The Functions Of Gap Junctional Intercellular Communication On The Osteogenic Differentiation Of Rat Bone Marrow Stromal Cells

Objective To investigate the functions of connexin43(Cx43) on the osteogenic differentiation of rat bone marrow stromal cells(BMSCs) in vitro.Methods The BMSCs were taken out from the femur and tibia of the male Sprague-Dawley rat by the whole bone marrow culture and purified by adhesive- screening method, and then were treated in the L-DMEM. When the passage culture reached the third generation we divided the BMSCs into three groups, ie,controled group(CG), induced group(IG) and experimental group(EG). At our experiment, dexamethasone,β-glycerophosphate and Vitamin C were combined used as the osteogenic inductor while 18α-glycyrrhetinic acid as the blocker of gap junctional intercellular communcation(GJIC). L-DMEM, L-DMEM+ osteogenic inductor, L-DMEM+ osteogenic inductor+AGA were added into CG, IG and EG, respectively. During the culture, the capability of proliferation and alkaline phosphatase(ALP) relative activity of different groups of cells were tested at various day. At 7th day, Cx43 was allocated by fluorescent technique. And semi-quantitative RT-PCR analysis were used for measuring the expression of Cx43, osteocalcin(OC), sialoprotein(SA) mRNA at the same day. At 10th day, we explored the function of GJIC by scrape-loading and dye transfer method. Then at 21st day, alizarin red S staining was used to observe the mineralization nodules among the matrix.Results At day 7 and 9, the proliferated ability of 3 groups sequenced as: EG