Maisonneuve-Rosemont Hospital Research Center, Montreal, Québec, Canada.
Invest Ophthalmol Vis Sci. 2012 Sep 19;53(10):6343-54. doi: 10.1167/iovs.12-9708.
We evaluated the in vivo functionality of a corneal endothelium tissue-engineered using corneal endothelial cells from human patients with Fuchs endothelial corneal dystrophy (FECD).
A total of 15 healthy cats underwent full-thickness corneal transplantation. All transplants were of xenogeneic human origin and all grafts but two were tissue-engineered. In seven animals the graft corneal endothelium was tissue-engineered using cultured corneal endothelial cells from humans with FECD (TE-FECD). Two control animals were grafted with an endothelium engineered using cultured endothelial cells from normal eye bank corneas (TE-normal). Two controls received a native full-thickness corneal transplant, and four other controls were grafted with the stromal carrier only (without endothelial cells). Outcome parameters included graft transparency (0, opaque to 4, clear), pachymetry, optical coherence tomography, endothelial cell morphometry, transmission electron microscopy (TEM), and immunostaining of function-related proteins.
Seven days after transplantation, 6 of 7 TE-FECD grafts, all TE-normal grafts, and all normal native grafts were clear (transparency score >3), while all carriers-only grafts were opaque (score <1). The mean pachymetry was 772 ± 102 μm for TE-FECD, 524 ± 11 μm for TE-normal, 555 ± 48 for normal native, and 1188 ± 223 μm for carriers only. TEM showed subendothelial loose fibrillar material deposition in all TE-FECD grafts. The TE endothelium expressed Na(+)-K(+)/ATPase and Na(+)/HCO3(-).
Restoration of transparency and corneal thickness demonstrated that the TE-FECD grafts were functional in vivo. This novel FECD seven-day living model suggests a potential role for tissue engineering leading to FECD cell rehabilitation.
我们评估了使用来自患有 Fuchs 内皮角膜营养不良(FECD)的患者的角膜内皮细胞构建的角膜内皮组织工程的体内功能。
共有 15 只健康猫接受了全层角膜移植。所有移植物均来自异种人源,除了两个之外,所有移植物均为组织工程化的。在 7 只动物中,使用来自患有 FECD 的人培养的角膜内皮细胞构建了组织工程化的移植物角膜内皮(TE-FECD)。两只对照动物接受了使用来自正常眼库角膜的培养的内皮细胞构建的组织工程化移植物(TE-正常)。两只对照动物接受了全厚角膜的天然移植物,另外四只对照动物仅接受了基质载体移植(无内皮细胞)。观察指标包括移植物透明度(0 级,完全不透明至 4 级,完全透明)、角膜厚度、光学相干断层扫描、内皮细胞形态计量学、透射电子显微镜(TEM)和功能相关蛋白的免疫染色。
移植后 7 天,7 个 TE-FECD 移植物中有 6 个、所有 TE-正常移植物和所有正常天然移植物均为透明(透明度评分>3),而所有仅基质载体移植物均为不透明(评分<1)。TE-FECD 的平均角膜厚度为 772±102μm,TE-正常为 524±11μm,正常天然为 555±48μm,仅基质载体为 1188±223μm。TEM 显示所有 TE-FECD 移植物的亚内皮层均有疏松纤维状物质沉积。TE 内皮细胞表达 Na(+)-K(+)/ATP 酶和 Na(+)/HCO3(-)。
透明度和角膜厚度的恢复表明 TE-FECD 移植物在体内具有功能。这种新型 FECD 7 天活体模型表明组织工程可能对 FECD 细胞康复具有潜在作用。