The Experts below are selected from a list of 273 Experts worldwide ranked by ideXlab platform
Weidong Tian - One of the best experts on this subject based on the ideXlab platform.
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development of immortalized hertwig s epithelial root sheath cell lines for cementum and Dentin Regeneration
Stem Cell Research & Therapy, 2019Co-Authors: Sicheng Zhang, Guoqing Chen, Weihua Guo, Zirui Zhang, Weidong TianAbstract:Hertwig’s epithelial root sheath (HERS) is important in guiding tooth root formation by differentiating into cementoblasts through epithelial–mesenchymal transition (EMT) and inducing odontoblastic differentiation of dental papilla through epithelial–mesenchymal interaction (EMI) during the tooth root development. Thus, HERS cells are critical for cementum and Dentin formation and might be a potential cell source to achieve tooth root Regeneration. However, limited availability and lifespan of primary HERS cells may represent an obstacle for biological investigation and therapeutic use of tooth tissue engineering. Therefore, we constructed, characterized, and tested the functionality of immortalized cell lines in order to produce a more readily available alternative to HERS cells. Primary HERS cells were immortalized via infection with lentivirus vector containing the gene encoding simian virus 40 Large T Antigen (SV40LT). Immortalized HERS cell subclones were isolated using a limiting dilution method, and subclones named HERS-H1 and HERS-C2 cells were isolated. The characteristics of HERS-H1 and HERS-C2 cells, including cell proliferation, ability of epithelial–mesenchymal transformation and epithelial–mesenchymal interaction, were determined by CCK-8 assay, immunofluorescence staining, and real-time PCR. The cell differentiation into cementoblast-like cells or periodontal fibroblast-like cells was confirmed in vivo. And the inductive influence of the cell lines on dental papilla cells (DPCs) was also confirmed in vivo. HERS-H1 and HERS-C2 cells share some common features with primary HERS cells such as epithelial-like morphology, positive expression of CK14, E-Cadherin, and Vimentin, and undergoing EMT in response to TGF-beta. HERS-C2 cells showed the EMT characteristics and could differentiate into cementum-forming cells in vitro and generate cementum-like tissue in vivo. HERS-H1 could induce the differentiation of DPCs into odontoblasts in vitro and generation of Dentin-like tissue in vivo. We successfully isolated and characterized novel cell lines representing two key features of HERS cells during the tooth root development and which were useful substitutes for primary HERS cells, thereby providing a biologically relevant, unlimited cell source for studies on cell biology, developmental biology, and tooth root Regeneration.
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Development of immortalized Hertwig’s epithelial root sheath cell lines for cementum and Dentin Regeneration
BMC, 2019Co-Authors: Sicheng Zhang, Guoqing Chen, Weihua Guo, Zirui Zhang, Weidong TianAbstract:Abstract Background Hertwig’s epithelial root sheath (HERS) is important in guiding tooth root formation by differentiating into cementoblasts through epithelial–mesenchymal transition (EMT) and inducing odontoblastic differentiation of dental papilla through epithelial–mesenchymal interaction (EMI) during the tooth root development. Thus, HERS cells are critical for cementum and Dentin formation and might be a potential cell source to achieve tooth root Regeneration. However, limited availability and lifespan of primary HERS cells may represent an obstacle for biological investigation and therapeutic use of tooth tissue engineering. Therefore, we constructed, characterized, and tested the functionality of immortalized cell lines in order to produce a more readily available alternative to HERS cells. Methods Primary HERS cells were immortalized via infection with lentivirus vector containing the gene encoding simian virus 40 Large T Antigen (SV40LT). Immortalized HERS cell subclones were isolated using a limiting dilution method, and subclones named HERS-H1 and HERS-C2 cells were isolated. The characteristics of HERS-H1 and HERS-C2 cells, including cell proliferation, ability of epithelial–mesenchymal transformation and epithelial–mesenchymal interaction, were determined by CCK-8 assay, immunofluorescence staining, and real-time PCR. The cell differentiation into cementoblast-like cells or periodontal fibroblast-like cells was confirmed in vivo. And the inductive influence of the cell lines on dental papilla cells (DPCs) was also confirmed in vivo. Results HERS-H1 and HERS-C2 cells share some common features with primary HERS cells such as epithelial-like morphology, positive expression of CK14, E-Cadherin, and Vimentin, and undergoing EMT in response to TGF-beta. HERS-C2 cells showed the EMT characteristics and could differentiate into cementum-forming cells in vitro and generate cementum-like tissue in vivo. HERS-H1 could induce the differentiation of DPCs into odontoblasts in vitro and generation of Dentin-like tissue in vivo. Conclusions We successfully isolated and characterized novel cell lines representing two key features of HERS cells during the tooth root development and which were useful substitutes for primary HERS cells, thereby providing a biologically relevant, unlimited cell source for studies on cell biology, developmental biology, and tooth root Regeneration
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treated Dentin matrix paste as a novel pulp capping agent for Dentin Regeneration
Journal of Tissue Engineering and Regenerative Medicine, 2017Co-Authors: Jinlong Chen, Weihua Guo, Bo Yang, Caiyun Cui, Xiangchen Qiao, Weidong TianAbstract:Regenerating Dentin and preserving pulp vitality are the two key targets for the treatment of dental pulp exposure. Calcium hydroxide (CH), the widely used capping agent, may induce potential tunnel defect in reparative Dentin and cause inflammation or even necrosis in pulp tissues. This study aimed to produce a novel pulp capping agent with better bioactivities. Treated Dentin matrix (TDM) paste (TDMP) was fabricated consisting of TDM powder and aqueous TDM extract. The chemical and biological characteristics of TDMP were investigated, and its effect on the odontogenic differentiation of dental pulp stem cells explored at gene and protein level; the therapeutic effect for pulp exposure in miniature swine was further verified. TDMP possessed better biocompatibility with neutral pH value, significantly promoted the proliferation of dental pulp stem cells, and enhanced the gene and protein expressions of alkaline phosphatase, bone sialoprotein, Dentin sialoprotein etc., compared with CH. In vivo pulp capping using TDMP presented the formation of continuous reparative Dentin bridge thicker and denser than CH group. Moreover, pulp tissues under TDMP capping sites showed relatively slight angiectasis than those induced by CH. TDMP could achieve both Dentin Regeneration and vital pulp conservation, and might serve as a feasible substitute for CH in dental pulp repair procedure. Copyright © 2017 John Wiley & Sons, Ltd.
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human treated Dentin matrix as a natural scaffold for complete human Dentin tissue Regeneration
Biomaterials, 2011Co-Authors: Weihua Guo, Bo Yang, Lijuan Guo, Lei Sheng, Gang Chen, Qing Zou, Dan Xie, Yali Chen, Weidong TianAbstract:An essential aspect of tooth tissue engineering is the identification of suitable scaffolding materials to support cell growth and tissue Regeneration. Treated Dentin matrix (TDM) from a rat has recently been shown to be a suitable scaffold for rat Dentin Regeneration. However, due to species-specific differences, it remains unclear whether a similar fabrication method can be extended to human TDM and human Dentin Regeneration. Therefore, this present study explored the biological response to a human TDM (hTDM) created using a modified Dentin treatment method. Various biological characteristics, including cell proliferation, cell migration, cell viability, and cytotoxity were investigated. To assess the inductive capacity of hTDM, dental follicle cells (DFCs) were combined with hTDM and were implanted in vivo for 8 weeks in a mouse model. The resulting grafts were studied histologically. The results showed hTDM released Dentinogenic factors, indicating that hTDM could play a sustained role in odontogenesis. DFC attachment, growth, viability, and cytotoxicity on the surface of hTDM showed a notable improvement over those on calcium phosphate controls. Most importantly, in vivo hTDM induced and supported Regeneration of complete Dentin tissues, which expressed Dentin markers DSP and DMP-1. As cells in and around the regenerated Dentin were positive for human mitochondria, implanted DFCs and hTDM were responsible for the regenerated Dentin tissues. In conclusion, hTDM is indicated as an ideal biomaterial for human Dentin Regeneration.
Misako Nakashima - One of the best experts on this subject based on the ideXlab platform.
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trophic effects and regenerative potential of mobilized mesenchymal stem cells from bone marrow and adipose tissue as alternative cell sources for pulp Dentin Regeneration
Cell Transplantation, 2015Co-Authors: Masashi Murakami, K. Iohara, Yuki Hayashi, Yohei Osako, Yujiro Hirose, Misako NakashimaAbstract:Dental pulp stem cell (DPSC) subsets mobilized by granulocyte-colony-stimulating factor (G-CSF) are safe and efficacious for complete pulp Regeneration. The supply of autologous pulp tissue, however, is very limited in the aged. Therefore, alternative sources of mesenchymal stem/progenitor cells (MSCs) are needed for the cell therapy. In this study, DPSCs, bone marrow (BM), and adipose tissue (AD)-derived stem cells of the same individual dog were isolated using G-CSF-induced mobilization (MDPSCs, MBMSCs, and MADSCs). The positive rates of CXCR4 and G-CSFR in MDPSCs were similar to MADSCs and were significantly higher than those in MBMSCs. Trophic effects of MDPSCs on angiogenesis, neurite extension, migration, and antiapoptosis were higher than those of MBMSCs and MADSCs. Pulp-like loose connective tissues were regenerated in all three MSC transplantations. Significantly higher volume of regenerated pulp and higher density of vascularization and innervation were observed in response to MDPSCs compared to MBMSC and MADSC transplantation. Collagenous matrix containing Dentin sialophosphoprotein (DSPP)-positive odontoblast-like cells was the highest in MBMSCs and significantly higher in MADSCs compared to MDPSCs. MBMSCs and MADSCs, therefore, have potential for pulp Regeneration, although the volume of regenerated pulp tissue, angiogenesis, and reinnervation, were less. Thus, in conclusion, an alternative cell source for dental pulp/Dentin Regeneration are stem cells from BM and AD tissue.
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side population cells isolated from porcine dental pulp tissue with self renewal and multipotency for Dentinogenesis chondrogenesis adipogenesis and neurogenesis
Stem Cells, 2006Co-Authors: K. Iohara, M. Ito, Atsushi Tomokiyo, Li Zheng, Kenji Matsushita, Misako NakashimaAbstract:Dental pulp has the potential to form Dentin as a regenerative response to caries. This Regeneration is mediated by stem/progenitor cells. Thus, stem cell therapy might be of potential utility in induction of reparative Dentin. We isolated side population (SP) cells from dental pulp based on the exclusion of the DNA binding dye Hoechst 33342 by flow cytometry and compared its self-renewal capacities and multipotency with non-SP cells and primary pulp cells. The cumulative cell number of the SP cells was greater than the non-SP cells and primary pulp cells. Bmi1 was continuously expressed in SP cells, suggesting longer proliferative lifespan and self-renewal capacity of SP cells. Next, the maintenance of the multilineage differentiation potential of pulp SP cells was investigated. Expression of type II collagen and aggrecan confirmed chondrogenic conversion (30%) of SP cells. SP cells expressed peroxisome proliferator-activated receptor γ and adaptor protein 2, showing adipogenic conversion. Expression of mRNA and proteins of neurofilament and neuromodulin confirmed neurogenic conversion (90%). These results demonstrate that pulp SP cells maintain multilineage differentiation potential. We further examined whether bone morphogenetic protein 2 (BMP2) could induce differentiation of pulp SP cells into odontoblasts. BMP2 stimulated the expression of Dentin sialophosphoprotein (Dspp) and enamelysin in three-dimensional pellet cultures. Autogenous transplantation of the Bmp2-supplemented SP cells on the amputated pulp stimulated the reparative Dentin formation. Thus, adult pulp contains SP cells, which are enriched for stem cell properties and useful for cell therapy with BMP2 for Dentin Regeneration.
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gene therapy for Dentin Regeneration with bone morphogenetic proteins
Current Gene Therapy, 2006Co-Authors: Misako Nakashima, K. Iohara, Li ZhengAbstract:Recent advances in stem cell biology and gene therapy technology have provided the great potential of adult stem cells for therapeutic use in Regeneration of lost tissue due to diseases including cancer, trauma, and even caries. Dental pulp tissues harbor mesenchymal stem/progenitor cells and have potential to regenerate and/or repair Dentin-pulp complex after injury such as caries. There are two main methods, in vivo and ex vivo gene therapy. In in vivo gene therapy the healing potential of pulp tissue is enhanced by genes inducing Dentin directly applied on the exposed/amputated dental pulp. In ex vivo gene therapy, pulp stem/progenitor cells transfected with some therapeutically proven genes to induce differentiation into odontoblasts which are transplanted on the exposed/ amputated pulp. In the inflamed pulp under deep caries or trauma, possibly due to the limited supply of pulp stem/progenitor cells, it might be useful to apply cell-based ex vivo gene therapy compared to in vivo gene therapy. Before clinical use of ex vivo gene therapy for Dentin Regeneration in endodontics, there is a need for establishment of isolation, identification and expansion of the pulp stem cells. A safe and efficient gene delivery system also needs to be optimized. In this review we provide an overview of our current knowledge in the biology and function of adult pulp stem cells. This is followed by a discussion of the challenges of translating basic cellular and molecu lar biology of differentiation of pulp stem cells to safe and efficient gene therapy for Dentin Regeneration.
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bone morphogenetic proteins in Dentin Regeneration for potential use in endodontic therapy
Cytokine & Growth Factor Reviews, 2005Co-Authors: Misako NakashimaAbstract:The human dentition is indispensable for nutrition and physiology. The teeth have evolved for mastication of food. Caries is a common dental problem in which the Dentin matrix is damaged. When the caries is deep and the dental pulp is exposed, the pulp has to be removed in many cases, resulting ultimately in loss of the tooth. Therefore, the Regeneration of Dentin-pulp complex is the long-term goal of operative dentistry and endodontics. The key elements of Dentin Regeneration are stem cells, morphogens such as bone morphogenetic proteins (BMPs) and a scaffold of extracellular matrix. The dental pulp has stem/progenitor cells that have the potential to differentiate into Dentin-forming odontoblasts in response to BMPs. Pulpal wound healing consists of stem/progenitor cells release from dental pulp niche after noxious stimuli such as caries, migration to the injured site, proliferation and differentiation into odontoblasts. There are two main strategies for pulp therapy to regenerate Dentin: (1) in vivo method of enhancing the natural healing potential of pulp tissue by application of BMP proteins or BMP genes, (2) ex vivo method of isolation of stem/progenitor cells, differentiation with BMP proteins or BMP genes and transplantation to the tooth. This review summarizes recent advances in application of BMPs for Dentin Regeneration and possible use in endodotic therapy.
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Dentin Regeneration by dental pulp stem cell therapy with recombinant human bone morphogenetic protein 2
Journal of Dental Research, 2004Co-Authors: K. Iohara, Misako Nakashima, A. Nakasima, M. Ishikawa, M. Ito, Akifumi AkamineAbstract:Regenerative medicine is based on stem cells, signals, and scaffolds. Dental pulp tissue has the potential to regenerate Dentin in response to noxious stimuli, such as caries. The progenitor/stem cells are responsible for this Regeneration. Thus, stem cell therapy has considerable promise in Dentin Regeneration. Culture of porcine pulp cells, as a three-dimensional pellet, promoted odontoblast differentiation compared with monolayers. The expression of Dentin sialophosphoprotein (Dspp) and enamelysin/matrix metalloproteinase 20 (MMP20) mRNA confirmed the differentiation of pulp cells into odontoblasts and was stimulated by the morphogenetic signal, bone morphogenetic protein 2 (BMP2). Based on the in vitro experiments, an in vivo evaluation of pulp progenitor/stem cells in the dog was performed. The autogenous transplantation of the BMP2-treated pellet culture onto the amputated pulp stimulated reparative Dentin formation. In conclusion, BMP2 can direct pulp progenitor/stem cell differentiation into odontoblasts and result in Dentin formation.
Weihua Guo - One of the best experts on this subject based on the ideXlab platform.
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development of immortalized hertwig s epithelial root sheath cell lines for cementum and Dentin Regeneration
Stem Cell Research & Therapy, 2019Co-Authors: Sicheng Zhang, Guoqing Chen, Weihua Guo, Zirui Zhang, Weidong TianAbstract:Hertwig’s epithelial root sheath (HERS) is important in guiding tooth root formation by differentiating into cementoblasts through epithelial–mesenchymal transition (EMT) and inducing odontoblastic differentiation of dental papilla through epithelial–mesenchymal interaction (EMI) during the tooth root development. Thus, HERS cells are critical for cementum and Dentin formation and might be a potential cell source to achieve tooth root Regeneration. However, limited availability and lifespan of primary HERS cells may represent an obstacle for biological investigation and therapeutic use of tooth tissue engineering. Therefore, we constructed, characterized, and tested the functionality of immortalized cell lines in order to produce a more readily available alternative to HERS cells. Primary HERS cells were immortalized via infection with lentivirus vector containing the gene encoding simian virus 40 Large T Antigen (SV40LT). Immortalized HERS cell subclones were isolated using a limiting dilution method, and subclones named HERS-H1 and HERS-C2 cells were isolated. The characteristics of HERS-H1 and HERS-C2 cells, including cell proliferation, ability of epithelial–mesenchymal transformation and epithelial–mesenchymal interaction, were determined by CCK-8 assay, immunofluorescence staining, and real-time PCR. The cell differentiation into cementoblast-like cells or periodontal fibroblast-like cells was confirmed in vivo. And the inductive influence of the cell lines on dental papilla cells (DPCs) was also confirmed in vivo. HERS-H1 and HERS-C2 cells share some common features with primary HERS cells such as epithelial-like morphology, positive expression of CK14, E-Cadherin, and Vimentin, and undergoing EMT in response to TGF-beta. HERS-C2 cells showed the EMT characteristics and could differentiate into cementum-forming cells in vitro and generate cementum-like tissue in vivo. HERS-H1 could induce the differentiation of DPCs into odontoblasts in vitro and generation of Dentin-like tissue in vivo. We successfully isolated and characterized novel cell lines representing two key features of HERS cells during the tooth root development and which were useful substitutes for primary HERS cells, thereby providing a biologically relevant, unlimited cell source for studies on cell biology, developmental biology, and tooth root Regeneration.
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Development of immortalized Hertwig’s epithelial root sheath cell lines for cementum and Dentin Regeneration
BMC, 2019Co-Authors: Sicheng Zhang, Guoqing Chen, Weihua Guo, Zirui Zhang, Weidong TianAbstract:Abstract Background Hertwig’s epithelial root sheath (HERS) is important in guiding tooth root formation by differentiating into cementoblasts through epithelial–mesenchymal transition (EMT) and inducing odontoblastic differentiation of dental papilla through epithelial–mesenchymal interaction (EMI) during the tooth root development. Thus, HERS cells are critical for cementum and Dentin formation and might be a potential cell source to achieve tooth root Regeneration. However, limited availability and lifespan of primary HERS cells may represent an obstacle for biological investigation and therapeutic use of tooth tissue engineering. Therefore, we constructed, characterized, and tested the functionality of immortalized cell lines in order to produce a more readily available alternative to HERS cells. Methods Primary HERS cells were immortalized via infection with lentivirus vector containing the gene encoding simian virus 40 Large T Antigen (SV40LT). Immortalized HERS cell subclones were isolated using a limiting dilution method, and subclones named HERS-H1 and HERS-C2 cells were isolated. The characteristics of HERS-H1 and HERS-C2 cells, including cell proliferation, ability of epithelial–mesenchymal transformation and epithelial–mesenchymal interaction, were determined by CCK-8 assay, immunofluorescence staining, and real-time PCR. The cell differentiation into cementoblast-like cells or periodontal fibroblast-like cells was confirmed in vivo. And the inductive influence of the cell lines on dental papilla cells (DPCs) was also confirmed in vivo. Results HERS-H1 and HERS-C2 cells share some common features with primary HERS cells such as epithelial-like morphology, positive expression of CK14, E-Cadherin, and Vimentin, and undergoing EMT in response to TGF-beta. HERS-C2 cells showed the EMT characteristics and could differentiate into cementum-forming cells in vitro and generate cementum-like tissue in vivo. HERS-H1 could induce the differentiation of DPCs into odontoblasts in vitro and generation of Dentin-like tissue in vivo. Conclusions We successfully isolated and characterized novel cell lines representing two key features of HERS cells during the tooth root development and which were useful substitutes for primary HERS cells, thereby providing a biologically relevant, unlimited cell source for studies on cell biology, developmental biology, and tooth root Regeneration
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treated Dentin matrix paste as a novel pulp capping agent for Dentin Regeneration
Journal of Tissue Engineering and Regenerative Medicine, 2017Co-Authors: Jinlong Chen, Weihua Guo, Bo Yang, Caiyun Cui, Xiangchen Qiao, Weidong TianAbstract:Regenerating Dentin and preserving pulp vitality are the two key targets for the treatment of dental pulp exposure. Calcium hydroxide (CH), the widely used capping agent, may induce potential tunnel defect in reparative Dentin and cause inflammation or even necrosis in pulp tissues. This study aimed to produce a novel pulp capping agent with better bioactivities. Treated Dentin matrix (TDM) paste (TDMP) was fabricated consisting of TDM powder and aqueous TDM extract. The chemical and biological characteristics of TDMP were investigated, and its effect on the odontogenic differentiation of dental pulp stem cells explored at gene and protein level; the therapeutic effect for pulp exposure in miniature swine was further verified. TDMP possessed better biocompatibility with neutral pH value, significantly promoted the proliferation of dental pulp stem cells, and enhanced the gene and protein expressions of alkaline phosphatase, bone sialoprotein, Dentin sialoprotein etc., compared with CH. In vivo pulp capping using TDMP presented the formation of continuous reparative Dentin bridge thicker and denser than CH group. Moreover, pulp tissues under TDMP capping sites showed relatively slight angiectasis than those induced by CH. TDMP could achieve both Dentin Regeneration and vital pulp conservation, and might serve as a feasible substitute for CH in dental pulp repair procedure. Copyright © 2017 John Wiley & Sons, Ltd.
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human treated Dentin matrix as a natural scaffold for complete human Dentin tissue Regeneration
Biomaterials, 2011Co-Authors: Weihua Guo, Bo Yang, Lijuan Guo, Lei Sheng, Gang Chen, Qing Zou, Dan Xie, Yali Chen, Weidong TianAbstract:An essential aspect of tooth tissue engineering is the identification of suitable scaffolding materials to support cell growth and tissue Regeneration. Treated Dentin matrix (TDM) from a rat has recently been shown to be a suitable scaffold for rat Dentin Regeneration. However, due to species-specific differences, it remains unclear whether a similar fabrication method can be extended to human TDM and human Dentin Regeneration. Therefore, this present study explored the biological response to a human TDM (hTDM) created using a modified Dentin treatment method. Various biological characteristics, including cell proliferation, cell migration, cell viability, and cytotoxity were investigated. To assess the inductive capacity of hTDM, dental follicle cells (DFCs) were combined with hTDM and were implanted in vivo for 8 weeks in a mouse model. The resulting grafts were studied histologically. The results showed hTDM released Dentinogenic factors, indicating that hTDM could play a sustained role in odontogenesis. DFC attachment, growth, viability, and cytotoxicity on the surface of hTDM showed a notable improvement over those on calcium phosphate controls. Most importantly, in vivo hTDM induced and supported Regeneration of complete Dentin tissues, which expressed Dentin markers DSP and DMP-1. As cells in and around the regenerated Dentin were positive for human mitochondria, implanted DFCs and hTDM were responsible for the regenerated Dentin tissues. In conclusion, hTDM is indicated as an ideal biomaterial for human Dentin Regeneration.
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the use of Dentin matrix scaffold and dental follicle cells for Dentin Regeneration
Biomaterials, 2009Co-Authors: Weihua Guo, Xiaojun Zhang, Chunmei Wang, Yuan Liu, Yalei Zhou, Jing Zhou, Manjing Zhang, Zhihong Deng, Yan JinAbstract:Scaffold and inductive microenvironment are the two most important factors for Dentin Regeneration. They have been addressed with hydroxyapatite, tricalcium phosphate, polyglycolic acid, calcined bovine bone, and collagen, among other things. However, as of yet, no scaffold and inductive microenvironment combination has been shown to contribute to the Regeneration of complete and prefabricated-shaped Dentin tissues that include Dentin, preDentin and odontoblasts. To test the supporting and inductive effects of treated Dentin matrix (TDM) on complete and prefabricated-shaped Dentin Regeneration, dental follicle cells (DFCs) were seeded onto TDM and further incubated for 1 and 2 weeks in vitro and for 2 and 4 weeks in vivo. The results show that in vitro, in addition to Dentin sialoprotein (DSP) and Dentin matrix protein 1 (DMP1) (regarded as identifying markers of odontoblasts), DFCs induced by TDM expressed osteocalcin, bone sialoprotein, type I collagen, osteopontin, osteonectin and alkaline phosphatase (all expressed by odontoblasts), and that complete and prefabricated-shaped Dentin was successfully regenerated. Most importantly, it was found that in vivo TDM supports and induces Regeneration of complete and prefabricated-shaped Dentin, and regenerated Dentin expresses DSP and DMP1, which are identifying Dentin markers. Taken together, these results suggest that, for Dentin Regeneration, TDM is a suitable scaffold and inductive microenvironment and DFCs are a suitable cell type. The combination of TDM and DFCs may constitute a promising approach for future clinical Dentin Regeneration.
Anibal R Diogenes - One of the best experts on this subject based on the ideXlab platform.
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evaluation of a commercially available hyaluronic acid hydrogel restylane as injectable scaffold for dental pulp Regeneration an in vitro evaluation
Journal of Endodontics, 2017Co-Authors: Vanessa Chrepa, Obadah Austah, Anibal R DiogenesAbstract:Abstract Introduction Regenerative endodontic procedures (REPs) are viable alternatives for treating immature teeth, yet these procedures do not predictably lead to pulp-Dentin Regeneration. A true bioengineering approach for dental pulp Regeneration requires the incorporation of a scaffold conducive with the Regeneration of the pulp-Dentin complex. Several materials have been proposed as scaffolds for REPs; nonetheless, the majority are not eligible for immediate clinical chairside use. Thus, the aim of this study was to evaluate Restylane, a Food and Drug Administration-approved hyaluronic acid–based gel, as possible scaffold for REPs. Methods Stem cells of the apical papilla (SCAP) were cultured either alone or in mixtures with either Restylane or Matrigel scaffolds. Groups were cultured in basal culture medium for 6, 24, and 72 hours, and cell viability was assessed. For the mineralizing differentiation experiments, groups were cultured in differentiation medium either for 7 days and processed for alkaline phosphatase activity or for 14 days and processed for gene expression by using quantitative reverse-transcription polymerase chain reaction. SCAP in basal medium served as control. Results Cell encapsulation in either Restylane or Matrigel demonstrated reduced cell viability compared with control. Nonetheless, cell viability significantly increased in the Restylane group in the course of 3 days, whereas it decreased significantly in the Matrigel group. Restylane promoted significantly greater alkaline phosphatase activity and upregulation of Dentin sialophosphoprotein, Dentin matrix acidic phosphoprotein-1, and matrix extracellular phosphoglycoprotein, compared with control. Conclusions A Food and Drug Administration-approved hyaluronic acid–based injectable gel promoted SCAP survival, mineralization, and differentiation into an odontoblastic phenotype and may be a promising scaffold material for REPs.
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evaluation of a commercially available hyaluronic acid hydrogel restylane as injectable scaffold for dental pulp Regeneration an in vitro evaluation
Journal of Endodontics, 2017Co-Authors: Vanessa Chrepa, Obadah Austah, Anibal R DiogenesAbstract:Abstract Introduction Regenerative endodontic procedures (REPs) are viable alternatives for treating immature teeth, yet these procedures do not predictably lead to pulp-Dentin Regeneration. A true bioengineering approach for dental pulp Regeneration requires the incorporation of a scaffold conducive with the Regeneration of the pulp-Dentin complex. Several materials have been proposed as scaffolds for REPs; nonetheless, the majority are not eligible for immediate clinical chairside use. Thus, the aim of this study was to evaluate Restylane, a Food and Drug Administration-approved hyaluronic acid–based gel, as possible scaffold for REPs. Methods Stem cells of the apical papilla (SCAP) were cultured either alone or in mixtures with either Restylane or Matrigel scaffolds. Groups were cultured in basal culture medium for 6, 24, and 72 hours, and cell viability was assessed. For the mineralizing differentiation experiments, groups were cultured in differentiation medium either for 7 days and processed for alkaline phosphatase activity or for 14 days and processed for gene expression by using quantitative reverse-transcription polymerase chain reaction. SCAP in basal medium served as control. Results Cell encapsulation in either Restylane or Matrigel demonstrated reduced cell viability compared with control. Nonetheless, cell viability significantly increased in the Restylane group in the course of 3 days, whereas it decreased significantly in the Matrigel group. Restylane promoted significantly greater alkaline phosphatase activity and upregulation of Dentin sialophosphoprotein, Dentin matrix acidic phosphoprotein-1, and matrix extracellular phosphoglycoprotein, compared with control. Conclusions A Food and Drug Administration-approved hyaluronic acid–based injectable gel promoted SCAP survival, mineralization, and differentiation into an odontoblastic phenotype and may be a promising scaffold material for REPs.
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Hypoxia Modulates the Differentiation Potential of Stem Cells of the Apical Papilla
Journal of Endodontics, 2014Co-Authors: Julie Vanacker, Anibal R Diogenes, P. De Berdt, Julian Leprince, Caroline Bouzin, Aiswarya Viswanath, Amandine Everard, Patrice D. Cani, Olivier Feron, A. Des RieuxAbstract:Introduction: Stem cells from the apical papilla (SCAP) are a population of mesenchymal stem cells likely involved in regenerative endodontic procedures, and of potential use as therapeutic agents in other tissues. In these situations, SCAP are exposed to hypoxic conditions either within a root canal devoid of an adequate blood supply or in a scaffold material immediately after implantation. However, the effect of hypoxia on SCAP proliferation and differentiation is largely unknown. Therefore, the objective of this study was to evaluate the effect of hypoxia on SCAP fate. Methods: SCAP were cultured under normoxia (21%O2) or hypoxia (1%O2), in basal or differentiation media. Cellular proliferation, gene expression, differentiation and protein secretion were analyzed by live imaging, qRT-PCR, cellular staining and ELISA, respectively. Results: Hypoxia had no effect on SCAP proliferation, but it evoked the upregulation of genes specific for osteogenic differentiation (RUNX2, ALP, TGF β1), neuronal differentiation (CNP, SNAIL, NSE, GDNF, NT3), and angiogenesis (VEGFA, VEGFB). Hypoxia also increased the sustained production of VEGFa by SCAP. Moreover, hypoxia augmented the neuronal differentiation of SCAP in presence of differentiation exogenous factors as detected by the up-regulation of NSE, VEGFB and GDNF and the expression of neuronal markers (PanF and NeuN). Conclusion: This study demonstrates that hypoxia induces spontaneous differentiation of SCAP into osteogenic and neurogenic lineages while maintaining the release of the pro-angiogenic factor VEGFa. This highlights the potential of SCAP to promote pulp-Dentin Regeneration. Moreover, SCAP may represent potential therapeutic agents for neurodegenerative conditions due to their robust differentiation potential.
K. Iohara - One of the best experts on this subject based on the ideXlab platform.
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trophic effects and regenerative potential of mobilized mesenchymal stem cells from bone marrow and adipose tissue as alternative cell sources for pulp Dentin Regeneration
Cell Transplantation, 2015Co-Authors: Masashi Murakami, K. Iohara, Yuki Hayashi, Yohei Osako, Yujiro Hirose, Misako NakashimaAbstract:Dental pulp stem cell (DPSC) subsets mobilized by granulocyte-colony-stimulating factor (G-CSF) are safe and efficacious for complete pulp Regeneration. The supply of autologous pulp tissue, however, is very limited in the aged. Therefore, alternative sources of mesenchymal stem/progenitor cells (MSCs) are needed for the cell therapy. In this study, DPSCs, bone marrow (BM), and adipose tissue (AD)-derived stem cells of the same individual dog were isolated using G-CSF-induced mobilization (MDPSCs, MBMSCs, and MADSCs). The positive rates of CXCR4 and G-CSFR in MDPSCs were similar to MADSCs and were significantly higher than those in MBMSCs. Trophic effects of MDPSCs on angiogenesis, neurite extension, migration, and antiapoptosis were higher than those of MBMSCs and MADSCs. Pulp-like loose connective tissues were regenerated in all three MSC transplantations. Significantly higher volume of regenerated pulp and higher density of vascularization and innervation were observed in response to MDPSCs compared to MBMSC and MADSC transplantation. Collagenous matrix containing Dentin sialophosphoprotein (DSPP)-positive odontoblast-like cells was the highest in MBMSCs and significantly higher in MADSCs compared to MDPSCs. MBMSCs and MADSCs, therefore, have potential for pulp Regeneration, although the volume of regenerated pulp tissue, angiogenesis, and reinnervation, were less. Thus, in conclusion, an alternative cell source for dental pulp/Dentin Regeneration are stem cells from BM and AD tissue.
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side population cells isolated from porcine dental pulp tissue with self renewal and multipotency for Dentinogenesis chondrogenesis adipogenesis and neurogenesis
Stem Cells, 2006Co-Authors: K. Iohara, M. Ito, Atsushi Tomokiyo, Li Zheng, Kenji Matsushita, Misako NakashimaAbstract:Dental pulp has the potential to form Dentin as a regenerative response to caries. This Regeneration is mediated by stem/progenitor cells. Thus, stem cell therapy might be of potential utility in induction of reparative Dentin. We isolated side population (SP) cells from dental pulp based on the exclusion of the DNA binding dye Hoechst 33342 by flow cytometry and compared its self-renewal capacities and multipotency with non-SP cells and primary pulp cells. The cumulative cell number of the SP cells was greater than the non-SP cells and primary pulp cells. Bmi1 was continuously expressed in SP cells, suggesting longer proliferative lifespan and self-renewal capacity of SP cells. Next, the maintenance of the multilineage differentiation potential of pulp SP cells was investigated. Expression of type II collagen and aggrecan confirmed chondrogenic conversion (30%) of SP cells. SP cells expressed peroxisome proliferator-activated receptor γ and adaptor protein 2, showing adipogenic conversion. Expression of mRNA and proteins of neurofilament and neuromodulin confirmed neurogenic conversion (90%). These results demonstrate that pulp SP cells maintain multilineage differentiation potential. We further examined whether bone morphogenetic protein 2 (BMP2) could induce differentiation of pulp SP cells into odontoblasts. BMP2 stimulated the expression of Dentin sialophosphoprotein (Dspp) and enamelysin in three-dimensional pellet cultures. Autogenous transplantation of the Bmp2-supplemented SP cells on the amputated pulp stimulated the reparative Dentin formation. Thus, adult pulp contains SP cells, which are enriched for stem cell properties and useful for cell therapy with BMP2 for Dentin Regeneration.
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gene therapy for Dentin Regeneration with bone morphogenetic proteins
Current Gene Therapy, 2006Co-Authors: Misako Nakashima, K. Iohara, Li ZhengAbstract:Recent advances in stem cell biology and gene therapy technology have provided the great potential of adult stem cells for therapeutic use in Regeneration of lost tissue due to diseases including cancer, trauma, and even caries. Dental pulp tissues harbor mesenchymal stem/progenitor cells and have potential to regenerate and/or repair Dentin-pulp complex after injury such as caries. There are two main methods, in vivo and ex vivo gene therapy. In in vivo gene therapy the healing potential of pulp tissue is enhanced by genes inducing Dentin directly applied on the exposed/amputated dental pulp. In ex vivo gene therapy, pulp stem/progenitor cells transfected with some therapeutically proven genes to induce differentiation into odontoblasts which are transplanted on the exposed/ amputated pulp. In the inflamed pulp under deep caries or trauma, possibly due to the limited supply of pulp stem/progenitor cells, it might be useful to apply cell-based ex vivo gene therapy compared to in vivo gene therapy. Before clinical use of ex vivo gene therapy for Dentin Regeneration in endodontics, there is a need for establishment of isolation, identification and expansion of the pulp stem cells. A safe and efficient gene delivery system also needs to be optimized. In this review we provide an overview of our current knowledge in the biology and function of adult pulp stem cells. This is followed by a discussion of the challenges of translating basic cellular and molecu lar biology of differentiation of pulp stem cells to safe and efficient gene therapy for Dentin Regeneration.
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Dentin Regeneration by dental pulp stem cell therapy with recombinant human bone morphogenetic protein 2
Journal of Dental Research, 2004Co-Authors: K. Iohara, Misako Nakashima, A. Nakasima, M. Ishikawa, M. Ito, Akifumi AkamineAbstract:Regenerative medicine is based on stem cells, signals, and scaffolds. Dental pulp tissue has the potential to regenerate Dentin in response to noxious stimuli, such as caries. The progenitor/stem cells are responsible for this Regeneration. Thus, stem cell therapy has considerable promise in Dentin Regeneration. Culture of porcine pulp cells, as a three-dimensional pellet, promoted odontoblast differentiation compared with monolayers. The expression of Dentin sialophosphoprotein (Dspp) and enamelysin/matrix metalloproteinase 20 (MMP20) mRNA confirmed the differentiation of pulp cells into odontoblasts and was stimulated by the morphogenetic signal, bone morphogenetic protein 2 (BMP2). Based on the in vitro experiments, an in vivo evaluation of pulp progenitor/stem cells in the dog was performed. The autogenous transplantation of the BMP2-treated pellet culture onto the amputated pulp stimulated reparative Dentin formation. In conclusion, BMP2 can direct pulp progenitor/stem cell differentiation into odontoblasts and result in Dentin formation.