Department of Biomedical Engineering, School of Engineering, Tufts University, 4 Colby St Medford, MA 02155, USA.
Biomaterials. 2010 Dec;31(34):8953-63. doi: 10.1016/j.biomaterials.2010.08.017.
RGD-coupled silk protein-biomaterial lamellar systems were prepared and studied with human cornea fibroblasts (hCFs) to match functional requirements. A strategy for corneal tissue engineering was pursued to replicate the structural hierarchy of human corneal stroma within thin stacks of lamellae-like tissues, in this case constructed from scaffolds constructed with RGD-coupled, patterned, porous, mechanically robust and transparent silk films. The influence of RGD-coupling on the orientation, proliferation, ECM organization, and gene expression of hCFs was assessed. RGD surface modification enhanced cell attachment, proliferation, alignment and expression of both collagens (type I and V) and proteoglycans (decorin and biglycan). Confocal and histological images of the lamellar systems revealed that the bio-functionalized silk human cornea 3D constructs exhibited integrated corneal stroma tissue with helicoidal multi-lamellar alignment of collagen-rich and proteoglycan-rich extracellular matrix, with transparency of the construct. This biomimetic approach to replicate corneal stromal tissue structural hierarchy and architecture demonstrates a useful strategy for engineering human cornea. Further, this approach can be exploited for other tissue systems due to the pervasive nature of such helicoids in most human tissues.
RGD 偶联丝蛋白生物材料层状体系的制备及与人角膜成纤维细胞（hCFs）的研究，以满足功能要求。采用角膜组织工程策略，在薄的层状组织叠片中复制人角膜基质的结构层次，在这种情况下，使用 RGD 偶联、图案化、多孔、机械坚固和透明的丝膜构建支架来构建。评估了 RGD 偶联对 hCFs 的取向、增殖、细胞外基质组织和基因表达的影响。RGD 表面修饰增强了细胞的附着、增殖、排列以及 I 型和 V 型胶原和蛋白聚糖（decorin 和 biglycan）的表达。层状体系的共聚焦和组织学图像显示，生物功能化的丝质人角膜 3D 构建体表现出整合的角膜基质组织，具有富含胶原和蛋白聚糖的细胞外基质的螺旋状多层层状排列，并且构建体具有透明性。这种复制角膜基质组织结构层次和结构的仿生方法展示了一种用于工程化人角膜的有用策略。此外，由于这种螺旋结构在大多数人体组织中普遍存在，因此该方法可用于其他组织系统。
