摘要
目的 观察人骨形态发生蛋白-2（BMP-2）基因转染脂肪干细胞（ADSCs）复合脱钙骨基质材料（DBM）构建组织工程化骨修复骨缺损的效果.方法 按Urist描述方法制备DBM材料.体外分离培养兔ADSCs,以人BMP-2重组腺病毒载体（rAd5-BMP2-EGFP）转染第2代ADSCs,将转染后的ADSCs复合DBM材料复合培养7d.取31只新西兰大白兔建立兔桡骨15 mm长节段性骨缺损,分别植入rAd5-BMP2-EGFP转染的自体ADSCs复合DBM（A组,n=9）、自体ADSCs复合DBM（B组,n=9）、单纯DBM（C组,n=9）以及空白组（D组,n=4）,术后8周及24周通过大体观察、X线、组织学以及生物力学方法检测缺损修复.结果 人BMP-2重组腺病毒转染后ADSCs表达绿色荧光蛋白,细胞复合DBM培养扫描电镜观察证实大量细胞黏附支架材料.术后8、24周大体观察及组织学检测,A、B两组均有不同程度骨性修复,C组及D组无连续性骨性修复.术后8周X线评分A、B、C组分别为（9.12 ±0.47）、（6.73±0 39）、（2.00±0.10）分,3组间差异有统计学意义（P＜0.05,n=6）;术后24周X线评分A、B、C组分别为（11.50±0.31）、（9.00±0.48）、（2.10±0.10）分,3组间差异有统计学意义（P＜0.05,n=12）.生物力学检测植入后24周A组及B组最大载荷分别为（97.0±14.7）N及（76.2±9.6）N;抗压强度分别为（8.7±0.4） mPa及（7.0±0.6） mPa,弹性模量分别为（185.3±21.9） GPa及（159.8±15.7） GPa,A组生物力学性能优于B组（P＜0.05,n =6）.结论 以ADSCs为种子细胞复合DBM构建组织工程骨可以较好的修复节段性骨缺损,BMP-2基因增强组织工程骨可以改善修复效果.
Objective To evaluate the efficiency of adipose-derived stem cells transfected with adenovirus-mediated bone morphogenetic protein-2 （BMP-2）combined with demineralized bone matrix （DBM） on the repair of critical-sized radial defects in rabbits.Methods DBM of rabbit was prepared based on the Urist＇ s method.Rabbit adipose-derived stem cells （ADSCs） were isolated,cultured in vitro,and cells of passage 2 were transfected with recombinant adenovirus vector containing human bone morphogenetic protein 2 （rAd5-BMP2-EGFP）.These cells were coculmred with DBM and observed by scanning electron microscope （SEM）.Critical-size radial defects were made bilaterally in 31 rabbits,and individual sites were implanted with gene enhanced tissue-engineered bone （rAd-BMP2-EGFP-ADSCs/DBM,group A,n =9）,tissue-engineered bone without modification （ADSCs/DBM,group B,n =9）,single DBM scaffold （DBM,group C,n =9） or no implants （group D,n =4）.New bone formation was evaluated at 8 and 24 weeks by X-ray examination,gross samples observation and histological observation.The biomechanical test were also performed 24 weeks postoperation.Results After transfection,the BMP-2 expression was detected by fluorescence microscope indirectly.Scanning electron microscope （SEM） demonstrated integration of ADSCs with the DBM scaffold.8 and 24 weeks after implantation,gross samples observation,histological observation showed that there were new bones formed and the bone defects were repaired successfully to some extent in group A and B,but continuous bony repair was not obtained in group C and D.The X-ray evaluated scores of group A,B and C were 9.12 ±0.47,6.73 ±0.39,and 2.00 ±0.10 at 8 weeks post operation （P〈0.05,n=6）,and were 11.5 ±0.31,9.00±0.48,2.10±0.10 at 24 weeks post operation （P 〈0.05,n =12）,there was significant difference hetween A,B and C.24 weeks after implantation,biomechanical analysis showed that the maximum loads of group A and B were （97.0 ± 14.7） N and （76.2 ± 9.6） N,the
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第11期2509-2511,共3页
Chinese Journal of Experimental Surgery
关键词
骨缺损修复
组织工程
基因转染
脂肪干细胞
骨形态发生蛋白-2
Bone defects repair
Tissue engineering
Gene transfection
Adipose-derived stern cells
Bone morphogenetic protein-2
