozkurt, Erdogan E. Hakki, Rahime M. NohutcuAbstract:Background: Bone morphogenetic protein (BMP)-7 is a potent bone-inducing factor and was shown to promote periodontal regeneration in vivo and in vitro; however, to our knowledge, the specific effect of BMP-7 on Cementoblasts has not been defined. We aimed to investigate the effects of BMP-7 on Cementoblasts, which are cells responsible for tooth root–cementum formation. We hypothesized that BMP-7 would regulate mineralized tissue–associated genes in Cementoblasts and influence the expression profile of genes associated with Cementoblast extracellular matrix (ECM) and cell adhesion molecules (CAMs).Methods: A murine immortalized Cementoblast cell line (OCCM.30) was cultured with and without 50 ng/ml BMP-7. After 72 hours, total RNA was isolated, and mRNA levels for bone/cementum markers, including bone sialoprotein (BSP), osteocalcin (OCN), osteopontin (OPN), and runt-related transcription factor-2 (Runx2), were investigated by real-time quantitative reverse transcription-polymerase chain reaction (Q-PCR)....
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wnt signaling inhibits Cementoblast differentiation
2010Co-Authors: Eiji Nemoto, Martha J. Somerman, Yohei Koshikawa, Sousuke Kanaya, Masahiro Tsuchiya, Masato Tamura, Hidetoshi ShimauchiAbstract:Wnt signaling has been implicated in increased bone formation by controlling mesenchymal stem cell or osteoblastic cell functions; however the role of Wnt signaling on cementogenesis has not been examined. Exposure to Wnt3a inhibited the expression of the osteocalcin (OCN) gene. This effect was accompanied by decreased gene expression of Runx2. Pretreatment with Dickkopf-1 attenuated the suppressive effects of Wnt3a on mRNA expression of Runx2 and OCN on Cementoblasts. These findings suggest that canonical Wnt signaling inhibits Cementoblast differentiation via regulation of expression of Runx2. Elucidating the role of Wnt in controlling Cementoblast function will provide new tools needed to improve on existing periodontal regeneration therapies.
Zhengguo Cao - One of the best experts on this subject based on the ideXlab platform.
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mir 146a 5p attenuates il 1β induced il 6 and il 1β expression in a Cementoblast derived cell line
Oral Diseases, 2020Co-Authors: Jiawen Pan, Zhengguo Cao, Yunru Hao, Chen Zhang, Jiaqi ZhuAbstract:Objective miR-146a is widely induced during the immune response. However, little is known about the biogenesis, function and mechanism of miR-146a in Cementoblasts during the pathogenesis of periodontitis. This study aimed to investigate the effects of miR-146a in murine Cementoblast-derived OCCM-30 cells following IL-1β stimulation. Materials and methods OCCM-30 cells were cultured and exposed to IL-1β. IL-6, IL-1β and TNF-α, and miR-146a-5p expression was assessed by qRT-PCR. Mimics/inhibitors were transiently transfected into cells to determine the function of miR-146a-5p. Signalling pathways including p38 MAPK, ERK1/2 and NF-κB were studied by using specific inhibitors. The indicated proteins were measured by Western blot analysis and ELISA. Results In IL-1β-stimulated OCCM-30 cells, the expression levels of miR-146a-5p along with IL-6 and IL-1β increased in a time-dependent manner. The ERK1/2, p38 MAPK and NF-κB pathway were activated upon IL-1β stimulation. Blocking the NF-κB pathway decreased IL-6, IL-1β and miR-146a-5p expression. The overexpression of miR-146a-5p reduced IL-6 and IL-1β expression, while the inhibition of miR-146a-5p increased IL-6 and IL-1β expression in IL-1β-treated OCCM-30 cells. miR-146a-5p attenuated IL-6 and IL-1β expression via the IRAK1/TRAF6 pathway. Conclusion This study suggested that miR-146a-5p attenuates IL-1β-induced inflammatory factors in Cementoblast-derived cell line.
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Irisin promotes Cementoblast differentiation via p38 MAPK pathway
Oral diseases, 2020Co-Authors: Jiaqi Zhu, Zhengguo Cao, Yunlong Wang, Yunru Hao, Jiawen PanAbstract:Irisin is a newly identified exercise-induced myokine which can affect glucose metabolism and cortical bone mass and strength. However, the influence of irisin on Cementoblasts remains largely unknown. An immortalized mouse Cementoblast cell line OCCM-30 was used in this study. Cementoblast differentiation markers and PGC-1α in cells cultured with mineral induction medium were evaluated by qRT-PCR. Cementoblast mineralization was evaluated by alizarin red staining. Differentiation markers and the activity of p38 MAPK pathway under irisin stimulation were assessed by qRT-PCR or Western blot analysis. p38 MAPK pathway inhibitor SB203580 or p38 siRNA was used to further identify the regulatory mechanism. Cell proliferation treated with irisin was examined by CCK-8 method. The expression of Runx2, osterix, ALP, and PGC-1α was up-regulated consistently under mineral induction. The formation of mineralized nodules was increased by irisin. Runx2, osterix, ALP, and osteocalcin were obviously up-regulated under irisin stimulation as well as the activity of p38 MAPK pathway. When pretreated with SB203580 or p38 siRNA before irisin stimulation, the irisin-induced differentiation was distinctly suppressed. OCCM-30 cell proliferation was enhanced when treated with high-dose irisin for long time. Irisin can promote the differentiation of Cementoblasts via p38 MAPK pathway. © 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. All rights reserved.
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irisin promotes Cementoblast differentiation via p38 mapk pathway
Oral Diseases, 2020Co-Authors: Jiaqi Zhu, Zhengguo Cao, Yunlong Wang, Yunru Hao, Jiawen PanAbstract:Objective Irisin is a newly identified exercise-induced myokine which can affect glucose metabolism and cortical bone mass and strength. However, the influence of irisin on Cementoblasts remains largely unknown. Material and methods An immortalized mouse Cementoblast cell line OCCM-30 was used in this study. Cementoblast differentiation markers and PGC-1α in cells cultured with mineral induction medium were evaluated by qRT-PCR. Cementoblast mineralization was evaluated by alizarin red staining. Differentiation markers and the activity of p38 MAPK pathway under irisin stimulation were assessed by qRT-PCR or Western blot analysis. p38 MAPK pathway inhibitor SB203580 or p38 siRNA was used to further identify the regulatory mechanism. Cell proliferation treated with irisin was examined by CCK-8 method. Results The expression of Runx2, osterix, ALP, and PGC-1α was up-regulated consistently under mineral induction. The formation of mineralized nodules was increased by irisin. Runx2, osterix, ALP, and osteocalcin were obviously up-regulated under irisin stimulation as well as the activity of p38 MAPK pathway. When pretreated with SB203580 or p38 siRNA before irisin stimulation, the irisin-induced differentiation was distinctly suppressed. OCCM-30 cell proliferation was enhanced when treated with high-dose irisin for long time. Conclusion Irisin can promote the differentiation of Cementoblasts via p38 MAPK pathway.
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MiR-361-3p/Nfat5 Signaling Axis Controls Cementoblast Differentiation:
Journal of dental research, 2019Co-Authors: Haiqing Liao, Chenxi Jiang, H Liu, H. Sun, Xiaoxuan Wang, Junbo Xiang, Zhengguo CaoAbstract:The development of periodontal tissue is a complex process, including Cementoblast proliferation and differentiation. Emerging reports suggest that microRNAs (miRNAs) play crucial roles in gene regulatory networks governing numerous biological processes. However, how miRNAs modulate Cementoblast proliferation and differentiation remains largely unknown. In a previous study, we performed miRNA microarray profiling to fully reveal the expression patterns of miRNAs involved in Cementoblast differentiation. We focused on miR-361-3p, which decreased during Cementoblast differentiation. Overexpression of miR-361-3p resulted in decreased Cementoblast differentiation, whereas the functional inhibition of miR-361-3p yielded the opposite effect. The bioinformatics approach identified nuclear factor of activated T-cell 5 (Nfat5) as a potential target of miR-361-3p, which was further verified by dual luciferase assay. Meanwhile, the expression pattern of Nfat5 was verified both in vitro and in vivo. Furthermore, knockdown of Nfat5 mimicked the inhibitory effect of overexpressing miR-361-3p in Cementoblasts. Moreover, multiple signaling pathways, including the Erk1/2, JNK, p38, PI3K-Akt, and NF-κB pathways, were notably activated, and the Wnt/s-catenin pathway was blocked by downregulation of Nfat5 or forced expression of miR-361-3p in Cementoblast differentiation. Finally, the complementary approach demonstrated that miR-361-3p regulated Cementoblast differentiation via or partially via Erk1/2 and PI3K-Akt. Overall, our study elucidated that the JNK, p38, NF-κB, and Wnt/s-catenin pathways act as balancing players in the miR-361-3p/Nfat5 signaling axis during Cementoblast differentiation.
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CXXC5 Mediates P. gingivalis-suppressed Cementoblast Functions Partially via MAPK Signaling Network.
International journal of biological sciences, 2019Co-Authors: Xiaoxuan Wang, Chenxi Jiang, H Liu, Haiqing Liao, Yi Guo, Zhengguo CaoAbstract:Porphyromonas (P.) gingivalis associates tightly with periodontal diseases and it is also a dominant pathogen of periapical periodontitis. However, the influence of P. gingivalis on Cementoblasts, root surface cells pivotal in the apical areas, and the possible involvement of other molecules remain largely elusive. CXXC5 is a nuclear protein that regulates gene expression as well as cell growth, differentiation, and apoptosis. In this study, P. gingivalis repressed the mineralization capacity of Cementoblasts by inducing inflammatory reactions and inhibiting cell differentiation. Intriguingly, the expression of CXXC5 decreased in P. gingivalis-treated OCCM-30 cells and apical periodontitis models but gradually increased during mineralization. Furthermore, RNA interference of CXXC5 significantly inhibited Cementoblast differentiation, represented by decline of bone-associated markers Osterix, osteocalcin (OCN), and alkaline phosphatase (ALP). CXXC5 overexpression facilitated differentiation, and therefore attenuated the P. gingivalis-repressed effects on OCCM-30 cells. In addition, Erk1/2, p38, and PI3K-Akt were inactivated by silencing CXXC5 and activated upon its overexpression, whereas Wnt/β-catenin exhibited an opposite trend. The employment of specific inhibitors revealed that the CXXC5-dependent promotions of Cementoblast differentiation were partially abrogated by p38 and PI3K-Akt inhibitors but were exacerbated by inhibiting Erk1/2. Overall, our experiment demonstrated a novel function of CXXC5 in the regeneration of impaired cementum caused by P. gingivalis invasion and suggested that MAPK signaling network balances the facilitation effects of CXXC5 in Cementoblast differentiation.
Takashi Takata - One of the best experts on this subject based on the ideXlab platform.
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Identification of regulatory mRNA and microRNA for differentiation into Cementoblasts and periodontal ligament cells
Journal of periodontal research, 2020Co-Authors: Tomoyuki Iwata, Masae Kitagawa, Noriyoshi Mizuno, Takayoshi Nagahara, Eri Kaneda-ikeda, Mikihito Kajiya, Katsuhiro Takeda, Minami Yoshioka, Ryoichi Yagi, Takashi TakataAbstract:OBJECTIVE Periodontitis causes periodontal tissue destruction and results in physiological tooth dysfunction. Therefore, periodontal regeneration is ideal therapy for periodontitis. Mesenchymal stem cells (MSCs) are useful for periodontal regenerative therapy as they can differentiate into periodontal cells; however, the underlying regulatory mechanism is unclear. In this study, we attempted to identify regulatory genes involved in periodontal cell differentiation and clarify the differentiation mechanism for effective periodontal regenerative therapy. BACKGROUND The cementum and periodontal ligament play important roles in physiological tooth function. Therefore, cementum and periodontal ligament regeneration are critical for periodontal regenerative therapy. Mesenchymal stem cell transplantation can be a common periodontal regenerative therapy because these cells have multipotency and self-renewal ability, which induces new cementum or periodontal ligament formation. Moreover, MSCs can differentiate into Cementoblasts. Cementoblast- or periodontal ligament cell-specific proteins have been reported; however, it is unclear how these proteins are regulated. MicroRNA (miRNA) can also act as a key regulator of MSC function. Therefore, in this study, we identified regulatory genes involved in Cementoblast or periodontal cell differentiation and commitment. METHODS Human MSCs (hMSCs), Cementoblasts (HCEM), and periodontal ligament cells (HPL cells) were cultured, and mRNA or miRNA expression was evaluated. Additionally, Cementoblast-specific genes were overexpressed or suppressed in hMSCs and their expression levels were investigated. RESULTS HCEM and HPL cells expressed characteristic genes, of which we focused on ets variant 1 (ETV1), miR-628-5p, and miR-383 because ETV1 is a differentiation-related transcription factor, miR-628-5p was the second-highest expressed gene in HCEM and lowest expressed gene in HPL cells, and miR-383 was the highest expressed gene in HCEM. miR-628-5p and miR-383 overexpression in hMSCs regulated ETV1 mRNA expression, and miR-383 overexpression downregulated miR-628-5p expression. Moreover, miR-383 suppression decreased miR-383 expression and enhanced ETV1 mRNA expression, but miR-383 suppression also decreased miR-628-5p. Furthermore, silencing of ETV1 expression in hMSCs regulated miR-628-5p and miR-383 expression. Concerning periodontal cell commitment, miR-628-5p, miR-383, and ETV1 regulated the expression of HCEM- or HPL cell-related genes by adjusting the expression of these miRNAs. CONCLUSION HCEM and HPL cells show characteristic mRNA and miRNA profiles. In particular, these cells have specific miR-383, miR-628-5p, and ETV1 expression patterns, and these genes interact with each other. Therefore, miR-383, miR-628-5p, and ETV1 are key genes involved in cementogenesis or HPL cell differentiation.
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the c terminus of amelogenin enhances osteogenic differentiation of human Cementoblast lineage cells
Journal of Periodontal Research, 2017Co-Authors: Ryo Kunimatsu, Takashi Takata, Mutsumi Miyauchi, Yuki Yoshimi, Naoto Hirose, Tetsuya Awada, Li Zhu, Pamela Denbesten, Kotaro TanimotoAbstract:Background and Objectives Amelogenin proteins are the major constituent of developing extracellular enamel matrix and are believed to have an exclusively epithelial origin. Recent studies have suggested that amelogenins might induce the differentiation and maturation of various cells, including Cementoblast lineage cells. However, the residues comprising the active site of amelogenin remain unclear. The purpose of this study was to identify the active site region of amelogenin by studying the effects of amelogenin fragments on the osteogenic differentiation of Cementoblasts. Material and Methods Amelogenin fragments lacking the C-terminus (rh163) and N-terminus (rh128) and a fragment consisting of the C-terminal region of rh174 (C11 peptide) were synthesized and purified. Human Cementoblast lineage cells were cultured in osteogenic differentiation medium and treated with 0, 10, 100 or 1000 ng/mL of rh163, rh128 or C11 peptide. The mRNA levels of bone markers were examined by real-time polymerase chain reaction analysis. Alkaline phosphatase activity and calcium deposition were also determined. Mineralization was evaluated by alizarin red staining. Results The osteogenic differentiation of human Cementoblast lineage cells was significantly enhanced by treatment with rh128 or C11 peptide, whereas rh163 had no significant effect as compared with untreated controls. Conclusions The C-terminus of amelogenin promotes the osteogenic differentiation of human Cementoblast lineage cells, indicating the possible utility of C11 peptide in periodontal tissue regeneration.
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The C-terminus of amelogenin enhances osteogenic differentiation of human Cementoblast lineage cells.
Journal of periodontal research, 2016Co-Authors: Ryo Kunimatsu, Takashi Takata, Mutsumi Miyauchi, Yuki Yoshimi, Naoto Hirose, Tetsuya Awada, Li Zhu, Pamela Denbesten, Kotaro TanimotoAbstract:Amelogenin proteins are the major constituent of developing extracellular enamel matrix and are believed to have an exclusively epithelial origin. Recent studies have suggested that amelogenins might induce the differentiation and maturation of various cells, including Cementoblast lineage cells. However, the residues comprising the active site of amelogenin remain unclear. The purpose of this study was to identify the active site region of amelogenin by studying the effects of amelogenin fragments on the osteogenic differentiation of Cementoblasts. Amelogenin fragments lacking the C-terminus (rh163) and N-terminus (rh128) and a fragment consisting of the C-terminal region of rh174 (C11 peptide) were synthesized and purified. Human Cementoblast lineage cells were cultured in osteogenic differentiation medium and treated with 0, 10, 100 or 1000 ng/mL of rh163, rh128 or C11 peptide. The mRNA levels of bone markers were examined by real-time polymerase chain reaction analysis. Alkaline phosphatase activity and calcium deposition were also determined. Mineralization was evaluated by alizarin red staining. The osteogenic differentiation of human Cementoblast lineage cells was significantly enhanced by treatment with rh128 or C11 peptide, whereas rh163 had no significant effect as compared with untreated controls. The C-terminus of amelogenin promotes the osteogenic differentiation of human Cementoblast lineage cells, indicating the possible utility of C11 peptide in periodontal tissue regeneration. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
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Acemannan induces Cementoblast proliferation, differentiation, extracellular matrix secretion, and mineral deposition
Journal of Medicinal Plants Research, 2012Co-Authors: Dusida Sahawat, Polkit Sangvanich, Suparaporn Kanthasuwan, Takashi Takata, Masae KitagawaAbstract:Acemannan, a polysaccharide extract from Aloe vera gel, has been reported to induce oral wound healing and dentin regeneration in vitro and in vivo. However, the regenerative effect of acemannan on Cementoblasts, cementum forming cells, has not been defined. In this study, we hypothesized acemannan would induce Cementoblast proliferation, differentiation, extracellular matrix and growth factor secretion, and mineral deposition. An immortalized human Cementoblast cell line (HCEM-2) was treated with acemannan in vitro. The de novo DNA synthesis, alkaline phosphatase activity, collagen type I and vascular endothelial growth factor, and osteopontin expression were determined by [ 3 H] thymidine incorporation, biochemical assay, enzyme linked immunosorbent assay, and western blotting, respectively. In vitro mineral nodule formation was determined on day 21 of incubation using alizarin red staining. Our study revealed acemannan at concentrations of 2, 4, and 8 mg/ml significantly stimulated Cementoblast proliferation approximately 2.5 fold. Acemannan also significantly enhanced alkaline phosphatase activity at day 3 and 6 of incubation. Expression of type I collagen, vascular endothelial growth factor, and osteopontin were significantly increased compared with the untreated group. At day 21, acemannan treatment markedly induced mineral deposition compared with the untreated group. These results suggest acemannan may be a possible therapeutic agent for cementum regeneration.
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f spondin regulates the differentiation of human Cementoblast like hcem cells via bmp7 expression
Biochemical and Biophysical Research Communications, 2012Co-Authors: Masae Kitagawa, Mutsumi Miyauchi, Yoshimitsu Abiko, Takashi TakataAbstract:Abstract Cementum plays an important role in the attachment of connective tissue to the root surface. However, the detailed mechanism of cementum formation has not yet been clarified. We previously established human Cementoblast-like cell lines (HCEM) and human periodontal ligament cell lines (HPL) by infection of hTERT gene. Using those cell lines, we compared the gene expression of them and identified F-spondin as a Cementoblast specific gene. In this study, to clarify the role of F-spondin in the differentiation of periodontal ligament cells to Cementoblasts, we compared the gene expression of F-spondin-overexpressed HPL (HPL-spondin) cells with HPL parent cells. We found that several genes expressed higher level in HPL-spondin cells than in HPL cells, such as heparin sulfate 6-sulfotranferase, calcitonin-related polypeptide beta, bone morphogenetic proteins 7 (BMP7), BMP2 and BMP8B. Among those genes, we focused on BMP7 and examined the interaction between F-spondin and BMP7, because BMP7 was reported to enhance Cementoblast function. Moreover, we further examined the effect of BMP7 peptide on the expression of mineralization-associated genes in HCEM cells. RT-PCR and real-time PCR analyses showed that HPL-spondin expressed BMP7, but not HPL cells. And BMP7 and phospho-Smad1/5/8 protein production were detected in HPL-spondin by Western blot. siSPON1 inhibited expression of type I collagen, runt-related transcription factor 2 (RUNX2) and bone sialoprotein (BSP) mRNA in HCEM cells. And the mineralization tended to be decreased in siSPON1 treated cells by ALZ staining and the quantification analysis. Moreover, we examined the effect of BMP7 peptide on the gene expressions of HCEM cells by RT-PCR. Increase of the osteopontin and BSP mRNA was observed in BMP7 treated HCEM cells. These findings indicate that F-spondin regulates the differentiation of HCEM cells via BMP7 expression.
Weixiong Xiao - One of the best experts on this subject based on the ideXlab platform.
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Article Multi-Walled Carbon Nanotubes Promote Cementoblast Differentiation and Mineralization through the TGF-β/Smad Signaling Pathway
2016Co-Authors: Zhimin Zhu, Weixiong XiaoAbstract:Abstract: Excretion of cementum by Cementoblasts on the root surface is a process indispensable for the formation of a functional periodontal ligament. This study investigated whether carboxyl group-functionalized multi-walled carbon nanotubes (MWCNT-COOH) could enhance differentiation and mineralization of mammalian Cementoblasts (OCCM-30) and the possible signaling pathway involved in this process. Cementoblasts were incubated with various doses of MWCNT-COOH suspension. Cell viability was detected, and a scanning electron microscopy (SEM) observed both the nanomaterials and the growth of cells cultured with the materials. Alizarin red staining was used to investigate the formation of calcium deposits. Real-time PCR and western blot were used to detect Cementoblast differentiation and the underlying mechanisms through the expression of the osteogenic genes and the downstream effectors of the TGF-β/Smad signaling. The results showed that 5 µg/mL MWCNT-COOH had the most obvious effects on promoting differentiation without significant toxicity. Alp, Ocn, Bsp, Opn, Col1 and Runx2 gene expression was up-regulated. Smad2 and Smad3 mRNA was up-regulated, while Smad7 was first down-regulated on Day 3 and later up-regulated on Day 7. The elevated level
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Multi-walled carbon nanotubes promote Cementoblast differentiation and mineralization through the TGF-β/Smad signaling pathway.
International journal of molecular sciences, 2015Co-Authors: Zhimin Zhu, Weixiong XiaoAbstract:Excretion of cementum by Cementoblasts on the root surface is a process indispensable for the formation of a functional periodontal ligament. This study investigated whether carboxyl group-functionalized multi-walled carbon nanotubes (MWCNT-COOH) could enhance differentiation and mineralization of mammalian Cementoblasts (OCCM-30) and the possible signaling pathway involved in this process. Cementoblasts were incubated with various doses of MWCNT-COOH suspension. Cell viability was detected, and a scanning electron microscopy (SEM) observed both the nanomaterials and the growth of cells cultured with the materials. Alizarin red staining was used to investigate the formation of calcium deposits. Real-time PCR and western blot were used to detect Cementoblast differentiation and the underlying mechanisms through the expression of the osteogenic genes and the downstream effectors of the TGF-β/Smad signaling. The results showed that 5 µg/mL MWCNT-COOH had the most obvious effects on promoting differentiation without significant toxicity. Alp, Ocn, Bsp, Opn, Col1 and Runx2 gene expression was up-regulated. Smad2 and Smad3 mRNA was up-regulated, while Smad7 was first down-regulated on Day 3 and later up-regulated on Day 7. The elevated levels of phospho-Smad2/3 were also confirmed by western blot. In sum, the MWCNT-COOH promoted Cementoblast differentiation and mineralization, at least partially, through interactions with the TGF-β/Smad pathway.
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multi walled carbon nanotubes promote Cementoblast differentiation and mineralization through the tgf β smad signaling pathway
International Journal of Molecular Sciences, 2015Co-Authors: Zhimin Zhu, Weixiong XiaoAbstract:Excretion of cementum by Cementoblasts on the root surface is a process indispensable for the formation of a functional periodontal ligament. This study investigated whether carboxyl group-functionalized multi-walled carbon nanotubes (MWCNT-COOH) could enhance differentiation and mineralization of mammalian Cementoblasts (OCCM-30) and the possible signaling pathway involved in this process. Cementoblasts were incubated with various doses of MWCNT-COOH suspension. Cell viability was detected, and a scanning electron microscopy (SEM) observed both the nanomaterials and the growth of cells cultured with the materials. Alizarin red staining was used to investigate the formation of calcium deposits. Real-time PCR and western blot were used to detect Cementoblast differentiation and the underlying mechanisms through the expression of the osteogenic genes and the downstream effectors of the TGF-β/Smad signaling. The results showed that 5 µg/mL MWCNT-COOH had the most obvious effects on promoting differentiation without significant toxicity. Alp, Ocn, Bsp, Opn, Col1 and Runx2 gene expression was up-regulated. Smad2 and Smad3 mRNA was up-regulated, while Smad7 was first down-regulated on Day 3 and later up-regulated on Day 7. The elevated levels of phospho-Smad2/3 were also confirmed by western blot. In sum, the MWCNT-COOH promoted Cementoblast differentiation and mineralization, at least partially, through interactions with the TGF-β/Smad pathway.
Haiqing Liao - One of the best experts on this subject based on the ideXlab platform.
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MiR-361-3p/Nfat5 Signaling Axis Controls Cementoblast Differentiation:
Journal of dental research, 2019Co-Authors: Haiqing Liao, Chenxi Jiang, H Liu, H. Sun, Xiaoxuan Wang, Junbo Xiang, Zhengguo CaoAbstract:The development of periodontal tissue is a complex process, including Cementoblast proliferation and differentiation. Emerging reports suggest that microRNAs (miRNAs) play crucial roles in gene regulatory networks governing numerous biological processes. However, how miRNAs modulate Cementoblast proliferation and differentiation remains largely unknown. In a previous study, we performed miRNA microarray profiling to fully reveal the expression patterns of miRNAs involved in Cementoblast differentiation. We focused on miR-361-3p, which decreased during Cementoblast differentiation. Overexpression of miR-361-3p resulted in decreased Cementoblast differentiation, whereas the functional inhibition of miR-361-3p yielded the opposite effect. The bioinformatics approach identified nuclear factor of activated T-cell 5 (Nfat5) as a potential target of miR-361-3p, which was further verified by dual luciferase assay. Meanwhile, the expression pattern of Nfat5 was verified both in vitro and in vivo. Furthermore, knockdown of Nfat5 mimicked the inhibitory effect of overexpressing miR-361-3p in Cementoblasts. Moreover, multiple signaling pathways, including the Erk1/2, JNK, p38, PI3K-Akt, and NF-κB pathways, were notably activated, and the Wnt/s-catenin pathway was blocked by downregulation of Nfat5 or forced expression of miR-361-3p in Cementoblast differentiation. Finally, the complementary approach demonstrated that miR-361-3p regulated Cementoblast differentiation via or partially via Erk1/2 and PI3K-Akt. Overall, our study elucidated that the JNK, p38, NF-κB, and Wnt/s-catenin pathways act as balancing players in the miR-361-3p/Nfat5 signaling axis during Cementoblast differentiation.
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CXXC5 Mediates P. gingivalis-suppressed Cementoblast Functions Partially via MAPK Signaling Network.
International journal of biological sciences, 2019Co-Authors: Xiaoxuan Wang, Chenxi Jiang, H Liu, Haiqing Liao, Yi Guo, Zhengguo CaoAbstract:Porphyromonas (P.) gingivalis associates tightly with periodontal diseases and it is also a dominant pathogen of periapical periodontitis. However, the influence of P. gingivalis on Cementoblasts, root surface cells pivotal in the apical areas, and the possible involvement of other molecules remain largely elusive. CXXC5 is a nuclear protein that regulates gene expression as well as cell growth, differentiation, and apoptosis. In this study, P. gingivalis repressed the mineralization capacity of Cementoblasts by inducing inflammatory reactions and inhibiting cell differentiation. Intriguingly, the expression of CXXC5 decreased in P. gingivalis-treated OCCM-30 cells and apical periodontitis models but gradually increased during mineralization. Furthermore, RNA interference of CXXC5 significantly inhibited Cementoblast differentiation, represented by decline of bone-associated markers Osterix, osteocalcin (OCN), and alkaline phosphatase (ALP). CXXC5 overexpression facilitated differentiation, and therefore attenuated the P. gingivalis-repressed effects on OCCM-30 cells. In addition, Erk1/2, p38, and PI3K-Akt were inactivated by silencing CXXC5 and activated upon its overexpression, whereas Wnt/β-catenin exhibited an opposite trend. The employment of specific inhibitors revealed that the CXXC5-dependent promotions of Cementoblast differentiation were partially abrogated by p38 and PI3K-Akt inhibitors but were exacerbated by inhibiting Erk1/2. Overall, our experiment demonstrated a novel function of CXXC5 in the regeneration of impaired cementum caused by P. gingivalis invasion and suggested that MAPK signaling network balances the facilitation effects of CXXC5 in Cementoblast differentiation.
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MicroRNA‐181b‐5p modulates tumor necrosis factor‐α‐induced inflammatory responses by targeting interleukin‐6 in Cementoblasts
Journal of cellular physiology, 2019Co-Authors: Xiaoxuan Wang, Chenxi Jiang, Haiqing Liao, Junbo Xiang, Hualing Sun, Huan Liu, Zhengguo CaoAbstract:Tooth cementum is a bone-like mineralized tissue and serves as a microbial barrier against invasion and destruction. Cementum is also responsible for tooth stability and defending pulp from outside stimuli, which is formed by Cementoblasts. Although it is crucial for periodontal and periapical diseases, the mechanisms underlying the pathophysiological changes of Cementoblasts and their inflammatory responses remain unclear. MiR-181b is found to modulate vascular inflammation and endotoxin tolerance. In this study, miR-181b-5p was downregulated in tumor necrosis factor-α (TNF-α)-stimulated Cementoblasts, whereas proinflammatory molecules increased. The mouse periapical lesions have similar results, which imitate an inflammatory environment for Cementoblasts in vivo. The bioinformatics analysis and dual luciferase reporter assay suggested that miR-181b-5p targeted interleukin-6 (IL-6). Overexpressing miR-181b-5p negatively regulated IL-6 and proinflammatory chemokine. Western blot analysis and luciferase activity reporter assay verified that miR-181b-5p weakened the NF-κB activity. Hence, miR-181b-5p moderated proinflammatory chemokine production by targeting IL-6 in Cementoblasts and NF-κB signaling pathway was involved. Furthermore, miR-181b-5p promoted Cementoblast apoptosis, which may enhance the resolution of inflammation. Overall, our data revealed that miR-181b-5p was a negative regulator of TNF-α-induced inflammatory responses in Cementoblasts.
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Downregulation of angiopoietin-like protein 2 inhibits Cementoblast differentiation partially by activating the ERK1/2 signaling pathway.
American journal of translational research, 2019Co-Authors: Chenxi Jiang, H Liu, H. Sun, Xiaoxuan Wang, Haiqing Liao, Zhengguo CaoAbstract:Angiopoietin-like protein 2 (ANGPTL2) is abundantly expressed in adipose tissue, is associated with tissue homeostasis, and promotes osteoblast and chondrocyte differentiation. In teeth, cementum, a thin layer of mineralized tissue that is formed by Cementoblasts, covers the entire root surface and is a vital component of periodontium. The Cementoblasts regulate the deposition and mineralization of the cementum matrix. However, the effects of ANGPTL2 on Cementoblast differentiation have not been studied. The objective of this study was to elucidate the role of ANGPTL2 during Cementoblast differentiation and determine its underlying mechanisms. Our results showed that the expression levels of ANGPTL2 gradually increased during Cementoblast differentiation. After ANGPTL2 was knocked down using short-hairpin RNA, the levels of the osteogenic markers osterix (OSX), alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin (OCN) decreased. In addition, ALP activity and the number of calcified nodules were dramatically reduced compared with those in the negative control. Interestingly, the ERK1/2 signaling pathway was activated after ANGPTL2 knockdown. Treatment with PD98059, the inhibitor of the ERK1/2 signaling pathway, partially rescued the decreased differentiation capability of Cementoblast caused by ANGPTL2 downregulation. Collectively, ANGPTL2 knockdown inhibited Cementoblast differentiation partially by activating the ERK1/2 signaling pathway. These findings suggest that ANGPTL2 was indispensable in Cementoblast differentiation.
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downregulation of angiopoietin like protein 2 inhibits Cementoblast differentiation partially by activating the erk1 2 signaling pathway
American Journal of Translational Research, 2019Co-Authors: Chenxi Jiang, H Liu, H. Sun, Xiaoxuan Wang, Haiqing Liao, Zhengguo CaoAbstract:Angiopoietin-like protein 2 (ANGPTL2) is abundantly expressed in adipose tissue, is associated with tissue homeostasis, and promotes osteoblast and chondrocyte differentiation. In teeth, cementum, a thin layer of mineralized tissue that is formed by Cementoblasts, covers the entire root surface and is a vital component of periodontium. The Cementoblasts regulate the deposition and mineralization of the cementum matrix. However, the effects of ANGPTL2 on Cementoblast differentiation have not been studied. The objective of this study was to elucidate the role of ANGPTL2 during Cementoblast differentiation and determine its underlying mechanisms. Our results showed that the expression levels of ANGPTL2 gradually increased during Cementoblast differentiation. After ANGPTL2 was knocked down using short-hairpin RNA, the levels of the osteogenic markers osterix (OSX), alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin (OCN) decreased. In addition, ALP activity and the number of calcified nodules were dramatically reduced compared with those in the negative control. Interestingly, the ERK1/2 signaling pathway was activated after ANGPTL2 knockdown. Treatment with PD98059, the inhibitor of the ERK1/2 signaling pathway, partially rescued the decreased differentiation capability of Cementoblast caused by ANGPTL2 downregulation. Collectively, ANGPTL2 knockdown inhibited Cementoblast differentiation partially by activating the ERK1/2 signaling pathway. These findings suggest that ANGPTL2 was indispensable in Cementoblast differentiation.