In vitro model of the closed eye environment and material-induced inflammation

炎症是细胞对生物医学设备(如隐形眼镜、支架和血管移植)反应的组成部分。尽管进行了多年的研究，我们还没有设计出能够与人体相互作用而不会引起并发症的材料。要设计与人类生物系统兼容的材料，首先必须了解细胞与材料是如何相互作用的。此外，需要适当的实验模型来测试细胞在适当模拟生物研究系统的环境中的反应。因为眼睛是一个很难建模的环境，所以在这一领域做的研究很少。尤其是中性粒细胞，即炎性白血球，在长时间闭眼(3小时或更长时间)时会侵入眼前部。这些中性粒细胞由于其炎性细胞的性质，有可能造成重大损害，因为它们拥有旨在对抗感染和异物的炎性化合物武器库。然而，尽管在闭眼环境中可以在数十万人中发现它们，但它们对正常炎症刺激的反应和反应能力似乎有限。这些细胞的行为是由于它们与其他细胞的相互作用还是由于泪液中存在特定的化合物？使用体外机制方法，这项研究资助提案旨在更好地了解所涉及的细胞机制。在不同的细胞培养条件下，中性粒细胞将与已知存在于泪液中的不同蛋白质孵育，并将被允许与角膜和结膜细胞相互作用。在模拟眼睛前面的环境中，特定的蛋白质、炎性化合物和细胞在中性粒细胞对炎性刺激做出反应的能力中所起的作用将被确定。这些实验不仅使我们能够建立闭眼环境的真实体外模型，而且还使我们能够表征炎症细胞的反应。此外，使用这个体外模型，我们将确定异物，如隐形眼镜，对中性粒细胞炎症反应的影响。最终，这项研究计划将创造关于眼睛环境中炎症的细胞机制的新知识，以及这些机制如何与生物材料的存在引起的炎症反应有关。总之，这项研究计划将有助于确定关键的分子途径，这些途径将代表制造商开发更好的药物和与眼睛更相容的材料的新研究战略。该计划将培训四名研究生，使用与细胞生物学、体外模型和材料相互作用相关的方法学进行跨学科研究。学生将获得的专业知识将使他们能够加入迅速崛起的、至关重要的加拿大生物医学和生物技术行业。

Inflammation is an integral part of the cell response to biomedical devices, such as contact lens, stent and vascular graft. Despite many years of research, we have yet to design materials that are able to interact with the human body without causing complications. To design materials that are compatible with human biological systems, one must first understand how cells interact with materials. Furthermore, appropriate experimental models are needed to test the cell response in an environment that appropriately mimics the biological system of study. Because the eye is a difficult environment to model, little research has been done in this area. This is especially true for neutrophils, inflammatory white blood cells, which invade the front of the eye during prolonged eye closure (3 hours or more). These neutrophils, due to their nature as inflammatory cells, have the potential to create significant damage as they possess an arsenal of inflammatory compounds aimed at fighting off infection and foreign materials. However, while they can be found in the hundreds of thousands in the closed-eye environment, they appear to have a limited ability to react and respond to normal inflammatory stimulus. Is the behavior of these cells due to their interactions with other cells or due to the presence of specific compounds in the tears? Using a mechanistic in vitro approach, this research grant proposal aims to gain a better understanding of the cellular mechanisms involved. Using various cell culture conditions, neutrophils will be incubated with different proteins known to be present in tears and will be allowed to interact with corneal and conjunctival cells. The role that specific proteins, inflammatory compounds and cells play in the neutrophil’s ability to respond to inflammatory stimulus in an environment that mimics the front of the eye will be determined. These experiments will not only enable us to develop a true in vitro model of the closed-eye environment but will also enable us to characterize the inflammatory cell response. Furthermore, using this in vitro model, we will determine the effect that a foreign material such as a contact lens has on the neutrophil inflammatory response. Ultimately, this research program will create new knowledge on the cellular mechanisms of inflammation in the ocular environment, but also on how these can relate to the inflammatory response induced by the presence of a biomaterial. All together, this research program will help identify key molecular pathways that will represent novel research strategies for manufacturers to develop better pharmaceuticals and materials that are more compatible with the eye. This program will train four graduate students in interdisciplinary research using methodology related to cell biology, in vitro models and interaction with materials. The expertise the students will gain will allow them to join the rapidly emerging and critically important Canadian biomedical and biotechnology industry.