摘要
目的选择观察细胞和组织工程血管支架黏附的最佳方法。方法用生物可降解材料聚β羟基丁酯（PHB）作为支架材料,运用盐析方法制成实心和不同孔径的膜片;将培养的第3代血管平滑肌细胞种植于膜片上,于培养的第1、3、7、11、14天,采用倒置显微镜下观察、苏木素-伊红（HE）染色、甲苯胺蓝染色、扫描电镜检查及MTT法检测材料上细胞附着和生长情况。结果倒置显微镜下无法观察附着于材料上的细胞;HE和扫描电镜标本制备过程中细胞丢失多,不能为材料提供准确的信息;甲苯胺蓝染色可快速观察材料上细胞附着情况;MTT法可定量测定材料上的细胞数。经比较发现,150～200μm孔径的PHB膜片上血管平滑肌细胞附着和生长最佳。结论甲苯胺蓝染色和MTT比色法是观察细胞在材料上黏附和生长的最佳方法。
Objective To choose an optimal method for observing cells adhesion on tissue engineering vessel scaffold.Methods Scaffolds of various pore caliber were made by salt-eluting method using biodegradable poly-β-hydroxybutyrate （PHB）.Vascular smooth muscle cells were implanted in the scaffolds of various pore caliber. Scaffolds were examined by phase-contrast microscope,HE staining,toluidine blue staining,scanning electronic microscope and MTT method on culturing days of 1,3,7,11,14 for finding how cells grew. Results The cells in scaffolds could not be seen under phase-contrast microscope.HE staining and scanning electronic microscope could not provide any accurate information due to cells loss during specimen preparation.Toluidine blue staining was a rapid method for observing cells adhesion on the scaffolds,and MTT method could quantitatively measure cells number on the scaffolds.Scaffolds with 150～200μm pore caliber showed the most number of the cells.Conclusion Toluidine blue staining and MTT method are the optimal methods for observing cells growth on the scaffolds.
出处
《江苏医药》
CAS
CSCD
北大核心
2008年第5期484-486,共3页
Jiangsu Medical Journal
关键词
血管组织工程
生物可降解材料
Vascular tissue engineering
Biodegradable material
