The Study Of Expression Of Microcapsulated BMSC's BMP-2 In Calcium Phosphate Cement

ObjectiveTo study the growth of bone marrow-derived mesenchymal stem cells (BMSCs). In Calcium Phosphate Cement and the amount of BMP-2 expressed by BMSCs and it's biological activation,which provide some theoretical confidence in the feasibility of animal experiment that compound the CPC and microcapsulated BMSC with target gene, then plant it into the bone-defected modleMethod1) Divide the 12-holes-cultivating-board into groupA,B,C,and every each group include 4 holes2) Mix the CPC powder with salt and purified water in proportion of 1:1:0.5 ,then dispacth them to the 8 holes of group A and group B averagely, after it solidify, bath it with purified water, after the salt of the mixture dissolved,the space previously token by the salt become little holes in the CPC solid,then we get a bigger interspace modle3) Tag the group A as experiment group and dispatch some microcapsulated BMSCs with target gene into the 4 holes of it.Group B for blank ,the 4 holes of it take in some microcapsule with no BMSC . The group C contained 4 holes with nothing in it,which take in some microcapsulated BMSCs with target gene. 4) And then dispatch 1ml culture medium with 200ug/ml DOX which serve as the exutory of Tet-on system to every one of the 12 holes,5) exchane the culture medium in 3,6,9,12day,and extract the culture medium which is in the every one of the 12 holes for ELISA to evaluate the concentration of BMP-2 respectively.6) Finally section the CPC solid which is in the 8 holes of the board into slice for observation under optic microscope to evaluate ossification respectivelyResult:1) Result:group A and group C have exprssed BMP-2 in significant concentration,and there is on significance in the difference betweem the amount of BMP-2 expressed by group A and group C. and there is no significant amount of BMP-2 expressed by group B. some collagen and osteoid can be seen through optic microscope in the slice sectioned from group A and there is no significant change in the slice that sectioned from group B2) Conclusion:Microcapsulated BMSC survived in the CPC multi-holes modle and expressed significant amount of BMP-2 and induced ossification...