The Influences Of Osteoking On The Growth And Differentiation Of HBMSC

Objective(s):By studying the Osteoking for the cultured human bone marrow mesenchymal stem cells,to explore Osteoking in the treatment of osteoporosis and osteoporotic fracture bone formation mechanism.Osteoking directly acts on human bone marrow mesenchymal stem cells,then the corresponding changes occoured between the optimal concentration and human bone marrow mesenchymal stem cells.For further researching of Osteoking,promoting osteoporosis and bone formation mechanism research of osteoporotic fracture.Methods:Through the establishment of the human bone marrow mesenchymal stem cell line,giving Osteoking.Firstly,screening Osteoking cellular level to stimulate thehuman bone marrow mesenchymal stem cells between the optimal dilution of doses.At different concentrations of Osteoking to act on human bone marrow mesenchymal stem cells,finding the proliferation of growth.Then according to the growth of bone marrow mesenchymal stem cells from proliferation,observing 3th weeks and 4th weeks after continuous cultured.Osteoking acts on human bone marrow mesenchymal stem cells by different stimuli.The human bone marrow mesenchymal stem cells have multiple differentiation and the abilities of differentiation potential,can be conducive to osteoporosis and bone formation of osteoporotic fracture direction of cells.For example,osteoblasts,osteoclast,fat cell.Training after 3 weeks?4 weeks,using conventional dyeing method and immune cell histochemical method on the optimum dilution concentration Osteoking to act on human bone marrow mesenchymal stem cells to differentiate between qualitative research.Results:Experimental results show that the Osteoking dilution 16 times,its stimulus on the human bone marrow mesenchymal stem cell proliferation effect is most obvious,there was a statistical difference(P = 0.032,<0.05).Dilute concentration too small with Osteoking to join in the cell culture,will inhibit the growth of human bone marrow mesenchymal stem cell proliferation,and kill human bone marrow mesenchymal stem cells;Dilute concentrations greater than 16 times with Osteoking to act on human bone marrow mesenchymal stem cells,proliferation effect is 16 times dilution Osteoking is abate,but overall is still to promote proliferation.Through the human bone marrow mesenchymal stem cells' proliferation curve,the results showed that after 24 h' training,the number of the human bone marrow mesenchymal stem cells of Osteoking and 1-34 parathyroid hormone hole is lower than the control group;During 24-48 h after the treatment,the OSsteoking and PTH group cells proliferation,far higher than the control group.After 3 d' training,the Osteoking group cell proliferation rate droped,PTH group no obvious proliferation cells;but the Osteoking cells' number is higher than the PTH group cells' number.After 5 days' training,the Osteoking groups of cells begin to be less than the control cells.Angiohyalinosis the normal morphological cells of hBMSC in photoscope are the long fusiform,the distribution is more uniform' the parallel arrangement is spiral or vortex,the nucleus is clear,the nuclear plasma transparency is high.After four weeks of continuous induction of 16 times of the healing agent,the presence of alizarin red stain was not apparent in the presence of the obvious eosinophilic cells,and the visible nodules were not visible to the naked eye;The oil red O has a large number of tiny round particles(fatty blisters)in the field of view.After the azocine stain,the cell skeleton was blue,but it did not see the visible oval,smaller cartilaginous cells.Under fluorescent microscopy,the negative control group had only the blue dye and the unspecific display of the cell skeleton;A large number of osteoblast cells were marked by red fluorescent markers in the hBMSC.A single osteoblast is a fusiform cell,and the cell skeleton is clearly visible,the cytoplasm is rich,the nucleus is large and blue.Fat cells specificity immune fluorescent dye,a large number of cells specificity coloring,cytoskeleton and the nucleus is clearly visible,occasionally seen bright,round,not shaded area of the smaller(lipid drops cavity);No chondrocytes are found to be specific.Conclusion(s):<1>.After in vitro cultivation of the human bone marrow mesenchymal stem cells built,the dilution with 16 times' normal concentration of Osteoking can promote human bone marrow mesenchymal stem cell proliferation.But high dose of Osteoking kills the human bone marrow stem cells.<2>.The Osteoking acts on human bone marrow mesenchymal stem cells with 3 weeks,promoting human bone marrow mesenchymal stem cells into fat cells,osteoblasts;After 4 weeks,the division was more pronounced;But the human bone marrow mesenchymal stem cells did not show differentiation in the direction of the chondrocytes...