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Effects of cold atmospheric plasma (CAP) on ß-defensins, inflammatory cytokines, and apoptosis-related molecules in keratinocytes in vitro and in vivo.

Arndt S, Landthaler M, Zimmermann JL, Unger P, Wacker E, Shimizu T, Li YF, Morfill GE, Bosserhoff AK, Karrer S - PLoS ONE (2015)

Bottom Line: In summary, our results suggest that a 2 min application of CAP induces gene expression of key regulators important for inflammation and wound healing without causing proliferation, migration or cell death in keratinocytes.The induction of ß-defensins in keratinocytes describes an absolutely new plasma strategy.Activation of antimicrobial peptides supports the well-known antibacterial effect of CAP treatment, whereas the mechanism of ß-defensin activation by CAP is not investigated so far.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology, University Regensburg, D-93042 Regensburg, Germany.

ABSTRACT
Cold atmospheric plasma (CAP) has been gaining increasing interest as a new approach for the treatment of skin diseases or wounds. Although this approach has demonstrated promising antibacterial activity, its exact mechanism of action remains unclear. This study explored in vitro and in vivo whether CAP influences gene expression and molecular mechanisms in keratinocytes. Our results revealed that a 2 min CAP treatment using the MicroPlaSter ß in analogy to the performed clinical studies for wound treatment induces expression of IL-8, TGF-ß1, and TGF-ß2. In vitro and in vivo assays indicated that keratinocyte proliferation, migration, and apoptotic mechanisms were not affected by the CAP treatment under the applied conditions. Further, we observed that antimicrobial peptides of the ß-defensin family are upregulated after CAP treatment. In summary, our results suggest that a 2 min application of CAP induces gene expression of key regulators important for inflammation and wound healing without causing proliferation, migration or cell death in keratinocytes. The induction of ß-defensins in keratinocytes describes an absolutely new plasma strategy. Activation of antimicrobial peptides supports the well-known antibacterial effect of CAP treatment, whereas the mechanism of ß-defensin activation by CAP is not investigated so far.

No MeSH data available.


Related in: MedlinePlus

Expression of IL-8, TGF-ß1, and TGF-ß2 after the CAP treatment.(a, b, c) mRNA expression analysis of IL-8, TGF-ß1, and TGF-ß2 in human keratinocytes was performed 6 h, 24 h, 48 h, and 72 h after CAP treatment for 2 min by LightCycler 1.2 technology. (d, e, f) IL-8 secretion by human keratinocytes was analyzed 24 h, 48 h, and 72 h after CAP treatment for 2 min by FlowCytomix and TGF-ß1, and TGF-ß2 secretion by ELISA Technology (g) mRNA expression of murine TGF-ß1 (mTGF-ß1) and murine TGF-ß2 (mTGF-ß2) in epidermal skin was analyzed after 5x CAP treatment for 2 min and was compared to corresponding placebo control. *p < 0.05.
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pone.0120041.g001: Expression of IL-8, TGF-ß1, and TGF-ß2 after the CAP treatment.(a, b, c) mRNA expression analysis of IL-8, TGF-ß1, and TGF-ß2 in human keratinocytes was performed 6 h, 24 h, 48 h, and 72 h after CAP treatment for 2 min by LightCycler 1.2 technology. (d, e, f) IL-8 secretion by human keratinocytes was analyzed 24 h, 48 h, and 72 h after CAP treatment for 2 min by FlowCytomix and TGF-ß1, and TGF-ß2 secretion by ELISA Technology (g) mRNA expression of murine TGF-ß1 (mTGF-ß1) and murine TGF-ß2 (mTGF-ß2) in epidermal skin was analyzed after 5x CAP treatment for 2 min and was compared to corresponding placebo control. *p < 0.05.

Mentions: For analysis, keratinocytes were exposed to CAP for 2 min or remained untreated and were further incubated for 24 h before supernatants were collected and processed for the simultaneous detection of the relative expression levels of 36 human cytokines and acute phase proteins. Surprisingly, we observed that only interleukin 8 (IL-8) out of 36 analyzed cytokines was significantly up-regulated by the CAP treatment for 2 min (fold change (CAP/ctr.) 3.01; p value 0.0467). Array results concerning IL-8 induction were verified on mRNA level at different time points (6 h, 24 h, 48 h, and 72 h) after the CAP treatment (Fig. 1a) and on protein level determined by FlowCytomix Technology 24 h, 48 h, and 72 h after exposure (Fig. 1d). FlowCytomix results confirmed a significant IL-8 induction 24 h after CAP treatment. On mRNA level a significant induction was obtained after 48 h and 72 h, whereas a non-significant induction was already seen 24 h after CAP treatment. A dual effect of plasma could lead to a fast secretion of preformed IL-8 protein within 24 h on one hand and also to a production of new IL-8 on the other hand, which could explain the later IL-8 mRNA levels.


Effects of cold atmospheric plasma (CAP) on ß-defensins, inflammatory cytokines, and apoptosis-related molecules in keratinocytes in vitro and in vivo.

Arndt S, Landthaler M, Zimmermann JL, Unger P, Wacker E, Shimizu T, Li YF, Morfill GE, Bosserhoff AK, Karrer S - PLoS ONE (2015)

Expression of IL-8, TGF-ß1, and TGF-ß2 after the CAP treatment.(a, b, c) mRNA expression analysis of IL-8, TGF-ß1, and TGF-ß2 in human keratinocytes was performed 6 h, 24 h, 48 h, and 72 h after CAP treatment for 2 min by LightCycler 1.2 technology. (d, e, f) IL-8 secretion by human keratinocytes was analyzed 24 h, 48 h, and 72 h after CAP treatment for 2 min by FlowCytomix and TGF-ß1, and TGF-ß2 secretion by ELISA Technology (g) mRNA expression of murine TGF-ß1 (mTGF-ß1) and murine TGF-ß2 (mTGF-ß2) in epidermal skin was analyzed after 5x CAP treatment for 2 min and was compared to corresponding placebo control. *p < 0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4359157&req=5

pone.0120041.g001: Expression of IL-8, TGF-ß1, and TGF-ß2 after the CAP treatment.(a, b, c) mRNA expression analysis of IL-8, TGF-ß1, and TGF-ß2 in human keratinocytes was performed 6 h, 24 h, 48 h, and 72 h after CAP treatment for 2 min by LightCycler 1.2 technology. (d, e, f) IL-8 secretion by human keratinocytes was analyzed 24 h, 48 h, and 72 h after CAP treatment for 2 min by FlowCytomix and TGF-ß1, and TGF-ß2 secretion by ELISA Technology (g) mRNA expression of murine TGF-ß1 (mTGF-ß1) and murine TGF-ß2 (mTGF-ß2) in epidermal skin was analyzed after 5x CAP treatment for 2 min and was compared to corresponding placebo control. *p < 0.05.
Mentions: For analysis, keratinocytes were exposed to CAP for 2 min or remained untreated and were further incubated for 24 h before supernatants were collected and processed for the simultaneous detection of the relative expression levels of 36 human cytokines and acute phase proteins. Surprisingly, we observed that only interleukin 8 (IL-8) out of 36 analyzed cytokines was significantly up-regulated by the CAP treatment for 2 min (fold change (CAP/ctr.) 3.01; p value 0.0467). Array results concerning IL-8 induction were verified on mRNA level at different time points (6 h, 24 h, 48 h, and 72 h) after the CAP treatment (Fig. 1a) and on protein level determined by FlowCytomix Technology 24 h, 48 h, and 72 h after exposure (Fig. 1d). FlowCytomix results confirmed a significant IL-8 induction 24 h after CAP treatment. On mRNA level a significant induction was obtained after 48 h and 72 h, whereas a non-significant induction was already seen 24 h after CAP treatment. A dual effect of plasma could lead to a fast secretion of preformed IL-8 protein within 24 h on one hand and also to a production of new IL-8 on the other hand, which could explain the later IL-8 mRNA levels.

Bottom Line: In summary, our results suggest that a 2 min application of CAP induces gene expression of key regulators important for inflammation and wound healing without causing proliferation, migration or cell death in keratinocytes.The induction of ß-defensins in keratinocytes describes an absolutely new plasma strategy.Activation of antimicrobial peptides supports the well-known antibacterial effect of CAP treatment, whereas the mechanism of ß-defensin activation by CAP is not investigated so far.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology, University Regensburg, D-93042 Regensburg, Germany.

ABSTRACT
Cold atmospheric plasma (CAP) has been gaining increasing interest as a new approach for the treatment of skin diseases or wounds. Although this approach has demonstrated promising antibacterial activity, its exact mechanism of action remains unclear. This study explored in vitro and in vivo whether CAP influences gene expression and molecular mechanisms in keratinocytes. Our results revealed that a 2 min CAP treatment using the MicroPlaSter ß in analogy to the performed clinical studies for wound treatment induces expression of IL-8, TGF-ß1, and TGF-ß2. In vitro and in vivo assays indicated that keratinocyte proliferation, migration, and apoptotic mechanisms were not affected by the CAP treatment under the applied conditions. Further, we observed that antimicrobial peptides of the ß-defensin family are upregulated after CAP treatment. In summary, our results suggest that a 2 min application of CAP induces gene expression of key regulators important for inflammation and wound healing without causing proliferation, migration or cell death in keratinocytes. The induction of ß-defensins in keratinocytes describes an absolutely new plasma strategy. Activation of antimicrobial peptides supports the well-known antibacterial effect of CAP treatment, whereas the mechanism of ß-defensin activation by CAP is not investigated so far.

No MeSH data available.


Related in: MedlinePlus