Decellularizing corneal stroma using N2 gas
Decellularizing corneal stroma using N2 gas
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发表时间：
2008-05
期刊：
影响因子：
2.2
通讯作者：
S. Amano;Naoki Shimomura;Seiichi Yokoo;K. Araki-Sasaki;S. Yamagami
中科院分区：
文献类型：
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作者：
S. Amano;Naoki Shimomura;Seiichi Yokoo;K. Araki-Sasaki;S. Yamagami
目的探讨利用N2气分离猪角膜基质的新方法的有效性，以及将脱细胞的猪角膜基质用于兔角膜移植模型的可行性。方法将猪眼角膜置于试管中，从液氮中取氮气倒入管内，冷冻眼角膜，使管内缺氧。将试管盖紧后，在室温下保存7天，并对猪角膜进行组织学检查。将经上述方法处理的猪角膜间质盘置入兔角膜间质袋中，观察6个月。结果hechst 33342染色、苏木精染色和伊红染色均显示猪角膜间质在氮气中培养1周后细胞核较少。末端脱氧核苷酸转移酶介导的dUTP缺口末端标记（TUNEL）实验显示，在氮气中培养3天的猪角膜基质中有许多阳性细胞核。将脱细胞的猪角膜基质移植到兔角膜基质袋中，移植后6个月仍保持透明。结论该方法在不降低角膜透明度的情况下使角膜基质脱细胞。
Purpose To examine the efficacy of a novel method of decellularizing porcine corneal stroma using N2 gas from liquid N2 and the feasibility of using decellularized porcine corneal stroma in a corneal transplantation model in rabbits. Methods Porcine corneas were placed in a tube, and N2 gas from liquid N2 was poured into the tube to freeze the corneas and make the inside of the tube hypoxic. After fastening the cap firmly, the tube was kept at room temperature for seven days, and the porcine corneas were examined histologically. A porcine corneal stromal disk treated with the aforementioned method was inserted into a pocket of rabbit corneal stroma and observed for six months. Results Hoechst 33342 and hematoxylin and eosin staining both showed few cellular nuclei in the porcine corneal stroma incubated in N2 gas for one week. A terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay showed many positively stained nuclei in the porcine corneal stroma incubated in N2 gas for three days. The porcine corneal stroma that was decellularized and transplanted into a rabbit corneal stromal pocket remained clear for six months after transplantation. Conclusions This method using N2 gas decellularizes corneal stroma without reducing corneal transparency.