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干燥脱水保存鸵鸟脱细胞角膜基质载体材料的细胞毒性
作者姓名:刘先宁  朱秀萍  吴 洁  王莉芳  银 勇
作者单位:1陕西省眼科研究所,陕西省西安市 710002;2陕西省食品药品检验所,陕西省西安市 710002
基金项目:陕西省科技统筹创新工程计划项目(2012KTCQ03-11)。
摘    要:背景:陕西省眼科研究所通过前期研究发现鸵鸟角膜具有开发成为人角膜材料替代品的优势。 目的:判断鸵鸟角膜基质支架材料对细胞的潜在毒性作用。 方法:采用细胞培养方法,将干燥脱水法保存的鸵鸟脱细胞角膜基质载体支架制备成浸提液与L-929细胞共同培养,采用MTT比色法评价其作用1,2,3 d,对细胞生长和增殖的影响。 结果与结论:干燥脱水法保存的鸵鸟脱细胞角膜基质载体支架浸提液组培养1,3,5 d,对实验细胞毒性反应级别均为1级。参照《中华人民共和国国家标准GB/T16886.5-2003》进行细胞毒性试验,供试品组见少量细胞呈圆形,疏松贴壁,无胞浆内颗粒,偶见细胞溶解。结合MTT法分析结果表明,采用干燥脱水法保存鸵鸟脱细胞角膜基质载体材料,细胞毒性为1级,为合格材料。

关 键 词:组织构建  组织构建细胞学实验  组织工程角膜  角膜  鸵鸟  脱细胞角膜基质  支架  干燥脱水  保存  细胞毒性试验  省级基金  
收稿时间:2013-01-03

Cytotoxicity of dehydrated ostrich acellular corneal stroma as a carrier material
Authors:Liu Xian-ning  Zhu Xiu-ping  Wu Jie  Wang Li-fang  Yin Yong
Institution:1Shaanxi Institute of Ophthalmology, Xi’an  710002, Shaanxi Province, China; 2Shaanxi Institute for Food and Drug Control, Xi’an  710002, Shaanxi Province, China
Abstract:BACKGROUND: Previous studies from Shaanxi Institute of Ophthalmology have shown that ostrich cornea has the advantages to be developed into the alternatives of human corneal material.  OBJECTIVE:To determine the potential toxic effects of ostrich corneal stromal scaffold on cells. METHODS: Cell culture methods were used to culture L-929 cells in the extracts of ostrich acellular corneal stroma which was dried and dehydrated. 3-(4,5)-Dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide assay was used to evaluate the growth and proliferation of cells after cultured for 1, 2 and 3 days. RESULTS AND CONCLUSION:After the cells were cultured in the extracts of ostrich acellular corneal stroma subjected to dryness and dehydration for 1, 3 and 5 days, and the toxicity level of cultured cells was graded as level 1. The cytotoxicity test was conducted according to the “National Standard of the People's Republic of China GB/T16886.5-2003”. After cultured in the extracts of ostrich acellular corneal stroma, a small number of cells were round in shape and loosely adherent without intracytoplasmic granules, and cell lysis could be observed occasionally. The results of 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide assay showed that the ostrich acellular corneal stromal scaffold which was dried and dehydrated had level 1 of cytotoxicity and could be considered as a qualified material.
Keywords:tissue construction  cytology experiment in tissue construction  tissue-engineered cornea  cornea  ostrich  acellular corneal stroma  scaffold  dehydration  preservation  cytotoxicity test  provincial grants-supported paper  
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