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Junqi Ling - One of the best experts on this subject based on the ideXlab platform.
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nkd2 promotes the differentiation of Dental Follicle stem progenitor cells into osteoblasts
International Journal of Molecular Medicine, 2018Co-Authors: Chanchan Chen, Jianying Zhang, Junqi Ling, Yuluan HouAbstract:Dental Follicle stem/progenitor cells have the potential to undergo osteogenesis. naked cuticle homolog 2 (Nkd2) is a signal‑inducible feedback antagonist of the canonical Wnt signaling pathway. The purpose of the present study was to investigate the function of Nkd2 in the differentiation of Dental Follicle stem/progenitor cells (DFSCs) into osteoblasts. Immunohistochemistry, reverse transcription‑quantitative polymerase chain reaction and western blotting were employed to detect Nkd2 expression in rat DFSCs. In addition, rat DFSCs (rDFSCs) were transfected with small interfering RNAs to examine the effect of Nkd2 on the differentiation of these cells into osteoblasts. Furthermore, the function of Nkd2 in the Wnt/β‑catenin pathway in rDFSCs was investigated using β‑catenin/T‑cell factor luciferase activity assays and western blotting. It was revealed that the expression of Nkd2 was upregulated during the differentiation of rDFSCs into osteoblasts. Furthermore, osteoblast differentiation ability and Wnt/β‑catenin pathway activity were significantly decreased in Nkd2‑silenced rDFSCs compared with the si‑NC group (P<0.05 and P<0.001, respectively). The results suggest that Nkd2 promotes the differentiation of rDFSCs into osteoblasts through Wnt/β‑catenin signaling.
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Nkd2 promotes the differentiation of Dental Follicle stem/progenitor cells into osteoblasts.
International Journal of Molecular Medicine, 2018Co-Authors: Chanchan Chen, Jianying Zhang, Junqi Ling, Yu DuAbstract:: Dental Follicle stem/progenitor cells have the potential to undergo osteogenesis. naked cuticle homolog 2 (Nkd2) is a signal‑inducible feedback antagonist of the canonical Wnt signaling pathway. The purpose of the present study was to investigate the function of Nkd2 in the differentiation of Dental Follicle stem/progenitor cells (DFSCs) into osteoblasts. Immunohistochemistry, reverse transcription‑quantitative polymerase chain reaction and western blotting were employed to detect Nkd2 expression in rat DFSCs. In addition, rat DFSCs (rDFSCs) were transfected with small interfering RNAs to examine the effect of Nkd2 on the differentiation of these cells into osteoblasts. Furthermore, the function of Nkd2 in the Wnt/β‑catenin pathway in rDFSCs was investigated using β‑catenin/T‑cell factor luciferase activity assays and western blotting. It was revealed that the expression of Nkd2 was upregulated during the differentiation of rDFSCs into osteoblasts. Furthermore, osteoblast differentiation ability and Wnt/β‑catenin pathway activity were significantly decreased in Nkd2‑silenced rDFSCs compared with the si‑NC group (P
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naked cuticle homolog 2 positively regulates the osteogenic differentiation of rat Dental Follicle cells
Chinese journal of stomatology, 2017Co-Authors: Junqi Ling, Chanchan Chen, Jingjing Quan, Yu DuAbstract:Objective To examine the expression of naked cuticle homolog 2 (Nkd2) in the process of root development and osteogenic differentiation of Dental Follicle cells of rat (rDFC), in order to explore the molecular mechanisms of Nkd2 on the osteoblast differentiation of rDFCs. Methods Immunohistochemical analysis was used to detect the expression of Nkd2 in the base Dental Follicle of the mandibular first molar of rat at 1, 3, 5, 7, 9, 11 and 13 days postnatal. Mineralization nodule formation of rDFCs was detected by alizarin red staining and cetylpyridine. The change of Nkd2 during osteogenic differentiation of rDFCs was evaluated by Western blotting and the associations between Nkd2 and osteogenic cytokines of alkaline phosphatase (ALP), Runt-related transcription factor-2 (RUNX2) and osteocalcin (OCN) were examined. The rDFCs were transfected with small interfering RNA (siRNA) to knock down the expression of Nkd2 and Western blotting and quantitative real-time PCR (qPCR) were adopted to explore the effects of Nkd2 on osteogenic differentiation by detecting variations of Nkd2 and osteogenic factors ALP, RUNX2, OCN among silencing group (Si), negative control RNA group (Nc) and mock control group (Mock), respectively. Results The expression of Nkd2 in the base Dental Follicle of the mandibular first molar of rat was time dependent. Mineralization nodules of rDFCs and absorbance of cetylpyridine after osteogenic induction increased gradually (the absorbances of cetylpyridine were 0 week: 0.017±0.005, 1 week: 0.702±0.044, 2 weeks: 1.812±0.531, 3 weeks: 2.767±0.253, respectively). Results of Western blotting showed that Nkd2 (1.60±0.23) of mineralization group was significantly higher than that of control group (1) (P<0.05) at the early stage of osteogenic differentiation along with the expression of other osteogenic factors. The protein and mRNA of Nkd2 and osteogenic factors were significantly decreased in Si group compared with Nc and Mock groups (P<0.05), and no changes between Nc and Mock groups were observed. The changes of protein in Si, Nc and Mock groups were Nkd2: 0.42±0.10, 1.12±0.07, 1, ALP: 0.70±0.15, 1.11±0.14, 1, RUNX2: 0.58±0.08, 0.93±0.08, 1 and OCN: 0.64±0.06, 0.99±0.02, 1, respectively. The mRNA variances in Si, Nc and Mock groups were Nkd2: 0.39±0.05, 0.96±0.10, 1, ALP: 0.15±0.13, 1.01±0.07, 1, RUNX2: 0.39±0.31, 0.97±0.13, 1, OCN: 0.17±0.08, 1.08±0.21, 1, respectively. Conclusions Nkd2 participates in the root development process in rat and may acts as a positive role in the early stage of osteogenic differentiation of rDFCs in rat. Key words: Dental Follicle; Naked cuticle homolog 2; Dental Follicle cells; Osteogenic differentiation
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Naked cuticle homolog 2 positively regulates the osteogenic differentiation of rat Dental Follicle cells
Chinese journal of stomatology, 2017Co-Authors: Junqi Ling, Chanchan Chen, Jingjing Quan, Yu DuAbstract:Objective To examine the expression of naked cuticle homolog 2 (Nkd2) in the process of root development and osteogenic differentiation of Dental Follicle cells of rat (rDFC), in order to explore the molecular mechanisms of Nkd2 on the osteoblast differentiation of rDFCs. Methods Immunohistochemical analysis was used to detect the expression of Nkd2 in the base Dental Follicle of the mandibular first molar of rat at 1, 3, 5, 7, 9, 11 and 13 days postnatal. Mineralization nodule formation of rDFCs was detected by alizarin red staining and cetylpyridine. The change of Nkd2 during osteogenic differentiation of rDFCs was evaluated by Western blotting and the associations between Nkd2 and osteogenic cytokines of alkaline phosphatase (ALP), Runt-related transcription factor-2 (RUNX2) and osteocalcin (OCN) were examined. The rDFCs were transfected with small interfering RNA (siRNA) to knock down the expression of Nkd2 and Western blotting and quantitative real-time PCR (qPCR) were adopted to explore the effects of Nkd2 on osteogenic differentiation by detecting variations of Nkd2 and osteogenic factors ALP, RUNX2, OCN among silencing group (Si), negative control RNA group (Nc) and mock control group (Mock), respectively. Results The expression of Nkd2 in the base Dental Follicle of the mandibular first molar of rat was time dependent. Mineralization nodules of rDFCs and absorbance of cetylpyridine after osteogenic induction increased gradually (the absorbances of cetylpyridine were 0 week: 0.017±0.005, 1 week: 0.702±0.044, 2 weeks: 1.812±0.531, 3 weeks: 2.767±0.253, respectively). Results of Western blotting showed that Nkd2 (1.60±0.23) of mineralization group was significantly higher than that of control group (1) (P
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Wnt5a regulates Dental Follicle stem/progenitor cells of the periodontium
Stem cell research & therapy, 2014Co-Authors: Lusai Xiang, Mo Chen, Bin Cai, Xinchun Zhang, Chen Zhou, Chenglin Wang, Jeremy J. Mao, Junqi LingAbstract:Dental Follicle gives rise to one or several tissues of the periodontium including the periodontal ligament, cementum and/or alveolar bone. Whether Wnt5a is expressed in the postnatal periodontium or regulates Dental Follicle stem/progenitor cells is unknown. Dental Follicle stem/progenitor cells were isolated from postnatal day 1 (p1) to p11 from rat mandibular first molars. Immunolocalization mapped Wnt5a expression in the alveolar bone, periodontal ligament, and the developing ameloblast and odontoblast layers. Mononucleated and adherent cells were isolated from p7 Dental Follicle. Wnt5a was overexpressed in Dental Follicle stem/progenitor cells to study their proliferation, osteogenic differentiation and migration behavior, with subpopulations of native Dental Follicle stem/progenitor cells as controls, using real-time PCR (Taqman), Lenti-viral transfection, Western blotting and immunofluorescence. Wnt5a was expressed consistently in p1 to p11 rat peridontium. Native, p7 Dental Follicle stem/progenitor cells had modest ability to mineralize in the tested 14 days. Even in chemically defined osteogenesis medium, Dental Follicle stem/progenitor cells only showed modest mineralization. Upon addition of 300 ng/mL Wnt5a protein in osteogenesis medium, Dental Follicle stem/progenitor cells displayed mineralization that was still unremarkable. Chemically induced or Wnt5a-induced mineralization of Dental Follicle cells only occurred sparsely. Combination of Wnt5a with 100 ng/mL BMP2 finally prompted Dental Follicle stem/progenitor cells to produce robust mineralization with elevated expression of Runx2, alkaline phosphatase, collagen 1α1 and osteocalcin. Thus, native Dental Follicle stem/progenitor cells or some of their fractions may be somewhat modest in mineralization. Strikingly, Wnt5a protein significantly augmented RANKL ligand, suggesting putative regulatory roles of Dental Follicle stem/progenitor cells for the monocyte/osteoclast lineage and potential involvement in alveolar bone remodeling and/or resorption. P-Jnk1/2 was activated in Wnt5a overexpressed Dental Follicle cells; conversely, exposure to SP600125, a c-Jun N-terminal kinase (JNK) inhibitor attenuated Runx2, collagen 1α1 and osteocalcin expression either in the presence or absence of Wnt5a. Wnt5a overexpression in Dental Follicle stem/progenitor cells significantly reduced their proliferation rates, but robustly augmented their migration capacity. These findings provide a glimpse of Wnt5a’s putative roles in Dental Follicle stem/progenitor cells and the periodontium with implications in periodontal disease, tooth eruption, Dental implant bone healing and orthodontic tooth movement.
Gary E. Wise - One of the best experts on this subject based on the ideXlab platform.
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Hypoxia promotes growth of stem cells in Dental Follicle cell populations
Journal of biomedical science and engineering, 2011Co-Authors: Yuntao Dai, Gary E. Wise, Shaomian YaoAbstract:Adult stem cells (ASC) have been found in many tis-sues and are of great therapeutic potential due to their capability of differentiation. However, ASC comprise only a small fraction of the tissues. In order to use ASC for therapeutic purposes, it is important to obtain relatively pure stem cells in large quantities. Current methods for stem cell purification are mainly based on marker-dependent cell sorting techniques, which have various technical difficulties. In this study, we have attempted to develop novel conditions to favor the growth of the Dental Follicle stem cells (DFSC) such that the resultant cell populations are enriched in stem cells. Specifically, a heterogeneous Dental Follicle cell (H-DFC) population containing stem cells and homogenous non-stem cell Dental Follicle cell population were cultured at 1% or 5% hypoxic conditions. Only the heterogeneous population could increase proliferation in the hypoxic condition whereas the homogenous DFC did not change their proliferation rate. In addition, when the resultant cells from the heterogonous population were subjected to differentiation, they appeared to have a higher capacity of adipogenesis and osteogenesis as compared to the controls grown in the normal at-mosphere (normoxic condition). These hypoxia- treated cells also express higher levels of some stem cell markers. Together, these data suggest that stem cells are enriched by culturing the heterogeneous cell populations in a reduced O2 condition.
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tnf alpha upregulates expression of bmp 2 and bmp 3 genes in the rat Dental Follicle implications for tooth eruption
Connective Tissue Research, 2010Co-Authors: Shaomian Yao, Veronica Prpic, Fenghui Pan, Gary E. WiseAbstract:The Dental Follicle appears to regulate both the alveolar bone resorption and bone formation needed for tooth eruption. Tumor necrosis factor-alpha (TNF-alpha) gene expression is maximally upregulated at postnatal day 9 in the rat Dental Follicle of the first mandibular molar, a time that correlates with rapid bone growth at the base of the tooth crypt, as well as a minor burst of osteoclastogenesis. TNF-alpha expression is correlated with the expression of bone morphogenetic protein-2 (BMP-2), a molecule expressed in the Dental Follicle that can promote bone formation. Because BMP-2 signaling may be augmented by bone morphogenetic protein-3 (BMP-3), our objective in this study was to determine if the Dental Follicle expresses BMP-3 and if TNF-alpha stimulates the Dental Follicle cells to express BMP-2 and BMP-3. Dental Follicles were collected from different postnatal ages of rat pups. Dental Follicle cells were incubated with TNF-alpha to study its dosage and time-course effects on gene expression of BMP-2 and BMP-3, as determined by real-time RT-PCR. Next, immunostaining was conducted to confirm if the protein was synthesized and ELISA of the conditioned medium was conducted to determine if BMP-2 was secreted. We found that BMP-3 expression is correlated with the expression of TNF-alpha in the Dental Follicle and TNF-alpha significantly increased BMP-2 and BMP-3 expression in vitro. Immunostaining and ELISA showed that BMP-2 and BMP-3 were synthesized and secreted. This study suggests that TNF-alpha can upregulate the expression of bone formation genes that may be needed for tooth eruption.
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differentiation of stem cells in the Dental Follicle
Journal of Dental Research, 2008Co-Authors: Shaomian Yao, Fenghui Pan, V Prpic, Gary E. WiseAbstract:The Dental Follicle (DF) differentiates into the periodontal ligament. In addition, it may be the precursor of other cells of the periodontium, including osteoblasts and cementoblasts. We hypothesized that stem cells may be present in the DF and be capable of differentiating into cells of the periodontium. Stem cells were identified in the DF of the rat first mandibular molar by Hoechst staining, alkaline phosphatase staining, and expression of side-population stem cell markers. These cells were shown to be able to differentiate into osteoblasts/cementoblasts, adipocytes, and neurons. Treating the DF cell population with doxorubicin, followed by incubation in an adipogenesis medium, suggested that the adipocytes originated from stem cells. Thus, a possibly puripotent stem cell population is present in the rat DF.
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a dna microarray analysis of chemokine and receptor genes in the rat Dental Follicle role of secreted frizzled related protein 1 in osteoclastogenesis
Bone, 2007Co-Authors: Dawen Liu, Gary E. WiseAbstract:The Dental Follicle, a loose connective tissue sac that surrounds the unerupted tooth, appears to regulate the osteoclastogenesis needed for eruption; i.e., bone resorption to form an eruption pathway. Thus, DNA microarray studies were conducted to determine which chemokines and their receptors were expressed chronologically in the Dental Follicle, chemokines that might attract osteoclast precursors. In the rat first mandibular molar, a major burst of osteoclastogenesis occurs at day 3 with a minor burst at day 10. The results of the microarray confirmed our previous studies showing the gene expression of molecules such as CSF-1 and MCP-1 in the Dental Follicle cells. Other new genes also were detected, including secreted frizzled-related protein-1 (SFRP-1), which was found to be downregulated at days 3 and 9. Using rat bone marrow cultures to conduct in vitro osteoclastogenic assays, it was demonstrated that SFRP-1 inhibited osteoclast formation in a concentration-dependent fashion. However, with increasing concentrations of SFRP-1, the number of TRAP-positive mononuclear cells increased suggesting that SFRP-1 inhibits osteoclast formation by inhibiting the fusion of mononuclear cells (osteoclast precursors). Co-culturing bone marrow mononuclear cells and Dental Follicle cells demonstrated that the Dental Follicle cells were secreting a product(s) that inhibited osteoclastogenesis, as measured by counting of TRAP-positive osteoclasts. Adding an antibody either to SFRP-1 or OPG partially restored osteoclastogenesis. Adding both anti-SFRP-1 and anti-OPG fully negated the inhibitory effect of the Follicle cells upon osteoclastogenesis. These results strongly suggest that SFRP-1 and OPG, both secreted by the Dental Follicle cells, use different pathways to exert their inhibitory effect on osteoclastogenesis. Based on these in vitro studies of osteoclastogenesis, it is likely that the downregulation of SFRP-1 gene expression in the Dental Follicle at days 3 and 9 is a contributory factor in allowing the major and minor bursts of osteoclastogenesis to occur. Thus, inhibition of SFRP-1 gene expression in combination with inhibition of OPG gene expression likely are critical events in enabling alveolar bone resorption to occur such that teeth will erupt.
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regional differences of expression of bone morphogenetic protein 2 and rankl in the rat Dental Follicle
European Journal of Oral Sciences, 2006Co-Authors: Gary E. Wise, Shaomian YaoAbstract:Tooth eruption requires alveolar bone resorption and bone formation. The coronal half of the Dental Follicle probably mediates the bone resorption seen in the coronal region of the alveolar bony crypt, and the basal half of the Follicle mediates bone growth in the basal region. We hypothesized that the expression of a gene for bone resorption - receptor activator of nuclear factor kappa B ligand (RANKL) - would be higher in the coronal than in the basal region of the Follicle. Conversely, the level of expression of bone morphogenetic protein-2 (BMP-2), a gene for bone formation, would be higher in the basal region. Results obtained using laser-capture microdissection and real-time reverse transcription-polymerase chain reaction (RT-PCR) confirmed the hypothesis. Scanning electron micrographs of the bony crypt showed that the coronal area of the crypt was scalloped in appearance (bone resorption), whereas the basal area was trabecular (bone formation). Thus, the differences in bone activity at opposite poles of the crypt appear to be caused by differences in the regional expression of genes in the Dental Follicle and suggest a molecular mechanism whereby the Dental Follicle could regulate both the alveolar bone resorption and formation needed for eruption.
Lusai Xiang - One of the best experts on this subject based on the ideXlab platform.
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Wnt5a regulates Dental Follicle stem/progenitor cells of the periodontium
Stem cell research & therapy, 2014Co-Authors: Lusai Xiang, Mo Chen, Bin Cai, Xinchun Zhang, Chen Zhou, Chenglin Wang, Jeremy J. Mao, Junqi LingAbstract:Dental Follicle gives rise to one or several tissues of the periodontium including the periodontal ligament, cementum and/or alveolar bone. Whether Wnt5a is expressed in the postnatal periodontium or regulates Dental Follicle stem/progenitor cells is unknown. Dental Follicle stem/progenitor cells were isolated from postnatal day 1 (p1) to p11 from rat mandibular first molars. Immunolocalization mapped Wnt5a expression in the alveolar bone, periodontal ligament, and the developing ameloblast and odontoblast layers. Mononucleated and adherent cells were isolated from p7 Dental Follicle. Wnt5a was overexpressed in Dental Follicle stem/progenitor cells to study their proliferation, osteogenic differentiation and migration behavior, with subpopulations of native Dental Follicle stem/progenitor cells as controls, using real-time PCR (Taqman), Lenti-viral transfection, Western blotting and immunofluorescence. Wnt5a was expressed consistently in p1 to p11 rat peridontium. Native, p7 Dental Follicle stem/progenitor cells had modest ability to mineralize in the tested 14 days. Even in chemically defined osteogenesis medium, Dental Follicle stem/progenitor cells only showed modest mineralization. Upon addition of 300 ng/mL Wnt5a protein in osteogenesis medium, Dental Follicle stem/progenitor cells displayed mineralization that was still unremarkable. Chemically induced or Wnt5a-induced mineralization of Dental Follicle cells only occurred sparsely. Combination of Wnt5a with 100 ng/mL BMP2 finally prompted Dental Follicle stem/progenitor cells to produce robust mineralization with elevated expression of Runx2, alkaline phosphatase, collagen 1α1 and osteocalcin. Thus, native Dental Follicle stem/progenitor cells or some of their fractions may be somewhat modest in mineralization. Strikingly, Wnt5a protein significantly augmented RANKL ligand, suggesting putative regulatory roles of Dental Follicle stem/progenitor cells for the monocyte/osteoclast lineage and potential involvement in alveolar bone remodeling and/or resorption. P-Jnk1/2 was activated in Wnt5a overexpressed Dental Follicle cells; conversely, exposure to SP600125, a c-Jun N-terminal kinase (JNK) inhibitor attenuated Runx2, collagen 1α1 and osteocalcin expression either in the presence or absence of Wnt5a. Wnt5a overexpression in Dental Follicle stem/progenitor cells significantly reduced their proliferation rates, but robustly augmented their migration capacity. These findings provide a glimpse of Wnt5a’s putative roles in Dental Follicle stem/progenitor cells and the periodontium with implications in periodontal disease, tooth eruption, Dental implant bone healing and orthodontic tooth movement.
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wnt5a regulates Dental Follicle stem progenitor cells of the periodontium
Stem Cell Research & Therapy, 2014Co-Authors: Lusai Xiang, Mo Chen, Bin Cai, Xinchun Zhang, Chen Zhou, Chenglin Wang, Jeremy J. Mao, Junqi LingAbstract:Dental Follicle gives rise to one or several tissues of the periodontium including the periodontal ligament, cementum and/or alveolar bone. Whether Wnt5a is expressed in the postnatal periodontium or regulates Dental Follicle stem/progenitor cells is unknown. Dental Follicle stem/progenitor cells were isolated from postnatal day 1 (p1) to p11 from rat mandibular first molars. Immunolocalization mapped Wnt5a expression in the alveolar bone, periodontal ligament, and the developing ameloblast and odontoblast layers. Mononucleated and adherent cells were isolated from p7 Dental Follicle. Wnt5a was overexpressed in Dental Follicle stem/progenitor cells to study their proliferation, osteogenic differentiation and migration behavior, with subpopulations of native Dental Follicle stem/progenitor cells as controls, using real-time PCR (Taqman), Lenti-viral transfection, Western blotting and immunofluorescence. Wnt5a was expressed consistently in p1 to p11 rat peridontium. Native, p7 Dental Follicle stem/progenitor cells had modest ability to mineralize in the tested 14 days. Even in chemically defined osteogenesis medium, Dental Follicle stem/progenitor cells only showed modest mineralization. Upon addition of 300 ng/mL Wnt5a protein in osteogenesis medium, Dental Follicle stem/progenitor cells displayed mineralization that was still unremarkable. Chemically induced or Wnt5a-induced mineralization of Dental Follicle cells only occurred sparsely. Combination of Wnt5a with 100 ng/mL BMP2 finally prompted Dental Follicle stem/progenitor cells to produce robust mineralization with elevated expression of Runx2, alkaline phosphatase, collagen 1α1 and osteocalcin. Thus, native Dental Follicle stem/progenitor cells or some of their fractions may be somewhat modest in mineralization. Strikingly, Wnt5a protein significantly augmented RANKL ligand, suggesting putative regulatory roles of Dental Follicle stem/progenitor cells for the monocyte/osteoclast lineage and potential involvement in alveolar bone remodeling and/or resorption. P-Jnk1/2 was activated in Wnt5a overexpressed Dental Follicle cells; conversely, exposure to SP600125, a c-Jun N-terminal kinase (JNK) inhibitor attenuated Runx2, collagen 1α1 and osteocalcin expression either in the presence or absence of Wnt5a. Wnt5a overexpression in Dental Follicle stem/progenitor cells significantly reduced their proliferation rates, but robustly augmented their migration capacity. These findings provide a glimpse of Wnt5a’s putative roles in Dental Follicle stem/progenitor cells and the periodontium with implications in periodontal disease, tooth eruption, Dental implant bone healing and orthodontic tooth movement.
Weidong Tian - One of the best experts on this subject based on the ideXlab platform.
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Comparative study on differentiation of cervical-loop cells and Hertwig’s epithelial root sheath cells under the induction of Dental Follicle cells in rat
Scientific Reports, 2018Co-Authors: Jie Chen, Guoqing Chen, Ye Tian, Weidong TianAbstract:Cervical loop cells (CLC) and Hertwig’s epithelial root sheath (HERS) cells are believed to play critical roles in distinct developmental patterns between rodent incisors and molars, respectively. However, the differences in differentiation between CLC and HERS cells, and their response to inductions from Dental Follicle cells, remain largely unknown. In present study, CLC and HERS cells, as well as incisor Dental Follicle (IF) cells and molar Dental Follicle (MF) cells were isolated from post-natal 7-day rats. IF and MF cell derived conditioned medium (CM) was obtained for induction of CLC and HERS cells. In vitro experiments, we found that, under the induction of Dental Follicle cell derived CM, CLC cells maintained the epithelial polygonal-shapes and formed massive minerals, while part of HERS cells underwent shape transformation and generated granular minerals. CLC cells expressed higher enamel-forming and mineralization related genes, while HERS cells showed opposite expression patterns of BMP2, BMP4, AMBN and AMGN. In vivo, CLC cells generated enamel-like tissues while HERS cells formed cementum-periodontal ligament-like structures. Taken together, CLC and HERS cells present distinct differentiation patterns under the inductions from Dental Follicle cells.
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comparative study on differentiation of cervical loop cells and hertwig s epithelial root sheath cells under the induction of Dental Follicle cells in rat
Scientific Reports, 2018Co-Authors: Jie Chen, Guoqing Chen, Ye Tian, Weidong TianAbstract:Cervical loop cells (CLC) and Hertwig’s epithelial root sheath (HERS) cells are believed to play critical roles in distinct developmental patterns between rodent incisors and molars, respectively. However, the differences in differentiation between CLC and HERS cells, and their response to inductions from Dental Follicle cells, remain largely unknown. In present study, CLC and HERS cells, as well as incisor Dental Follicle (IF) cells and molar Dental Follicle (MF) cells were isolated from post-natal 7-day rats. IF and MF cell derived conditioned medium (CM) was obtained for induction of CLC and HERS cells. In vitro experiments, we found that, under the induction of Dental Follicle cell derived CM, CLC cells maintained the epithelial polygonal-shapes and formed massive minerals, while part of HERS cells underwent shape transformation and generated granular minerals. CLC cells expressed higher enamel-forming and mineralization related genes, while HERS cells showed opposite expression patterns of BMP2, BMP4, AMBN and AMGN. In vivo, CLC cells generated enamel-like tissues while HERS cells formed cementum-periodontal ligament-like structures. Taken together, CLC and HERS cells present distinct differentiation patterns under the inductions from Dental Follicle cells.
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Cells isolated from cryopreserved Dental Follicle display similar characteristics to cryopreserved Dental Follicle cells.
Cryobiology, 2017Co-Authors: Hefeng Yang, Jingjing Sun, Weihua Guo, Jinglong Chen, Weidong TianAbstract:Dental Follicle tissue is a promising resource of mesenchymal stem cells for cytotherapeutic approaches and tissue engineering applications. There are two procedures for banking of human Dental Follicle stem cells have been reported. Conventional method requires cell isolation, expansion and immediate cryopreservation. Whereas Dental Follicle stem cells can be isolated from cryopreserved Dental Follicle fragments. The aim of this study was to compare the characteristics of Dental Follicle cells isolated from cryopreserved fragments (DFCs-CF) with Dental Follicle cells recovered from cryopreserved cells (DFCs-CC). Dental Follicle fragments obtained after mechanical disaggregation were divided into two parts, with one part maintained in culture, while another part underwent cryopreservation. Dental Follicle fragments and Dental Follicle cells from fresh tissue were stored in liquid nitrogen for 3 months. After thawing, the isolation, morphology, proliferation, cell cycle, colony-forming-unit ability, stemness-related marker expression, apoptosis, and multi-lineage differentiation potential of DFCs-CF were tested compared with DFCs-CC. DFCs-CF expressed mesenchymal stem cells marker, proliferated well, showed similar levels of mRNA for stemness- and apoptosis-related genes and exhibited the capacity of multi-lineage differentiation similar to those of DFCs-CC. These results imply that cryopreservation of Dental Follicle fragments is an effective banking method for isolation of Dental Follicle cells.
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cytoskeletal binding proteins distinguish cultured Dental Follicle cells and periodontal ligament cells
Experimental Cell Research, 2016Co-Authors: Ye Tian, Guoqing Chen, Weihua Guo, Yaling Yang, Weidong TianAbstract:Human Dental Follicle cells (DFCs) and periodontal ligament cells (PDLCs) derived from the ectomesenchymal tissue, have been shown to exhibit stem/progenitor cell properties and the ability to induce tissue regeneration. Stem cells in Dental Follicle differentiate into cementoblasts, periodontal ligament fibroblasts and osteoblasts, these cells form cementum, periodontal ligament and alveolar bone, respectively. While stem cells in Dental Follicle are a precursor to periodontal ligament fibroblasts, the molecular changes that distinguish cultured DFCs from PDLCs are still unknown. In this study, we have compared the immunophenotypic features and cell cycle status of the two cell lines. The results suggest that DFCs and PDLCs displayed similar features related to immunophenotype and cell cycle. Then we employed an isobaric tag for relative and absolute quantitation (iTRAQ) proteomics strategy to reveal the molecular differences between the two cell types. A total of 2138 proteins were identified and 39 of these proteins were consistently differentially expressed between DFCs and PDLCs. Gene ontology analyses revealed that the protein subsets expressed higher in PDLCs were related to actin binding, cytoskeletal protein binding, and structural constituent of muscle. Upon validation by real-time PCR, western blotting, and immunofluorescence staining. Tropomyosin 1 (TPM1) and caldesmon 1 (CALD1) were expressed higher in PDLCs than in DFCs. Our results suggested that PDLCs display enhanced actin cytoskeletal dynamics relative to DFCs while DFCs may exhibit a more robust antioxidant defense ability relative to PDLCs. This study expands our knowledge of the cultured DFCs and PDLCs proteome and provides new insights into possible mechanisms responsible for the different biological features observed in each cell type.
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comparative study of human Dental Follicle cell sheets and periodontal ligament cell sheets for periodontal tissue regeneration
Cell Transplantation, 2013Co-Authors: Shujuan Guo, Weihua Guo, Yi Ding, Jian Gong, Qing Zou, Dan Xie, Yali Chen, Weidong TianAbstract:Periodontal ligament cell (PDLC) sheets have been shown to contribute to periodontal tissue regeneration. Dental Follicle cells (DFCs), acknowledged as the precursor cells of PDLCs, have demonstrat...
Mo Chen - One of the best experts on this subject based on the ideXlab platform.
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Wnt5a regulates Dental Follicle stem/progenitor cells of the periodontium
Stem cell research & therapy, 2014Co-Authors: Lusai Xiang, Mo Chen, Bin Cai, Xinchun Zhang, Chen Zhou, Chenglin Wang, Jeremy J. Mao, Junqi LingAbstract:Dental Follicle gives rise to one or several tissues of the periodontium including the periodontal ligament, cementum and/or alveolar bone. Whether Wnt5a is expressed in the postnatal periodontium or regulates Dental Follicle stem/progenitor cells is unknown. Dental Follicle stem/progenitor cells were isolated from postnatal day 1 (p1) to p11 from rat mandibular first molars. Immunolocalization mapped Wnt5a expression in the alveolar bone, periodontal ligament, and the developing ameloblast and odontoblast layers. Mononucleated and adherent cells were isolated from p7 Dental Follicle. Wnt5a was overexpressed in Dental Follicle stem/progenitor cells to study their proliferation, osteogenic differentiation and migration behavior, with subpopulations of native Dental Follicle stem/progenitor cells as controls, using real-time PCR (Taqman), Lenti-viral transfection, Western blotting and immunofluorescence. Wnt5a was expressed consistently in p1 to p11 rat peridontium. Native, p7 Dental Follicle stem/progenitor cells had modest ability to mineralize in the tested 14 days. Even in chemically defined osteogenesis medium, Dental Follicle stem/progenitor cells only showed modest mineralization. Upon addition of 300 ng/mL Wnt5a protein in osteogenesis medium, Dental Follicle stem/progenitor cells displayed mineralization that was still unremarkable. Chemically induced or Wnt5a-induced mineralization of Dental Follicle cells only occurred sparsely. Combination of Wnt5a with 100 ng/mL BMP2 finally prompted Dental Follicle stem/progenitor cells to produce robust mineralization with elevated expression of Runx2, alkaline phosphatase, collagen 1α1 and osteocalcin. Thus, native Dental Follicle stem/progenitor cells or some of their fractions may be somewhat modest in mineralization. Strikingly, Wnt5a protein significantly augmented RANKL ligand, suggesting putative regulatory roles of Dental Follicle stem/progenitor cells for the monocyte/osteoclast lineage and potential involvement in alveolar bone remodeling and/or resorption. P-Jnk1/2 was activated in Wnt5a overexpressed Dental Follicle cells; conversely, exposure to SP600125, a c-Jun N-terminal kinase (JNK) inhibitor attenuated Runx2, collagen 1α1 and osteocalcin expression either in the presence or absence of Wnt5a. Wnt5a overexpression in Dental Follicle stem/progenitor cells significantly reduced their proliferation rates, but robustly augmented their migration capacity. These findings provide a glimpse of Wnt5a’s putative roles in Dental Follicle stem/progenitor cells and the periodontium with implications in periodontal disease, tooth eruption, Dental implant bone healing and orthodontic tooth movement.
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wnt5a regulates Dental Follicle stem progenitor cells of the periodontium
Stem Cell Research & Therapy, 2014Co-Authors: Lusai Xiang, Mo Chen, Bin Cai, Xinchun Zhang, Chen Zhou, Chenglin Wang, Jeremy J. Mao, Junqi LingAbstract:Dental Follicle gives rise to one or several tissues of the periodontium including the periodontal ligament, cementum and/or alveolar bone. Whether Wnt5a is expressed in the postnatal periodontium or regulates Dental Follicle stem/progenitor cells is unknown. Dental Follicle stem/progenitor cells were isolated from postnatal day 1 (p1) to p11 from rat mandibular first molars. Immunolocalization mapped Wnt5a expression in the alveolar bone, periodontal ligament, and the developing ameloblast and odontoblast layers. Mononucleated and adherent cells were isolated from p7 Dental Follicle. Wnt5a was overexpressed in Dental Follicle stem/progenitor cells to study their proliferation, osteogenic differentiation and migration behavior, with subpopulations of native Dental Follicle stem/progenitor cells as controls, using real-time PCR (Taqman), Lenti-viral transfection, Western blotting and immunofluorescence. Wnt5a was expressed consistently in p1 to p11 rat peridontium. Native, p7 Dental Follicle stem/progenitor cells had modest ability to mineralize in the tested 14 days. Even in chemically defined osteogenesis medium, Dental Follicle stem/progenitor cells only showed modest mineralization. Upon addition of 300 ng/mL Wnt5a protein in osteogenesis medium, Dental Follicle stem/progenitor cells displayed mineralization that was still unremarkable. Chemically induced or Wnt5a-induced mineralization of Dental Follicle cells only occurred sparsely. Combination of Wnt5a with 100 ng/mL BMP2 finally prompted Dental Follicle stem/progenitor cells to produce robust mineralization with elevated expression of Runx2, alkaline phosphatase, collagen 1α1 and osteocalcin. Thus, native Dental Follicle stem/progenitor cells or some of their fractions may be somewhat modest in mineralization. Strikingly, Wnt5a protein significantly augmented RANKL ligand, suggesting putative regulatory roles of Dental Follicle stem/progenitor cells for the monocyte/osteoclast lineage and potential involvement in alveolar bone remodeling and/or resorption. P-Jnk1/2 was activated in Wnt5a overexpressed Dental Follicle cells; conversely, exposure to SP600125, a c-Jun N-terminal kinase (JNK) inhibitor attenuated Runx2, collagen 1α1 and osteocalcin expression either in the presence or absence of Wnt5a. Wnt5a overexpression in Dental Follicle stem/progenitor cells significantly reduced their proliferation rates, but robustly augmented their migration capacity. These findings provide a glimpse of Wnt5a’s putative roles in Dental Follicle stem/progenitor cells and the periodontium with implications in periodontal disease, tooth eruption, Dental implant bone healing and orthodontic tooth movement.