Experimental Study Of On Treatment Early Avascular Necrosis Of Femoral Head In Rabbit With BMSCs Take CCPC As Support Mortieral

Objective: The BMSCs were isolated, cultured and proliferated skilled in the grade of experiment and have commenced using in the clinical. There is not suitable support material. To improve the curative effect of BMSCs, this experiment to compare the curative effect of rabbits avascular necrosis of the femoral head which the CCPC and autoallergic cancellated bone as support material.Methods: 1. The preparation of the group and experimental animals: Breed all animals of prepare for operation a week early to make them adapt to the environment and make all the animals disease-free. All the animals wait for the operation without eating and drinking 24 hours. 24 adult New Zealand rabbits were divided into two groups randomly. 12 rabbits of Group A take the CCPC as support material. 12 rabbits of Group B take the utoallergic cancellated bone as support material. The specimens were obtained 2, 4, 6 and 8 weeks after operation. The BMSCs were isolated, cultured and proliferated by cultivating whole bone marrow which was draw-off from the posterior superior iliac spine of rabbits before 1 week of the operation. The bone marrow were plant into 6-hole cell culture plate, culture fluid is 20% fetal bovine serum DMEM, put into cell incubator, the density of go down to posterity is 1×106 . The third generation make into cell suspension, group A is infiltrate the cell suspension into CCPC, group B is into gelatin sponge. Then put them into cell incubator for 24 hours, in the operation group A put the CCPC into the femoral head defect area, Group B put the gelatin sponge which admixed with the autoallergic cancellated bone into the femoral head defect area. 3. Operation method: bilateral femoral head of all animals will be operated. Prone position, 40mg/kg of sodium pentothal to intraperitoneal injection of anesthesia. Conventional disinfection, back-lateral incision of the hip, and exposed the junction of femoral head and neck, not cut round ligament, not hip dislocation ,and protect the surrounding tissue, femoral head were refrigerated with liquefided nitrogen for 3 minutes. Bore from the back of collum femoris into femoral head to reach to the base of arthrodial cartilage after it rewarmming, with broach which diameter is 3.5mm, then the group A embedding the combination of CCPC and BMSCs, the group B embedding the combination of the pill of autoallergic cancellated bone, suture wound. 4. H.E dyeing: Cut the femoral head at the basement which be preserved in 10% formalin, marked, and put them into 8% formic acid solution for three times to demineralized, which need two weeks. Dewatering. Transparent. Steeping wax. Repair the specimen slice 5μm. Waterbath. Stretched preparation. Bake slice. H.E dyeing : Fuse wax. Hydration with alcohol. Dyeing with Urolen blue. Acidation. Return orchid. Dyeing with eosin. Hydration with alcohol again. Transparent. Mounting with resin. 5. X-ray and Hsitology observation: X-ray were used in observation the changing of bone density of the bore hole area, Light microscope were used in observation the change of bone trabecular of the bore hole area.6.Statistical analysis: Statistical analysis by SSPS 11.5 make t-analysis of the percentage of new bone trabecular areas.Results: 1. X-ray examination: group A, at 2 weeks there is a high density area in the femoral head embedding CCPC part, bone trabecula structure were normal. At 4 weeks the density of the high density area in femoral head are degrade, bone trabecula structure were normal. At 6 weeks there is new bone trabecula which in the femoral head embedding CCPC part, the other parts density is higher and the bone trabecula is confused. At 8 weeks there is mature bone trabecula in the femoral head embedding CCPC part and its density become to normal, the other parts density are much higher and the bone trabecula is confused, cystic degeneration emerge. Group B at 2 weeks there is a low density area in the femoral head embedding autoallergic cancellated bone part, the other parts bone trabecula structure were normal. At 4 weeks the density of the low density area of femoral head is increased, the other parts bone trabecula structure was normal. At 6 weeks there new bone trabecula which in the femoral head embedding autoallergic cancellated bone part, the other parts density are higher and the bone trabecula is confused. At 8 weeks there is mature bone trabecula in the femoral head operated part and its density become to normal, the other parts density is much higher and the bone trabecula is confused, cystic degeneration emerge. In General, Group A the density of the femoral head bore hole part become normal from high for embedding CCPC, at last the mature bone trabecula emerge. Group B the density of the femoral head bore hole part become normal from low for embedding utoallergic cancellated bone. at last the mature bone trabecula emerge too.2. Group A : At 2 weeks there are abound osteoblast in the bore hole part, the borderline of the defect area had some osteoid and newly formed bone At 4 weeks there is lats of osteoid and newly formed bone in the defect area. At 6 weeks there is some mature bone trabecula and bone marrow emerge in the defect area. At 8 weeks the bone trabecula become mature and lots of bone marrow emerge in the defect area. Group B: At 2 weeks there is lots of osteoid and newly formed bone at the borderline, but the centre of the defect area is filled with neutrophil cell and macrophage. At 4 weeks the newly formed bone can not be filled with all defect area and some specimens the centre of defect area have little bone trabecula, the others have not. At 6 weeks the situation is similar with 4 weeks, the bone trabecula is much mature and the marrow emerged. At 8 weeks the defect area is filled with bone trabecula and bone marrow. The other part of the femoral head change is similar with Group A.3. Statistical analysis Bone histomorphometry examination shows that both A, B Group bone tissue repair is prosperity, at last form the mature bone trabecula, and the result is similar. At 8 weeks the area percentage of newly formed bone trabecula in the defect area of Group A is 0.804650±0.0022681, Group B is 0.801267±0.0040373. The Statistical analysis shows that the data have homoscedasticity. In generally it is indiscriminately. Compare Group A with Group B P=0.842>0.05, not statistically significant.Conclusions:1. The combination of BMSCs and CCPC has strongly osteogenesis without cell inducing factor.2. The combination of BMSCs and CCPC has the same osteogenesis with the combination of BMSCs as autoallergic cancellated bone. 3. CCPC is a satisfactory material of BMSCs.