采用微流控牙芯片和牙龈等效物对银氨氟化氢的细胞毒性进行表征。

Characterization of silver diamine fluoride cytotoxicity using microfluidic tooth-on-a-chip and gingival equivalents.

机构信息

Faculty of Dentistry, National University of Singapore, Singapore.

出版信息

Dent Mater. 2022 Aug;38(8):1385-1394. doi: 10.1016/j.dental.2022.06.025. Epub 2022 Jun 29.

DOI:10.1016/j.dental.2022.06.025

PMID:35778310

Abstract

OBJECTIVE

This study aims to characterize the cytotoxicity potential of silver diamine fluoride (SDF) on dental pulp stem cells (DPSC) and gingival equivalents.

METHODS

DPSC cultured on 96-well plates was exposed directly to SDF (0.0001-0.01%) and cell viability (IC) quantified. Effect of SDF on DPSC viability under flow (with dentin barrier) conditions was evaluated using a custom-designed microfluidic "tooth-on-a-chip". Permeability of dentin discs (0.5-1.5 mm thickness) was evaluated using lucifer yellow permeation assay. Dentin discs were treated with 38% SDF (up to 3 h), and cell viability (live/dead assay) of the DPSC cultured in the inlet (unexposed) and outlet (exposed) regions of the pulp channel was evaluated. To assess the mucosal corrosion potential, gingival equivalents were treated with 38% SDF for 3 or 60 min (OECD test guideline 431) and characterized by MTT assay and histomorphometric analysis.

RESULTS

DPSC exposed directly to SDF showed a dose-dependent reduction in cell viability (IC: 0.001%). Inlet channels (internal control) of the tooth-on-a-chip exposed to PBS and SDF-exposed dentin discs showed> 85% DPSC viability. In contrast, the outlet channels of SDF-exposed dentin discs showed a decreased viability of< 31% and 0% (1.5 and ≤1.0 mm thick dentin disc, respectively) (p < 0.01). The gingiva equivalents treated with SDF for 3 and 60 min demonstrated decreased epithelial integrity, loss of intercellular cohesion and corneal layer detachment with significant reduction in intact epithelial thickness (p < 0.05).

SIGNIFICANCE

SDF penetrated the dentin (≤1 mm thick) inducing significant death of the pulp cells. SDF also disrupted gingival epithelial integrity resulting in mucosal corrosion.

摘要

目的

本研究旨在表征银胺氟化物（SDF）对牙髓干细胞（DPSC）和牙龈等同物的细胞毒性潜力。

方法

将 DPSC 培养在 96 孔板上，直接暴露于 SDF（0.0001-0.01%）并定量细胞活力（IC）。使用定制的微流控“牙片上的牙”评估 SDF 在流动（有牙本质屏障）条件下对 DPSC 活力的影响。使用荧光素黄渗透测定法评估 0.5-1.5mm 厚牙本质盘的渗透性。用 38%的 SDF 处理牙本质盘（长达 3 小时），并评估培养在牙髓通道入口（未暴露）和出口（暴露）区域的 DPSC 的细胞活力（死活测定）。为了评估粘膜腐蚀性潜力，用 38%的 SDF 处理牙龈等同物 3 或 60 分钟（OECD 测试指南 431），并用 MTT 测定和组织形态计量分析进行表征。

结果

直接暴露于 SDF 的 DPSC 显示细胞活力呈剂量依赖性降低（IC：0.001%）。牙片上的微流控入口通道（内部对照）暴露于 PBS 和 SDF 暴露的牙本质盘显示>85%的 DPSC 活力。相比之下，SDF 暴露的牙本质盘的出口通道显示活力降低<31%和 0%（分别为 1.5 和≤1.0mm 厚牙本质盘）（p<0.01）。用 SDF 处理 3 和 60 分钟的牙龈等同物显示上皮完整性降低，细胞间连接丧失和角膜层脱落，完整上皮厚度显著减少（p<0.05）。