Comparison of cytokine response to anti-inflammatory modulators

Event Abstract Back to Event Comparison of cytokine response to anti-inflammatory modulators Alda Diaz Perez1, 2, Tina M. Poseno2, Jeannine M. Durdik1, 3 and Julie A. Stenken1, 2 1 University of Arkansas, Cell and Molecular Biology, United States 2 University of Arkansas, Department Chemistry and Biochemistry, United States 3 University of Arkansas, Department of Biological Sciences, United States Statement Purpose: Macrophages (Mф) are plastic cells that display different phenotypic changes in response to chemical cues (cytokines, pattern receptors, and hormones) in the microenvironment. There is a significant interest in modulating the phenotype of these cells for improved outcomes for biomaterials implants[1]. While there are many potential modulators to use, how they compare between in vitro outputs and in vivo outcomes is poorly understood. The objective of this study is to compare the in vitro and in vivo cytokine response from macrophages to the macrophage modulator resolvin D1 (RvD1) using cell culture and subcutaneous sponge implants. Methods: Peritoneal macrophage (PM) Cell Culture: Peritoneal macrophages were extracted from the peritoneal cavity of male Sprague-Dawley rats and then the cells were centrifuged at 1100 RPM. Cells were incubated in media (which contained 15 v/v % fetal bovine serum, 1 v/v % of 100U/mL penicillin, and 84 v/v % of F-12K) for 2 hours to allow the Mфs to attach to the surface. Mф were cultured at 5.5 x 105 cells/ mL per well. PMs were induced with lipopolysaccharide (LPS) at 50 ng/ mL, and then 5 min later, the cells were treated with RvD1 at 1 µM, 300 nM, 100 nM, 30 nM and 10 nM and then incubated for 24 hrs. Wound fluid from sponge: Autoclaved polyvinyl alcohol (PVA) sponges (in 0.9 wt% NaCl) were soaked in 1 nM RvD1 for approximately 5 min, then placed subcutaneously (four sponges) into the dorsal subcutaneous space of male Sprague Dawley rats. Sponges were removed 24 hours post-implantation. Wound fluid from the sponges and supernatant from the cultured PMs were quantified for TNF-α and IL-6 using ELISA. Results and Discussion: A B C Figure 1 IL-6 and TNF-α concentration in cell culture and TNF-α in sponge experiments: (A and B) PMs compared with LPS showed significance differences ( ) using ANOVA and Tukey post-hoc (p≤0.05) N=3. (C) TNF-α from sponge fluid. For most of the RvD1 treatments an approximate 60% decrease in TNF-α concentration was observed after 24 hours incubation (Figure 1A). Interestingly, RvD1 (1 µM) did not show significantly decrease TNF-α or IL-6 (Figure 1B) compared to LPS treatment alone. In Figure 1C, RvD1 caused roughly a 45% decrease in TNF-α concentration in vivo in a sponge model. Predicting in vivo macrophage activation properties is not well described in the literature. This work serves as an initial step toward elucidating efficacious modulators for macrophage activation in biomaterials contexts. While the use of LPS in the cell culture experiments promotes a macrophage phenotype that would not be expected in vivo, the ability to modulate the macrophage phenotype after LPS stimulation provides a means to compare and contrast in vitro vs in vivo responses. NIH EB 014404References:[1] Daley, J. M., Brancato, S. K., Thomay, A. A., Reichner, J. S., & Albina, J. E. (2010). The phenotype of murine wound macrophages. J Leukoc Biol, 87(1), 59-67. Keywords: cytokine, Drug delivery, biomaterial Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Fibrosis and biomaterials Citation: Diaz Perez A, Poseno TM, Durdik JM and Stenken JA (2016). Comparison of cytokine response to anti-inflammatory modulators. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02760 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Alda Diaz Perez Tina M Poseno Jeannine M Durdik Julie A Stenken Google Alda Diaz Perez Tina M Poseno Jeannine M Durdik Julie A Stenken Google Scholar Alda Diaz Perez Tina M Poseno Jeannine M Durdik Julie A Stenken PubMed Alda Diaz Perez Tina M Poseno Jeannine M Durdik Julie A Stenken Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.