Effect of Gujia capsules on promoting vascular endothelial growth factor expression, angiogenesis and osteogenic differentiation in femoral defects
LIU Han-tao,ZHAO Liang-hu,LIU Cai
Orthopedics Department of Panzhihua University Affiliated Hospital, Panzhihua617000, China
Abstract:
Objective To investigate the mechanisms of Gujia capsules (GJ) in promoting intertrochanteric fractures and femoral defects healing. Methods Cells were divided into Control group, Vehicle group, 10 μg/mL GJ group, 50 μg/mL GJ group, 100 μg/mL GJ group. Bone marrow mesenchymal stem cells (BMSCs) were induced to differentiate into osteoblasts and vascular endothelial cells in vitro. The proliferation ability of BMSCs were determined by CCK-8 assay. The expression levels of osteoblasts differentiation markers ALP, OCN and RUNX2; the vascular endothelial cell differentiation markers VEGF and Ang-2 were determined by Western blot. An OVX-induced osteoporosis and femoral defects mouse model was established. 18 mice were randomly divided into three groups: In vivo control group, Model group and GJ treatment group, 6 mice in each. Bone mineral density (BMD) and bone volume fraction (BV/TV) in the right proximal femoral defect were measured by Micro-CT. HE staining and histopathology were used to analyze the effects of GJ on the healing of femoral defects. The expression of VEGF in the right proximal femoral defects were determined by immunohistochemical staining. Results Compared with Induction group, BMSCs proliferation ability was enhanced after 100 μg/mL GJ treatment (P <0.05). The relative expression levels of ALP, OCN, RUNX2, VEGF and Ang-2 were increased (P <0.05) in 100 μg/mL GJ treatment group cells. Compared with 10 μg/mL GJ group and 50 μg/mL GJ group, the relative expression levels of ALP, OCN, RUNX2, VEGF and Ang-2 were increased (P <0.05) in 100 μg/mL GJ treatment group cells. Compared with In vivo control group, in Model group the degree of intrachondral ossification was lower, and collagen deposition and differentiation of osteoblasts and chondrocytes were rare. BMD values, BV/TV values and IHC-VEGF relative optical densities were decreased (P <0.05). Compared with Model group, in GJ treatment group, the degree of intrachondral ossification was higher, with a large amount of collagen deposition and differentiation of osteoblasts and chondrocytes. BMD values, BV/TV values and IHC-VEGF relative optical densities were increased (P <0.05). Conclusion GJ could promote angiogenesis, osteogenic differentiation and enhance bone healing ability by up-regulating VEGF expression in femoral defects.
