The formation of a tissue-engineered cornea using plastically compressed collagen scaffolds and limbal stem cells.
The formation of a tissue-engineered cornea using plastically compressed collagen scaffolds and limbal stem cells.
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DOI：
10.1007/978-1-62703-432-6_9
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发表时间：
2013
影响因子：
--
通讯作者：
S. Mi;C. Connon
中科院分区：
文献类型：
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作者：
S. Mi;C. Connon
胶原蛋白具有优异的生物相容性，可生物降解，并且具有低免疫原性。因此，该蛋白质是用于形成用于治疗用途的角膜支架的非常合适的底物。然而，常规胶原蛋白凝胶的高度水合性质导致凝胶结构脆弱且难以操作。在这一章中，我们描述了一种新的方法，培养角膜缘上皮细胞（LEC）的压缩胶原支架。压缩的胶原支架可以使用一种不依赖于细胞的过程快速构建，该过程产生具有可控微观特征的致密和机械强度高的胶原结构。我们将角膜基质细胞包埋在胶原凝胶中，随后压缩并涂覆层粘连蛋白。由此产生的构建体支持LEC的生理形态和分层。分化LEC的特异性标志物细胞角蛋白3（CK 3）和未分化LEC的标志物细胞角蛋白14（CK 14）的表达在压缩胶原构建体和主要常规支架剥脱羊膜（AM）上扩张的LEC中相似。因此，我们证明，层粘连蛋白包被的，压缩的胶原蛋白凝胶含有角膜基质细胞可以支持LEC的扩展，分层和分化的程度，是裸露的AM相媲美。我们的新型压缩胶原/角膜基质细胞结构具有用作组织工程人工角膜的潜力。
Collagen has excellent biocompatibility, is biodegradable, and possesses low immunogenicity. Therefore, this protein is a very suitable substrate for the formation of a corneal scaffold for therapeutic use. The highly hydrated nature of conventional collagen gels, however, results in a gel that is structurally weak and difficult to manipulate. In this chapter, we describe a novel method to cultivate limbal epithelial cells (LEC) on a compressed collagen scaffold. The compressed collagen scaffold can be rapidly constructed using a cell-independent process, which produces dense and mechanically strong collagen constructs with controllable microscale features.We embedded corneal keratocytes in a collagen gel, which we subsequently compressed and coated with laminin. The resulting construct supported the physiological morphology and stratification of LEC. The expression of a specific marker for differentiated LEC, cytokeratin 3 (CK3), and a marker for undifferentiated LEC, cytokeratin 14 (CK14), were similar in LEC expanded on both the compressed collagen construct and the leading conventional scaffold, denuded amniotic membrane (AM). We therefore demonstrate that a laminin-coated, compressed collagen gel containing keratocytes can support LEC expansion, stratification, and differentiation to a degree that is comparable to denuded AM. Our novel compressed collagen/keratocyte construct has potential for use as a tissue-engineered artificial cornea.