Research On Intensified Beta-tricalcium Phosphate/Platelet-rich Plasma Gel Multi-scaffold

Objective: Discussion on preparing method, mechanical properties , Toxicity and biocompatibility of intensified beta-tricalcium phosphate/ platelet-rich plasma gel multi-scaffold,which regard as Scaffold materials in tissue engineering of bone.Methods : 1.Ordinary beta-tricalcium phosphate and intensified beta-tricalcium phosphate.are prepared 2.The preparation of PRP by two step centrifugation and the preparation of intensified beta-tricalcium phosphate/platelet-rich plasma gel multi-scaffold.3.The determination of the mechanical properties of intensified beta-tricalcium phosphate/ platelet-rich plasma gel multi-scaffold 4. Beagle's bone marrow mesenchymal stem cells is isolated and cultured in vitro ,and purified and expanded to obtain seed cells which seed beta-tricalcium phosphate/ platelet-rich plasma gem multi-scaffold. 5. Adhesion rate and cytotoxicity is measured in beagle's bone marrow mesenchymal stem cells compositing with beta-tricalcium phosphate/platelet-rich plasma gel multi-scaffold. 6. The pore size of the composite material and cells'gowth is observed by Electron Microscope.Results: 1. It has 143.3±31.9×10~9/L plasma in whole blood and 1239.3±124.0×10~9/L plasma in PRP.2.The max-load of beta-tricalcium phosphate/platelet-rich plasma gel multi-scaffold is 945.64±86.38N and it's compressive strength is 13.06±0.47MPa; The max-load of ordinary beta-tricalcium phosphate is110.11±14.26N and it's compressive strength is 1.61±0.23MPa. The compressive strength of beta-tricalcium phosphate/platelet-rich plasma gel multi-scaffold has statistic difference compared with ordinary beta-tricalcium phosphate(P<0.01).3.The adhesion ratio of the 3rd passage cells were 50±2%,80±1%,92±2% and 96±1% in the intensified beta-tricalcium phosphate/platelet-rich plasma gel multi-scaffold at 4,8,12,24h.4.The RGR in the intensified beta-tricalcium phosphate/platelet-rich plasma gel multi-scaffold were all exceeded than 1.0 and the cytotoxicity of this material was considered innocuity in 2d,4d,8d.5.Beagle's bone marrow mesenchymal stem cells composite with beta-tricalcium phosphate/platelet-rich plasma gel multi-scaffold by electron microscope observation: The material surface is the granulated outward appearance, loose porous, between the hole communicates mutually;the pure intensified beta-tricalcium material's aperture size is 100～400um different, the average size of which is 314.24±156.17um ;the pore size is 1～10um, the depth varies. PRP is widespread packing and well-distributing in material hole. beagle's BMSCs vaccination material is observed 2 weeks later, the cell stretches out breaks out, each other is connected, the cover crevice, and crawls in the crevice to grow. Some cells obviously secret Extracellular Matrix. After induced, the massive cells has vigorous growth.Conclusion:1.The mechanical properties intensified of beta-tricalcium phosphate/platelet-rich plasma gel multi-scaffold has approximately surpassed ordinary Cancellous Bone's strength.2.The intensified beta-tricalcium phosphate/platelet-rich plasma gel multi-scaffold has non-toxic,and the biological compatibility is well;cells can grow well on the material,which is ideal scaffold materials in tissue engineering of bone.