The Role of Protein Change (Cellular Protein Loss and Denaturation) in Determining Outcomes of Heating, Cryotherapy and Irreversible Electroporation
2018-04
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The Role of Protein Change (Cellular Protein Loss and Denaturation) in Determining Outcomes of Heating, Cryotherapy and Irreversible Electroporation
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2018-04
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Atrial fibrillation currently affects millions of people in the US alone. Focal therapy is an increasingly attractive treatment for atrial fibrillation that avoids the debilitating effects of drugs for disease control. Perhaps the most widely used focal therapy for atrial fibrillation (AF) is heat-based radiofrequency (heating), although cryotherapy (cryo) is rapidly replacing it due to a reduction in side effects and positive clinical outcomes. A third focal therapy, irreversible electroporation (IRE), is also being considered in some settings. This study was designed to help guide treatment thresholds and compare mechanism of action across heating, cryo, and IRE. Testing was undertaken on HL-1 cells, a well-established cardiomyocyte cell line, to assess injury thresholds for each treatment method. Cell viability, as assessed by Hoechst and PI staining, was found to be minimal after exposure to temperatures ≤-40 °C (cryo), ≥60 °C (heating), and when field strengths ≥1500 V/cm (IRE) were used. Viability was then correlated to protein denaturation fraction (PDF) as assessed by Fourier Transform Infrared (FTIR) spectroscopy, and protein loss fraction (PLF) as assessed by Bicinchoninic Acid (BCA) assay after the three treatments. These protein changes were assessed both in the supernatant and the pellet of cell suspensions post treatment. We found that dramatic viability loss (≥50%) correlated strongly with ≥12% protein change (PLF, PDF or a combination of the two) in every focal treatment. These studies help in defining both cellular thresholds and protein-based mechanisms of action that can be used to improve focal therapy application for atrial fibrillation.
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University of Minnesota M.S.M.E. thesis. April 2018. Major: Mechanical Engineering. Advisor: John Bischof. 1 computer file (PDF); vi, 48 pages.
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Liu, Feng. (2018). The Role of Protein Change (Cellular Protein Loss and Denaturation) in Determining Outcomes of Heating, Cryotherapy and Irreversible Electroporation. Retrieved from the University Digital Conservancy, https://hdl.handle.net/11299/201723.
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