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光固化时间对应用于永生化成牙本质细胞系的修复性树脂复合材料细胞毒性的影响。

Effects of light-curing time on the cytotoxicity of a restorative resin composite applied to an immortalized odontoblast-cell line.

作者信息

de Souza Costa Carlos Alberto, Hebling Josimeri, Hanks Carl Thomas

机构信息

University of São Paulo State/UNESP, School of Dentistry, Araraquara, Brazil.

出版信息

Oper Dent. 2003 Jul-Aug;28(4):365-70.

Abstract

This in vitro study evaluated the cytotoxic effects of a restorative resin composite applied to an immortalized odontoblast-cell line (MDPC-23). Seventy-two round resin discs (2-mm thick and 4 mm in diameter) were light-cured for 20 or 40 seconds and rinsed, or not, with PBS and culture medium. The resin discs were divided into four experimental groups: Group 1: Z-100/20 seconds; Group 2: Z-100/20 seconds/rinsed; Group 3: Z-100/40 seconds; Group 4: Z-100/40 seconds/rinsed. Circular filter paper was used as a control material (Group 5). The round resin discs and filter papers were placed in the bottom of wells of four 24-well dishes (18 wells for each experimental and control group). MDPC-23 cells (30,000 cells/cm2) were plated in the wells and allowed to incubate for 72 hours. The zone of inhibition around the resin discs was measured under inverted light microscopy; the MTT assay was carried out for mitochondrial respiration and cell morphology was measured under SEM. The scores obtained from inhibition zone and MTT assay were analyzed with the Kruskal-Wallis followed by Dunnett tests. In Groups 1, 2, 3 and 4, the thickness of the inhibition zone was 1,593 +/- 12.82 microm, 403 +/- 15.49 microm, 1,516 +/- 9.81 microm and 313 +/- 13.56 microm, respectively. There was statistically significant difference among the experimental and control groups at the 0.05 level of significance. The MTT assay demonstrated that the resin discs of the experimental groups 1, 2, 3 and 4 reduced the cell metabolism by 83%, 40.1%, 75.5% and 24.5%. Only between the Groups 2 and 4 was there no statistically significant difference for mitochondrial respiration. Close to the resin discs, the MDPC-23 cells exhibited rounded shapes, with only a few cellular processes keeping the cells attached to the substrate or, even disruption of plasma membrane. Adjacent to the inhibition zone, the cultured cells exhibited multiple fine cellular processes on the cytoplasmic membrane organized in epithelioid nodules, similar to the morphology observed to the control group. Based on the results, the authors may conclude that the Z-100 resin composite light cured for 20 seconds was more cytopathic to MDPC-23 cells than Z-100 light cured for 40 seconds. The cytotoxic effects of the resin discs decreased after rinsing them with PBS and culture medium. This was confirmed by MTT assay and upon evaluation of the inhibition zone, which was narrower following rinsing of the resin discs.

摘要

这项体外研究评估了一种修复性树脂复合材料应用于永生化成牙本质细胞系(MDPC - 23)的细胞毒性作用。将72个圆形树脂盘(2毫米厚,直径4毫米)光固化20秒或40秒,并用或不用磷酸盐缓冲液(PBS)和培养基冲洗。树脂盘分为四个实验组:第1组:Z - 100/20秒;第2组:Z - 100/20秒/冲洗;第3组:Z - 100/40秒;第4组:Z - 100/40秒/冲洗。圆形滤纸用作对照材料(第5组)。将圆形树脂盘和滤纸放置在四个24孔培养板孔的底部(每个实验组和对照组18个孔)。将MDPC - 23细胞(30,000个细胞/平方厘米)接种到孔中并孵育72小时。在倒置光学显微镜下测量树脂盘周围的抑制区;进行MTT试验以检测线粒体呼吸,并在扫描电子显微镜下测量细胞形态。从抑制区和MTT试验获得的分数用Kruskal - Wallis检验,随后进行Dunnett检验分析。在第1、2、3和4组中,抑制区的厚度分别为1,593±12.82微米、403±15.49微米、1,516±9.81微米和313±13.56微米。在0.05显著性水平下,实验组和对照组之间存在统计学显著差异。MTT试验表明,第1、2、3和4实验组的树脂盘使细胞代谢分别降低了83%、40.1%、75.5%和24.5%。仅第2组和第4组之间线粒体呼吸没有统计学显著差异。靠近树脂盘处,MDPC - 23细胞呈现圆形,只有少数细胞突起使细胞附着于基质,甚至细胞膜破裂。在抑制区附近,培养的细胞在细胞质膜上呈现多个精细的细胞突起,组织成上皮样结节,类似于对照组观察到的形态。基于这些结果,作者可以得出结论,光固化20秒的Z - 100树脂复合材料对MDPC - 23细胞的细胞病变作用比光固化40秒的Z - 100更强。用PBS和培养基冲洗树脂盘后,其细胞毒性作用降低。MTT试验以及对抑制区的评估证实了这一点,树脂盘冲洗后抑制区变窄。

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