Department of Orthopedics, Huizhou Third People's Hospital, Guangzhou Medical University, Huizhou, Guangdong, China.
Department of Orthopedics, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, Guangzhou, Guangdong, China.
Braz J Med Biol Res. 2020 Dec 9;54(2):e9944. doi: 10.1590/1414-431X20209944. eCollection 2020.
The aim of this study was to inhibit adipogenic differentiation by transfecting two growth factors, platelet-derived growth factor (PDGF-BB) and bone morphogenic protein 2 (BMP-2), into modified rat bone marrow mesenchymal stem cells (rBMSCs) and then compounded with platelet-rich plasma (PRP). To achieve rBMSCs, the osteoporosis model of rats was established, and then the rBMSCs from the rats were isolated and identified. Co-transfection of rBMSCs with PDGF-BB-GFP and BMP-2 and detection of PDGF-BB/BMP-2 expression in transfected BMSCs was assessed by qRT-PCR and western blot, respectively. Moreover, the effect of the two growth factors transfection of rBMSCs on adipogenic differentiation was evaluated by oil red O staining and western blot, respectively. Finally, construction of the two growth factors transfection of rBMSCs compounded with PRP and detection of adipogenic differentiation were assessed by oil red O staining, CCK-8, and western blot, respectively. In vitro studies revealed that the two growth factors transfection of rBMSCs compounded with PRP promoted cell viability and inhibited adipogenic differentiation and could be promising for inhibiting adipogenic differentiation.
本研究旨在通过将两种生长因子（血小板衍生生长因子（PDGF-BB）和骨形态发生蛋白 2（BMP-2））转染到改良的大鼠骨髓间充质干细胞（rBMSCs）中，并与富血小板血浆（PRP）复合，来抑制成脂分化。为了获得 rBMSCs，建立了大鼠骨质疏松模型，然后从大鼠中分离和鉴定 rBMSCs。通过 qRT-PCR 和 Western blot 分别评估 rBMSCs 中转染的 PDGF-BB-GFP 和 BMP-2 的共转染以及转染的 BMSCs 中 PDGF-BB/BMP-2 的表达。此外，通过油红 O 染色和 Western blot 分别评估两种生长因子转染 rBMSCs 对成脂分化的影响。最后，通过油红 O 染色、CCK-8 和 Western blot 分别评估构建 rBMSCs 中转染的两种生长因子复合 PRP 以及检测成脂分化的情况。体外研究表明，rBMSCs 中转染的两种生长因子复合 PRP 促进细胞活力并抑制成脂分化，有望抑制成脂分化。
