(275) Interaction among keratinocytes, fibroblasts and CD163-overexpressing macrophages for a more efficient wound healing process

Abstract

Peripheral neuroimmune interactions play an important role in the development of pain and wound healing process after surgery. We hypothesize that the over-expression of the scavenger receptor gene CD163 in human macrophages will promote an M2 cellular phenotype that will promote a more efficient wound healing process. We will test this hypothesis following these aims: 1) Induce CD163 in macrophages using a nanoparticle that target these cells specifically; 2) Promote a more efficient wound healing process with CD163 overexpressing macrophages using an in vitro assay; and 3) Determine that the observed effects are achieved specifically by the overexpression of CD163. For this purpose, we used an in vitro wound healing scratch assay with primary human keratinocytes and fibroblasts co-cultured with macrophages overexpressing CD163 (or control groups) in the presence or absence of antibodies, anti-CD163 antibody [RM3/1] or its isotype control IgG1. Microscopy, imaging analysis, gene transfection using mannosylated nanoparticle (mPEI) for targeting specifically macrophages and RT-qPCR were performed. We have previously shown that the induction of CD163 in macrophages successfully promotes an anti-inflammatory phenotype. We confirmed the over-expression of CD163 in M1 macrophages at 48 and 72 hours after transfection. Using an in vitro scratch assay we also observed that the addition of CD163-blocking antibody, but not isotype control, blocked the efficient and faster wound healing process induced by CD163-overexpressing macrophages. Moreover, the interaction among keratinocytes, fibroblasts and CD163-overexpressing macrophages resulted in an increased CD163 gene expression in these macrophages. CD163 seems to play a critical role in the induction of wound healing by promoting an anti-inflammatory phenotype in macrophages. We further postulate that this approach could promote a more efficient wound healing process following major surgeries, which would reduce the incidence of chronic postoperative pain. Supported by Rita-Allen-Foundation & American-Pain-Society 2011-Pain-grant (EAR-S); National-Institutes-of-Health, NIGMS, R15GM109333 (EAR-S); 2015-PRSI-program (PCSP); Presbyterian-College-School-of-Pharmacy (DF,AA) and CAPES-Foundation: Ministry-of-Education-of-Brazil (PDSE:BEX10794/14-0).