Ozone mediators effect on "in vitro" scratch wound closure.

Area of Science:

• Biomedical Science
• Wound Healing Research
• Oxidative Stress Mechanisms

Background:

• Low-dose ozone therapy shows benefits in wound healing, primarily due to bactericidal and pro-oxidant effects.
• Ozone's effects are mediated by oxidative molecules like 4-hydroxynonenal (HNE) and hydrogen peroxide (H2O2), formed from reactions with biological systems.
• The precise cellular mechanisms behind ozone's positive impact on wound closure remain largely uninvestigated.

Purpose of the Study:

• To investigate the effects of varying non-toxic doses of ozonated saline on an in vitro wound scratch model using human keratinocytes.
• To identify the key molecular mediators responsible for the wound healing benefits of ozonated saline.

Main Methods:

• Utilized an in vitro wound scratch model with human keratinocytes.
• Administered ozonated saline at concentrations from 2 to 300 μM.
• Assessed cell proliferation using BrdU assay and PCNA protein levels.
• Investigated the roles of HNE and H2O2, individually and in combination, to replicate ozonated saline effects.
• Analyzed Nrf2 pathway activation via nuclear translocation and HO1 gene expression.

Main Results:

• Ozonated saline significantly improved in vitro wound healing by enhancing keratinocyte proliferation.
• Neither HNE nor H2O2 alone significantly improved wound closure, but their combination demonstrated a positive effect.
• Nrf2 pathway activation was observed, evidenced by nuclear translocation of Nrf2 and increased HO1 gene expression.

Conclusions:

• Ozonated saline effectively promotes wound closure in vitro by stimulating keratinocyte proliferation.
• The beneficial effects of ozonated saline on wound healing are attributed to a combination of molecules, with HNE and H2O2 playing crucial roles.
• Activation of the Nrf2 pathway is involved in the cellular response to ozonated saline treatment.