{"title":"The Biosafety of Periodontal Ligament Fibroblasts Induced by Cold Atmospheric Plasma in vitro: Cytotoxicity, Genotoxicity and Mechanisms","authors":"Xiaojiao Zhang, Yinglong Li, Ruonan Ma, Zhitong Chen, Ruixue Wang, Zuomin Wang","doi":"10.1007/s11090-024-10482-8","DOIUrl":null,"url":null,"abstract":"<div><p>Cold atmospheric plasma (CAP) has been reported as a promising technique in dentistry. The biosafety and mechanisms on periodontal tissue are especially important in clinical practice. This study aims to evaluate the cytotoxicity, genotoxicity and mechanisms of human periodontal ligament fibroblasts (hPDLFs) induced by CAP. The antimicrobial effect on <i>Porphyromonas gingivalis (P. gingivalis)</i> was evaluated using the colony-forming unit methods (CFU). Human periodontal ligament fibroblasts were treated with CAP for variable times (1, 2, 4, 8, and 16 min). CCK-8 assays were performed to detect cell viability and flow cytometry was performed to measure the cell cycle and apoptosis. Cell migration ability was determined by scratch assays. The physicochemical properties of plasma activated medium were evaluated, including the pH values, H<sub>2</sub>O<sub>2</sub> and NO levels. DCF-DA staining and analysis were performed to evaluate intracellular ROS levels inside cells using flow cytometry. Immunofluorescence of DNA double-strand breaks (DSBs) marker, phosphorylated γH2AX was used to establish the genotoxicity of plasma. The results showed that CAP has a significant inhibition effect of <i>P. gingivalis</i> after 6 min treatment. It has significantly dose-dependent effects that ranging from increasing cell proliferation to inducing apoptosis. A low doses (1, 2, 4, and 8 min) could enhance the cell proliferation. A high dose (16 min) resulted in the inhibition of cell proliferation (<i>p</i> < 0.01). It is also shown that these effects are primarily due to the formation of reactive oxygen and nitrogen species (RONS).</p></div>","PeriodicalId":734,"journal":{"name":"Plasma Chemistry and Plasma Processing","volume":"44 4","pages":"1635 - 1653"},"PeriodicalIF":2.6000,"publicationDate":"2024-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Plasma Chemistry and Plasma Processing","FirstCategoryId":"5","ListUrlMain":"https://link.springer.com/article/10.1007/s11090-024-10482-8","RegionNum":3,"RegionCategory":"物理与天体物理","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"ENGINEERING, CHEMICAL","Score":null,"Total":0}
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Abstract
Cold atmospheric plasma (CAP) has been reported as a promising technique in dentistry. The biosafety and mechanisms on periodontal tissue are especially important in clinical practice. This study aims to evaluate the cytotoxicity, genotoxicity and mechanisms of human periodontal ligament fibroblasts (hPDLFs) induced by CAP. The antimicrobial effect on Porphyromonas gingivalis (P. gingivalis) was evaluated using the colony-forming unit methods (CFU). Human periodontal ligament fibroblasts were treated with CAP for variable times (1, 2, 4, 8, and 16 min). CCK-8 assays were performed to detect cell viability and flow cytometry was performed to measure the cell cycle and apoptosis. Cell migration ability was determined by scratch assays. The physicochemical properties of plasma activated medium were evaluated, including the pH values, H2O2 and NO levels. DCF-DA staining and analysis were performed to evaluate intracellular ROS levels inside cells using flow cytometry. Immunofluorescence of DNA double-strand breaks (DSBs) marker, phosphorylated γH2AX was used to establish the genotoxicity of plasma. The results showed that CAP has a significant inhibition effect of P. gingivalis after 6 min treatment. It has significantly dose-dependent effects that ranging from increasing cell proliferation to inducing apoptosis. A low doses (1, 2, 4, and 8 min) could enhance the cell proliferation. A high dose (16 min) resulted in the inhibition of cell proliferation (p < 0.01). It is also shown that these effects are primarily due to the formation of reactive oxygen and nitrogen species (RONS).
期刊介绍：
Publishing original papers on fundamental and applied research in plasma chemistry and plasma processing, the scope of this journal includes processing plasmas ranging from non-thermal plasmas to thermal plasmas, and fundamental plasma studies as well as studies of specific plasma applications. Such applications include but are not limited to plasma catalysis, environmental processing including treatment of liquids and gases, biological applications of plasmas including plasma medicine and agriculture, surface modification and deposition, powder and nanostructure synthesis, energy applications including plasma combustion and reforming, resource recovery, coupling of plasmas and electrochemistry, and plasma etching. Studies of chemical kinetics in plasmas, and the interactions of plasmas with surfaces are also solicited. It is essential that submissions include substantial consideration of the role of the plasma, for example, the relevant plasma chemistry, plasma physics or plasma–surface interactions; manuscripts that consider solely the properties of materials or substances processed using a plasma are not within the journal’s scope.
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