The Study On The Effect Of ShengJiYe-MSCs-DMB Composite Repairing Rabbit Radial Bone Defect

Bone defect is common in clinic, which caused by many kinds of diseases, and it'shard to treat. The present treatment for the bone defects includes autogenous andallogenous bone transplantation, the former is restricted in amount and may causedistress, the latter can arouse the immunological rejection and other complications. Sincethe American Science Foundation introduced the concept of "Tissue Engineering" in1987, the study on tissue engineering have received noticeable achievement, it is possibleto use the tissue-engineering bone to repair the bone defect and it becomes one trend inclinic. According to Chinese traditional herb ShengJiYe can cure infectious open bonefracture, this experiment study the influence of ShengJiYe on MSCs' proliferation anddifferentiation in vivo.Aim: 1 To establish and perfect the method to isolate and culture MSCs, and getmore seed cells possibly. 2 Put the characteristic Chinese herb ,which can cure infectiousopen fracture, in MSCs' proliferation and differentiation in vivo. Explore new applicationof Chinese herb medicine in tissue-engineering bone, and deepen our knowledge onChinese herb medicine's action.Method: 1 Get bone marrow from rabbits' tibias, obtain MSCs by density gradientcentrifugation and plastic-adherence, then cultivate and amplify them. 2 Put fibrin sealantasnd dematrix bone (DMB)as carrier of MSCs,construct tissue- engineering bone ofMSCs-DMB composite as blank group,and MSCs-BMP-DMB composite,MSCs-ShengJiYe-DMB, MSCs- ShengJiYe- BMP-DMB as experimental group to repairrabbit 10mm segmental radial defect.3 The efficiency of osteogenesis was evaluation byradiography, histology, immuno- histochemical staining, tetracycline fluorescencetest, Von Kossa staining and BMD analysis.Result: 1 MSCs can be isolated by means of density-gradient-centrifugation andtheirs adherence to plastic. The primary generation can be cultured for 13-14 days. Afterpassaged, the MSCs look fibroblastoid, growing in the form of fingerprint and can proliferate. 2 The radiographic and histologic evaluation show: BMP-ShengJiYe has thestrongest power to form the new bone callus and make them mature; BMP has the secondcapability to form and mature new bone callus, and ShengJiYe has the lesser ability toinduce osteogenesis. The pure scaffold-MSCs form and mature new bone callus slowliest.3 Histologic analysis confirrn the mechanism of osteogenesis are both intramembranousand endochondral ossification, and the new-born woven bone grow along the scaffold. 4BrdU labelled specimen 4 weeks post operation show MSCs take part in repairing bonedefect. 5 Tetracycline fluorescence test reveal the volumn and forlning rate of new bornbone: BMP-ShengJiYe group＞BMP group＞ShengJiYe group＞blank group.6 ColⅠimmunohistochemical staining show much new bone matrix, and experimentalgroup have more than blank group. 7 Von Kossa staining display new born boneexist near the fracture and scaffold.Conclusion: 1 Isolate the layer of mono-nucleus cells in the rabbit bone marrow bymeans of density-gradient-centrifugation and plastic-adherence, and the obtained MSCsare pure and can proliferate. 2 In vivo, ShengJiYe has the ability to make MSCsproliferate and to induce MSCs differentiating to osteoblasts. And ShengJiYe canenhance BMP's ossification. 3 Dematrix bone have natural porous structure, goodbiocompatibility and biodegradability, can be used to as the scaffold oftissue-engineering bone. DMB-ShengJiYe composite have good osteoconduction andosteoinduction. 4 Fibrin sealant can ensure the viability of the cells in the scaffold ascell-carrier. 5 At present, we can't confirm the definite components acting on MSCs'proliferation and differentiation, We should further study the single medicine in ShenJiYeand the effective components of the medicine complex, and explore the mechanism ofMSCs' proliferation and differentiation to osteoblast.