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Sterilized, freeze-dried amniotic membrane : a useful substrate for ocular surface reconstruction.

机译：无菌，冷冻干燥的羊膜：用于眼表重建的有用基质。

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PURPOSE. To examine the feasibility of using sterilized, freeze-dried amniotic membrane (FD-AM) as a substrate for cultivating autologous corneal epithelial cells for ocular surface reconstruction. METHODS. Human AM deprived of amniotic epithelial cells by incubation with EDTA was freeze dried, vacuum packed, and sterilized with -irradiation. The resultant FD-AM was characterized for its physical, biological, and morphologic properties by stretch stress tests, immunohistochemistry, electron microscopy, and cell culture. In addition, 3 weeks after an ocular surface injury, the conjunctivalized corneal surfaces of eyes in eight rabbits were surgically reconstructed by transplantation of autologous cultivated corneal epithelial cells on FD-AM. RESULTS. A stretch stress test revealed no significant differences between sterilized FD-AM and cryopreserved AM. Immunohistochemistry for several extracellular matrix molecules and electron microscopic analysis of FD-AM revealed that the process of drying and irradiation did not affect its biological and morphologic properties. The corneal epithelial cells cultivated on FD-AM had four to five stratified, well-differentiated cell layers. Corneas that were grafted with the cultivated corneal epithelial cells on FD-AM were clear and were all epithelialized at 10 days after surgery. CONCLUSIONS. The sterilized, freeze-dried AM retained most of the physical, biological, and morphologic characteristics of cryopreserved AM; consequently, it is a useful biomaterial for ocular surface reconstruction.
机译：目的。为了检验使用无菌，冻干羊膜（FD-AM）作为基质培养自体角膜上皮细胞进行眼表重建的可行性。方法。将通过与EDTA一起温育而被剥夺羊膜上皮细胞的人AM冷冻干燥，真空包装，并用放射线灭菌。通过拉伸应力测试，免疫组织化学，电子显微镜和细胞培养来表征所得的FD-AM的物理，生物学和形态学特性。另外，在眼表面损伤后3周，通过将自体培养的角膜上皮细胞移植到FD-AM上，通过手术重建了八只兔子的眼结膜化的角膜表面。结果。拉伸应力测试显示无菌FD-AM与冷冻保存的AM之间无显着差异。几种细胞外基质分子的免疫组织化学和FD-AM的电子显微镜分析表明，干燥和辐射过程不影响其生物学和形态学特性。在FD-AM上培养的角膜上皮细胞具有4至5个分层且分化良好的细胞层。在FD-AM上移植了培养的角膜上皮细胞的角膜是透明的，并且在术后10天全部上皮化。结论。灭菌，冷冻干燥的AM保留了冷冻保存的AM的大部分物理，生物学和形态特征。因此，它是用于眼表重建的有用的生物材料。

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