摘要
目的观察聚乳酸支架材料与角膜上皮样细胞的体外生物相容性。方法通过热压法与流延法分别制作聚乳酸支架材料。复苏培养前期实验获得的角膜上皮样细胞,免疫荧光化学法鉴定,将第三代角膜上皮样细胞分别种植于上述2种聚乳酸支架材料,显微镜下观察此两种支架材料上细胞的存活与生长情况;采用伊红染色观察支架材料上细胞数量与形态,噻唑蓝法检测细胞活力。结果热压法聚乳酸支架材料为纤维交织立体结构,而流延法材料为实性结构,无孔隙。细胞种植于支架材料后,细胞种植4d后镜下可见热压法聚乳酸支架材料组大量细胞存活,且紧贴着支架材料生长并向支架材料内伸展,伊红染色可见材料上大量着色的细胞;流延法组细胞存活极少,伊红染色仅见极少细胞附着在材料上。结论热压法聚乳酸支架材料具备纤维孔隙结构,支持角膜上皮样细胞的存活与生长,具备与角膜上皮样细胞有较好的生物相容性,有进一步开发角膜上皮样细胞移植支架的前景。
Objective This study aimed to investigate the biocompatibility of different spatial structure of polylactic acid scaffolds to corneal epithelial-like cells in vitro. Methods Polylactic acid scaffolds were made respectively by hot pressing （fiber membrane made with electrostatic spinning, and then hot pressing）, and tape-casting methods. After resuscitation of corneal epithelial cells, the third generation of cells were identified by immunohistoehemistry, then were grown in the above two kinds of polylactie acid scaffolds. Cell viability was detected by MTT. And the survival and growth of cells were observed under microscope. Results Polylactic acid scaffold made in hot pressing method was fiber interwoven stereo structure, and that made in tape-casting was a solid structure with no pore. MTT values were lower at 48 h and 96 h, respectively, in tape-casting-made polylactic acid scaffold than that in hot pressing-made polylactic acid scaffold. The microscope observation and eosin staining found that the cells survival better in hot pressing-made polylactic acid scaffold than that in tape-casting-made polylactic acid scaffold. Conclusion The hot pressing-made polylactic acid scaffold with fiber pore structure, were suitable for epithelial-like cells survival and growth. There was the further development of scaffold for corneal epitbolioid-like cells transplantation.
出处
《四川解剖学杂志》
2015年第1期1-5,共5页
Sichuan Journal of Anatomy
基金
四川省教育厅重点科研项目(15ZA0266)
四川省科技厅应用基础研究资助项目(2011JYZ035)
四川省科技创新苗子工程项目(2014-102)
教育部人才培养能力提高项目(项目代码:J1103604)
关键词
聚乳酸
生物支架材料
角膜上皮样细胞
生物相容性
Polylactic acid（PLA） scaffolds
corneal epithelial-like cells
biocompatibility
