Combination Use Of Bone Tissue Engineering And Guided Bone Regeneration With N-HA/CS/CMC To Repair Segmental Radial Bone Defect In Rabbits

ObjectiveTo study the feasibility for repairing segmental defects with single orcombination of n-HA/CS/CMC composite membrane and n-HA/CS/CMCtri-component composite. To provide the rational for the clinical applicationof the new n-HA/CS/CMC composite membrane and n-HA/CS/CMCtri-component composite, and implore the effects on the repairing bonedefects with combined application of guided bone regeneration membraneand tissue engineering bone.MethodsExperiment in vitro: BMSCs were isolated by density gradientcentrifugation from the bone marrow of New Zealand rabbits, and theosteoblasts were differentiated from BMSCs by bone induction. The growthstates and growth curve of BMSCs and osteoblast cells were detected andcompared. The osteoblasts were identified using alkaline phosphatase andcalcium steining. Then, the BMSCs and osteoblasts were inoculated with n-HA/CS/CMC tri-component porous composite and n-HA/PA66by aconcentration of1.0×105/ml, and cultured in vitro. The growth state andadhesion of cells were observed, and the growth and proliferation of cellswere examined by MTT, and the adhesion between cells and material wereobserved by HE staining.Experiment in vivo: The experimental model of10mm bone segmentaldefect were produced in New Zealand white rabbits with both radius, andwere divided into group A, group B and group C according to transplantmaterials. Group A was divided into two groups, the group AL and GroupBL, and the segmental defect on the left side of the former group wasimbedded with n-HA/CS/CMC+BMSCs which was coated with amembrane of n-HA/CS/CMC, while the segmental defect on the right side ofthe latter group was imbedded with n-HA/CS/CMC+osteoblasts, which wascoated with a membrane of n-HA/CS/CMC. Group B: the segmental defecton the left side of the group BL was imbedded withn-HA/CS/CMC+BMSCs+BMSCs without membrane coated, while thesegmental defect on the right side of the group BR was imbedded withn-HA/PA66+BMSCs with a membrane of n-HA/CS/CMC coated. Group C:the segmental defect on the left side of the group CL was coateded with amembrane of n-HA/CS/CMC, while the segmental defect on the right side ofthe group CR taken as the control group was imbedded with nothing.Postoperative gross observation, X-ray evaluation, histological examination and concentration tests of BMP and VEGF in the implanting region wascarried out.ResultsExperiment in vitro: MSCs and osteoblasts presented a good adhesionand growth in the n-HA/CS/CMC tri-component porous composite andn-HA/PA66.Experiment in vivo: according to gross observation, X-ray evaluation,histological examination and concentration tests of BMP and VEGF, newbone formed and increased as time went on in the radial segmental defect inall the rabbits from group A, group B and group CL4weeks postoperatively.In addition, new bone formation developed earlier and the regionalconcentration of the osteoinductive factor was much higher in the group AL,group AR and group BR than group BL and group CL.ConclusionsExperiment in vitro: n-HA/CS/CMC tri-component porous compositecould be considered as ideal carriers for constructing tissue engineering bonein bone defects repairing study.Experiment in vivo: The tissue engineering bone constructed byBMSCs or osteoblasts and n-HA/CS/CMC tri-component porous compositecan be taken as repairing material for the segmental bone defects. Then-HA/CS/CMC composite membrane can guide bone regeneration. And thecombined use of n-HA/CS/CMC n-HA/CS/CMC tri-component porous composite tissue engineering bone and n-HA/CS/CMC membrane canenhance bone defects repairing.