Microscope Mountable Bioreactor

Lower Back Pain is an ailment that affects nearly 80% of the world population. Many factors can contribute to the development of this disease, including degeneration of the intervertebral disc (IVD), specifically in discogenic chronic back pain (DCBP). A promising drug target for the treatment of DCBP is an ion channel in intervertebral disc cells, called transient receptor potential channel 6 (TRPC6). This channel is critical in regulating tissue homeostasis with the regulation of mechanical and osmotic stress of the IVD. TRPC6 has been shown to have increased expression in degraded IVDs. This channel has been shown in various studies to be activated by physical cues, such as cyclic strain and compression, which is extremely relevant in the IVD, specifically in the annulus fibrosis (AF) that maintains mechanical stability in the IVD, as these cues are experienced in daily life. Activation of TRPC6 has shown upward trends in inflammation in other cell types in various studies, including lung tissue. Modulation of TRPC6 activity can be assessed with calcium influx in cells by marking calcium with an immunofluorescent staining. The goal of this project is to design and create a working prototype of a microscope-mounted bioreactor that has the ability to stretch IVD cells that have been seeded onto polydimethylsiloxane (PDMS) chambers that are compressed at a controlled strain and strain rate. Calcium will be marked in the cells and the calcium influx will be measured, giving us a much better understanding of TRPC6 activation in IVD cells. We will show our completed design and our proposed future improvements.