利用表没食子儿茶素没食子酸酯开发用于牙科的生物相容性银纳米粒子。

Developing biocompatible silver nanoparticles using epigallocatechin gallate for dental use.

机构信息

Faculty of Dentistry, The University of Hong Kong, Hong Kong, China.

School of Stomatology, Shenzhen University Health Science Center, Shenzhen, China.

出版信息

Arch Oral Biol. 2019 Jun;102:106-112. doi: 10.1016/j.archoralbio.2019.03.022. Epub 2019 Mar 28.

DOI:10.1016/j.archoralbio.2019.03.022

PMID:30999064

Abstract

OBJECTIVE

To develop silver nanoparticles (AgNPs) using epigallocatechin gallate (EGCG) and evaluate its biocompatibility and inhibition effect on Streptococcus mutans biofilm growth.

DESIGN

AgNPs were synthesized using EGCG as a reducing agent. Cytotoxicity was assessed using half-maximal inhibitory concentration (IC) against human gingival fibroblast (HGF-1) and stem cells from human exfoliated deciduous teeth (SHED). Antibacterial properties were evaluated with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against S. mutans. Dentine blocks were treated with AgNPs, silver nitrate (AgNO), or water before being incubated with S. mutans. The kinetics, morphology and viability of the biofilm at different time points were assessed by colony-forming units (CFUs), scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM), respectively. Lactic acid and polysaccharide production of the biofilm were also investigated.

RESULTS

Spherical AgNPs with diameter 17 ± 7 nm were developed. The IC of AgNPs and AgNO against HGF-1 were 44.88 ± 11.39 μg/mL and 11.53 ± 6.96 μg/mL, respectively (p < 0.001), whereas those against SHED were 68.02 ± 24.48 μg/mL and 9.54 ± 6.63 μg/mL, respectively (p = 0.02). The MIC of AgNPs and AgNO were 32.22 ± 7.34 μg/mL and 48.89 ± 15.11 μg/mL, respectively (p = 0.01), whereas their MBC was 63.33 ± 11.73 μg/mL and 85.00 ± 20.77 μg/mL, respectively (p = 0.02). Log CFUs of the AgNPs group were the lowest among the groups (p < 0.001). SEM and CLSM found a confluent biofilm in AgNO and water groups but not in AgNPs group. Biofilms in AgNPs group was revealed with lowest level of acidic acid and polysaccharides production (p < 0.001).

CONCLUSION

This study developed biocompatible AgNPs which inhibited the growth of a cariogenic biofilm.

摘要

目的

使用表没食子儿茶素没食子酸酯（EGCG）合成银纳米粒子（AgNPs），并评价其生物相容性及对变异链球菌生物膜生长的抑制作用。

设计

以 EGCG 为还原剂合成 AgNPs。通过对人牙龈成纤维细胞（HGF-1）和人脱落乳牙牙髓干细胞（SHED）的半数最大抑制浓度（IC）评估细胞毒性。采用最低抑菌浓度（MIC）和最低杀菌浓度（MBC）评估 AgNPs 对变异链球菌的抑菌效果。用 AgNPs、硝酸银（AgNO）或水对牙本质块进行处理，然后与变异链球菌孵育。采用菌落形成单位（CFUs）、扫描电子显微镜（SEM）和共聚焦激光扫描显微镜（CLSM）分别在不同时间点评估生物膜的动力学、形态和活力。还研究了生物膜的乳酸和多糖的产生。

结果

成功制备出直径为 17±7nm 的球形 AgNPs。AgNPs 和 AgNO 对 HGF-1 的 IC 分别为 44.88±11.39μg/mL 和 11.53±6.96μg/mL（p<0.001），而对 SHED 的 IC 分别为 68.02±24.48μg/mL 和 9.54±6.63μg/mL（p=0.02）。AgNPs 和 AgNO 的 MIC 分别为 32.22±7.34μg/mL 和 48.89±15.11μg/mL（p=0.01），MBC 分别为 63.33±11.73μg/mL 和 85.00±20.77μg/mL（p=0.02）。与其他组相比，AgNPs 组的 log CFUs 最低（p<0.001）。SEM 和 CLSM 发现，AgNO 和水组形成了融合生物膜，而 AgNPs 组则没有。AgNPs 组生物膜产生的酸性物质和多糖最少（p<0.001）。