Li Qing, Wang Hongmei, Dai Zhenye, Cao Yichen, Jin Chuanyu
Qingdao Chunghao Tissue Engineering Co, Ltd, Qingdao, Shangdong Province, China.
Cornea. 2017 Nov;36(11):1343-1351. doi: 10.1097/ICO.0000000000001319.
To construct an acellular porcine corneal stroma (aPCS) as a human corneal stroma alternative and to further explore its biomechanical properties.
A combination of DNA-RNA enzymes and ultrasound technology was used to strip the native porcine corneal cells. The microstructure of aPCS was observed by H&E staining, DAPI staining, and α-Gal tests. The mechanical properties were detected by a tension machine. Cytotoxicity of aPCS was measured by the MTT assay. The subcutaneous embedding experiment in rats was also used to detect immunity and degradation. The aPCS was transplanted into the rabbit cornea by lamellar keratoplasty, general observations were made at 3 days, 1 week, 1 month, and 3 months after implantation, respectively.
The microstructure and mechanical properties of aPCS were not damaged during the decellularization process. The aPCS extracts had no significant cytotoxicity on human corneal stroma cells. Moreover, the subcutaneous embedding experiment in rats demonstrated that aPCS could not be degraded and induced no immune reaction in and around the transplanted discs. More important is that the aPCS reconstructed normal corneal stroma and maintained corneal transparency and thickness, with almost no neovascularization and inflammation at 3 months after surgery.
The aPCS prepared in this study had good biocompatibility, safety, and low antigenicity, which has great potential for corneal disease treatment.
构建脱细胞猪角膜基质（aPCS）作为人角膜基质替代品，并进一步探究其生物力学特性。
采用DNA-RNA酶与超声技术相结合的方法去除猪角膜中的天然细胞。通过苏木精-伊红染色、4',6-二脒基-2-苯基吲哚染色及α-半乳糖测试观察aPCS的微观结构。使用拉力机检测其力学性能。采用MTT法检测aPCS的细胞毒性。通过大鼠皮下植入实验检测其免疫性和降解情况。将aPCS通过板层角膜移植术植入兔角膜，分别在植入后3天、1周、1个月和3个月进行大体观察。
脱细胞过程中aPCS的微观结构和力学性能未受损。aPCS提取物对人角膜基质细胞无明显细胞毒性。此外，大鼠皮下植入实验表明，aPCS在移植盘及其周围不会降解，也不会引发免疫反应。更重要的是，aPCS重建了正常的角膜基质，维持了角膜的透明度和厚度，术后3个月几乎没有新生血管形成和炎症反应。
本研究制备的aPCS具有良好的生物相容性、安全性和低抗原性，在角膜疾病治疗方面具有巨大潜力。
