摘要
目的:探讨同种异体骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)体外分离培养后种植到孔径和孔隙率不同且复合不同胶原和生长因子的PLGA支架上再种植到大鼠肌袋内构建组织工程化骨软骨复合组织的可行性。方法:制作复合BMP-2和Ι型胶原PLGA支架与复合bFGF、TGF-β1和Ⅱ型胶原PLGA支架后把两者用生物蛋白胶粘合形成复合支架,把体外培养扩增的BMSCs种植到复合支架上,植入实验组A组、对照组B组、空白组C组SD大鼠肌袋内,于术后4、8、12周取材,分别行大体观察、HE染色、甲苯胺蓝染色、Ⅱ型胶原免疫组化、ALP染色、扫描电镜、透视电镜观察。结果:实验组大体标本成骨区呈白色类骨样组织,质硬,成软骨区呈乳白色类软骨样组织,质较硬,两区融合;扫描电镜观察可见成骨细胞、破骨细胞及软骨细胞,透视电镜尚可见细胞浆内有大量的线粒体和内质网,并见大量胶原分泌,与对照组、空白组有明显区别,组织学评分表明A组较B、C两组差异具有统计学意义(P<0.05)。结论:BMSCs体外分离扩增后种植到孔径和孔隙率不同且复合不同胶原和生长因子的PLGA支架上再植入动物肌袋可构建骨软骨复合组织。
Aim：To explore the feasibility of constructing tissue engineered osteochondral composite in the rat muscle pouch through isolating and culturing of allogenic bone mesenchymal stem cells（BMSCs） culturing on the poly-lactide-co-glycolic acid（PLGA）scaffolds,which have different interstice ratio and different diameter and modified with different collagen and different cellular growth factor. Methods： Modified with the factor of BMP-2 and typeⅠcollagen PLGA scaffold and modified with the bFGF,TGF-β1 and typeⅡcollagen PLGA scaffold were made and adhered together using fabrin glue to form composite scaffold,transplanted the isolating and culturing of BMSCs to the composite scaffold,then planted the cellular-scaffold composite to the SD rat＇s muscle pouch of experiment group（Group A）,controlled group（Group B）,blank group（Group C）,4,8,12 weeks after operation,observed their general shapes,HE,Toluidine Blue,typeⅡcollagen,ALP staining,scan electron microscope and perspective electron microscope.Results： The experiment group＇s general shape： osteoblast area showed the white,hard,osteoid tissue,the cartilage area showed the ivory-white,less hard,chondroid tissue,the two areas syncretize;osteoblasts,osteoclasts and chondral cells were observed using the scan electron microscope,and with the perspective electron microscope,a great mass of mitochondrion and endoplasmic reticulum,and much of collagen were observed,this result was obvious difference from the controlled and blank group,histology evaluation showed that group A had the statistic difference compared with the group B and C.Conclusion：After isolation and expansion in vitro,the BMSCs can be cultured on the PLGA scaffolds which have different interstice ratio and different diameter and modified with different collagen and different cellular growth factor,and then plant the cellular-scaffold into the muscle pouch of animal,in this way we can construct the osteochondral composite tissue.
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
北大核心
2010年第2期168-173,共6页
Journal of Jinan University(Natural Science & Medicine Edition)
基金
国家863高技术研究发展计划资助项目(2007AA09Z440)
澳门医学科技研究基金项目(031/2009/A)
广州市科技计划项目(2008Z1-E411)
关键词
异体骨髓间充质干细胞
聚乙醇酸-乳酸共聚体
诱导
骨软骨复合组织
bone marrow mesenchymal stem cells（BMSCs）
poly-lactide-co-glycolide（PLGA）
induce
osteochondral composite tissue
