Wound healing in db/db mice with type 2 diabetes using non-contact exposure with an argon non-thermal atmospheric pressure plasma jet device

PLoS One. 2022 Oct 14;17(10):e0275602. doi: 10.1371/journal.pone.0275602. eCollection 2022.

Abstract

A non-thermal atmospheric pressure plasma jet (APPJ) may stimulate cells and tissues or result in cell death depending on the intensity of plasma at the target; therefore, we herein investigated the effects of non-thermal plasma under non-contact conditions on the healing of full-thickness wounds in diabetic mice (DM+ group) and normal mice (DM- group). A hydrogen peroxide colorimetric method and high performance liquid chromatography showed that APPJ produced low amounts of reactive oxygen and nitrogen species. Ten-week-old male C57BL/6j mice with normal blood glucose levels (DM- group) and 10-week-old male C57BLKS/J Iar-+Leprdb/+Leprdb mice (DM+ group) received two full-thickness cutaneous wounds (4 mm in diameter) on both sides of the dorsum. Wounds were treated with or without the plasma jet or argon gas for 1 minute and were then covered with a hydrocolloid dressing (Hydrocolloid), according to which mice were divided into the following groups: DM+Plasma, DM+Argon, DM+Hydrocolloid, DM-Plasma, DM-Argon, and DM-Hydrocolloid. Exudate weights, wound areas, and wound area ratios were recorded every day. Hematoxylin and eosin staining was performed to assess re-epithelialization and α-SMA immunohistological staining to evaluate the formation of new blood vessels. Non-thermal plasma under non-contact conditions reduced the production of exudate. Exudate weights were smaller in the DM+Plasma group than in the DM+Hydrocolloid and DM+Argon groups. The wound area ratio was smaller for plasma-treated wounds, and was also smaller in the DM+Plasma group than in the DM+Hydrocolloid and DM+Argon groups on days 1-21 (p<0.01). Wound areas were smaller in the DM-Plasma group than in the DM-Argon group until day 14 and differences were significant on days 1-5 (p<0.01). The percentage of re-epithelialization was significantly higher in the DM+Plasma group than in the DM+Argon and DM+Hydrocolloid groups (p<0.01). The number of new blood vessels that had formed by day 7 was significantly higher in the DM+Plasma group than in the DM+Hydrocolloid and DM+Argon groups (p<0.05). These results indicate that treatment with the current non-thermal plasma APPJ device under non-contact conditions accelerated wound healing in diabetic mice.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Argon
  • Blood Glucose
  • Colloids / pharmacology
  • Diabetes Mellitus, Experimental* / therapy
  • Diabetes Mellitus, Type 2* / therapy
  • Eosine Yellowish-(YS)
  • Hematoxylin
  • Hydrogen Peroxide
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Nitrogen
  • Oxygen
  • Plasma Gases* / pharmacology
  • Plasma Gases* / therapeutic use
  • Wound Healing

Substances

  • Blood Glucose
  • Colloids
  • Plasma Gases
  • Argon
  • Hydrogen Peroxide
  • Nitrogen
  • Oxygen
  • Eosine Yellowish-(YS)
  • Hematoxylin

Grants and funding

This work was supported by JSPS KAKENHI (Grant Number 18H03072 to TN and Grant Number 21H03219 to KM). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.