| For a long time, how to treat macro- piece bone defects is always a formidable problem to the osteopaths. If we had dealt those unsuitable, that would bring about mutilation, result in severe pain and thymo-burden to the patients. We found external injury and inflammation and tumor were the main causes to macro-piece bone defects by statistics. Autogenous bone transplantation is the most ideal method to treat this disease, but autogenous bone resources are limited, and this method would lead more pains to the patients. The same race variant bone resources are also limited, and maybe bring HB and AIDS and so on contagious diseases. The functions of artificial bones are not as good as the natural bones. So to search more ideal substitute bones to repair macro-piece bone defects becomes the hot spot in the domains of medical and biomaterial sciences. The method by means of heterogeneity bones combined with autogenous cells cograft macro-piece bone defects supplys a new way for osteopaths. At the first part of the experiments, we prepared the inbred line little-ear pig of BanNa os integumentales and cultivated and appraised the MSCs of the rabbits in vitro, as well as combined MSCs with inbred line little-ear pig of BanNa os integumentales. At the second part, we explored the feasibility and the bone formation superiority of the mesenchymal stem cells combined with inbred line little-ear pig of BanNa os integumentale and BMP-2 on repairing macro-piece bone defects of rabbits, and judged the worth and applied prospect of the inbred line little-ear pig of BanNa in the domain of bone transplantion .Thus provided experimental evidences for the inbred line little-ear pig of BanNa os integumentales to treat macro- piece bone defects in the future.Part I The experiment about prepared the donator bones, cultivated and appraised the MSCs of the rabbits , combined MSCs with the donator bones.Objective To explore the suitable part bones of the inbred line little-ear pig for donator bones, and prepared them. Cultivated and appraised the MSCs of the rabbits at extraorgan, as well as combined MSCs with inbred line little-ear pig of BanNa os integumentales, thus provided ideal materials for the Part II. Methods 1.We dissected all month ages inbred line little-ear pigs of BanNa, to find how old pig's bones were suitable to adult rabbits.2. Obtain 40 fibulas of the pigs ,got rid of the periost and the marrow , prepared more 70 os integumentale donator ,and all those donators were about 1.5×0.3×0.4cm,then bored some holes in them. 3. Cleaned those donator bones some times by distilled water, then defated and deproteined. Freeze-dryed them by freeze dryer, at the last,sterilized those bones. 4. MSCs of the rabbits were cultured and amplificated, then combined the MSCs with inbred line little-ear pig of BanNa os integumentales in vitro. Results 1.We found the fibula of six-seven month ages pig of BanNa was suitable to the adult rabbit's radial by anatomy. 2. Cultured the MSCs of those 35 rabbits in vitro successfully. 3. Combined the MSCs with inbred line little-ear pig of BanNa os integumentales in vitro successfully, prepared ideal transplanted data. Conclusion 1. The fibula of six-seven month ages pig of BanNa was suitable to the adult rabbit's radial . 2. The MSCs of the rabbits can mix together with the inbred line little-ear pig of BanNa os integumentales , and grow well. We found the MSCs had adhered the surfaces and the holes of the bones by SEM.Part II The Experimental study of MSCs combined with donator bones and BMP-2 on repairing macro-piece bone defects of rabbit.Objective To explore the feasibility of the mesenchymal stem cells combined with inbred line little-ear pig of BanNa os integumentale and BMP-2 on repairing macro-piece bone defects of rabbits.Methods There are 35 adult big-ear rabbit of Japan (all male).A 1.5 centimeter segmental defect was created in the both mid-upper part of the radial extremities of those adult rabbits.The left defects bones of the adult rabbits were implanted into composited xenogeeic os integumentales and BMP-2 ,those rabbits are exoerimental group. And those bone grafts were defatting and deproteinization,then were cultured with MSCs after degerm .The right defets of the adult rabbits were implanted merely defatting and deproteinization os integumentales of little-ear pig of BanNA .Those rabbits are control group.The bone defets of another 5 adult rabbits were not filled with anything ,they are blank group. The repair capability of the defects was assessed by physical ,X-ray, histology,bone Mineral density exaMinations 4,8 ,12and 16 weeks after operation . Results 12 weeks after operation , the defects were repaired completely and its bone Mineral density is near to the normal bone in experimental group , and after 16 weeks the defects were completely repaired. The bone Mineral density measurement after 4,8,12and1 6 weeks postpoperatively at all sites of bone defects revealed that the results of the experimental group were significantly higher than their controls(BMC P〈0.05,BMD P〈0.01). the defects were repaired slowly and the quantity of new bone formation is less in control group , the defects of blank group were not repaired . Conclusion 1.It is satisfaction that the MSCs combined with inbred line little-ear pig os integumentale and BMP-2 on repairing macro-piece bone defects. 2.MSCs composited with inbred line little-ear pig bone and BMP-2 on repairing segmental bone defects is obviously superior to the single xenogeneic bone in quantity and quality of new bone formation . 3. Inbred line pig os integumentale is superordinary vicariousness support material in bone tissue project . That supply a new and dependable method to treat bone defective. |
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