Low back pain affects nearly 85% of the population of the United States, and is associated with a huge socioeconomic burden. One of the leading causes of low back pain is Intervertebral Disc (IVD) degeneration, which results in an ingrowth of blood vessels and neurites, as well as a sensitization of those neurites. Previous research has indicated that both Aggrecan plays a role in regulating angiogenesis in the IVD, and Aggrecan has also been shown to inhibit neurogenesis and sensitize neurites. However, it is unclear which components of Aggrecan control its inhibitory effects. The goal of this study was to determine how intact and degraded Chondroitin Sulfate groups, the dominant side-chains of Aggrecan, affect angiogenesis in vitro, as well as the sensitization of neurites. Human Umbilical Vein Endothelial Cells (HUVECs) were cultured with either intact or degraded Chondroitin Sulfate-A (CSA), Chondroitin Sulfate-B (CSB), or Chondroitin Sulfate-C (CSC) at a concentration of either 10 µg/ml or 100µg/ml for 16 hours during a tubular formation assay. The tubular assay was quantified for total tubular length using the Angiogenesis Analyzer plugin for Image J. The assay showed that no inhibition occurred for any of the groups, and that pro-angiogenic effects were observed for the 100µg/ml concentration of degraded CSC. The protocol developed to investigate the effect of Aggrecan on neurite sensitization involved optimizing the methodology for extracting Dorsal Root Ganglia (DRGs) cells from mice for use as a primary cell model. The protocol developed is based on literature, but optimizes for simplicity, time, and reduction in the cost of surgical instruments. This protocol achieves these goals, and attains high viability for DRG cell clusters, but fails to consistently retrieve viable cells from the intact DRGs This study sought to investigate the role chondroitin sulfate side chains play in angiogenesis, and it found evidence that the degraded factors can have pro-angiogenic effects. The study also sought to develop a protocol that could be used to aggrecan in the role of sensitization, and it improved the speed and simplicity of existing procedures.No embargoAcademic Major: Biomedical Engineerin
