Department of Integrative Medical Biology, Anatomy, Umeå University, Umeå, 90187, Sweden.
Department of Clinical Sciences, Ophthalmology, Umeå University, Umeå, 90187, Sweden.
Adv Healthc Mater. 2017 Mar;6(5). doi: 10.1002/adhm.201601238. Epub 2016 Dec 27.
The optimal functionality of the native corneal stroma is mainly dependent on the well-ordered arrangement of extracellular matrix (ECM) and the pressurized structure. In order to develop an in vitro corneal model, it is crucial to mimic the in vivo microenvironment of the cornea. In this study, the influence of surface topography and mechanical strain on keratocyte phenotype and ECM formation within a biomimetic 3D corneal model is studied. By modifying the surface topography of materials, it is found that patterned silk fibroin film with 600 grooves mm optimally supports cell alignment and ECM arrangement. Furthermore, treatment with 3% dome-shaped mechanical strain, which resembles the shape and mechanics of native cornea, significantly enhances the expression of keratocyte markers as compared to flat-shaped strain. Accordingly, a biomimetic 3D corneal model, in the form of a collagen-modified, silk fibroin-patterned construct subjected to 3% dome-shaped strain, is created. Compared to traditional 2D cultures, it supports a significantly higher expression of keratocyte and ECM markers, and in conclusion better maintains keratocyte phenotype, alignment, and fusiform cell shape. Therefore, the novel biomimetic 3D corneal model developed in this study serves as a useful in vitro 3D culture model to improve current 2D cultures for corneal studies.
天然角膜基质的最佳功能主要取决于细胞外基质（ECM）的有序排列和受压结构。为了开发体外角膜模型，模拟角膜的体内微环境至关重要。在这项研究中，研究了表面形貌和机械应变对仿生 3D 角膜模型中角膜细胞表型和 ECM 形成的影响。通过修饰材料的表面形貌，发现具有 600 个凹槽 mm 的图案化丝素蛋白膜能最佳地支持细胞排列和 ECM 排列。此外，与平面应变相比，类似于天然角膜形状和力学的 3%穹顶形机械应变处理显著增强了角膜细胞标志物的表达。因此，创建了一种仿生 3D 角膜模型，其形式为经胶原修饰、丝素蛋白图案化构建体，并施加 3%穹顶形应变。与传统的 2D 培养相比，它支持更高水平的角膜细胞和 ECM 标志物的表达，并且更好地维持了角膜细胞表型、排列和梭形细胞形状。因此，本研究开发的新型仿生 3D 角膜模型可用作改进当前用于角膜研究的 2D 培养的有用体外 3D 培养模型。
