title: Characterization of a novel standardized human three-dimensional skin wound healing model using non-sequential fractional ultrapulsed CO2 laser treatments.
abstract: OBJECTIVE
  At present, there is no standardized in vitro human skin model for wound healing. Therefore, our aim was to establish and characterize an in vitro/ex vivo three-dimensional (3D) wound healing model, which we employed to analyze the effects of dexpanthenol on wound healing and gene regulation.
  
  
  METHODS
  The novel human 3D skin wound healing model using scaffold and collagen 3D organotypic skin equivalents was irradiated with a non-sequential fractional ultrapulsed CO2 laser. These standardized injured full-thickness skin equivalents enable qRT-PCR, microarray, and histological studies analyzing the effect of topically or systemically applied compounds on skin wound healing.
  
  
  RESULTS
  These human laser-irradiated skin models were found to be appropriate for in vitro wound healing analysis. Topical treatment of skin wounds with a 5% dexpanthenol water-in-oil emulsion or two different 5% dexpanthenol oil-in-water emulsions clearly enhanced wound closure compared to laser-irradiated untreated control models. To find out whether this positive effect is caused by the active substance dexpanthenol, laser-irradiated skin models were cultured in calciumpantothenate containing medium (20 μg/ml) compared to skin equivalents cultured without calciumpantothenate. 3D models cultured in calciumpantothenate revealed considerably faster wound closure compared to the control models. Quantitative RT-PCR studies showed enhanced mRNA expression of MMP3, IL1α, keratin-associated protein 4-12 (KRTAP4-12), and decreased expression of S100A7 in laser-irradiated skin models cultured in medium containing calciumpantothenate.
  
  
  CONCLUSIONS
  This novel standardized human 3D skin wound healing model proves useful for topical pharmacological studies on wound healing and reveals new insights into molecular mechanisms of dexpanthenol-mediated effects on wound healing. In addition, these novel 3D model systems can be used to monitor ex vivo effects of various laser systems on gene expression and morphology of human skin.
authors.affiliation: Department of Dermatology and Allergology, RWTH Aachen University, Aachen, Germany.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Marquardt
authors.forename: Philipp M
authors.initials: PM
authors.lastname: Amann
authors.forename: Ruth
authors.initials: R
authors.lastname: Heise
authors.forename: Katharina
authors.initials: K
authors.lastname: Czaja
authors.forename: Timm
authors.initials: T
authors.lastname: Steiner
authors.forename: Hans F
authors.initials: HF
authors.lastname: Merk
authors.forename: Claudia
authors.initials: C
authors.lastname: Skazik-Voogt
authors.forename: Jens M
authors.initials: JM
authors.lastname: Baron
mesh_terms: D002478:Cells, Cultured; D062109:Dermatologic Surgical Procedures / Q000379:methods; D006801:Humans; D054020:Lasers, Gas / Q000627:therapeutic use*; D008954:Models, Biological; D010205:Pantothenic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D014945:Wound Healing / Q000187:drug effects* / Q000235:genetics
chemical_list: D010205:Pantothenic Acid; C007288:dexpanthenol
keywords: Bepanthen; calciumpantothenate; full-thickness organotypic skin equivalents; human skin reconstitution model; pharmacogical skin model