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Plgf-1 Contained In Normal Wound Myofibroblast-derived Microvesicles Stimulates Collagen Production By Dermal Fibroblasts
Syrine Arif.
Université Laval, Québec, QC, Canada.

Syrine Arif*, Sébastien Larochelle*, Véronique J. Moulin* *Centre de recherche en organogénèse expérimentale de l’Université Laval/LOEX - Centre de recherche du CHU de Québec-Université Laval, QC, Canada.
Myofibroblasts are differentiated cells from fibroblasts that appear during the last stages of healing. They can communicate with other cells using secreted factors but also using microvesicles (MVs). Our aim is to evaluate the effect of MVs produced by myofibroblasts from normal wound (Wmyo) on skin fibroblasts and to determine the signaling molecule responsible for these effects. To model a cell to cell communication between Wmyo and fibroblasts, we studied fibroblasts uptake of fluorescent Wmyo-derived MVs by flow cytometry. We analyzed cytokines distribution in MV samples by a multiplex ELISA detecting 45 cytokines. Fibroblasts cells were then cultured in presence of different concentration of MVs or a selected cytokine for 5 days. Following the treatments, parameters linked to the ECM were studied including pro-collagen I production assayed by ELISA. Finally, fibroblasts were treated with MVs preincubated with a neutralizing antibody for PLGF-1 before to evaluate collagen production.In the presence of fluorescent MVs, a large portion of fibroblast cells demonstrated fluorescence shift with a fold change of 2.96 ±031 versus untreated fibroblasts (N= 2). Cytokine array analysis showed that MVs samples contained a high amount of PLGF-1 (57.53 pg/ug proteins in MVs ± 13.11, N= 6). Cutaneous fibroblasts treated with MVs (10µg protein/ml) or PLGF-1 (10ng/ml) significantly simulated pro-collagen I level production with a fold change of 1.80±0.18 for MVs and, 2.07±0.18 for PLGF-1 (N=3) versus untreated cells. Finally, the neutralization of PLGF-1 in MVs samples effectively neutralized the production of procollagen I (N=3) by fibroblasts. Wmyo derived-MVs stimulate fibroblast collagen production during healing possibly through PLGF-1 signaling.

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