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

Abstract

The aim of the present study was to evaluate the proliferation and osteogenic differentiation ability of gingiva-derived 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 runt-related transcription factor (RUNX)2 mRNA and protein expression were evaluated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) 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 third-generation gingival stem cells exhibited the highest proliferative capacity, and the CCK-8 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 RT-qPCR 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.

Effects of Concentrated Growth Factor on the Proliferation, Migration, and Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells: An in vitro study

“…Our results further revealed that with the concentrations of CGF from 5-10%, signi cant promotion effects on the proliferation and migration of BMSCs in a dose-dependent manner were shown. However, higher concentrations of CGF extract do not guarantee stronger promoting effects, even will have inhibitory effects on the cultured cells [19,35,36]. In this article, the effect of a 20% CGF extract was inferior to those of the 10% group, which is consistent with other's results [19,37,38].…”

“…First, CGF extracts contains not only growth factors that favor osteogenic but also unosteogenic cytokines [39]. At high concentrations, the inhibitory effects of the in ammatory cytokines in CGF might play a dominant role [19]. Second, it has previously been proposed that pH value is affected by the change in platelet count, which may negatively affect cell proliferation [40].…”

“…ALP is reported as a marker of osteoblast differentiation [41][42][43]. As the induction time increases and the osteogenic differentiation of cells progresses, its activity gradually increases [19]. The osteogenic activity of BMP-2 can be detected by measuring ALP activity [44,45].…”

“…BMSCs have played a key role in cell-based therapy and tissue engineering due to the multiple differentiation potential of them in resent years [19][20][21]. BMSCs are the preferred candidate cell resource for bone tissue engineering and cell-based therapies because they can differentiate into bone-forming osteoblasts and play a crucial role in bone regeneration [22].…”

Human Oral Stem Cells, Biomaterials and Extracellular Vesicles: A Promising Tool in Bone Tissue Repair

“…Recently, Gingiva-derived MSCs (GMSCs), a novel type of multipotent MSCs showing potential self-renewing multipotent differentiation into several cell lineages and immunomodulatory capacities, has been described [34,35]. GMSCs attract an increasing consideration for their easy isolation, stable phenotype, normal karyotype, high proliferative capacity and telomerase activity after long-term culture [36]. GMSCs and PDLSCs are considered to be optimal candidates for tissue engineering and cell-based therapies [37].…”