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一种新型血清电泳法制备脱细胞角膜基质作为人工角膜支架

A novel serum: Electrophoresis method to prepare acellular corneal matrix as an artificial corneal scaffold.

作者信息

Li Qing, Xie Cuicui, Wang Hongmei, Zhang Fenghua, Mu Lanlan

机构信息

Qingdao Chunghao Tissue Engineering Co., Ltd., Qingdao, China.

出版信息

Int J Artif Organs. 2020 Feb;43(2):127-136. doi: 10.1177/0391398819869941. Epub 2019 Sep 6.

Abstract

INTRODUCTION

The aim of this study was to develop a novel decellularization method in order to obtain an ideal scaffold with good biocompatibility.

METHODS

The porcine corneas were treated with human serum for 5 days or serum-electrophoresis respectively. The electrophoresis (100 V/cm) was performed in sterilized buffer containing 40-mM tris base, 18-mM glacial acetic acid, and antibiotics for 1 h at 4°C. The properties of artificial corneal scaffolds were characterized by morphological and histological examinations. The biocompatibility and biological safety were examined by subcutaneous implant test and lamellar keratoplasty.

RESULTS AND CONCLUSIONS

The transparency and appearance of serum-electrophoresis acellular porcine corneal matrix were better than serum acellular porcine corneal matrix. DNA and α-gal in serum-electrophoresis acellular porcine corneal matrix were more efficiently removed than those in serum acellular porcine corneal matrix (p < 0.05). The subcutaneous and corneal implantation experiments showed serum-electrophoresis acellular porcine corneal matrix had better biocompatibility compared to serum acellular porcine corneal matrix (p < 0.01). This novel serum-electrophoresis decellularization method may be valuable for preparation of xenogenic corneal tissue for clinical application.

摘要

引言

本研究的目的是开发一种新型去细胞方法,以获得具有良好生物相容性的理想支架。

方法

将猪角膜分别用人血清处理5天或进行血清电泳。电泳(100 V/cm)在含有40 mM三羟甲基氨基甲烷碱、18 mM冰醋酸和抗生素的无菌缓冲液中于4°C进行1小时。通过形态学和组织学检查对人工角膜支架的特性进行表征。通过皮下植入试验和板层角膜移植术检查生物相容性和生物安全性。

结果与结论

血清电泳去细胞猪角膜基质的透明度和外观优于血清去细胞猪角膜基质。血清电泳去细胞猪角膜基质中的DNA和α-半乳糖比血清去细胞猪角膜基质中的去除效率更高(p < 0.05)。皮下和角膜植入实验表明,与血清去细胞猪角膜基质相比,血清电泳去细胞猪角膜基质具有更好的生物相容性(p < 0.01)。这种新型血清电泳去细胞方法对于制备用于临床应用的异种角膜组织可能具有重要价值。

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