P‐glycoprotein overexpression in bone marrow–derived multipotent stromal cells decreases the risk of steroid‐induced osteonecrosis in the femoral head

P‐glycoprotein (P‐gp) plays a role in steroid‐induced osteonecrosis of the femoral head (ONFH), but the underlying mechanism remains unknown. We hypothesized that P‐gp overexpression can prevent ONFH by regulating bone marrow–derived multipotent stromal cell (BMSC) adipogenesis and osteogenesis. BMSCs from Sprague–Dawley rats were transfected with green fluorescent protein (GFP) or the multidrug resistance gene 1 (MDR1) encoding GFP and P‐gp. Dexamethasone was used to induce BMSC differentiation. Adipogenesis was determined by measuring peroxisome proliferator‐activated receptor (PPAR‐γ) expression and the triglyceride level. Osteogenesis was determined by measuring runt‐related transcription factor 2 (Runx2) expression and alkaline phosphatase activity. For in vivo experiments, rats were injected with saline, BMSCs expressing GFP (GFP‐BMSCs) or BMSCs expressing GFP‐P‐gp (MDR1‐GFP‐BMSCs). After dexamethasone induction, adipogenesis was determined by measuring PPAR‐γ expression and fatty marrow, whereas osteogenesis was detected by measuring Runx2 expression, trabecular parameters and the mineral apposition rate, followed by evaluation of the incidence of ONFH. Overexpression of P‐gp in BMSCs resulted in markedly decreased expression of adipogenic markers and increased expression of osteogenic markers. Compared with rats injected with saline, rats injected with GFP‐BMSCs showed reduced ONFH, and the injected GFP‐positive BMSCs attached to trabecular surfaces and exhibited an osteoblast‐like morphology. Compared with the rats injected with BMSCs expressing GFP alone, rats injected with BMSCs overexpressing GFP and P‐gp showed lower adipocytic variables, higher osteogenic variables and lower incidence of ONFH. Overexpression of P‐gp inhibited BMSC adipogenesis and promoted osteogenesis, which reduced the incidence of steroid‐induced ONFH.