Novel in vitro dynamic corneal model with online mechanical characterisation for pharmaceutical screening and tissue engineering applications

兔眼经常被用来测试治疗角膜疾病的新药或新疗法。圆锥角膜就是这种疾病的一个例子，它的机械性能会随着时间的推移而变化。人们对利用动物进行眼科研究的担忧与日俱增，但缺乏用足够的组织模型来取代这些实验的努力。为了减少或消除使用动物模型来了解角膜疾病的生物学和治疗，必须开发更多与生理相关的体外模型。角膜的一个特征与某些疾病直接相关，那就是角膜基质或组织的机械完整性。然而，到目前为止，没有一种3D组织模型能够评估或利用角膜的机械性能作为体外药物测试的结果衡量标准。在这项提议中，我们建议开发一种新的在线机械特性表征工具，该工具可以在无菌条件下以非破坏性的在线方式随着时间的推移监控活组织的构建。这一设置是新发展的一部分，涉及建立用于角膜组织模型的生物反应器，以促进在长期培养期间的生长。这一新的方法使体外组织的机械性能能够用作生物标记物。在成功地建立了方法学之后，我们将使用这个体外模型来筛选药物和治疗圆锥角膜的新疗法。我们的3D模型中的药物疗效将与现有的动物模型数据进行比较。功效将被定义为保持或改善机械完整性。

The rabbit eye is frequently used to test new drugs or new therapies for corneal disease. One example of such as disease is keratoconus where the mechanical properties change over time. Concerns over the use of animals for eye research has been growing but efforts to replace these experiments with adequate tissue models have been lacking. To reduce or eliminate the use of animal models for understanding the biology and treatment of corneal disease, more physiologically relevant in vitro models must be developed. One feature of the cornea which is directly linked to some disease conditions is the mechanical integrity of the corneal stroma or tissue. However, to date, none of the 3D tissue models are able to assess or utilise the mechanical properties of the cornea as an outcome measure of drug testing in vitro. In this proposal, we propose to develop a novel on-line characterisation tool for mechanical properties, which can monitor viable tissue construction over time in a non-destructive and on-line manner under sterile conditions. This set up is part of new developments involving the establishment of a bioreactor for corneal tissue models which facilitates growth over a prolonged culture period. This novel methodology enables the mechanical property of the in vitro tissue to be used as a biomarker. After successfully establishing the methodologies, we shall use this in vitro model for the screening of pharmaceutical drugs and new therapies for keratoconus. Efficacy of drugs in our 3D models will be compared with existing data using animal models. Efficacy will be defined as maintained or improved mechanical integrity.