Construction of tissue-engineered cornea composed of amniotic epithelial cells and acellular porcine cornea for treating corneal alkali burn
Construction of tissue-engineered cornea composed of amniotic epithelial cells and acellular porcine cornea for treating corneal alkali burn
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DOI：
10.1016/j.biomaterials.2013.05.045
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发表时间：
2013-09-01
期刊：
影响因子：
14
通讯作者：
Jin, Yan
中科院分区：
文献类型：
--
作者：
Luo, Hailang;Lu, Yongbo;Jin, Yan
虽然脱细胞角膜已被报道是同种异体角膜移植治疗角膜损伤的潜在替代品，但由于炎症和新生血管的存在，严重的角膜损伤难以修复。由于羊膜上皮细胞（AECs）的抗炎和抗血管生成特性，羊膜作为眼表重建中的移植物的使用已变得广泛。本研究旨在构建由脱细胞猪角膜（APC）和AEC组成的组织工程化角膜（TEC），用于修复严重的角膜损伤。从所制备的APC中完全去除角膜细胞，并且保持天然猪角膜（NPC）的微观结构、机械性能和稳定性。在体外，MU和流式细胞术分析表明，APC没有负面影响细胞活力和凋亡。在体内，角膜袋和皮下移植证明APC不能触发接受的免疫应答。从人羊膜分离的AEC具有增殖潜力，并且在6代之前呈现健康的形态。在APC表面培养7天后，AEC分层为5-6层。我们发现AEC重建了被脱细胞过程破坏的基底膜。ELISA结果显示，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，IL 6，Fas-L和PDEF。最后，板层角膜移植术治疗碱烧伤的结果表明，移植的TEC是透明的，并完全接种到宿主角膜。然而，移植的APC由于宿主排斥而降解。因此，我们的结论是，TEC的AEC和APC组成的严重角膜损伤的修复具有很大的潜力。(C)2013爱思唯尔有限公司保留所有权利。
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, MU 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. (C) 2013 Elsevier Ltd. All rights reserved.