The Effects of an Autotaxin Inhibitor on an Arthritic Environment Modeled In Human Dermal Fibroblasts Exposed to Tumor Necrosis Factor-É‘
School Name
Hamilton Career Center
Grade Level
11th Grade
Presentation Topic
Cell and Molecular Biology
Presentation Type
Non-Mentored
Abstract
The purpose of this study was to determine if an autotaxin inhibitor could slow or reduce the inflammatory response in fibroblast cells and potentially be used as a treatment for rheumatoid arthritis and other chronic inflammatory diseases. For this study, HA130 was selected as the inhibitor because of its specificity to the autotaxin enzyme, reversible nature, and lack of effects on other essential pathways. To model an arthritic environment in vitro, tumor necrosis factor alpha, one of the major inflammatory cytokines found in the RA patient synovium, was chosen. TNFa is known to increase cell proliferation and viability. For this reason, an MTS assay was chosen to quantitatively measure the results as a color change occurs in the presence of formazan, a byproduct of the compound tetrazolium's reduction; this reaction only takes place in active, proliferating cells. Human dermal fibroblasts were cultured in standard media. Upon reaching confluency, the HDFs were passaged into a 12 well plate. The experiment involved 4 groups: the control group, exposed to standard media only, grew in wells 1A, 1B, and 1C; the TNFa group, exposed to TNFa for 24 hours, grew in wells 2A, 2B, and 2C; the TNFa/HA130 group, exposed to TNFa media for 24 hours and HA130 media for 24 hours, grew in wells 3A, 3B, and 3C; and the HA130 group, exposed to HA130 media for 24 hours, grew in wells 4A, 4B, and 4C. Using a student t-test, the results of the MTS assay were found not to be statistically significant. However, further research modifying this experiment could prove autotaxin inhibitors as a promising treatment option for rheumatoid arthritis.
Recommended Citation
Booker, McKenna, "The Effects of an Autotaxin Inhibitor on an Arthritic Environment Modeled In Human Dermal Fibroblasts Exposed to Tumor Necrosis Factor-É‘" (2020). South Carolina Junior Academy of Science. 244.
https://scholarexchange.furman.edu/scjas/2020/all/244
Start Date
3-28-2020 10:00 AM
Presentation Format
Written Only
Group Project
No
The Effects of an Autotaxin Inhibitor on an Arthritic Environment Modeled In Human Dermal Fibroblasts Exposed to Tumor Necrosis Factor-É‘
The purpose of this study was to determine if an autotaxin inhibitor could slow or reduce the inflammatory response in fibroblast cells and potentially be used as a treatment for rheumatoid arthritis and other chronic inflammatory diseases. For this study, HA130 was selected as the inhibitor because of its specificity to the autotaxin enzyme, reversible nature, and lack of effects on other essential pathways. To model an arthritic environment in vitro, tumor necrosis factor alpha, one of the major inflammatory cytokines found in the RA patient synovium, was chosen. TNFa is known to increase cell proliferation and viability. For this reason, an MTS assay was chosen to quantitatively measure the results as a color change occurs in the presence of formazan, a byproduct of the compound tetrazolium's reduction; this reaction only takes place in active, proliferating cells. Human dermal fibroblasts were cultured in standard media. Upon reaching confluency, the HDFs were passaged into a 12 well plate. The experiment involved 4 groups: the control group, exposed to standard media only, grew in wells 1A, 1B, and 1C; the TNFa group, exposed to TNFa for 24 hours, grew in wells 2A, 2B, and 2C; the TNFa/HA130 group, exposed to TNFa media for 24 hours and HA130 media for 24 hours, grew in wells 3A, 3B, and 3C; and the HA130 group, exposed to HA130 media for 24 hours, grew in wells 4A, 4B, and 4C. Using a student t-test, the results of the MTS assay were found not to be statistically significant. However, further research modifying this experiment could prove autotaxin inhibitors as a promising treatment option for rheumatoid arthritis.
