Abstract
Ablative fractional laser treatment leads to a loss of matrix metalloproteinase-3 (MMP-3) expression; therefore, in the present in vitro study, we addressed the role of MMP-3 and its regulation by calcium pantothenate in wound healing processes at the molecular level. Utilizing confocal laser microscopy, we investigated MMP-3 protein expression in fractional ablative CO2 laser-irradiated skin models. In addition, we established full-thickness 3D skin models using fibroblasts and keratinocytes with a MMP-3 knockdown that were irradiated with a fractional ablative Er:YAG laser to set superficial injuries with standardized dimensions and minimal thermal damage to the surrounding tissue. We revealed an upregulation of MMP-3 protein expression in laser-irradiated skin models receiving aftercare treatment with calcium pantothenate. Skin models with MMP-3 knockdown exhibited a slower wound closure after laser treatment compared to controls. Gene expression profiling detected an MMP-3 knockdown-dependent upregulation of cytokines and chemokines (e.g. IL-36B, CXCL17, IL-37, CXCL5), antimicrobial peptides (e.g., S100A7, S100A12), epidermal crosslinking enzymes (TGM5), and differentiation markers (e.g., LOR, KRT1, FLG2). We also detected a downregulation of cathepsin V and MMP-10, both of which play a prominent role in wound healing processes. After fractional ablative laser injury, an aftercare treatment with calcium pantothenate accelerated wound closure in MMP-3 expressing models faster than in MMP-3 knockdown models. Our data substantiate a major role of MMP-3 in wound healing processes after ablative laser treatments. For the first time, we could show that calcium pantothenate exerts its wound healing-promoting effects at least partly via MMP-3.
Highlights
Following injury, the process of wound healing is broadly classified into three different phases, namely inflammation, proliferation, and remodeling [1]
We detected that the addition of calcium pantothenate or dexpanthenol to laser-injured 3D skin models resulted in a significantly faster wound closure compared to control models, which was associated with the increased expression of matrix metalloproteinase-3 (MMP-3) [9]
matrix metalloproteinases (MMPs)-3 appears to play an important role in wound healing, our previous data showed a downregulation of MMP-3 after laser irradiation of 3D skin models that could be rescued by calcium pantothenate treatment [8, 9]
Summary
The process of wound healing is broadly classified into three different phases, namely inflammation, proliferation, and remodeling [1]. We detected that the addition of calcium pantothenate or dexpanthenol to laser-injured 3D skin models resulted in a significantly faster wound closure compared to control models, which was associated with the increased expression of MMP-3 [9].
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