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Comparison of in vivo immune responses following transplantation of vascularized and non-vascularized human dermo-epidermal skin substitutes

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Pediatric Surgery International Aims and scope Submit manuscript

Abstract

Purpose

Autologous bio-engineered dermo-epidermal skin substitutes (DESS) represent an alternative therapeutic option for a definitive treatment of skin defects in human patients. Largely, the interaction of host immune cells with transplanted DESS is considered to be essential for the granulation tissue formation, graft take, and its functionality. The aim of this study was to compare the spatiotemporal distribution and density of host-derived monocytes/macrophages and granulocytes in vascularized (vascDESS) versus non-vascularized DESS (non-vascDESS) in a rat model.

Methods

Keratinocytes and the stromal vascular fraction (SVF) were derived from human skin or human adipose tissue, respectively. Human SVF containing both endothelial and mesenchymal/stromal progenitors was used to develop a vascularized collagen type I-based dermal component in vitro. The donor-matched, monolayer-expanded adipose stromal cells lacking endothelial cells were used as a negative control. Subsequently, human keratinocytes were seeded on top of hydrogels to build dermo-epidermal skin grafts. After transplantation onto full-thickness skin wounds on the back of immuno-incompetent rats, grafts were excised and analyzed after 1 and 3 weeks. The expression of distinct inflammatory cell markers specific for host-derived monocytes/macrophages (CD11b, CD68) or granulocytes (HIS48) was analyzed by immunofluorescence microscopy.

Results

All skin grafts were infiltrated by host-derived monocytes/macrophages (CD11b+, CD68+) and granulocytes (HIS48+) between 1–3 week post-transplantation. When compared to non-vascDESS, the vascDESS showed an increased granulocyte infiltration at all time points analyzed with the majority of cells scattered throughout the whole dermal part. Whereas a moderate number of rat monocytes/macrophages (CD11b+, CD68+) were found in vascDESS at 1 week, only a few cells were detected in non-vascDESS. We observed a time-dependent decrease of monocytes/macrophages in all transplants at 3 weeks.

Conclusions

These results demonstrate a distinct spatiotemporal distribution of monocytes/macrophages as well as granulocytes in our transplants that closely resemble the one observed during physiological wound healing. The differences identified between vascDESS and non-vascDESS may indicate that human endothelial cells lining blood capillaries of vascDESS accelerate infiltration of monocytes and leukocytes.

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Acknowledgements

This work was financially supported by the EU-FP6 project EuroSTEC (soft tissue engineering for congenital birth defects in children: contract: LSHB-CT-2006-037409), by the EU-FP7 project EuroSkinGraft (FP7/2007-2013: Grant Agreement Nr 279024), by the EU-FP7 (MultiTERM, Grant Agreement Nr 238551), and the Clinical Research Priority Programs (CRPP) of the Faculty of Medicine of the University of Zurich. We are particularly grateful to the Gaydoul Foundation and the sponsors of “DonaTissue” (Thérèse Meier and Robert Zingg) for their generous financial support and interest in our work.

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Authors and Affiliations

  1. Tissue Biology Research Unit, University Children’s Hospital Zurich, Zurich, Switzerland

    Agnes S. Klar, Thomas Biedermann & Ernst Reichmann

  2. Department of Surgery, University Children’s Hospital Zurich, Steinwiesstrasse 75, 8032, Zurich, Switzerland

    Martin Meuli

  3. Children’s Research Center, University Children’s Hospital Zurich, Zurich, Switzerland

    Agnes S. Klar, Thomas Biedermann, Ernst Reichmann & Martin Meuli

  4. Department of Plastic, Reconstructive, Aesthetical, and Hand Surgery, Kantonsspital Aarau, Aarau, Switzerland

    Claudia Simmen-Meuli

Authors
  1. Agnes S. Klar
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  2. Thomas Biedermann
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  3. Claudia Simmen-Meuli
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  4. Ernst Reichmann
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  5. Martin Meuli
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Correspondence to Martin Meuli.

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Klar, A.S., Biedermann, T., Simmen-Meuli, C. et al. Comparison of in vivo immune responses following transplantation of vascularized and non-vascularized human dermo-epidermal skin substitutes. Pediatr Surg Int 33, 377–382 (2017). https://doi.org/10.1007/s00383-016-4031-x

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  • DOI: https://doi.org/10.1007/s00383-016-4031-x

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