State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, 54 Xian Lie Nan Road, Guangzhou 510060, PR China.
Biomaterials. 2011 Nov;32(31):7812-21. doi: 10.1016/j.biomaterials.2011.07.012. Epub 2011 Jul 23.
To investigate the feasibility of using acellular porcine limbal stroma for limbal stem cell microenvironment reconstruction. Limbal reconstruction was performed in rabbit partial limbal defect models. Rabbits were randomly divided into four groups: acellular porcine limbal stroma, de-epithelized rabbit limbal autograft stroma, de-epithelized porcine limbal stroma and acellular porcine corneal stroma transplantation groups. In both the acellular porcine limbal stroma and de-epithelized rabbit limbal autograft stroma groups, cornea transparency and epithelium integrity were sustained and graft rejection was not observed. The basal epithelial cells of the grafts showed the K3+/P63+/Ki67+ phenotype at postoperative month 1, but it returned to K3-/P63+/Ki67+(phenotype characteristic of limbal epithelium) by postoperative months 3 and 6. In the de-epithelized porcine limbal stroma group, acute and serious immune rejection occurred by postoperative days 8-10. The basal epithelial cells of the grafts showed the K3+/P63+/Ki67+ phenotype at postoperative month 1. In the acellular porcine corneal stroma group, there were some new vessel invasion into the peripheral cornea and mild corneal opacity. The basal epithelial cells of the grafts showed the K3+/P63+/Ki67+ phenotype at postoperative months 1, 3, and 6. In conclusion, acellular porcine limbal stroma possessed very low immunogenicity, retained a good original limbal ECM microenvironment, and thus the reconstructed rabbit limbal microenvironment maintained limbal epithelial stem cell stemness and proliferation.
为了研究使用去细胞猪角膜缘基质重建角膜缘干细胞微环境的可行性，我们进行了兔部分角膜缘缺损模型的角膜缘重建。兔子被随机分为四组：去细胞猪角膜缘基质组、去上皮兔自体角膜缘基质组、去上皮猪角膜缘基质组和去细胞猪角膜基质组。在去细胞猪角膜缘基质组和去上皮兔自体角膜缘基质组中，角膜透明度和上皮完整性得以维持，且未观察到移植物排斥反应。术后 1 个月，移植物的基底上皮细胞呈 K3+/P63+/Ki67+表型，但在术后 3 个月和 6 个月时恢复为 K3-/P63+/Ki67+（具有角膜缘上皮特征的表型）。在去上皮猪角膜缘基质组中，术后 8-10 天发生急性和严重的免疫排斥反应。术后 1 个月，移植物的基底上皮细胞呈 K3+/P63+/Ki67+表型。在去细胞猪角膜基质组中，有一些新生血管侵入周边角膜，角膜轻度混浊。术后 1、3、6 个月，移植物的基底上皮细胞呈 K3+/P63+/Ki67+表型。总之，去细胞猪角膜缘基质具有极低的免疫原性，保留了良好的原始角膜缘 ECM 微环境，因此重建的兔角膜缘微环境维持了角膜缘上皮干细胞的干性和增殖能力。
