Modulation of proliferation and differentiation of gingivaderived mesenchymal stem cells by concentrated growth factors: Potential implications in tissue engineering for dental regeneration and repair

The aim of the present study was to evaluate the proliferation and osteogenic differentiation ability of gingivaderived mesenchymal stem cells (GMSCs) cultured with different concentrations of concentrated growth factors (CGF). GMSCs were isolated from gingival connective tissues and characterized by flow cytometry, immunofluorescence staining and immunohistochemical staining. Cell proliferation activity was determined by the MTT assay, and the effect of CGF on MCSCs was detected with the Cell Counting Kit (CCK)8 assay. Mineralization induction was evaluated by alkaline phosphatase (ALP)positive cell staining and mineralized nodule formation assay. Dentin matrix acidic phosphoprotein (DMP)1, dentin sialophosphoprotein (DSPP), bone morphogenetic protein (BMP)2 and runtrelated transcription factor (RUNX)2 mRNA and protein expression were evaluated by reverse transcriptionquantitative polymerase chain reaction (RTqPCR) analysis and western blotting. The flow cytometry, immunofluorescence staining and immunohistochemical staining results indicated that the cultured cells were GMSCs. The MTT assay results revealed that the thirdgeneration gingival stem cells exhibited the highest proliferative capacity, and the CCK8 results indicated that 10% CGF achieved the most prominent promotion of GMSC proliferation. ALP activity analysis and mineralized nodule assay demonstrated that CGF may successfully induce osteogenic differentiation of GMSCs, whereas RTqPCR and western blot analyses demonstrated that CGF is involved in the differentiation of GMSCs by regulating the expression of DMP1, DSPP, BMP2 and RUNX2 (P<0.05). In conclusion, CGF were demonstrated to promote the proliferation and osteogenic differentiation of GMSCs. Therefore, CGF may be applied in tissue engineering for tooth regeneration and repair.