Limits...
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

Detection of apoptotic cells in CAP and placebo treated murine skin tissue.Apoptotic cells were analyzed using a fluorometric TUNEL assay system for detection of fragmented DNA as indicator of apoptotic events. Representative photomicrographs are presented of placebo and CAP treated skin. Arrowheads exemplarily indicate apoptotic cells. Number of TUNEL positive cells were counted in three different fields and averaged. Placebo treatment was set to 100% ± SD. Magnification as indicated.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4359157&req=5

pone.0120041.g002: Detection of apoptotic cells in CAP and placebo treated murine skin tissue.Apoptotic cells were analyzed using a fluorometric TUNEL assay system for detection of fragmented DNA as indicator of apoptotic events. Representative photomicrographs are presented of placebo and CAP treated skin. Arrowheads exemplarily indicate apoptotic cells. Number of TUNEL positive cells were counted in three different fields and averaged. Placebo treatment was set to 100% ± SD. Magnification as indicated.

Mentions: To analyze whether apoptosis is affected by CAP in keratinocytes, we used a human apoptosis array which detects 35 apoptosis-related proteins and searched for candidates differentially expressed 24 h after CAP treatment for 2 min. In analogy to our result obtained for fibroblasts [20] the evaluation of the array showed that these pro-apoptotic or anti-apoptotic factors were not influenced by the CAP treatment in keratinocytes (S1 Table). Additionally, we analyzed the skin of the treated mice for the appearance of apoptotic cells. Here, we observed that the number of apoptotic cells in the treated skin area did not differ between 5x CAP or 5x placebo treatment for 2 min (Fig. 2a, b). These results suggest that apoptotic mechanisms are not activated in keratinocytes by the CAP treatment under used treatment conditions.


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)

Detection of apoptotic cells in CAP and placebo treated murine skin tissue.Apoptotic cells were analyzed using a fluorometric TUNEL assay system for detection of fragmented DNA as indicator of apoptotic events. Representative photomicrographs are presented of placebo and CAP treated skin. Arrowheads exemplarily indicate apoptotic cells. Number of TUNEL positive cells were counted in three different fields and averaged. Placebo treatment was set to 100% ± SD. Magnification as indicated.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0120041.g002: Detection of apoptotic cells in CAP and placebo treated murine skin tissue.Apoptotic cells were analyzed using a fluorometric TUNEL assay system for detection of fragmented DNA as indicator of apoptotic events. Representative photomicrographs are presented of placebo and CAP treated skin. Arrowheads exemplarily indicate apoptotic cells. Number of TUNEL positive cells were counted in three different fields and averaged. Placebo treatment was set to 100% ± SD. Magnification as indicated.
Mentions: To analyze whether apoptosis is affected by CAP in keratinocytes, we used a human apoptosis array which detects 35 apoptosis-related proteins and searched for candidates differentially expressed 24 h after CAP treatment for 2 min. In analogy to our result obtained for fibroblasts [20] the evaluation of the array showed that these pro-apoptotic or anti-apoptotic factors were not influenced by the CAP treatment in keratinocytes (S1 Table). Additionally, we analyzed the skin of the treated mice for the appearance of apoptotic cells. Here, we observed that the number of apoptotic cells in the treated skin area did not differ between 5x CAP or 5x placebo treatment for 2 min (Fig. 2a, b). These results suggest that apoptotic mechanisms are not activated in keratinocytes by the CAP treatment under used treatment conditions.

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