| Background: Dental implant has become a hot subject in various branches of stomatology.More and more people choose implant to repair dentition defect and dentition loss,but implantation in bone defect area has become a difficult problem in implant surgery.Guiding bone regeneration has become the key to solve this problem.Autogenous bone transplantation is the gold standard to guide bone regeneration,but it is directly related to the complications related to the acquisition of autogenous bone.Such as neurovascular injury,deep tissue infection,bleeding,and donor area pain occur from time to time,and patients’ acceptance of opening up the second operation area is low.Therefore,the selection of bone substitute materials has become the goal of implant researchers.at present,the use of xenogeneic bone extraction materials to repair bone defects is one of the more promising research directions.Objective:To establish an animal experimental model of bone defect and to study the effect of BPE on the repair of rat skull,so as to provide experimental basis for clinical use of xenogeneic BPE to repair bone defect.Method:Three are three parts:the preparation of materials,animal experiments and examination methods.1.Preparation of materials:(1)Preparation of decalcified bone matrix(DBM): fresh porcine tibia was ground into powder after acellular and decalcified treatment,and then freeze-dried.(2)Preparation of BPE: DBM was divided into two parts and two kinds of BPE were extracted by dialysis and cold acetone precipitation respectively.2.Animal experiment: a total of 30 8-week-old male SPF SD rats(provided by the animal experiment of Dalian Medical University)weighed300±20g.Among them,10 rats were randomly made with unilateral 6 mm round bone defect of the parietal bone.The defect side was not filled with material as the blank control group,and the other side was the normal group.The remaining 20 rats were made bilateral parietal bone defects and randomly filled with 4 different materials,each with 10 defect areas.The groups are as follows: 1 normal group: incision of soft tissue on the surface of skull without defect;(2)Defect group:The skull defect without filling material was prepared and sutured directly.(3)Bio-oss group:The skull defect was prepared,and the defect area was filled with Bio-oss bone powder,and the skin was sutured.(4)DBM group:The skull defect was prepared and the defect area was filled with DBM,suture skin.(5)Dialysis BPE group:the skull defect was prepared and the defect area was filled with BPE,suture skin.(6)Cold acetone BPE group: the skull defect was prepared,and the defect area was filled with cold acetone precipitated BPE,to suture the skin.The periosteum of the operation area was removed in all groups.Results: 1.Preparation of BPE:Both dialyzed BPE and cold acetone BPE are white flocculent and soft in texture.Cold acetone BPE is yellowish in color and looser in structure than dialyzed BPE.2.Gross observation of skull samples:The gross observation of the inner surface of the skull sample showed that the bone surface of the normal group was smooth,the tissue of the defect group was thin and sunken,and the skull defect was soft in the Bio-oss group,and the bone powder particles could still be seen on the surface of the skull defect in the defect group,and the bone powder particles could still be seen on the surface of the skull defect in the defect group,and the bone powder particles could still be seen on the surface.The general view of the inner surface of skull in DBM group,dialysis BPE group and cold acetone BPE group was similar,the tissue of defect area was thicker than that of control group,and the tissue of defect area in cold acetone BPE group was the thickest.3.CBCT image analysis:The results of CBCT showed that the skull of the normal group was continuous and intact;There was no obvious new bone around the bone defect area in the defect group;In the Bio-oss group,the defect area was irregular,the distribution of bone powder particles was blurred,and the edge of the bone powder particles was blurred;In the DBM group,dialysis BPE group and cold acetone BPE group,the edge of the defect area was irregular,and a small amount of new bone with different width could be seen,and the density was slightly lower than that of normal bone.4.HE staining of tissue sections:The results of HE staining showed that in the normal group,the internal and external plate of the skull was dense and continuous and the structure was intact,the defect area of the defect group was covered with a thin layer of fibrous tissue,and there was only a small amount of immature new bone between the defect area and the original bone.In the Bio-oss group,the fibrous tissue wrapped in the position of bone powder particles,red staining in the central area and a small amount of new bone formation in the periphery.In DBM group,immature new bone formed near the defect edge of host bone,which was more than that in defect group,while there were similar results in dialysis BPE group and cold acetone BPE group,and there were more immature bone formation in the edge of defect adjacent to host bone in dialysis BPE group,but there was a small amount of collagen deposition in the center of defect area in dialysis BPE group.5.Masson staining of tissue sections:The results of Masson trichromatic staining showed that the skulls of the normal group were deeply red and closely arranged with collagen fibers,while the histological results of the defect group showed that the defect area was covered with a thin layer of fibrous tissue,and there was a clear boundary between the defect and the original bone.In Bio-oss group,the position occupied by bone particles showed a gap,and the central defect area was occupied by bone powder particles.The fibrous tissue is wrapped in implanted bone powder particles,accompanied by a small amount of new bone formation.In the DBM group,there were a small amount of blue-stained new bone on the edge of the host bone;In the dialysis BPE repair group,there were more immature bone formation at the edge of the defect adjacent to the host bone and a small amount of collagen matrix deposition in the center of the defect;And there were more blue-stained new bone on the edge of the host bone in the cold acetone BPE group,but only a thin layer of fibrous tissue was covered in the center of the defect area.6.Statistical analysis of newborn bone area in defect area:The percentage of new bone area measured by Masson staining in each group was statistically analyzed by SPSS18.0.It was found that the percentage of new bone mass in each group was 18.42%±1.77% in Bio-oss group,23.78%±2.02% in DBM group,30.44%±1.56% in dialysis BPE group,20.34%±1.31% in cold acetone BPE group,and 10.57%±1.81% in each group.And there was statistical significance(P < 0.01).The new bone mass in dialysis BPE group was higher than that in DBM group(P < 0.01);The new bone mass in DBM group was higher than that in cold acetone BPE group(P <0.01);The new bone mass in dialysis BPE group was higher than that in Bio-oss group(P < 0.01);The new bone mass in cold acetone BPE group was higher than that in Bio-oss group(P < 0.05);The new bone mass in dialysis BPE group was higher than that in cold acetone BPE group(P < 0.01).Conclusion: 1.BPE has a certain ability of bone repair,but the effect of maintaining osteogenic space is poor.2.There are significant differences in the effect of BPE prepared by different methods on skull osteogenesis,which may be related to the different contents of osteogenic components in BPE. |
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