A rabbit anterior cornea replacement derived from acellular porcine cornea matrix, epithelial cells and keratocytes
A rabbit anterior cornea replacement derived from acellular porcine cornea matrix, epithelial cells and keratocytes
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DOI：
10.1016/j.biomaterials.2010.05.066
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发表时间：
2010-10-01
期刊：
影响因子：
14
通讯作者：
Wu, Xinyi
中科院分区：
文献类型：
--
作者：
Pang, Kunpeng;Du, Liqun;Wu, Xinyi
本研究的目的是以脱细胞猪角膜基质(APCM)为支架构建兔前角膜替代物。用0.5%(wt./vol.)处理的新鲜猪角膜制备支架。十二烷基硫酸钠(SDS)溶液，在4摄氏度的冷冻室中搅拌24小时。H&E和DAPI染色证实角膜上皮细胞被完全清除。基质结构和力学性能得到很好的保存。四甲基偶氮唑盐比色法测定提取物对兔角膜上皮细胞、角膜上皮细胞和内皮细胞无细胞毒性。此外，在动物植入后的6个月内，没有迹象表明移植的椎间盘内或周围发生了免疫反应。为构建兔前角膜替代物，将角膜基质细胞注入APCM中，在动态培养系统中培养7d，然后在基质构建表面培养角膜上皮细胞7d。该构建物的表型与正常兔角膜相似，上皮细胞中高表达细胞角蛋白3，间质细胞中高表达波形蛋白。这些结果表明，用十二烷基硫酸钠制备的APCM可能是一种适合于角膜组织工程的支架材料。(C)2010爱思唯尔有限公司。保留所有权利。
The aim of this study was to construct a rabbit anterior cornea replacement with an acellular porcine cornea matrix (APCM) as a scaffold. The scaffold was prepared from fresh porcine corneas which were treated with 0.5% (wt./vol.) sodium dodecyl sulfate (SDS) solution and stirred for 24 h in a 4 degrees C refrigeration chamber. The complete removal of corneal cells was confirmed by H&E and DAPI staining. The stroma structure and mechanical properties were well preserved. The extracts had no cytotoxicity to rabbit corneal keratocytes, epithelial and endothelial cells as determined by MTT assay. Moreover, there was no sign that an immune reaction occurred in or around the transplanted disks within 6 months of animal implantation. To construct a rabbit anterior cornea replacement, keratocytes were injected into APCM and cultured for 7 days in a dynamic culturing system, followed by culturing corneal epithelial cells on the stroma construct surface for another 7 days. The phenotype of the construct was similar to normal rabbit corneas, with high expression of cytokeratin 3 in the epithelial cell layer and expression of vimentin in the stromal cells. These results suggested that the APCM developed by using SDS might be a suitable scaffold for cornea tissue engineering. (C) 2010 Elsevier Ltd. All rights reserved.