Cardoso Miguel, Coelho Ana, Marto Carlos Miguel, Gonçalves Ana Cristina, Paula Anabela, Ribeiro Ana Bela Sarmento, Ferreira Manuel Marques, Botelho Maria Filomena, Laranjo Mafalda, Carrilho Eunice
Institute of Integrated Clinical Practice, Faculty of Medicine, University of Coimbra, 3000-075 Coimbra, Portugal.
Institute of Biophysics, Faculty of Medicine, University of Coimbra, 3000-548 Coimbra, Portugal.
Materials (Basel). 2021 Oct 27;14(21):6435. doi: 10.3390/ma14216435.
This study aimed to assess the cytotoxicity of commercially available adhesive strategies-etch-and-rinse (Adper Scotchbond 1 XT, 3M ESPE, St. Paul, MN, USA, SB1), self-etch (Clearfil SE Bond 2, Kuraray Noritake Dental Inc., Tokyo, Japan, CSE), and universal (Scotchbond Universal, 3M Deutschland GmbH, Neuss, Germany, SBU). MDPC-23 cells were exposed to adhesives extracts in different concentrations and exposure times. To access cell metabolic activity, viability, types of cell death, and cell cycle, the MTT assay, SRB assay, double labeling with annexin V and propidium iodide, and labeling with propidium iodide/RNAse were performed, respectively. Cultures were stained with May-Grünwald Giemsa for qualitative cytotoxicity assessment. The SB1, CSE, and SBU extracts determined a significant reduction in cell metabolism and viability. This reduction was higher for prolonged exposures, even for less concentrated extracts. CSE extracts significantly reduced the cell's metabolic activity at higher concentrations (50% and 100%) from 2 h of exposure. After 24 and 96 h, a metabolic activity reduction was verified for all adhesives, even at lower concentrations. These changes were dependent on the adhesive, its concentration, and the incubation time. Regarding cell viability, SBU extracts were the least cytotoxic, and CSE was significantly more cytotoxic than SB1 and SBU. The adhesives determined a reduction in viable cells and an increase in apoptotic, late apoptosis/necrosis, and necrotic cells. Moreover, on cultures exposed to SB1 and CSE extracts, a decrease in the cells in S and G2/M phases and an increase in the cells in G0/G1 phase was observed. Exposure to SBU led to an increase of cells in the S phase. In general, all adhesives determined cytotoxicity. CSE extracts were the most cytotoxic and were classified as having a higher degree of reactivity, leading to more significant inhibition of cell growth and destruction of the cell's layers.
本研究旨在评估市售粘结策略——酸蚀冲洗粘结剂（Adper Scotchbond 1 XT，美国明尼苏达州圣保罗3M ESPE公司，SB1）、自酸蚀粘结剂（Clearfil SE Bond 2，日本东京可乐丽诺瑞特牙科公司，CSE）和通用粘结剂（Scotchbond Universal，德国诺伊斯3M德国有限公司，SBU）的细胞毒性。将MDPC - 23细胞暴露于不同浓度和暴露时间的粘结剂提取物中。为了评估细胞代谢活性、活力、细胞死亡类型和细胞周期，分别进行了MTT法、SRB法、 annexin V和碘化丙啶双标记以及碘化丙啶/核糖核酸酶标记。用May - Grünwald Giemsa染色对培养物进行定性细胞毒性评估。SB1、CSE和SBU提取物均导致细胞代谢和活力显著降低。即使是浓度较低的提取物，长时间暴露后这种降低也更明显。CSE提取物在较高浓度（50%和100%）下，从暴露2小时起就显著降低了细胞的代谢活性。24小时和96小时后，即使在较低浓度下，所有粘结剂的代谢活性均有所降低。这些变化取决于粘结剂、其浓度和孵育时间。关于细胞活力，SBU提取物的细胞毒性最小，CSE的细胞毒性明显高于SB1和SBU。粘结剂导致活细胞减少，凋亡、晚期凋亡/坏死和坏死细胞增加。此外，在暴露于SB1和CSE提取物的培养物中，观察到S期和G2/M期细胞减少，G0/G1期细胞增加。暴露于SBU导致S期细胞增加。总体而言，所有粘结剂均具有细胞毒性。CSE提取物的细胞毒性最大，被归类为具有较高的反应性，导致对细胞生长的抑制更显著，细胞层破坏更严重。
