体外评估电子烟气溶胶、大麻烟雾和传统香烟烟雾对牙龈成纤维细胞/牙龈间充质干细胞特性的影响。
In vitro evaluation of the effect of the electronic cigarette aerosol, Cannabis smoke, and conventional cigarette smoke on the properties of gingival fibroblasts/gingival mesenchymal stem cells.
机构信息
Oral Medicine and Periodontology Department, Faculty of Dentistry, Cairo University, Cairo, Egypt.
Department of Surgery and Oral Medicine, National Research Centre, Cairo, Egypt.
出版信息
J Periodontal Res. 2022 Jan;57(1):104-114. doi: 10.1111/jre.12943. Epub 2021 Nov 8.
OBJECTIVE
The current study aimed to evaluate the effect of electronic cigarette (EC) aerosol, Cannabis, and conventional cigarettes smoke on gingival fibroblast/gingival mesenchymal stem cells' (GF/G-MSCs) of never smokers.
MATERIAL AND METHODS
Human GF/G-MSCs (n = 32) were isolated and characterized using light microscopy, flow cytometry, and multilineage differentiation ability. Following the application of aerosol/smoke extracts, GF/G-MSCs were evaluated for cellular proliferation; colony-forming units (CFU-F) ability; cellular viability (using the MTT assay); mitochondrial depolarization using JC-1 dye; and genes' expression of ATM, p21, Oct4, and Nanog.
RESULTS
Colony-forming units and viability (OD 450 nm) were significantly reduced upon exposure to Cannabis (mean ± SD; 5.5 ± 1.5; p < .00001, 0.47 ± 0.21; p < .05) and cigarettes smoke (2.3 ± 1.2 p < .00001, 0.59 ± 0.13, p < .05), while EC aerosol showed no significant reduction (10.8 ± 2.5; p = .05, 1.27 ± 0.47; p > .05) compared to the control group (14.3 ± 3, 1.33 ± 0.12). Significantly upregulated expression of ATM, Oct4, and Nanog (gene copies/GADPH) was noticed with Cannabis (1.5 ± 0.42, 0.82 ± 0.44, and 1.54 ± 0.52, respectively) and cigarettes smoke (1.52 ± 0.75, 0.7 ± 0.14, and 1.48 ± 0.79, respectively; p < .05), whereas EC aerosol caused no statistically significant upregulation of these genes compared to the control group (0.63 ± 0.1, 0.31 ± 0.12, and 0.64 ± 0.46, respectively; p > .05). The p21 gene was not significantly downregulated in EC aerosol (1.22 ± 0.46), Cannabis (0.71 ± 0.24), and cigarettes smokes (0.83 ± 0.54) compared to the control group (p = .053, analysis of variance).
CONCLUSION
Cannabis and cigarettes smoke induce DNA damage and cellular dedifferentiation and negatively affect the cellular proliferation and viability of GF/G-MSCs of never smokers, whereas EC aerosol showed a significantly lower impact on these properties.
目的
本研究旨在评估电子烟（EC）气溶胶、大麻和传统香烟烟雾对从不吸烟人群牙龈成纤维细胞/牙龈间充质干细胞（GF/G-MSCs）的影响。
材料与方法
使用光学显微镜、流式细胞术和多能分化能力对人 GF/G-MSCs（n=32）进行分离和鉴定。在应用气溶胶/烟雾提取物后，通过细胞增殖；集落形成单位（CFU-F）能力；细胞活力（MTT 检测）；使用 JC-1 染料检测线粒体去极化；以及 ATM、p21、Oct4 和 Nanog 基因的表达来评估 GF/G-MSCs。
结果
与对照组相比，暴露于大麻（平均±SD；5.5±1.5；p<0.00001，0.47±0.21；p<0.05）和香烟烟雾（2.3±1.2 p<0.00001，0.59±0.13，p<0.05）显著降低了 CFU-F 形成和活力（OD 450nm），而 EC 气溶胶则没有明显降低（10.8±2.5；p=0.05，1.27±0.47；p>0.05）。与对照组相比，大麻（基因拷贝数/GAPDH 分别为 1.5±0.42、0.82±0.44 和 1.54±0.52）和香烟烟雾（1.52±0.75、0.7±0.14 和 1.48±0.79；p<0.05）显著上调了 ATM、Oct4 和 Nanog 的表达，而 EC 气溶胶与对照组相比，这些基因的表达没有统计学意义的上调（0.63±0.1、0.31±0.12 和 0.64±0.46；p>0.05）。与对照组相比，p21 基因在 EC 气溶胶（1.22±0.46）、大麻（0.71±0.24）和香烟烟雾（0.83±0.54）中没有显著下调（p=0.053，方差分析）。
结论
大麻和香烟烟雾会导致 DNA 损伤和细胞去分化，并对从不吸烟人群的 GF/G-MSCs 的细胞增殖和活力产生负面影响，而 EC 气溶胶对这些特性的影响则明显较低。
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