An Ultra-thin Amniotic Membrane as Carrier in Corneal Epithelium Tissue-Engineering.
An Ultra-thin Amniotic Membrane as Carrier in Corneal Epithelium Tissue-Engineering.
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超薄羊膜作为角膜上皮组织工程的载体
DOI：
10.1038/srep21021
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发表时间：
2016-02-15
影响因子：
4.6
通讯作者：
Li W
中科院分区：
文献类型：
--
作者：
Zhang L;Zou D;Li S;Wang J;Qu Y;Ou S;Jia C;Li J;He H;Liu T;Yang J;Chen Y;Liu Z;Li W
羊膜作为角膜上皮组织载体广泛应用于角膜重建手术。然而，由于半透明的厚底层AM基质，工程化组织的透明度较低。为了克服这一缺点，我们开发了一种超薄AM（UAM），通过使用胶原酶IV剥离掉上皮剥脱AM（DAM）的一些基质。通过将基质变薄至约30 μm，其潮湿和干燥形式呈现无细胞的、光学透明的，其胶原蛋白框架变得致密和规则。角膜缘上皮细胞在UAM上扩增后形成的工程化兔角膜上皮细胞（RCEC）片层比在DAM上扩增的细胞片层更透明、更厚。此外，在UAM上扩增的组织工程细胞片中ΔNp63和ABCG 2基因表达高于在DAM上扩增的组织工程细胞片。此外，手术后2周，移植有基于UAM的细胞片的角膜显示出比移植有基于DAM的细胞片的角膜更高的透明度和更多的复层上皮。总之，在UAM上产生的组织工程化角膜上皮具有优选的结果，因为移植的组织比通过使用DAM获得的组织更透明并且更好地类似于天然组织的表型。UAM保存了致密的羊膜层，可能是一种理想的角膜上皮组织工程基质。
Amniotic membranes (AMs) are widely used as a corneal epithelial tissue carrier in reconstruction surgery. However, the engineered tissue transparency is low due to the translucent thick underlying AM stroma. To overcome this drawback, we developed an ultra-thin AM (UAM) by using collagenase IV to strip away from the epithelial denuded AM (DAM) some of the stroma. By thinning the stroma to about 30 μm, its moist and dry forms were rendered acellular, optically clear and its collagen framework became compacted and inerratic. Engineered rabbit corneal epithelial cell (RCEC) sheets generated through expansion of limbal epithelial cells on UAM were more transparent and thicker than those expanded on DAM. Moreover, ΔNp63 and ABCG2 gene expression was greater in tissue engineered cell sheets expanded on UAM than on DAM. Furthermore, 2 weeks after surgery, the cornea grafted with UAM based cell sheets showed higher transparency and more stratified epithelium than the cornea grafted with DAM based cell sheets. Taken together, tissue engineered corneal epithelium generated on UAM has a preferable outcome because the transplanted tissue is more transparent and better resembles the phenotype of the native tissue than that obtained by using DAM for this procedure. UAM preserves compact layer of the amniotic membrane and maybe an ideal substrate for corneal epithelial tissue engineering.