PDGFRβ+/c-kit+ pulp cells are odontoblastic progenitors capable of producing dentin-like structure in vitro and in vivo

TLDR

Successful pulp regeneration requires stem cells that can differentiate into odontoblasts and form pulp‑dentin‑like structures, and platelet‑derived growth factor (PDGF) is known to promote tissue repair and regeneration. The study aimed to identify a PDGF‑responsive subpopulation of dental pulp cells with dentin regeneration potential from 12 freshly extracted human third molars. PDGFRβ⁺/c‑kit⁺ pulp cells were isolated by flow cytometry, characterized for proliferation, odontoblastic gene expression, and mineralization in vitro, and GFP‑labeled cells were transplanted into empty root canals of nude rats. These cells proliferated faster than whole pulp cells, differentiated into odontoblasts with increased DMP1, DSPP, and osteocalcin expression, responded differently to BMP2 and PDGF‑BB, and formed globular dentin and pulp‑like tissue in vivo, demonstrating dentin‑like regenerative capacity.

Abstract

Successful pulp regeneration depends on identification of pulp stem cells capable of differentiation under odontoblastic lineage and producing pulp-dentinal like structure. Recent studies demonstrate that platelet-derived growth factor (PDGF) plays an important role in damage repair and tissue regeneration. The aim of this study was to identify a subpopulation of dental pulp cells responsive to PDGF and with dentin regeneration potential.Pulp tissues were isolated from 12 freshly extracted human impacted third molars. Pulp cells were sorted by their expression of PDGFRβ and stem cell marker genes via flow cytometry. For the selected cells, proliferation was analyzed by a colorimetric cell proliferation assay, differentiation was assessed by real time PCR detection the expression of odontoblast marker genes, and mineralization was evaluated by Alizarin Red S staining. GFP marked PDGFRβ+/c-kit+ pulp cells were transplanted into emptied root canals of nude rat lower left incisors. Pulp-dentinal regeneration was examined by immunohistochemistry.PDGFRβ+/c-kit+ pulp cells proliferated significantly faster than whole pulp cells. In mineralization media, PDGFRβ+/c-kit+ pulp cells were able to develop under odontoblastic linage as demonstrated by a progressively increased expression of DMP1, DSPP, and osteocalcin. BMP2 seemed to enhance whereas PDGF-BB seemed to inhibit odontoblastic differentiation and mineralization of PDGFRβ+/c-kit+ pulp cells. In vivo root canal transplantation study revealed globular dentin and pulp-like tissue formation by PDGFRβ+/c-kit+ cells.PDGFRβ+/c-kit+ pulp cells appear to have pulp stem cell potential capable of producing dentinal like structure in vitro and in vivo.