Surface molecules on HaCaT keratinocytes after interaction with non‐thermal atmospheric pressure plasma

Abstract

Non-thermal atmospheric-pressure plasmas have been developed that will be used in future for several purposes, e.g. medicine. Living tissues and cells are at the focus of plasma treatment, e.g. to improve wound healing, or induce apoptosis and growth arrest in tumour cells. Detailed investigations of plasma-cell interactions are needed. Cell surface adhesion molecules as integrins, cadherins or the EGFR (epidermal growth factor receptor) are of importance in wound healing and also for development of cancer metastasis. This study has focused on measurement of cell surface molecules on human HaCaT keratinocytes (human adult low calcium temperature keratinocytes) promoting adhesion, migration and proliferation as one important feature of plasma-cell interactions. HaCaT keratinocytes were treated with plasma by a surface dielectric barrier discharge in air. Cell surface molecules and induction of intracellular ROS (reactive oxygen species) were analysed by flow cytometry 24 h after plasma treatment. Besides a reduction of cell viability a significant down-regulation of E-cadherin and the EGFR expression occurred. The influence on α2- and β1-integrins was less pronounced, and expression of ICAM-1 (intercellular adhesion molecule 1) was unaffected. The extent of effects depended on the exposure time of cells to the plasma and the treatment regimen. Intracellular level of ROS detected by the fluorescent dye H2DCFDA (2',7'-dichlorodihydrofluorescein diacetate) increased by plasma treatment, but it was neither dependent on the treatment time nor related to the different treatment regimens. Two-dimensional cultures of HaCaT keratinocytes appear to be a suitable method of investigating plasma-cell interactions.

Differential Influence of Components Resulting from Atmospheric-Pressure Plasma on Integrin Expression of Human HaCaT Keratinocytes

“…Similar results for detachment of adherent cells or impaired adhesion after the treatment of different cell types with different plasma sources were reported by several investigators [20–22, 35, 36]. Viability of treated cells strongly depended on the plasma source, the treatment time/plasma dose, the working gas (e.g., air versus argon), or treatment regimen [23]. …”

Controlling plasma stimulated media in cancer treatment application

“…However, recently, the CAP stimulated media were found to be able to kill cancer cells as well as the direct CAP treatment did. 3,4 In contrast to conventional direct CAP treatment, which is more like a surgical strategy, using the CAP stimulated media to kill cancer cells is promising to be a pharmaceutics method. Glioblastoma is the most lethal brain cancer.…”

“…Despite the CAP stimulated media has been proved to be effective as well as the direct CAP treatment, 3 media was negligible, we just regulated the concentration of FBS in media. We performed CAP treatments on 100 ll of media in 96-well plate with distinct FBS volume concentration (0%, 10%, 20%, and 30%).…”

Aqueous Plasma Pharmacy: Preparation Methods, Chemistry, and Therapeutic Applications

“…Earlier Leibniz work, also with HaCaT cells, indicated that direct cell treatment in the presence of medium to reduce cell viability was accompanied by a significant downregulation of E-cadherin and epidermal growth factor receptor (EGFR), with a lesser impact on α 2 - and β 1 -integrins, and no impact on intercellular adhesion molecule 1 (ICAM-1). 190 Overall, this work informs the impact of plasma treatment on cell-surface adhesion molecules that are critical to wound healing.…”