Keratin films for ocular surface reconstruction
Keratin films for ocular surface reconstruction
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DOI：
10.1016/j.biomaterials.2011.01.052
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发表时间：
2011-05-01
期刊：
影响因子：
14
通讯作者：
Geerling, Gerd
中科院分区：
文献类型：
--
作者：
Reichl, Stephan;Borrelli, Maria;Geerling, Gerd
人羊膜（AM）经常被用作眼表重建的基质。它的缺点（例如，降低的透明度和生物力学强度、取决于供体的异质性）产生了对标准化替代物的需求。来自毛发或羊毛的角蛋白已被提议作为用于生产膜或细胞培养支架的合适材料。目前的研究是进行开发透明的，稳定的和可转移的薄膜的基础上，人发角蛋白，支持细胞的粘附和增殖。该膜通过多步骤程序进行工程化，包括角蛋白提取、中性和碱性透析、干燥和固化过程。通过SDS-PAGE、SEM和X射线衍射分析对角蛋白膜进行了研究。此外，溶胀和水的吸收的膜进行了研究，是拉伸强度和透光率（UV/维斯）。最后，在增殖研究中估计角膜上皮细胞在角蛋白膜和AM上的生长行为。此外，我们评估了胰蛋白酶消化过程中的接种效率和细胞脱离行为。成膜过程产生由纳米颗粒角蛋白结构组成的透明膜。通过改变蛋白质组成、添加软化剂或改变固化温度和时间，可以改变膜的特性。基于这些发现，开发了优化的方案。与AM相比，膜显示出改善的透光率和生物力学强度。此外，在膜上的细胞行为与在AM上发现的细胞行为相似。我们的结论是，角蛋白电影可能是一个新的，有前途的替代眼表重建。(C)2011爱思唯尔有限公司保留所有权利。
Human amniotic membrane (AM) is frequently used as a substrate for ocular surface reconstruction. Its disadvantages (e.g., reduced transparency and biomechanical strength, heterogeneity depending on donor) create the need for standardized alternatives. Keratin from hair or wool has been proposed as an appropriate material for producing films or cell cultivation scaffolds. The current study was performed to develop transparent, stable and transferable films based on human hair keratin that support cellular adhesion and proliferation. The films were engineered by a multi-step procedure including keratin extraction, neutral and alkaline dialysis, drying and a curing process. Keratin films were investigated by SDS-PAGE, SEM and X-ray analyses. Furthermore, swelling and water absorption of the films were studied, as were tensile strength and light transmission (UV/VIS). Finally, the growth behavior of corneal epithelial cells on the keratin films and AM was estimated in proliferation studies. In addition, we assessed the seeding efficiency and cell detachment behavior during trypsinization. The film-forming process resulted in transparent films composed of nanoparticulate keratin structures. The film characteristics could be varied by changing the protein composition, adding softening agents or varying the curing temperature and duration. Based on these findings, an optimized protocol was developed. The films showed improved light transmission and biomechanical strength in comparison to AM. Furthermore, cell behavior on the films was similar to that found on AM. We conclude that keratin films may represent a new, promising alternative for ocular surface reconstruction. (C) 2011 Elsevier Ltd. All rights reserved.