title: Myelinated and unmyelinated nerve fibers reinnervate tissue-engineered dermo-epidermal human skin analogs in an in vivo model.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Tissue Biology Research Unit, Department of Surgery, University Children's Hospital Zurich, Steinwiesstrasse 75, 8032, Zurich, Switzerland. thomas.biedermann@kispi.uzh.ch.|Children's Research Center, University Children's Hospital Zurich, Zurich, Switzerland. thomas.biedermann@kispi.uzh.ch.
authors.forename: T
authors.initials: T
authors.lastname: Biedermann
authors.affiliation: Tissue Biology Research Unit, Department of Surgery, University Children's Hospital Zurich, Steinwiesstrasse 75, 8032, Zurich, Switzerland.|Children's Research Center, University Children's Hospital Zurich, Zurich, Switzerland.
authors.forename: A S
authors.initials: AS
authors.lastname: Klar
authors.affiliation: Department of Surgery, University Children's Hospital Zurich, Zurich, Switzerland.|Children's Research Center, University Children's Hospital Zurich, Zurich, Switzerland.
authors.forename: S
authors.initials: S
authors.lastname: Böttcher-Haberzeth
authors.affiliation: Tissue Biology Research Unit, Department of Surgery, University Children's Hospital Zurich, Steinwiesstrasse 75, 8032, Zurich, Switzerland.|Children's Research Center, University Children's Hospital Zurich, Zurich, Switzerland.
authors.forename: E
authors.initials: E
authors.lastname: Reichmann
authors.affiliation: Department of Surgery, University Children's Hospital Zurich, Zurich, Switzerland.|Children's Research Center, University Children's Hospital Zurich, Zurich, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Meuli
mesh_terms: D000293:Adolescent; D000818:Animals; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D020405:Dermis / Q000294:innervation* / Q000637:transplantation; D004817:Epidermis / Q000294:innervation* / Q000637:transplantation; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D009413:Nerve Fibers, Myelinated / Q000637:transplantation*; D036421:Nerve Fibers, Unmyelinated / Q000637:transplantation*; D051381:Rats; D016038:Skin Transplantation / Q000379:methods; D017768:Skin, Artificial; D023822:Tissue Engineering / Q000379:methods*
keywords: Human skin substitutes; Peripheral nerve regeneration; Tissue engineering