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Tissue-engineered dermo-epidermal skin analogs exhibit de novo formation of a near natural neurovascular link 10 weeks after transplantation

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Abstract

Purpose

Human autologous tissue-engineered skin grafts are a promising way to cover skin defects. Clearly, it is mandatory to study essential biological dynamics after transplantation, including reinnervation. Previously, we have already shown that human tissue-engineered skin analogs are reinnervated by host nerve fibers as early as 8 weeks after transplantation. In this study, we tested the hypothesis that there is a de novo formation of a “classical” neurovascular link in tissue-engineered and then transplanted skin substitutes.

Methods

Keratinocytes, melanocytes, and fibroblasts were isolated from human skin biopsies. After expansion in culture, keratinocytes and melanocytes were seeded on dermal fibroblast-containing collagen type I hydrogels. These human tissue-engineered dermo-epidermal skin analogs were transplanted onto full-thickness skin wounds on the back of immuno-incompetent rats. Grafts were analyzed after 3 and 10 weeks. Histological sections were examined with regard to the ingrowth pattern of myelinated and unmyelinated nerve fibers into the skin analogs using markers such as PGP9.5, NF-200, and NF-160. Blood vessels were identified with CD31, lymphatic vessels with Lyve1. In particular, we focused on alignment patterns between nerve fibers and either blood and/or lymphatic vessels with regard to neurovascular link formation.

Results

3 weeks after transplantation, blood vessels, but no nerve fibers or lymphatic vessels could be observed. 10 weeks after transplantation, we could detect an ingrowth of myelinated and unmyelinated nerve fibers into the skin analogs. Nerve fibers were found in close proximity to CD31-positive blood vessels, but not alongside Lyve1-positive lymphatic vessels.

Conclusion

These data suggest that host-derived innervation of tissue-engineered dermo-epidermal skin analogs is initiated by and guided alongside blood vessels present early post-transplantation. This observation is consistent with the concept of a cross talk between neurovascular structures, known as the neurovascular link.

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References

  1. Hermanson A, Dalsgaard CJ (1987) Sensory reinnervation and sensibility in skin transplants. Med Biol 65(1):49–52

    CAS  PubMed  Google Scholar 

  2. Altun V, Hakvoort TE, van Zuijlen PP, van der Kwast TH, Prens EP (2001) Nerve outgrowth and neuropeptide expression during the remodeling of human burn wound scars. A 7-month follow-up study of 22 patients. Burns 27(7):717–722

    Article  CAS  PubMed  Google Scholar 

  3. Ward RS, Tucket RP, English KB, Johansson O, Saffle JR (2003) Substance P axons and sensory threshold increase in burn-graft human skin. J Surg Res 118:154–160

    Article  Google Scholar 

  4. Nedelec B, Hou Q, Sohbi I, Choiniüre M, Beauregard G, Dykes RW (2005) Sensory perception and neuroanatomical structures in normal and grafted skin of burn survivors. Burns 31:817–830

    Article  PubMed  Google Scholar 

  5. Anderson JR, Zorbas JS, Phillips JK, Harrison JL, Dawson LF, Bolt SE, Rea SM, Klatte JE, Paus R, Zhu B, Giles NL, Drummond PD, Wood FM, Fear MW (2010) Systemic decreases in cutaneous innervation after burn injury. J Investig Dermatol 130:1948–1951

    Article  CAS  PubMed  Google Scholar 

  6. Anderson JR, Fear MW, Phillips JK, Dawson LF, Wallace H, Wood FM, Rea SM (2011) A preliminary investigation of the reinnervation and return of sensory function in burn patients treated with INTEGRA®. Burns 37(7):1101–1108

    Article  PubMed  Google Scholar 

  7. Botchkarev VA, Eichmuller S, Johansson O, Paus R (1997) Hair cycle-dependent plasticity of skin and hair follicle innervation in normal murine skin. J Comp Neurol 386(3):379–395

    Article  CAS  PubMed  Google Scholar 

  8. Gagnon V, Larouche D, Parenteau-Bareil R, Gingras M, Germain L, Berthod F (2011) Hair follicles guide nerve migration in vitro and in vivo in tissue-engineered skin. J Investig Dermatol 131(6):1375–1378

    Article  CAS  PubMed  Google Scholar 

  9. Carmeliet P, Tessier-Lavigne M (2005) Common mechanisms of nerve and blood vessel wiring. Nature 436(7048):193–200

    Article  CAS  PubMed  Google Scholar 

  10. Carmeliet P (2003) Blood vessels and nerves: common signals, pathways and diseases. Nat Rev Genet 4(9):710–720

    Article  CAS  PubMed  Google Scholar 

  11. Glebova NO, Ginty DD (2005) Growth and survival signals controlling sympathetic nervous system development. Annu Rev Neurosci 28:191–222

    Article  CAS  PubMed  Google Scholar 

  12. Marko SB, Damon DH (2008) VEGF promotes vascular sympathetic innervation. Am J Physiol Heart Circ Physiol 294(6):H2646–H2652

    Article  CAS  PubMed  Google Scholar 

  13. Honma Y, Araki T, Gianino S, Bruce A, Heuckeroth R, Johnson E (2002) Artemin is a vascular-derived neurotropic factor for developing sympathetic neurons. Neuron 35(2):267–282

    Article  CAS  PubMed  Google Scholar 

  14. Long JB, Jay SM, Segal SS, Madri JA (2009) VEGF-A and Semaphorin3A: modulators of vascular sympathetic innervation. Dev Biol 334(1):119–132

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Chauvet S, Burk K, Mann F (2013) Navigation rules for vessels and neurons: cooperative signaling between VEGF and neural guidance cues. Cell Mol Life Sci 70(10):1685–1703

    Article  CAS  PubMed  Google Scholar 

  16. Biedermann T, Böttcher-Haberzeth S, Klar AS, Pontiggia L, Schiestl C, Meuli-Simmen C, Reichmann E, Meuli M (2013) Rebuild, restore, reinnervate: do human tissue engineered dermo-epidermal skin analogs attract host nerve fibers for innervation? Pediatr Surg Int 29(1):71–78

    Article  PubMed  Google Scholar 

  17. Biedermann T, Pontiggia L, Böttcher-Haberzeth S, Tharakan S, Braziulis E, Schiestl C, Meuli M, Reichmann E (2010) Human eccrine sweat gland cells can reconstitute a stratified epidermis. J Investig Dermatol 130(8):1996–2009

    Article  CAS  PubMed  Google Scholar 

  18. Böttcher-Haberzeth S, Biedermann T, Pontiggia L, Braziulis E, Schiestl C, Hendriks B, Eichhoff OM, Widmer DS, Meuli-Simmen C, Meuli M, Reichmann E (2013) Human eccrine sweat gland cells turn into melanin-uptaking keratinocytes in stratifying dermo-epidermal skin substitutes. J Investig Dermatol 133(2):316–324

    Article  PubMed  Google Scholar 

  19. Kiowski G, Biedermann T, Widmer DS, Civenni G, Burger C, Dummer R, Sommer L, Reichmann E (2012) Engineering melanoma progression in a humanized environment in vivo. J Investig Dermatol 132(1):144–153

    Article  CAS  PubMed  Google Scholar 

  20. Schneider J, Biedermann T, Widmer D, Montaño I, Meuli M, Reichmann E, Schiestl C (2009) Matriderm versus integra: a comparative experimental study. Burns 35(1):51–57

    Article  PubMed  Google Scholar 

  21. Böttcher-Haberzeth S, Biedermann T, Schiestl C, Hartmann-Fritsch F, Schneider J, Reichmann E, Meuli M (2012) Matriderm® 1 mm versus Integra® Single Layer 1.3 mm for one-step closure of full thickness skin defects: a comparative experimental study in rats. Pediatr Surg Int 28(2):171–177

    Article  PubMed  Google Scholar 

  22. Martin P, Lewis J (1989) Origins of the neurovascular bundle: interactions between developing nerves and blood vessels in embryonic chick skin. Int J Dev Biol 33(3):379–387

    CAS  PubMed  Google Scholar 

  23. Quaegebeur A, Lange C, Carmeliet P (2011) The neurovascular link in health and disease: molecular mechanisms and therapeutic implications. Neuron 71(3):406–424

    Article  CAS  PubMed  Google Scholar 

  24. Mukouyama YS, Shin D, Britsch S, Taniguchi M, Anderson DJ (2002) Sensory nerves determine the pattern of arterial differentiation and blood vessel branching in the skin. Cell 109(6):693–705

    Article  CAS  PubMed  Google Scholar 

  25. Ruiz de Almodovar C, Lambrechts D, Mazzone M, Carmeliet P (2009) Role and therapeutic potential of VEGF in the nervous system. Physiol Rev 89(2):607–648

    Article  CAS  PubMed  Google Scholar 

  26. Eichmann A, Thomas JL (2013) Molecular parallels between neural and vascular development. Cold Spring Harb Perspect Med 3(1):a006551

    Article  PubMed  Google Scholar 

  27. Hendrix S, Picker B, Liezmann C, Peters EM (2008) Skin and hair follicle innervation in experimental models: a guide for the exact and reproducible evaluation of neuronal plasticity. Exp Dermatol 17(3):214–227

    Article  PubMed  Google Scholar 

  28. Blais M, Parenteau-Bareil R, Cadau S, Berthod F (2013) Concise review: tissue-engineered skin and nerve regeneration in burn treatment. Stem Cells Transl Med 2(7):545–551

    Article  CAS  PubMed  Google Scholar 

  29. Caissie R, Gingras M, Champigny MF, Berthod F (2006) In vivo enhancement of sensory perception recovery in a tissue-engineered skin enriched with laminin. Biomaterials 27(15):2988–2993

    Article  CAS  PubMed  Google Scholar 

  30. Blais M, Grenier M, Berthod F (2009) Improvement of nerve regeneration in tissue-engineered skin enriched with Schwann cells. J Investig Dermatol 129(12):2895–2900

    Article  CAS  PubMed  Google Scholar 

Download references

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.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

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

    Thomas Biedermann, Agnieszka S. Klar, Sophie Böttcher-Haberzeth & Ernst Reichmann

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

    Sophie Böttcher-Haberzeth, Clemens Schiestl & Martin Meuli

Authors
  1. Thomas Biedermann
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  2. Agnieszka S. Klar
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  3. Sophie Böttcher-Haberzeth
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  4. Clemens Schiestl
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  5. Ernst Reichmann
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  6. Martin Meuli
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Corresponding author

Correspondence to Martin Meuli.

Additional information

T. Biedermann and A. S. Klar contributed equally to this paper.

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Biedermann, T., Klar, A.S., Böttcher-Haberzeth, S. et al. Tissue-engineered dermo-epidermal skin analogs exhibit de novo formation of a near natural neurovascular link 10 weeks after transplantation. Pediatr Surg Int 30, 165–172 (2014). https://doi.org/10.1007/s00383-013-3446-x

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