Niazi Fayez Hussain, Luddin Norhayati, Niazy Abdurahman, Mohamad Suharni, Harun Masitah Hayati, Noushad Mohammed, Ponnuraj Kannan Thirumulu
School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia.
Department of Restorative and Prosthetic Dentistry, College of Dentistry, Dar Al Uloom University, Riyadh 13314, Saudi Arabia.
Saudi Dent J. 2024 Nov;36(11):1432-1437. doi: 10.1016/j.sdentj.2024.08.009. Epub 2024 Aug 20.
This research assessed the mutagenicity and DNA damage of a novel type of nano-hydroxyapatite-silica glass ionomer cement (nano-HA-SiO-GIC) and a conventional GIC (cGIC) using Ames and Comet assays.
Cell viability was tested on human periodontal ligament fibroblasts (HPLFs) using 3.125 mg/ml, 6.25, 12.5, 25, 50, 100 and 200 mg/ml, on both types of GICs employing MTT assay. For the Comet assay, HPLFs were treated with IC, IC and IC of test materials and the tail moments were measured. In the Ames test, four genotypic variants of strains of (TA100, TA98, TA1537 and TA1535) and a strain of (WP2 uvrA) were employed. The material tested was extracted using sterile distilled water (0.2 g per ml) at 37 °C for 72 h. This was considered as 100 %, which was diluted to 50, 25, 12.5 and 6.25 % utilizing sterile distilled water. These five concentrations were incubated with the bacterial strains with and without metabolic activation (S9), along with appropriate positive controls. The number of revertant colonies was used to evaluate the outcome.
The highest cell viability (159.4 %) for nano-HA-SiO-GIC was noticed at 3.125 mg/ml, while the lowest (24.26 %) was observed at 200 mg per ml. IC, IC and IC values were 95.27, 51.4 and 20.1 mg/ml for cGIC, 106.9, 55.8 and 22.9 mg/ml for nano-HA-SiO-GIC, respectively. The IC of both test materials showed no significant DNA damage compared to that of the negative control based on the Comet assay. The plate treated with nano-HA-SiO-GIC showed less than double the average number of revertant colonies compared to that of negative control with regard to the Ames test.
It can be concluded that nano-HA-SiO-GIC is non-mutagenic based on the Ames test and did not cause DNA damage at the lowest concentration of IC based on the Comet assay.
本研究使用艾姆斯试验和彗星试验评估了一种新型纳米羟基磷灰石-二氧化硅玻璃离子水门汀(nano-HA-SiO-GIC)和传统玻璃离子水门汀(cGIC)的致突变性和DNA损伤情况。
使用MTT法,以3.125mg/ml、6.25mg/ml、12.5mg/ml、25mg/ml、50mg/ml、100mg/ml和200mg/ml的浓度检测两种类型玻璃离子水门汀对人牙周膜成纤维细胞(HPLFs)的细胞活力。对于彗星试验,用受试材料的半数抑制浓度(IC)、IC和IC处理HPLFs,并测量尾矩。在艾姆斯试验中,使用了四种基因型的鼠伤寒沙门氏菌菌株(TA100、TA98、TA1537和TA1535)和一种大肠杆菌菌株(WP2 uvrA)。受试材料在37℃下用无菌蒸馏水(每毫升0.2g)提取72小时。这被视为100%,然后用无菌蒸馏水将其稀释至50%、25%、12.5%和6.25%。这五个浓度与有或没有代谢激活剂(S9)的细菌菌株以及适当的阳性对照一起孵育。用回复突变菌落数来评估结果。
nano-HA-SiO-GIC在3.125mg/ml时细胞活力最高(159.4%),而在200mg/ml时最低(24.26%)。cGIC的半数抑制浓度(IC)、IC和IC值分别为95.27mg/ml、51.4mg/ml和20.1mg/ml,nano-HA-SiO-GIC的分别为106.9mg/ml、55.8mg/ml和22.9mg/ml。基于彗星试验,与阴性对照相比,两种受试材料的半数抑制浓度均未显示出明显的DNA损伤。在艾姆斯试验中,与阴性对照相比,用nano-HA-SiO-GIC处理的平板上回复突变菌落的平均数不到两倍。
基于艾姆斯试验可以得出结论,nano-HA-SiO-GIC无致突变性,并且基于彗星试验,在最低半数抑制浓度时不会引起DNA损伤。