摘要
目的评价组织工程骨膜体内成骨修复兔大段骨缺损及同种异体脱蛋白骨(deproteinized bone,DPB)作为组织工程骨膜辅助支架材料的效果。方法分离培养1月龄新西兰大白兔BMSCs,以成骨诱导培养液诱导成骨分化后,与猪小肠黏膜下层(small intestinal submucosa,SIS)复合构建组织工程骨膜,扫描电镜观察细胞与SIS复合情况。取新鲜同种异体骨段细密打孔后进行脱蛋白处理,制成DPB支架材料。将组织工程骨膜呈"外套状"包被DPB,并用可吸收缝线捆扎制备组织工程骨膜/DPB复合体。取4月龄清洁级新西兰大白兔48只,随机分成A、B、C、D 4组(n=12),切除左侧桡骨干3.5 cm长骨段制作大段骨缺损动物模型。A、B、C组分别于骨缺损处植入组织工程骨膜、DPB和组织工程骨膜/DPB复合体,D组骨缺损旷置。术后4、8、12周各组随机取4只动物行X线片观察及评分;术后8周各组随机处死4只动物,取骨缺损区标本行HE及Masson染色观察。结果扫描电镜观察构建的组织工程骨膜示SIS上黏附大量细胞。X线片检查示术后各时间点A、C组新生骨组织量明显多于B、D组。术后各时间点A、C组X线片评分显著高于B、D组,A组高于C组,B组高于D组,差异均有统计学意义(P<0.05)。组织学观察示A组骨缺损处为大量新生骨组织,新生骨组织中可见不规则髓腔样结构及丰富的血管腔;B组骨缺损处见植入的DPB,DPB降解不明显;C组骨缺损处见较多新生骨组织,新生骨组织中可见呈岛状分布的DPB,DPB降解明显;D组骨缺损处仅见少量胶原纤维。结论以SIS和BMSCs构建的组织工程骨膜可修复兔桡骨大段骨缺损;DPB不能对组织工程骨膜成骨起到理想的支架作用,组织工程骨膜辅助支架材料有待进一步探索。
Objective To evaluate the effect of tissue engineered periosteum on the repair of large diaphysis defect in rabbit radius, and the effect of deproteinized bone （DPB） as supporting scaffolds of tissue engineering periosteum. Methods Bone marrow mesenchymal stem ceils （BMSCs） were cultured from 1-month-old New Zealand Rabbit and osteogenetically induced into osteoblasts. Porcine small intestinal submucosa （SIS） scaffold was produced by decellular and a series mechanical and physiochemical procedures. Then tissue engineered periosteum was constructed by combining osteogenic BMSCs and SIS, and then the adhesion of cells to scaffolds was observed by scanning electron microscope （SEM）. Fresh allogeneic bone was drilled and deproteinized as DPB scaffold. Tissue engineered periosteum/DPB complex was constructed by tissue engineered periosteum and DPB. Tissue engineered periosteum was ＂coat-like＂ package the DPB, and bundled with absorbable sutures. Forty-eight New Zealand white rabbits （4-month-old） were randomly divided into 4 groups （groups A, B, C, and D, n=12）. The bone defect model of 3.5 cm in length in the left radius was created. Defect was repaired with tissue engineered periosteum in group A, with DPB in group B, with tissue engineered periosteum/DPB in group C; defect was untreated in group D. At 4, 8, and 12 weeks after operation, 4 rabbits in each group were observed by X-ray. At 8 weeks after operation, 4 rabbits of each group were randomly sacrificed for histological examination. Results SEM observation showed that abundant seeding cells adhered to tissue engineered periosteum. At 4, 8, and 12 weeks after operation, X-ray films showed the newly formed bone was much more in groups A and C than groups B and D. The X-ray film score were significantly higher in groups A and C than in groups B and D, in group A than in group C, and in group B than in group D （P 〈 0.05）. Histological staining indicated that there was a lot of newly formed bone in the defect space in group A, with abundant newly formed vessels and medullary cavity. While in group B, the defect space filled with the DPB, the degradation of DPB was not obvious. In group C, there was a lot of newly formed bone in the defect space, island-like DPB and obvious DPB degradation were seen in newly formed bone. In group D, the defect space only replaced by some connective tissue. Conclusion Tissue engineered periosteum constructed by SIS and BMSCs has the feasibility to repair the large diaphysis defect in rabbit. DPB isn＇t an ideal support scaffold of tissue engineering periosteum, the supporting scaffolds of tissue engineered periosteum need further exploration.
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2014年第4期511-516,共6页
Chinese Journal of Reparative and Reconstructive Surgery
基金
国家自然科学基金资助项目(30973064)~~
关键词
骨组织工程
组织工程骨膜
小肠黏膜下层
BMSCS
脱蛋白骨
兔
Bone tissue engineering Tissue engineered periosteum Small intestinal submucosa Bone marrow mesenchymal stem ceils Deproteinized bone Rabbit
