Comprehensive molecular characterization of microneedling therapy in a human three-dimensional skin model

Abstract

Background and objectives: Microneedling therapy is a widely used technique in dermatology. However, little is known about the underlying molecular effects of this therapy on extracellular matrix remodeling, wound healing, and inflammation. The aim of this study was to examine morphological and molecular changes caused by microneedling treatment in a standardized in vitro full-thickness 3D model of human skin.

Materials and methods: A microneedling device was used to treat full-thickness 3D skin models. Specimens were harvested at specified time points and qRT-PCR and microarray studies were performed. Frozen sections were examined histologically.

Results: Microneedling treatment caused morphological changes in the skin model resulting in an almost complete recovery of the epidermis five days after treatment. Microarray analysis identified an upregulation of genes that are associated with tissue remodeling and wound healing (e.g. COL3A1, COL8A1, TIMP3), epithelial proliferation and differentiation (KRT13, IGF1), immune cell recruitment (CCL11), and a member of the heat shock protein family (HSPB6). On the other hand, we detected a downregulation of pro-inflammatory cytokines (e.g. IL1α, IL1β, IL24, IL36γ, IL36RN), and antimicrobial peptides (e.g. S100A7A, DEFB4). These data were confirmed by independent RT-PCR analyses.

Conclusion: We present for the first time the direct molecular effects of microneedling therapy on epidermal keratinocytes and dermal fibroblasts using a standardized 3D skin model. Treatment resulted in histological alterations and changed the expression of various genes related to epidermal differentiation, inflammation, and dermal remodeling. This data suggests that skin microneedling plays a role in dermal remodeling, increases epidermal differentiation, and might also have a direct effect on collagen synthesis. These findings may increase our understanding of the molecular mechanisms of human skin repair induced by microneedling therapy and will allow comparisons with competing applications, such as ablative laser therapies.

Fig 1. Representative hematoxylin and eosin stained sections of 3D skin models zero and five days after microneedling treatment.

Untreated models are shown as controls. Magnification = 100 x, scale bar = 100 μm.

Fig 2. Gene expression profiling in microneedling-treated 3D skin models (microarray analysis).

(A) 3D skin models were harvested five days after microneedling treatment. Gene expression was measured using the Affymetrix Gene Chip Human Exon 2.0 ST array. (B) Gene ontology (GO) analysis of microarray results.

Fig 3. TaqMan real-time PCR analysis of four independent approaches displaying the expression of selected genes five days after microneedling treatment.Ctrl = control.

*p < 0.05, **p < 0.01.