犬骨髓间充质干细胞复合磷酸钙骨水泥修复骨缺损的实验研究  

Experimental study of canine bone marrow mesenchymal stem cells combined with calcium phosphate cement for repair of mandibular bone defects in Beagle dogs

作　　者：胡宜成 刘鑫[1] 沈际佳[2] 何家才[1] 陈乔尔[1] 

机构地区：[1]安徽医科大学口腔医学院·安徽医科大学附属口腔医院,安徽省口腔疾病研究中心实验室,安徽合肥230032 [2]安徽医科大学病原微生物教研室,安徽合肥230032

出　　处：《上海口腔医学》2014年第4期402-408,共7页Shanghai Journal of Stomatology

基　　金：国家自然科学基金(81371114);安徽高校省级自然科学研究重点项目(KJ2013A154)~~

摘　　要：目的:评价骨髓间充质干细胞复合磷酸钙骨水泥支架材料修复下颌骨缺损的效果。方法:分离培养犬骨髓间充质干细胞(BMSCs)。成骨诱导培养14 d后,分别采用茜素红染色与碱性磷酸酶染色,观察其诱导效果。将细胞与磷酸钙骨水泥(CPC)支架材料复合,用于动物实验。在4只Beagle犬的下颌骨每侧制作3处大小一定的骨缺损。随机将骨缺损分为3组进行处理:BMSCs-CPC组(移植复合种子细胞的支架材料)、CPC组(只移植支架材料)和空白组(不做任何处理)。分别于移植后第4、8周处死2只犬,行大体、X线、骨缺损修复区组织形态观察与计量分析。采用SPSS13.0软件包对数据进行统计学分析。结果:术后各组均有不同程度的骨再生。BMSCs-CPC组、CPC组中骨组织再生情况优于空白组。BMSCs-CPC组支架材料降解程度与新骨形成情况优于CPC组。术后第4、8周,BMSCs-CPC、CPC组新生骨面积百分比均显著高于空白对照组(P<0.05),BMSCs-CPC组新生骨面积百分比显著高于CPC组(P<0.01);BMSCs-CPC组中剩余支架材料面积百分比显著低于CPC组(P<0.01)。结论:BMSCs种子细胞复合CPC支架材料是一种有效的、促进新骨再生的骨缺损修复方法,有利于颌骨高度及宽度的保存。PURPOSE： To evaluate the effects of bone marrow mesenchymal stem cells （BMSCs） combined with calcium phosphate cement （CPC） scaffold for repair of mandibular defect in Beagle dogs. METHODS： BMSCs were isolated from Beagle dogs and cultured in DMEM plus 10% FBS. The induction effect was determined using alizarin red staining or alkaline phosphate staining at 14-day of culture. BMSCs were added to the CPC scaffold for animal experiments. In vivo, three critical size bone defects were surgically created in each side of the mandible. The bone defects were repaired with BMSCs-CPC （scaffolds with composite seeding cells）, CPC （scaffold alone） or no materials （blank group）. Two dogs were sacrificed at 4-week and 8-week after operation. Gross observation, X-ray imaging, histologic and histometric analyses were performed to evaluate the level of bone formation. RESULTS： Newly formed bones were detected within all defect sites after operation. The BMSCs-CPC group and CPC group showed increased bone formation compared with the blank group. The BMSCs-CPC group exhibited more bone formation and degradation of the material than the CPC group. The percentage of new bone in the BMSCs-CPC and CPC treated group were significantly higher than that in the control group （P〈0.05）, while the percentage of new bone in the BMSCs-CPC sites was higher than that in the CPC sites （P〈0.01）; the percentage of residual material in the BMSCs-CPC sites was lower than that in the CPC sites （P〈0.01） 4 weeks and 8 weeks after operation. CONCLUSIONS： Using the theory of tissue engineering, BMSCs composite CPC compound is an effective method in promoting new bone regeneration, which has a positive influence on the bone space preservation.

关 键 词：骨髓间充质干细胞 磷酸钙骨水泥 下颌骨缺损 BEAGLE犬 

分 类 号：R783[医药卫生—口腔医学]