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Article Dans Une Revue Journal of Investigative Dermatology Année : 2023

765 Cold atmospheric plasma-activated medium promotes wound healing by stimulation of human keratinocyte migration

Résumé

Cold Atmospheric Plasma (CAP) is an emerging technology that can generates a combination of Reactive Oxygen and Nitrogen Species (RONS) at low temperature, during interaction with liquid. CAP has been shown to improve skin wound healingin vivo, but the cell types and biological mechanisms involved are not well known. Our study aims to assess a possible effect on human keratinocytes of treatments with Plasma-Activated Medium (PAM). Human N/TERT-1 and primary keratinocytes in 2D culture were treated during 1h with calcium-containing PBS previously exposed to CAP for 10 to 120s. Burned human skin explants used as a 3D wound healing model were similarly treated. PBS exposed to helium gas alone was the control condition. We compared the keratinocyte viability (MTT and TUNEL assays), proliferation (cell counts and Ki67 expression) and differentiation (capacity to form 3D reconstructed epidermis). We also evaluated putative oxidative stress and mitochondrial dysfunctions. Effect on cell migration was assessed using both a 2D scratch assay and the 3D model. Generated RONS were quantified, and their involvement was investigated using acetylcysteine. Treatments of keratinocytes with PBS activated during 60s or more were cytotoxic, reduced keratinocyte migration, and induced oxidative stress. On the other side, treatments with PBS activated during 10-30s were not cytotoxic, did not affect keratinocyte proliferation and differentiation, did not induce any changes in mitochondria, but accelerated scratch closure by improving keratinocyte migration. Similarly, accelerated re-epithelialization (p < 0.005) was observed in the skin explant model. Whereas, micromolar concentrations of RONS were produced during the exposure of PBS to plasma, the PAM effects appeared to be independent of RONS. These results show that PAM improves wound healing through stimulation of keratinocyte migration, suggesting a novel therapeutic method for recalcitrant wounds.

Dates et versions

hal-04094669 , version 1 (11-05-2023)

Identifiants

Citer

A. Marches, S. Cousty, Nofel Merbahi, M. Simon. 765 Cold atmospheric plasma-activated medium promotes wound healing by stimulation of human keratinocyte migration. Journal of Investigative Dermatology, 2023, 143 (5), pp.S131. ⟨10.1016/j.jid.2023.03.774⟩. ⟨hal-04094669⟩
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