Bone mesenchymal stem cells co‑expressing VEGF and BMP‑6 genes to combat avascular necrosis of the femoral head

Abstract

The aim of the present study was to investigate the potential of bone mesenchymal stem cells (BMSCs) treated with a combination of vascular endothelial growth factor (VEGF) and bone morphogenetic protein-6 (BMP-6) genes for the treatment of avascular necrosis of the femoral head (ANFH). Rat BMSCs were isolated and purified using a density gradient centrifugation method. The purity and characteristics of the BMSCs were detected by cell surface antigens identification using flow cytometry. The experimental groups were administered with one of the following adeno-associated virus (AAV) vector constructs: AAV-green fluorescent protein (AAV-GFP), AAV-BMP-6, AAV-VEGF or AAV-VEGF-BMP-6. The expression of VEGF and BMP-6 was detected by reverse transcription-quantitative polymerase chain reaction, western blotting and ELISA assays. The effects of VEGF and BMP-6 on BMSCs were evaluated by angiogenic and osteogenic assays. The transfected BMSCs were combined with a biomimetic synthetic scaffold poly lactide-co-glycolide (PLAGA) and they were then subcutaneously implanted into nude mice. After four weeks, the implants were analyzed with histology and subsequent immunostaining to evaluate the effects of BMSCs on blood vessel and bone formation in vivo. In the AAV-VEGF-BMP-6 group, the expression levels of VEGF and BMP-6 were significantly increased and human umbilical vein endothelial cells tube formation was significantly enhanced compared with other groups. Capillaries and bone formation in the AAV-VEGF-BMP-6 group was significantly higher compared with the other groups. The results of the present study suggest that BMSCs expressing both VEGF and BMP-6 induce an increase in blood vessels and bone formation, which provides theoretical support for ANFH gene therapy.

A comparison of different bone graft materials in peri-implant guided bone regeneration

“…VEGF, which is expressed by endothelial cells, is one of the most important cytokines in angiogenesis and is associated with bone formation, mesenchymal condensation, cartilage formation, cartilage resorption, and blood vessel invasion. 28 In immunohistochemical examination, VEGF expression was detected in all groups at similar levels with no significant difference between the groups.…”

P53 and Parkin co-regulate mitophagy in bone marrow mesenchymal stem cells to promote the repair of early steroid-induced osteonecrosis of the femoral head

“…Effective treatment of early steroid-induced osteonecrosis of the femoral head (ONFH) is still a difficult and urgent problem in the field of orthopedics. Bone marrow mesenchymal stem cell (BMSCs) transplantation has been used to treat early steroid-induced ONFH [1][2][3] . Survival and stemness of BMSCs in the bone necrotic area are key to transplantation effectiveness [4][5][6][7] .…”

Shenfu Injection: A Famous Chinese Prescription That Promotes HCN4 Activity in Bone Marrow Mesenchymal Stem Cells

“…Initially, the isolated cells were identified according to the BMSCs' surface markers. As shown in Figure 1 , the possession of BMSC-like phenotypes by the isolated cells was confirmed by the positive rates of CD45, CD29, and CD44 (1.19%, 98.9%, and 98.19%, respectively) [ 21 , 22 ].…”

“…Following centrifugation, the cells were suspended in Dulbecco's modified eagle medium (DMEM, Gibco), supplemented with 20% fetal bovine serum (FBS; Hyclone) and cultured at 37°C in an incubator with 5% CO 2 . The surface markers of the BMSCs (CD45, CD29, and CD44) were identified to confirm the phenotype of the isolated cells [ 21 , 22 ].…”