Direct biological effects of fractional ultrapulsed CO2 laser irradiation on keratinocytes and fibroblasts in human organotypic full-thickness 3D skin models

  • L. Schmitt
  • S. Huth
  • P. M. Amann
  • Y. Marquardt
  • R. Heise
  • K. Fietkau
  • L. Huth
  • T. Steiner
  • F. Hölzle
  • J.M. Baron
Original Article

Abstract

Molecular effects of various ablative and non-ablative laser treatments on human skin cells—especially primary effects on epidermal keratinocytes and dermal fibroblasts—are not yet fully understood. We present the first study addressing molecular effects of fractional non-sequential ultrapulsed CO2 laser treatment using a 3D skin model that allows standardized investigations of time-dependent molecular changes ex vivo. While histological examination was performed to assess morphological changes, we utilized gene expression profiling using microarray and qRT-PCR analyses to identify molecular effects of laser treatment. Irradiated models exhibited dose-dependent morphological changes resulting in an almost complete recovery of the epidermis 5 days after irradiation. On day 5 after laser injury with a laser fluence of 100 mJ/cm2, gene array analysis identified an upregulation of genes associated with tissue remodeling and wound healing (e.g., COL12A1 and FGF7), genes that are involved in the immune response (e.g., CXCL12 and CCL8) as well as members of the heat shock protein family (e.g., HSPB3). On the other hand, we detected a downregulation of matrix metalloproteinases (e.g., MMP3), differentiation markers (e.g., LOR and S100A7), and the pro-inflammatory cytokine IL1α.

Overall, our findings substantiate the understanding of time-dependent molecular changes after CO2 laser treatment. The utilized 3D skin model system proved to be a reliable, accurate, and reproducible tool to explore the effects of various laser settings both on skin morphology and gene expression during wound healing.

Keywords

Fractional CO2 laser Organotypic skin equivalents Wound healing Gene regulation Biological effects Scar treatment Acne scars Skin rejuvenation 

Notes

Compliance with ethical standards

Conflict of interest

None.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional ethics committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Supplementary material

10103_2017_2409_MOESM1_ESM.pdf (878 kb)
Fig. S1 Gene expression in laser-irradiated 3D skin models (microarray analysis) that were cultured in nutrient deficient medium. 3D skin models were harvested 5 days after laser treatment and gene expression was measured using the Affymetrix® Gene Chip Human Exon 2.0 ST array. (PDF 877 kb)
10103_2017_2409_MOESM2_ESM.pdf (125 kb)
Fig. S2 Time dependent changes measured by TaqMan real-time PCR analysis on day 3 and 5 (models were cultured in nutrient deficient medium). (PDF 124 kb)

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Copyright information

© Springer-Verlag London Ltd., part of Springer Nature 2017

Authors and Affiliations

  • L. Schmitt
    • 1
  • S. Huth
    • 1
  • P. M. Amann
    • 1
  • Y. Marquardt
    • 1
  • R. Heise
    • 1
  • K. Fietkau
    • 1
  • L. Huth
    • 1
  • T. Steiner
    • 2
    • 3
  • F. Hölzle
    • 2
    • 3
  • J.M. Baron
    • 1
    • 3
  1. 1.Department of Dermatology and Allergology, Medical FacultyRWTH Aachen UniversityAachenGermany
  2. 2.Department of Oral and Maxillofacial Surgery, Medical FacultyRWTH Aachen UniversityAachenGermany
  3. 3.Interdisciplinary Center for Laser Medicine, Medical FacultyRWTH Aachen UniversityAachenGermany

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