摘要
目的 探索体外构建组织工程化人工血管的可行性。方法 通过酶消化法分离牛主动脉血管内皮细胞 ,培养、传代 ,纯化。采用交联胶原包埋处理聚羟基乙酸纤维无纺网 (PGA) ,将第 3～ 7代血管内皮细胞接种于上述材料上 ,通过特制的旋转装置 ,缓慢动态旋转培养 10天 ,使内皮细胞在管腔内表面附着生长 ,扫描电镜观察 ,采用 6 -酮 -前列腺素 - 1α放射免疫测试药盒测定管形材料中内皮细胞释放前列环素量。结果 培养血管内皮细胞 VIII因子相关抗原抗体染色呈阳性 ,内皮细胞在管腔内表面贴附良好 ,细胞之间融合成片 ,经 10天培养 ,形成较完整内膜层 ,覆盖率为 (91.2± 1.5 ) % ,前列环素生成率为 (4 .6± 0 .5 ) μg/ cm2 · min,与对照组比较有非常显著性差异 (P<0 .0 1)。结论 PGA+胶原支架适宜血管内皮细胞的生长 ,通过缓慢动态旋转培养技术 ,明显提高了血管内皮细胞的覆盖率 。
Objective To explore the feasibility of reconstructing tissue engineered vessel in vitro. Methods Bovine endothelial cells were isolated from calf thoracic aorta by enzyme digestion methods and subcultured and purified. The endothelial cells of the 3rd to 7th passages were seeded into the inner surface of tubular scaffold material by polyglycolic acid(PGA) coated with cross-linked collagen, and cultured in vitro for 10 days using dynamic rotation culture technique. Scanning electron microscopy was used to analyse the morphological characteristics, and prostacyclin released by endothelial cells was measured by radioimmunoassay of 6-keto-prostaglandin F1α. Results The Ⅷ factor staining of cultured endothelial cells was positive. The endothelial cells adhered well on the inner surface of tubular scaffold material with confluent monolayer covering(91.2±1.5)%. The endothelialized model released prostacyclin at a rate of (4.6±0.5) μg/cm 2·min. There was significant difference to control group ( P <0.05). Conclusion The PGA coating with collagen is an ideal scaffold for endothelial cells, the coverage rate is increased through dynamic rotation culture technique. It will lay a good foundation for architecture of a laminated structure of tissue engineered vessel.
出处
《中国修复重建外科杂志》
CAS
CSCD
2001年第2期109-112,共4页
Chinese Journal of Reparative and Reconstructive Surgery
基金
国家自然科学基金资助项目! (3 0 0 70 777)
关键词
组织工程
聚羟基乙酸
血管
旋转培养
血管模型
Tissue engineering Polyglycolic acid Vessel Rotation culture Foundation item: National Natural Science Foundation of China(30070777)
