Department of Ophthalmology, Seoul National University College of Medicine , Seoul, Korea.
Tissue Eng Part C Methods. 2009 Dec;15(4):635-45. doi: 10.1089/ten.TEC.2009.0022.
To investigate the propriety of decellularized porcine corneas as a source of lamellar corneal xenografts, we treated porcine corneas with (1) freezing, (2) three freezing-thawing, (3) hypertonic saline, (4) hyperosmolar glycerol, (5) trypsin/sodium dodecyl sulfate/Dispase, and (6) DNase/RNase. After processing, we examined the cells and collagen structures of the decellularized corneas using hematoxylin-eosin staining, terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay, and transmission electron microscopy. Cell viability was also assessed via organ culture. In addition, the outcomes of porcine anterior lamellar corneal xenografting were evaluated in rabbits. Graft integration and corneal thickness were assessed using anterior optical coherence tomography, and the corneas were histologically examined sequentially after transplantation. We found that porcine corneas treated with hypertonic saline-based decellularization had little immunogenicity with intact collagen structures. The porcine corneal xenografts decellularized with the hypertonic saline-based method were well integrated into the adjacent host tissues and remained clear in rabbit eyes for more than 6 months.
为了研究去细胞化猪角膜作为层状异种角膜移植物的适宜性,我们用(1)冷冻、(2)三次冷冻-解冻、(3)高渗盐水、(4)高渗甘油、(5)胰蛋白酶/十二烷基硫酸钠/分散酶和(6)DNase/RNase 处理猪角膜。处理后,我们使用苏木精-伊红染色、末端脱氧核苷酸转移酶介导的缺口末端标记(TUNEL)检测和透射电子显微镜检查去细胞化角膜的细胞和胶原结构。还通过器官培养评估细胞活力。此外,我们在兔中评估了猪前层状角膜异种移植的结果。使用前光学相干断层扫描评估移植物整合和角膜厚度,并在移植后依次对角膜进行组织学检查。我们发现,用高渗盐水去细胞化处理的猪角膜具有低免疫原性和完整的胶原结构。用高渗盐水法去细胞化的猪角膜异种移植物与相邻的宿主组织很好地整合在一起,在兔眼中保持透明状态超过 6 个月。