The dental pulp has irreplaceable roles in maintaining healthy teeth and its regeneration is a primary aim of regenerative endodontics.This study aimed to replicate the characteristics of dental pulp tissue by using c...The dental pulp has irreplaceable roles in maintaining healthy teeth and its regeneration is a primary aim of regenerative endodontics.This study aimed to replicate the characteristics of dental pulp tissue by using cranial neural crest(CNC)-like cells(CNCLCs);these cells were generated by modifying several steps of a previously established method for deriving NC-like cells from induced pluripotent stem cells(iPSCs).CNC is the anterior region of the neural crest in vertebrate embryos,which contains the primordium of dental pulp cells or odontoblasts.The produced CNCLCs showed approximately 2.5-12,000-fold upregulations of major CNC marker genes.Furthermore,the CNCLCs exhibited remarkable odontoblastic differentiation ability,especially when treated with a combination of the fibroblast growth factors(FGFs)FGF4 and FGF9.The FGFs induced odontoblast marker genes by 1.7-5.0-fold,as compared to bone morphogenetic protein 4(BMP4)treatment.In a mouse subcutaneous implant model,the CNCLCs briefly fated with FGF4+FGF9 replicated dental pulp tissue characteristics,such as harboring odontoblast-like cells,a dentin-like layer,and vast neovascularization,induced by the angiogenic self-assembling peptide hydrogel(SAPH),SLan.SLan acts as a versatile biocompatible scaffold in the canal space.This study demonstrated a successful collaboration between regenerative medicine and SAPH technology.展开更多
The cranial neural crest(CNC)cells play a vital role in craniofacial development and regeneration.They are multi-potent progenitors,being able to differentiate into various types of tissues.Both pre-migratory and post...The cranial neural crest(CNC)cells play a vital role in craniofacial development and regeneration.They are multi-potent progenitors,being able to differentiate into various types of tissues.Both pre-migratory and post-migratory CNC cells are plastic,taking on diverse fates by responding to different inductive signals.However,what sustains the multipotency ofCNCcells andderivatives remains largely unknown.In this study,we present evidence that FGF8 signaling is able to sustain progenitor status and multipotency of CNC-derived mesenchymal cells both in vivo and in vitro.We show that augmented FGF8 signaling in pre-migratory CNC cells prevents cell differentiation and organogenesis in the craniofacial region by maintaining their progenitor status.CNC-derived mesenchymal cells with Fgf8 overexpression or control cells in the presence of exogenous FGF8 exhibit prolonged survival,proliferation,and multi-potent differentiation capability in cell cultures.Remarkably,exogenous FGF8 also sustains the capability of CNC-derived mesenchymal cells to participate in organogenesis such as odontogenesis.Furthermore,FGF8-mediated signaling strongly promotes adipogenesis but inhibits osteogenesis of CNC-derived mesenchymal cells in vitro.Our results reveal a specific role for FGF8 in the maintenance of progenitor status and in fate determination of CNC cells,implicating a potential application in expansion and fate manipulation of CNC-derived cells in stem cell-based craniofacial regeneration.展开更多
基金supported by NIH grants,R01DE025885(E.S),R15EY029504(VAK)National Science Foundation NSF IIP 1903617(VAK).
文摘The dental pulp has irreplaceable roles in maintaining healthy teeth and its regeneration is a primary aim of regenerative endodontics.This study aimed to replicate the characteristics of dental pulp tissue by using cranial neural crest(CNC)-like cells(CNCLCs);these cells were generated by modifying several steps of a previously established method for deriving NC-like cells from induced pluripotent stem cells(iPSCs).CNC is the anterior region of the neural crest in vertebrate embryos,which contains the primordium of dental pulp cells or odontoblasts.The produced CNCLCs showed approximately 2.5-12,000-fold upregulations of major CNC marker genes.Furthermore,the CNCLCs exhibited remarkable odontoblastic differentiation ability,especially when treated with a combination of the fibroblast growth factors(FGFs)FGF4 and FGF9.The FGFs induced odontoblast marker genes by 1.7-5.0-fold,as compared to bone morphogenetic protein 4(BMP4)treatment.In a mouse subcutaneous implant model,the CNCLCs briefly fated with FGF4+FGF9 replicated dental pulp tissue characteristics,such as harboring odontoblast-like cells,a dentin-like layer,and vast neovascularization,induced by the angiogenic self-assembling peptide hydrogel(SAPH),SLan.SLan acts as a versatile biocompatible scaffold in the canal space.This study demonstrated a successful collaboration between regenerative medicine and SAPH technology.
基金This work was supported by the National Institutes of Health(R01DE24152 and R01DE14044 to Y.C.)the National Basic Research Program of China(2010CB944800)to W.T.the National Natural Science Foundation of China(81371134)to T.H.
文摘The cranial neural crest(CNC)cells play a vital role in craniofacial development and regeneration.They are multi-potent progenitors,being able to differentiate into various types of tissues.Both pre-migratory and post-migratory CNC cells are plastic,taking on diverse fates by responding to different inductive signals.However,what sustains the multipotency ofCNCcells andderivatives remains largely unknown.In this study,we present evidence that FGF8 signaling is able to sustain progenitor status and multipotency of CNC-derived mesenchymal cells both in vivo and in vitro.We show that augmented FGF8 signaling in pre-migratory CNC cells prevents cell differentiation and organogenesis in the craniofacial region by maintaining their progenitor status.CNC-derived mesenchymal cells with Fgf8 overexpression or control cells in the presence of exogenous FGF8 exhibit prolonged survival,proliferation,and multi-potent differentiation capability in cell cultures.Remarkably,exogenous FGF8 also sustains the capability of CNC-derived mesenchymal cells to participate in organogenesis such as odontogenesis.Furthermore,FGF8-mediated signaling strongly promotes adipogenesis but inhibits osteogenesis of CNC-derived mesenchymal cells in vitro.Our results reveal a specific role for FGF8 in the maintenance of progenitor status and in fate determination of CNC cells,implicating a potential application in expansion and fate manipulation of CNC-derived cells in stem cell-based craniofacial regeneration.
