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

Chen,Xiao; Chen,Yuhe; Hou,Yali; Song,Peng; Zhou,Minyue; Nie,Minhai; Liu,Xuqian

doi:10.3892/ijmm.2019.4172

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

Authors:
- Xiao Chen
- Yuhe Chen
- Yali Hou
- Peng Song
- Minyue Zhou
- Minhai Nie
- Xuqian Liu
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Affiliations: Department of Orthodontics, Mianyang Stomatological Hospital, Mianyang, Sichuan 621000, P.R. China, Laboratory of Oral and Maxillofacial Reconstruction and Regeneration, Southwest Medical University, Luzhou, Sichuan 646000, P.R. China, Department of Oral Pathology, College and Hospital of Stomatology, Hebei Medical University, Shijiazhuang, Hebei 050000, P.R. China
Published online on: April 24, 2019 https://doi.org/10.3892/ijmm.2019.4172
Pages: 37-46
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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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.

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