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Myelinated and unmyelinated nerve fibers reinnervate tissue-engineered dermo-epidermal human skin analogs in an in vivo model

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Abstract

Purpose

The clinical application of autologous tissue-engineered skin analogs is an important strategy to cover large skin defects. Investigating biological dynamics, such as reinnervation after transplantation, is essential to improve the quality of such skin analogs. Previously, we have examined that our skin substitutes are reinnervated by host peripheral nerve fibers as early as 8 weeks after transplantation. Here, we wanted to investigate the presence and possible differences regarding myelinated and unmyelinated host nerve fibers 15 weeks after the transplantation of light and dark human tissue-engineered skin analogs.

Methods

Human epidermal keratinocytes, melanocytes, and dermal fibroblasts were isolated from human light and dark skin biopsies. Keratinocytes and melanocytes were seeded on fibroblast-containing collagen type I hydrogels after expansion in culture. After additional culturing, the tissue-engineered dermo-epidermal skin analogs were transplanted onto full-thickness skin wounds created on the back of immuno-incompetent rats. Skin substitutes were excised and analyzed 15 weeks after transplantation. Histological sections were examined with regard to the ingrowth pattern of myelinated and unmyelinated nerve fibers into the skin analogs using markers, such as Substance P, NF200, and S100-Beta.

Results

We found myelinated and unmyelinated peripheral host nerve fibers 15 weeks after transplantation in the dermal part of our human skin substitutes. In particular, we identified large-diameter-myelinated Aβ- and Aδ-fibers, and small-diameter C-fibers. Furthermore, we observed myelinated nerves in close proximity to CD31-positive blood capillaries. In the long run, both types of ingrown host fibers showed an identical pattern in both light and dark skin analogs.

Conclusion

Our data suggest that myelinated and unmyelinated peripheral nerves reinnervate human skin substitutes in a long-term in vivo transplantation assay. Our tissue-engineered skin analogs attract A- and C-fibers to supply both light and dark skin analogs. Potentially, this process restores skin sensitivity and has, therefore, a significant relevance with regard to future application of autologous pigmented dermo-epidermal skin substitutes onto patients.

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Acknowledgments

This work was financially supported by the EU-FP7 project EuroSkinGraft (FP7/2007-2013: Grant Agreement No 279024), by the EU-FP7 (MultiTERM, Grant Agreement No 238551), and the Clinical Research Priority Programs (KFSP: From basic research to the clinic: Novel tissue-engineered skin grafts for Zurich) of the Faculty of Medicine of the University of Zurich. We are particularly grateful to the Fondation Gaydoul 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, Department of Surgery, University Children’s Hospital Zurich, Steinwiesstrasse 75, 8032, Zurich, Switzerland

    T. Biedermann, A. S. Klar & E. Reichmann

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

    S. Böttcher-Haberzeth & M. Meuli

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

    T. Biedermann, A. S. Klar, S. Böttcher-Haberzeth, E. Reichmann & M. Meuli

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  1. T. Biedermann
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Correspondence to T. Biedermann.

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Biedermann, T., Klar, A.S., Böttcher-Haberzeth, S. et al. Myelinated and unmyelinated nerve fibers reinnervate tissue-engineered dermo-epidermal human skin analogs in an in vivo model. Pediatr Surg Int 32, 1183–1191 (2016). https://doi.org/10.1007/s00383-016-3978-y

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