Experimental Study Of Zhx3Gene Transfected Bmscs And Osteogenesis In Vitro

ObjectiveTo investigate the influence of zinc fingers and homeoboxes3gene(ZHX3)transfected BMSCs and osteogenesis induced in vitro.MethodsTake the over expression of ZHX3gene lentivirus vector transfected into ratBMSCs. At the same time, We take the no-load virus transfect into BMSCs asnegative control group and use BMSCs without any treatment as blank controlgroup. Using fluorescent microscope to counting cell number and calculate therate of transfection. Western blot was used to detect the protein expression ofZHX3gene in the transected cells. In vitro osteogenic induct cellsafter21d take alkaline phosphates staining and alizarin red staining to observeosteogenic ability. Cartilage induct and adipogenic induct BMSCs respectively.Then find the difference of three group cells to investigate wheather ZHX3caneffect BMSCs on cartilage and adipogenic.Results(1) Cells express BMSCs phenotype which obtained by adherentmethod.(2) After transfection,ZHX3gene overexpression group and negativecontrol group express green fluorescence, blank control group do not express,and ZHX3gene expression in overexpression group is higher than the negativecontrol group.(3) Each group can find cells with expression of alkalinephosphatase-positive. Kaplow show that the score of overexpression group ishigher than the other two groups(P≤0.05). And mineralized nodules are visibleamong all groups, But the number and volume of mineralized nodules in overexpression group is larger than the others.（4） Red oil O staining afteradipogenic BMSCs appear light red，and a few cells appear red.but there is nothave difference between three guoups.So ZHX3gene does not effect BMSCs inadipogenic.And type Ⅱ collagen immunohistochemical staining atterchondrogenic all of three group cells expression typeⅡcollagen,however this nosignificance of statistics between three groups.Conclusion（1）ZHX3gene have a apposite expression when use the good titers oflentivirus.（2）ZHX3gene may promote BMSCs in vitro osteogenesis.