内皮祖细胞与纳米羟基磷灰石／胶原／聚乳酸复合物在修复兔骨缺损中的作用  

Promoting of angiogenesis and osteogenesis in radial critical bone defect regions of rabbits with nano- hydroxyapatite/collagen/PLA scaffolds plus endothelial progenitor cells

作　　者：吴东进[1] 郝爱华[4] 张程[1] 崔福斋[5] 王修文[1] 高春正[1] 周成军[2] 刘军莉[3] 谯勇[1] 马胜忠[1] 

机构地区：[1]山东大学第二医院脊柱外科,济南250033 [2]山东大学第二医院病理科,济南250033 [3]山东大学第二医院 分子生物学实验室,济南250033 [4]山东中医药大学附属医院放射科 [5]清华大学材料科学与工程系再生医学与仿生材料研究所

出　　处：《中华医学杂志》2012年第23期1630-1634,共5页National Medical Journal of China

基　　金：基金项目：山东省科技攻关课题（2009GG10002036）

摘　　要：目的探讨内皮祖细胞（EPC）与纳米羟基磷灰石／胶原／聚乳酸（nHAC／PLA）复合物在修复兔桡骨节段性骨缺损中促进血管新生及新骨形成的作用。方法从新西兰大兔骨髓分离、培养EPC并予以鉴定，将EPC与nHAC／PLA支架复合，以扫描电镜（SEM）观察EPC在支架内的生长状态。将36只新西兰大白兔随机数字表法分为3组，制作桡骨节段性骨缺损模型，分别植入EPC．nHAC／PLA复合物（A组，n=16）、nHAC／PLA支架材料（B组，n=16），其余4只（C组）不用任何内植物以作空白对照。分别于不同的时间点处死动物，收取桡骨并行放射学、组织学检查及血管密度计数。结果白骨髓分离的细胞为EPC，SEM显示其与nHAC／PLA复合后黏附、生长、增殖良好。A组骨缺损处的放射学评分[（4．3±0．5）比（3．0±0．0），均P〈0．05]、新骨形成区占骨缺损区的百分比[（96．0％±4．3％）比（34．0％±6．6％），均P〈0．05）]、微血管密度[（7．0±0．4）比（4．5±0．4），均P〈0．05）]均高于B组，差异有统计学意义，组织学显示A组的新骨形成及支架降解快于B组。C组无明显新骨形成。结论EPC—nHAC／PLA复合物能明显促进骨缺损处血管新生和新骨形成，提示其在应用组织工程骨修复大段骨缺损的再血管化和成骨方面有潜在的应用价值。Objective To explore the roles of nano-hydroxyapatite/collagen/PLA （nHAC/PLA） plus endothelial progenitor cells （EPCs） in repairing segmental bone defects of rabbit radius and enhancing angiogenesis and new bone formation. Methods EPCs isolated from New Zealand white rabbit bone marrow were cultured, identified and seeded into nHAC/PLA scaffolds. And the growth of EPCs in scaffolds was observed under scanning electron microscopy （ SEM ）. Thirty-six were randomly divided into 3 groups to establish segmental bone defect models in radii. Two groups were implanted with EPCs/scaffolds constructs （ group A, n = 16） and scaffolds alone （ group B, n = 16） respectively. The remaining four rabbits were used as negative control （ group C） and nothing was implanted. Animals were sacrificed at different timepoints and radii harvested to undergo radiological examination, histological examination and mierovessle density test. Results These cells isolated from bone marrow were confirmed as EPCs. SEM showed that EPCs attached to the nHAC/PLA scaffolds, grew and proliferated well. Animal experiments revealed that radiological scores （5w：2. 25 ±0. 50 vs 1.00 ±0. 00; 10w：2. 75 ±0. 50 vs 1.75 ±0. 50; 15w：4. 25 ±0. 50 vs 3.0 ± 0. 0 ; each P 〈 0.05 ）, percentage of new bone formation area in bone defect regions （ 5w ：29.0% ± 3.5% vs 8.1% ±0. 8% ; 10w：63.4% ± 5.5% vs 16. 6% ± 1.3% ; 15w：96. 0% ±4. 3% vs 34.0% ±6.6%; eachP〈0.05） and microvesseldensity （2w：13.5 ±0.9 vs4.3 ±1.0; 5w：9.8 ±0.7 vs4.8 ± 0. 3; 10w：7.0 ±0. 4 vs 4. 5 ±0. g; each P 〈0. 05） in group A were significantly higher than those in group B. No new bone formation occurred in group C. Conclusion The composite structure of EPCs-nHAC/PLA can enhance angiogenesis and new bone formation in segmental bone defects in rabbit radii. It may become a potential candidate of promoting revascularization of tissue engineering bone and repairing large bone defects.

关 键 词：修复外科手术 骨生成 组织疗法 干细胞 

分 类 号：R687.3[医药卫生—骨科学]