Department of Pharmaceutical Biology, University of Greifswald, Greifswald, Germany.
Cell Biol Int. 2012;36(12):1217-22. doi: 10.1042/CBI20120139.
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.
非热大气压等离子体已经发展到将用于未来的几个目的，例如医学。等离子体处理的重点是活组织和细胞，例如，以改善伤口愈合，或诱导肿瘤细胞凋亡和生长停滞。需要详细研究等离子体-细胞相互作用。细胞表面黏附分子如整合素、钙黏蛋白或表皮生长因子受体（EGF 受体）在伤口愈合中很重要，也是癌症转移发展的关键。本研究集中于测量人角质形成细胞（人成纤维细胞低钙温度角质形成细胞）表面促进黏附、迁移和增殖的细胞表面分子，作为等离子体-细胞相互作用的一个重要特征。通过空气表面电介质阻挡放电对 HaCaT 角质形成细胞进行等离子体处理。用流式细胞术分析细胞表面分子和细胞内 ROS（活性氧）的诱导，在等离子体处理后 24 小时。除了细胞活力降低外，E-钙黏蛋白和 EGFR 表达也明显下调。α2-和β1-整合素的影响较小，ICAM-1（细胞间黏附分子 1）的表达不受影响。影响的程度取决于细胞暴露于等离子体的时间和处理方案。用荧光染料 H2DCFDA（2'，7'-二氯二氢荧光素二乙酸酯）检测到的细胞内 ROS 水平通过等离子体处理而增加，但它既不依赖于处理时间，也与不同的处理方案无关。二维培养的 HaCaT 角质形成细胞似乎是研究等离子体-细胞相互作用的一种合适方法。
