The Development of Nanofabricated Platforms for the Study of Oxidized Lipid Systems
Reactive oxygen species, such as those caused by smoking, alcohol consumption, or environmental factors, can lead to the degradation of biological molecules like lipids. Oxidized lipids can alter the properties of cell membranes and potentially signal cell death resulting in problems such as cancer and atherosclerosis. CD36 is a protein found on some immune cells that is responsible for the recognition and binding of oxidized lipids and low-density lipoproteins. Learning more about the interaction between CD36 and oxidized moieties in the body is important to guide future clinical research towards new and much needed therapies. The extensive study of these interactions often necessitates robotics or time-consuming manual work which can be prohibitive to smaller research groups or fast-paced testing environments resulting in the slower progression of the field. Through the aid of microscale fabrication, new and easy to use platforms have been created for the high-throughput creation of multi-concentration arrays. The first method used microfluidic mixers to create concentration gradients of oxPL membranes which was analyzed through fluorescently labeled vesicles tagged with the CD36 protein. This work discovered a new target for CD36 that was confirmed by simulations of the lipid interactions. This directly demonstrates the ability of the platform to improve target-receptor screening for membrane supported systems. The second method uses grid arrays of lipids containing a light sensitive compound to generate controlled patterns of oxPL. This work expands previous labelled vesicle methods to analyze the behavior of immune cells as they respond to the oxPL patterns in real time. The incorporation of macrophages, a type of immune cell, into the grid system establishes more biologically accurate substrates while also allowing for real time analysis that is difficult to achieve via other methods. In conclusion a variety of different platforms have been established that will facilitate faster and more efficient research into protein-receptor studies allowing for larger and more complex systems, such as whole cell or multi-protein systems, to be assessed in the future.