中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (33): 6151-6154.doi: 10.3969/j.issn.1673-8225.2010.33.017

• 组织构建与生物活性因子 tissue construction and bioactive factors • 上一篇    下一篇

不同灭菌方式与羊膜上皮细胞活性：以组织形态及肝细胞生长因子和金属蛋白酶组织抑制剂1为标准的评价

杨雨昆，赵  敏   

  1. 重庆医科大学附属第一医院眼科，重庆市  400016
  • 出版日期:2010-08-13 发布日期:2010-08-13
  • 通讯作者: 赵敏，教授，重庆医科大学附属第一医院眼科，重庆市 400016 minzhao2002@yahoo.com.cn
  • 作者简介:杨雨昆★，男，1984年生，四川省宜宾市人，汉族，重庆医科大学在读硕士，主要从事角膜病、眼表面疾病方面的研究。
  • 基金资助:

    重庆市科技攻关项目(渝科成登字2007Y250)。

Different sterilization methods and amniotic membrane activity: Evaluation based on morphological structure, expressions of hepatocyte growth factor, as well as tissue inhibitor of metalloproteinase-1

Yang Yu-kun, Zhao Min   

  1. Department of Ophthalmology, First Affiliated Hospital of Chongqing University of Medical Sciences, Chongqing  400016, China
  • Online:2010-08-13 Published:2010-08-13
  • Contact: Zhao Min, Professor, Department of Ophthalmology, First Affiliated Hospital of Chongqing University of Medical Sciences, Chongqing 400016, China minzhao2002@yahoo.com.cn
  • About author:Yang Yu-kun★, Studying for master’s degree, Department of Ophthalmology, First Affiliated Hospital of Chongqing University of Medical Sciences, Chongqing 400016, China
  • Supported by:

    the Tackle Key Program in Science and Technology of Chongqing, No. 2007Y250*

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摘要：

背景：羊膜作为一种生物材料广泛运用于眼表疾病的治疗并取得积极疗效，现其灭菌方式有抗生素浸泡、过氧乙酸/乙醇混合液灭菌法、γ射线照射3种方式，但哪种方法灭菌效果最好，对羊膜损伤最小尚不清楚。
目的：探讨现有常用不同灭菌方式对羊膜生物活性的影响。
方法：取健康剖宫产产妇胎盘羊膜，分别以抗生素、过氧乙酸/乙醇混合液、γ射线、碘伏灭菌，苏木精-伊红染色观察羊膜上皮细胞形态，免疫荧光和RT-PCR检测羊膜上皮细胞肝细胞生长因子(hepatocyte growth factor，HGF)、金属蛋白酶组织抑制剂1(tissue inhibitor of metalloproteinase-1，TIMP-1)蛋白和mRNA的表达。
结果与结论：经过氧乙酸/乙醇混合液和抗菌药处理的羊膜上皮细胞形态完好，而经γ射线处理的细胞形态损害较大。经灭菌处理后，羊膜细胞的HGF和TIMP-1蛋白表达均有所下降(P < 0.05)，其表达量由高到低依次为过氧乙酸/乙醇混合液组、抗菌药组、碘伏组和γ射线组；HGF和TIMP-1的mRNA表达也有下降(P < 0.05)，其中γ射线处理组下降趋势更明显(P < 0.05)，其余3个处理组间差异无显著性意义(P > 0.05)。因此，就现行常用的灭菌方式而言，过氧乙酸/乙醇混合液对羊膜的损伤最小。

关键词: 羊膜, 灭菌, 肝细胞生长因子, 金属蛋白酶组织抑制剂1, 过氧乙酸/乙醇混合液

Abstract:

BACKGROUND: Amniotic membrane (AM) is widely used in treating ocular surface diseases and receives a positive curative effect. Different procedures, including antibiotics-antimyotics, peracetic acid-ethanol-sterilization (PES), γ-ray are applied for AM sterilization, it remains poorly understood which method is superior with little damage to the AM.
OBJECTIVE: To investigate the influence of different sterilizations on AM properties.
METHODS: AM obtained from healthy placenta was sterilized by 4 different procedures, including antibiotics-antimyotics, PES, γ-ray and Iodophors. Then the cell morphology was observed by haematoxylin-eosin staining, and the protein and mRNA expressions of hepatocyte growth factor (HGF) and tissue inhibitor of metalloproteases-1 (TIMP-1) were detected by immunohistochemistry and RT-PCR.
RESULTS AND CONCLUSION: The results revealed that the amniotic epithelium was badly destoried by the γ irradiation sterilization. Procedures with PES and antibiotics-antimyotics presented with completed tissue structures. The expression of HGF and TIMP-1 were decreased by the sequence of PES group, antibiotics-antimyotics group, Iodophors group and γ-ray group (P < 0.05). While the mRNA levels of HGF and TIMP-1 deceased in the sterilized group compared with the unsterilized group, the γ-ray group expressed least (P < 0.05) and the other three group showed no significant differences (P > 0.05). The PES was suggested as an alternative method for sterilization because this method influenced the properties of the amniontic epithelium least.

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