During the last stages of wound healing, myofibroblasts differentiate mainly from fibroblasts. Myofibroblasts from normal skin wounds (Wmyo) can communicate with its surrounding using secreted factors. They also have the capacity to produce microvesicles (MVs), a type of extracellular vesicles, as mediators of intercellular communication. MVs cargo are potentially capable of regulating the behavior of targeted cells and tissues. The aim of this study is to evaluate the effect of Wmyo-derived MVs on dermal fibroblasts and to determine the responsible signaling molecule. Microvesicles were obtained from culture media of myofibroblasts and characterized using protein quantification, dynamic light scattering and transmission electron microscopy. Uptake of fluorescent MVs in fibroblasts was assessed by flow cytometry. Cytokines concentrations were quantified in MV samples by a multiplex ELISA. Different concentration of MVs or a selected cytokine were used as treatments over fibroblasts culture for 5 days. Following the treatments, parameters linked to the extracellular matrix were studied. Lastly, the selected cytokine was neutralized within MVs before evaluating collagen production. We showed that Wmyo derived-MVs were internalized by dermal fibroblasts. Cytokine array analysis revealed that a large amount of placental growth factor 1 (PLGF-1) (0.88 ± 0.63 pg/μg proteins in MVs) could be detected in MVs samples. Cutaneous fibroblasts treated with MVs or PLGF-1 showed significantly stimulated procollagen I level production (Fold change of 1.80 ± 0.18 and 2.07 ± 0.18, respectively). Finally, the neutralization of PLGF-1 in MVs significantly inhibited the production of procollagen I by fibroblasts. Our study shows that Wmyo derived-MVs are involved in intercellular communication by stimulating collagen production by fibroblasts during wound healing. This effect is possibly attained through PLGF-1 signalling. These findings represent a promising opportunity to gain insight into how MVs and Wmyo may mediate the healing of a skin wound.
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The data that support the findings of this study are available from the corresponding author upon reasonable request.
Myofibroblasts from normal skin wounds
Placental growth factor 1
Transforming growth factor-ß
Alpha-Smooth Muscle Actin
Dulbecco’s Modified Eagle’s medium
Fetal bovine serum
Red Fluorescent Protein
Dynamic light scattering
PLGF-1 neutralizing antibodies
Lymphotoxin alpha/ tumor necrosis beta
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The authors thank Caroline Gilbert from the CHU of the Université Laval for the use of the Nanosizer® apparatus, Richard Janvier from IBIS of the Université Laval for the use of their TEM microscope and, Annie Karakeussian-Rimbaud from the CHUM for the handling of the multiplex ELISA device.
This work was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) (RGPIN2014–04404); les Fonds de recherche du Québec-Santé (FRQS) (Research Centre funding grant); the Quebec Cell, Tissue and Gene Therapy Network–ThéCell (a thematic network supported by FRQS).
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The authors declare that there are no conflicts of interest.
All procedures involving patients were reviewed and approved by the Research Ethical Committee of the Centre Hospitalier Universitaire (CHU) de Québec-Université Laval, 2014–04404.
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Arif, S., Larochelle, S. & Moulin, V.J. PLGF-1 contained in normal wound myofibroblast-derived microvesicles stimulated collagen production by dermal fibroblasts. J. Cell Commun. Signal. (2020). https://doi.org/10.1007/s12079-020-00572-5
- Wound healing