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Updated: Mar 9, 2026

Accessing the Cytotoxicity and Cell Response to Biomaterials
Published on: July 8, 2021
In vitro dentin barrier cytotoxicity testing of some dental restorative materials
1Dental Medical Devices Testing Center, Dental Materials Laboratory, Peking University School and Hospital of Stomatology, National Engineering Laboratory for Digital and Material Technology of Stomatology, Beijing Key Laboratory of Digital Stomatology, Beijing 100081, China.
Objectives:
To investigate the cytotoxicity of four dental restorative materials in three-dimensional (3D) L929 cell cultures using a dentin barrier test.
Methods:
The cytotoxicities of light-cured glass ionomer cement (Vitrebond), total-etching adhesive (GLUMA Bond5), and two self-etching adhesives (GLUMA Self Etch and Single Bond Universal) were evaluated. The permeabilities of human dentin disks with thicknesses of 300, 500, and 1000μm were standardized using a hydraulic device. Test materials and controls were applied to the occlusal side of human dentin disks. The 3D-cell scaffolds were placed beneath the dentin disks. After a 24-h contact with the dentin barrier test device, cell viabilities were measured by performing MTT assays. Statistical analysis was performed using the Mann-Whitney U test.
Results:
The mean (SD) permeabilities of the 300-μm, 500-μm, and 1000-μm dentin disks were 0.626 (0.214), 0.219 (0.0387) and 0.089 (0.028) μlmin-1cm-2cm H2O-1. Vitrebond was severely cytotoxic, reducing the cell viability to 10% (300-μm disk), 17% (500μm), and 18% (1000μm). GLUMA Bond5 reduced the cell viability to 40% (300μm), 83% (500μm), and 86% (1000μm), showing moderate cytotoxicity (300-μm) and non-cytotoxicity (500-μm and 1000-μm). Single Bond Universal and GLUMA Self Etch did not significantly reduce cell viability, regardless of the dentin thicknesses, which characterized them as non-cytotoxic.
Conclusions:
Cytotoxicity varied with the materials tested and the thicknesses of the dentin disks.
Clinical Significance:
The tested cytotoxicity of materials applied on 300-, 500-, and 1000-μm dentin disks indicates that the clinical use of the test materials (excepting self-etching adhesives) in deep cavities poses a potential risk of damage to the pulp tissues to an extent, depending on the thickness of the remaining dentin.
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