The Biosafety of Periodontal Ligament Fibroblasts Induced by Cold Atmospheric Plasma in vitro: Cytotoxicity, Genotoxicity and Mechanisms

IF 2.6 3区 物理与天体物理 Q3 ENGINEERING, CHEMICAL
Xiaojiao Zhang, Yinglong Li, Ruonan Ma, Zhitong Chen, Ruixue Wang, Zuomin Wang
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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).

Abstract Image

冷大气等离子体体外诱导牙周韧带成纤维细胞的生物安全性:细胞毒性、遗传毒性和机制
据报道,冷大气等离子体(CAP)是牙科领域一项很有前途的技术。在临床实践中,对牙周组织的生物安全性和机制尤为重要。本研究旨在评估 CAP 诱导的人牙周韧带成纤维细胞(hPDLFs)的细胞毒性、基因毒性和机制。使用菌落形成单位法(CFU)评估了对牙龈卟啉单胞菌(P. gingivalis)的抗菌效果。用 CAP 处理人类牙周韧带成纤维细胞的时间各不相同(1、2、4、8 和 16 分钟)。用 CCK-8 检测法检测细胞活力,用流式细胞仪测量细胞周期和细胞凋亡。细胞迁移能力通过划痕实验进行测定。评估了血浆活化培养基的理化性质,包括 pH 值、H2O2 和 NO 水平。使用流式细胞仪进行 DCF-DA 染色和分析,以评估细胞内的 ROS 水平。DNA双链断裂(DSBs)标记物磷酸化γH2AX的免疫荧光用于确定血浆的基因毒性。结果表明,6 分钟处理后,CAP 对牙龈脓胞有明显的抑制作用。它具有明显的剂量依赖性,从增加细胞增殖到诱导细胞凋亡。低剂量(1、2、4 和 8 分钟)可促进细胞增殖。高剂量(16 分钟)会抑制细胞增殖(p < 0.01)。研究还表明,这些影响主要是由于活性氧和氮物种(RONS)的形成。
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来源期刊
Plasma Chemistry and Plasma Processing
Plasma Chemistry and Plasma Processing 工程技术-工程:化工
CiteScore
5.90
自引率
8.30%
发文量
73
审稿时长
6-12 weeks
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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