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In vitro assessment of Neuronal PAS domain 2 mitigating compounds for scarless wound healing
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In vitro assessment of Neuronal PAS domain 2 mitigating compounds for scarless wound healing
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Journal Article
In vitro assessment of Neuronal PAS domain 2 mitigating compounds for scarless wound healing
2023
Overview
The core circadian gene Neuronal PAS domain 2 (
) is expressed in dermal fibroblasts and has been shown to play a critical role in regulating collagen synthesis during wound healing. We have performed high throughput drug screening to identify genes responsible for downregulation of
while maintaining cell viability. From this, five FDA-approved hit compounds were shown to suppress
expression in fibroblasts. In this study, we hypothesize that the therapeutic suppression of
by hit compounds will have two effects: (1) attenuated excessive collagen deposition and (2) accelerated dermal wound healing without hypertrophic scarring.
To test the effects of each hit compound (named Dwn1, 2, 3, 4, and 5), primary adult human dermal fibroblasts (HDFa) were treated with either 0, 0.1, 1, or 10 μM of a single hit compound. HDFa behaviors were assessed by picrosirius red staining and quantitative RT-PCR for
collagen synthesis, cell viability assay,
fibroblast-to-myofibroblast differentiation test, and cell migration assays.
Dwn1 and Dwn2 were found to significantly affect collagen synthesis and cell migration without any cytotoxicity. Dwn3, Dwn4, and Dwn5 did not affect collagen synthesis and were thereby eliminated from further consideration for their role in mitigation of gene expression or myofibroblast differentiation. Dwn1 also attenuated myofibroblast differentiation on HDFa.
Dwn1 and Dwn2 may serve as possible therapeutic agents for future studies related to skin wound healing.
Publisher
Frontiers Media SA,Frontiers Media S.A
Subject
