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01-02-2018 | Original Article

Characterization of M1 and M2 polarization of macrophages in vascularized human dermo-epidermal skin substitutes in vivo

Authors: Agnes S. Klar, Katarzyna Michalak-Mićka, Thomas Biedermann, Claudia Simmen-Meuli, Ernst Reichmann, Martin Meuli

Published in: Pediatric Surgery International | Issue 2/2018

Abstract

Aims and objectives

Vascularized bio-engineered human dermo-epidermal skin substitutes (vascDESS) hold promise for treating burn patients, including those with severe full-thickness wounds. We have previously shown that vascDESS promote wound healing by enhanced influx of macrophages and granulocytes. Immediately following transplantation, macrophages infiltrate the graft and differentiate into a pro-inflammatory (M1) or a pro-healing M2 phenotype. The aim of this study was to characterize the activation state of macrophages infiltrating skin transplants at distinct time points following transplantation.

Methods

Keratinocytes and the stromal vascular fraction (SVF) were derived from human skin or adipose tissue, respectively. Human SVF containing both endothelial and mesenchymal/stromal cells was used to generate vascularized dermal component in vitro, which was subsequently covered with human keratinocytes. Finally, vascDESS were transplanted on the back of immuno-incompetent rats, excised, and analyzed after 1 and 3 weeks using immunohistological techniques.

Results

A panel of markers of macrophage M1 (nitric oxide synthase: iNOS) and M2 (CD206) subclass was used. All skin grafts were infiltrated by both M1 and M2 rat macrophages between 1–3 weeks post-transplantation. CD68 (PG-M1) was used as a pan-macrophage marker. The number of CD68⁺CD206⁺ M2-polarized macrophages was higher in 3-week transplants as compared to early-stage transplants (1 week). In contrast, the number of CD68⁺iNOS⁺ M1 cells was markedly decreased in later stages in vivo.

Conclusions

Macrophages exhibit a heterogeneous and temporally regulated polarization during skin wound healing. Our results suggest that the phenotype of macrophages changes during healing from a more pro-inflammatory (M1) profile in early stages after injury, to a less inflammatory, pro-healing (M2) phenotype in later phases in vivo.

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Title: Characterization of M1 and M2 polarization of macrophages in vascularized human dermo-epidermal skin substitutes in vivo
Authors: Agnes S. Klar
Katarzyna Michalak-Mićka
Thomas Biedermann
Claudia Simmen-Meuli
Ernst Reichmann
Martin Meuli
Publication date: 01-02-2018
Publisher: Springer Berlin Heidelberg
Published in: Pediatric Surgery International / Issue 2/2018
Print ISSN: 0179-0358
Electronic ISSN: 1437-9813
DOI: https://doi.org/10.1007/s00383-017-4179-z

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Characterization of M1 and M2 polarization of macrophages in vascularized human dermo-epidermal skin substitutes in vivo

Abstract

Aims and objectives

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Aims and objectives

Methods

Results

Conclusions

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