Sharif Rabab, Priyadarsini Shrestha, Rowsey Tyler G, Ma Jian-Xing, Karamichos Dimitrios
Department of Cell Biology, University of Oklahoma Health Sciences Center.
Department of Ophthalmology/Dean McGee Eye Institute, University of Oklahoma Health Sciences Center.
J Vis Exp. 2018 Jan 24(131):56308. doi: 10.3791/56308.
Tissue engineering has gained substantial recognition due to the high demand for human cornea replacements with an estimated 10 million people worldwide suffering from corneal vision loss. To address the demand for viable human corneas, significant progress in three-dimensional (3D) tissue engineering has been made. These cornea models range from simple monolayer systems to multilayered models, leading to 3D full-thickness corneal equivalents. However, the use of a 3D tissue-engineered cornea in the context of in vitro disease models studied to date lacks resemblance to the multilayered 3D corneal tissue structure, function, and the networking of different cell types (i.e., nerve, epithelium, stroma, and endothelium). In addition, the demand for in vitro cornea tissue models has increased in an attempt to reduce animal testing for pharmaceutical products. Thus, more sophisticated models are required to better match systems to human physiological requirements, and the development of a model that is more relevant to the patient population is absolutely necessary. Given that multiple cell types in the cornea are affected by diseases and dystrophies, such as Keratoconus, Diabetic Keratopathy, and Fuchs, this model includes a 3D co-culture model of primary human corneal fibroblasts (HCFs) from healthy donors and neurons from the SH-SY5Y cell line. This allows us for the first time to investigate the interactions between the two cell types within the human corneal tissue. We believe that this model could potentially dissect the underlying mechanisms associated with the stromal-nerve interactions of corneal diseases that exhibit nerve damages. This 3D model mirrors the basic anatomical and physiological nature of the corneal tissue in vivo and can be used in the future as a tool for investigating corneal defects as well as screening the efficacy of various agents before animal testing.
由于全球估计有1000万人患有角膜视力丧失，对人类角膜替代品的需求很高，组织工程因此获得了广泛认可。为了满足对 viable 人类角膜的需求，三维（3D）组织工程取得了重大进展。这些角膜模型从简单的单层系统到多层模型，最终形成3D全层角膜等效物。然而，迄今为止，在体外疾病模型中使用3D组织工程角膜缺乏与多层3D角膜组织结构、功能以及不同细胞类型（即神经、上皮、基质和内皮）网络的相似性。此外，为了减少药品动物试验，对体外角膜组织模型的需求有所增加。因此，需要更复杂的模型来更好地使系统符合人类生理需求，开发与患者群体更相关的模型绝对必要。鉴于角膜中的多种细胞类型会受到疾病和营养不良的影响，如圆锥角膜、糖尿病性角膜病变和富克斯角膜内皮营养不良，该模型包括来自健康供体的原代人角膜成纤维细胞（HCF）与SH-SY5Y细胞系神经元的3D共培养模型。这使我们首次能够研究人类角膜组织内两种细胞类型之间的相互作用。我们相信，该模型可能有助于剖析与表现出神经损伤的角膜疾病的基质-神经相互作用相关的潜在机制。这种3D模型反映了体内角膜组织的基本解剖和生理特性，未来可作为研究角膜缺陷以及在动物试验前筛选各种药物疗效的工具。
