CAP is a room-temperature partially ionized gas that generates reactive oxygen and nitrogen species (RONS), ultraviolet radiation, and electric fields. These components allow CAP to regulate cell viability, proliferation, migration, and apoptosis—making it widely applicable in biomedical fields like antimicrobial therapy, wound healing, and cancer treatment.

However, the specific mechanisms by which CAP interacts with distinct cell types (e.g., normal somatic cells, cancer cells) and its regulatory effects on key signaling pathways remain incompletely understood. Clarifying these cell-level mechanisms is essential to translating CAP technology from basic research to clinical applications.

This study aims to systematically explore CAP’s biological effects on skin-related target cells and elucidate its underlying molecular mechanisms. It focuses on verifying CAP’s regulatory roles in cell proliferation, migration, and the expression of inflammatory/regulatory factors—providing experimental support for CAP’s use in skin disease treatment.

2. Experimental Methods

（1）Cell Culture: Human skin-derived target cells (e.g., keratinocytes) were cultured in DMEM medium supplemented with fetal bovine serum and antibiotics, maintained in a 5% CO₂ incubator at 37°C.

（2）CAP Treatment Protocol: A cold atmospheric plasma device (e.g., argon plasma jet) was used for treatment (standardized duration/dose for the experimental group), with an untreated group as the control.

（3）Detection Techniques:

• Molecular expression: qPCR (to measure mRNA levels of IL-8, TGF-β1, TGF-β2, HBD-2, MBD-2/3) and Western blot (to quantify corresponding protein levels).
• Cell proliferation: XTT assay (assessed at 24/48/72 h).
• Cell migration: Scratch wound assay (migration rate evaluated at 7 h, 12 h).
• In vivo validation: Mouse models were established, followed by qPCR detection of target factor mRNA levels.

3. Primary Results

4. Key Conclusions

（1）CAP can significantly upregulate the expression of regulatory factors (IL-8, TGF-β1/β2, HBD-2, MBD-2/3) in skin-related cells (at the mRNA or protein level), which may underpin CAP’s regulatory effects on cell function.

（2）Within the experimental treatment parameters, CAP does not alter the proliferation or migration of target cells—indicating it modulates factor expression without compromising basic cell viability (within a safe dose range).

（3）CAP’s regulatory effects on factor expression are consistent in both in vitro cell experiments and in vivo mouse models, confirming the reliability of the findings.

5. Research Significance

（1）Theoretical Significance: This study clarifies CAP’s regulatory effects on key factor expression in skin-related cells, expanding the theoretical framework of CAP-cell interactions.

（2）Experimental Basis: It provides specific experimental data (factor expression changes, cell function metrics) for CAP’s use in skin-related research, serving as a reference for future experimental design.

（3）Clinical Translation Value: By demonstrating that CAP modulates skin-related regulatory factor expression (without impairing basic cell viability), it lays groundwork for CAP’s clinical application in skin disease treatment (e.g., wound healing, inflammatory skin conditions).

References

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[3] Arndt S, Unger P, Berneburg M, Bosserhoff AK, Karrer S. Cold atmospheric plasma (CAP) activates angiogenesis-related molecules in skin keratinocytes, fibroblasts and endothelial cells and improves wound angiogenesis in an autocrine and paracrine mode. J Dermatol Sci. 2018;89:181–90.

[4] Hasse S, Duong Tran T, Hahn O, Kindler S, Metelmann HR, von Woedtke T, Masur K. Induction of proliferation of basal epidermal keratinocytes by cold atmospheric-pressure plasma. Clin Exp Dermatol. 2016;41:202–9. 

[5] Arndt S, Landthaler M, Zimmermann JL, Unger P, Wacker E, Shimizu T, Karrer S. Effects of cold atmospheric plasma (CAP) on ß-defensins, inflammatory cytokines, and apoptosis-related molecules in keratinocytes in vitro and in vivo. PLoS ONE. 2015;10(8): e0120041.

[6] Lou BS, Hsieh JH, Chen CM, Hou CW, Wu HY, Chou PY, Lai CH, Lee JW. Helium/argon-generated cold atmospheric plasma facilitates cutaneous wound healing. Front Bioeng Biotechnol. 2020;8:683

[7] Barton A, Wende K, Bundscherer L, Hasse S, Schmidt A, Bekeschus S, Masur K. Nonthermal plasma increases expression of wound healing related genes in a keratinocyte cell line. Plasma Med. 2013;3(3):287–98.