中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (30): 4449-4455.doi: 10.3969/j.issn.2095-4344.2016.30.006

• 材料生物相容性 material biocompatibility • 上一篇    下一篇

脱细胞猪角膜基质支架的制备及其生物相容性

霍银平，周利晓，孙成林   

  1. 郑州大学第五附属医院眼科，河南省郑州市  450052
  • 收稿日期:2016-04-27 出版日期:2016-07-15 发布日期:2016-07-15
  • 通讯作者: 周利晓，博士，主任医师，郑州大学第五附属医院眼科，河南省郑州市 450052
  • 作者简介:霍银平，女，1977年生，汉族，河南省开封市人，2007年郑州大学毕业，硕士，主治医师，主要从事眼底病白内障及眼表疾病研究。
  • 基金资助:

    河南省医学科技攻关计划项目(201503131)

Preparation of an acellular porcine corneal stroma scaffold and its biocompatibility

Huo Yin-ping, Zhou Li-xiao, Sun Cheng-lin   

  1. Department of Ophthalmology, the Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
  • Received:2016-04-27 Online:2016-07-15 Published:2016-07-15
  • Contact: Zhou Li-xiao, M.D., Chief physician, Department of Ophthalmology, the Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
  • About author:Huo Yin-ping, Master, Attending physician, Department of Ophthalmology, the Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
  • Supported by:

    the Medical Science and Technology Research Project of Henan, No. 201503131

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摘要：

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文题释义：
脱细胞角膜基质：角膜基质经脱细胞处理后，角膜上皮细胞被完全除去，胶原层变松散，基质内有少量残留细胞，经多次冻融后胶原层恢复原来结构，基质中残留细胞被除去，有微量核物质残留，经DNA- RNA酶处理后，残留核物质被除去，风干后角膜结构完整，胶原纤维排列整齐。
组织工程角膜载体支架材料：由于胶原、壳聚糖、纤维蛋白、透明质酸等载体支架均存在一定的局限性，和天然的角膜成分和结构存在差异，在临床应用中有一定限制。脱细胞角膜基质的成分和结构和天然的角膜比较相似，含有多种细胞因子，免疫原性比较低，可塑性比较好，机械强度比较好，是一种比较理想的组织工程角膜载体支架材料。


背景：研究发现多种生物材料可制备角膜基质支架且具有良好的生物相容性，而关于脱细胞猪角膜基质支架的制备和生物相容性研究不多。
目的：制备脱细胞猪角膜基质支架，探讨其与人角膜基质细胞的生物相容性。
方法：制备脱细胞猪角膜基质支架及其浸提液。取生长状态良好的人角膜基质细胞，分别以脱细胞猪角膜基质支架浸提液(实验组)、完全培养基培养(对照组)培养，培养后1，2，3 d，采用MTT法检测人角膜基质细胞的增殖；将人角膜基质细胞直接接种于脱细胞猪角膜基质支架上，接种1，3，7 d，采用免疫化学法测定人角膜基质细胞在支架内的生长情况。
结果与结论：①随着时间的增加，实验组与对照组人角膜基质细胞数量不断增加，两组不同时间点的吸光度值比较差异无显著性意义；②人角膜基质细胞在脱细胞猪角膜基质支架上生长良好，细胞整联蛋白β1、波形蛋白、乙醛脱氢酶3A1呈阳性表达，CD34、CDK2和K-Ras蛋白也呈阳性表达；③结果表明，脱细胞猪角膜基质支架无细胞毒性，与人角膜基质细胞具有良好的生物相容性。

ORCID: 0000-0002-8330-4397(周利晓)

关键词: 生物材料, 材料相容性, 脱细胞猪角膜基质支架, 生物相容性, 人角膜基质细胞, 整联蛋白β1, 波形蛋白, 乙醛脱氢酶3A1

Abstract:

BACKGROUND: Studies have found that a variety of biological materials can be used for preparing corneal stroma scaffolds that have good biocompatibility, but research on preparation and biocompatibility of the acellular porcine corneal stroma scaffold is little.
OBJECTIVE: To explore the preparation and biocompatibility of the acellular porcine corneal stroma scaffold.
METHODS: Acellular porcine corneal stroma scaffold and its extract were prepared. Well-grown human corneal stromal cells were selected and cultured in the extract of acellular porcine corneal stroma scaffold (experimental group) or in the complete medium (control group), respectively. After 1, 2 and 3 days of culture, the proliferation ability of human corneal stromal cells was detected by MTT assay. In the meanwhile, human corneal cells were directly seeded onto the acellular porcine corneal stroma scaffold, and then the cell growth on the scaffold was detected using immunochemical method.
RESULTS AND CONCLUSION: The number of human corneal stromal cells was in a rise with time in the two groups, and absorbance values had no significant difference between two groups at different time points of culture. Human corneal stromal cells grew well on the scaffold, and were positive for cell integrin β1, vimentin, aldehyde dehydrogenase 3A1, as well as CD34, CDK2 and K-Ras. These results show that the acellular porcine corneal stroma scaffold has no cytotoxicity, and has good biocompatibility.

 

Key words: Biocompatible Materials, Corneal Stroma, Tissue Engineering

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