Research and Development Center for Tissue Engineering, Fourth Military Medical University, Xi'an, Shaanxi 710032, China.
Biomaterials. 2013 Sep;34(28):6748-59. doi: 10.1016/j.biomaterials.2013.05.045. Epub 2013 Jun 10.
Although acellular corneas have been reported to be a potential substitute for allogeneic cornea transplantation to treat corneal injury, severe corneal injury is hard to repair due to inflammation and neovascularization. The use of the amniotic membrane as a graft in ocular surface reconstruction has become widespread because of the anti-inflammatory and anti-angiogenic properties of amniotic epithelial cells (AECs). Our objective was to construct a tissue-engineered cornea (TEC) composed of an acellular porcine cornea (APC) and AECs to repair severe corneal injury. Corneal cells were completely removed from the prepared APC, and the microstructure, mechanical properties, and stability of a natural porcine cornea (NPC) was maintained. In vitro, MTT and flow cytometry analyses showed that the APC did not negatively affect cell viability and apoptosis. In vivo, corneal pocket and subcutaneous transplantation demonstrated that the APC was incapable of trigging accepted immune response. AECs isolated from the human amniotic membrane have proliferation potential and present healthy morphology before 6 passages. After 7 days of culture on the surface of the APC, the AECs were stratified into 5-6 layers. We found that the AECs reconstituted the basement membrane that had been disrupted by the decellularization process. ELISA results showed that after culturing the TEC, the culture medium contained anti-inflammatory and anti-angiogenic growth factors, such as MIF, IL6, Fas-L, and PDEF. Finally, the results of lamellar keratoplasty to treat an alkali burn showed that the transplanted TEC was transparent and completely inoculated into the host cornea. However, the transplanted APC was degraded due to host rejection. Therefore, we conclude that a TEC composed of AECs and an APC holds great potential for the repair of severe corneal injury.
虽然去细胞角膜已被报道可作为同种异体角膜移植的替代品，用于治疗角膜损伤，但由于炎症和新生血管化，严重的角膜损伤仍然难以修复。由于羊膜上皮细胞 (AECs) 具有抗炎和抗血管生成特性，因此将羊膜用作眼表面重建的移植物已得到广泛应用。我们的目的是构建由去细胞猪角膜 (APC) 和 AECs 组成的组织工程角膜 (TEC)，以修复严重的角膜损伤。从制备好的 APC 中完全去除角膜细胞，保持天然猪角膜 (NPC) 的微观结构、机械性能和稳定性。体外 MTT 和流式细胞术分析表明 APC 不会对细胞活力和细胞凋亡产生负面影响。体内角膜囊和皮下移植表明 APC 不能引发可接受的免疫反应。从人羊膜中分离的 AECs 具有增殖潜力，在 6 代之前呈现出健康的形态。在 APC 表面培养 7 天后，AECs 分层为 5-6 层。我们发现 AECs 重建了去细胞化过程中断裂的基底膜。ELISA 结果表明，在培养 TEC 后，培养基中含有抗炎和抗血管生成的生长因子，如 MIF、IL6、Fas-L 和 PDEF。最后，用于治疗碱烧伤的板层角膜移植结果表明，移植的 TEC 是透明的，并且完全与宿主角膜融合。然而，由于宿主排斥反应，移植的 APC 被降解。因此，我们得出结论，由 AECs 和 APC 组成的 TEC 具有修复严重角膜损伤的巨大潜力。
