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Updated: Jun 2, 2026
Use of Frozen Tissue in the Comet Assay for the Evaluation of DNA Damage
Published on: March 24, 2020
Genotoxicity evaluation of dentine bonding agents by comet assay
1Faculty of Dentistry, Department of Conservative Dentistry and Endodontics, Gazi University, Ankara, Turkey.
Aim:
To evaluate the genotoxicity of four different adhesives, Clearfil SE Bond, SL Bond, i Bond and Clearfil Protect Bond and the primers of Clearfil SE Bond and Clearfil Protect Bond.
Methodology:
Genotoxicity assessment of the adhesives and primers was carried out in vitro in human lymphocytes at different elution concentrations, using the alkaline single-cell gel electrophoresis technique (comet assay). After the incubation of lymphocytes with varying volumes of the test agent, cells were embedded in a low-melting-point agarose suspension and then lysed in alkaline (pH>13) conditions. Electrophoresis was performed on the suspended lysed cells followed by visual analysis with staining of DNA. Fluorescence was than calculated to determine the extent of DNA damage using imaging software. Statistical comparison of the results was carried out by one-way analysis of variance (anova).
Results:
A significant increase (P<0.001) compared to untreated controls in DNA damage was observed with 'Clearfil Protect Bond' and 'Clearfil SE Bond' primer in human lymphocytes at concentrations of 2.5 and 5.0 mg mL(-1). Clearfil Protect Bond and Clearfil SE Bond adhesives induced significant (P < 0.001) DNA damage only at the higher concentration of 5.0 mg mL(-1) . No significant increase in DNA damage was observed with SL Bond and i Bond. No significant DNA damage was observed with any dentine bonding agents at the lower concentration of 1.25 mg mL(-1) .
Conclusions:
'Clearfil Protect Bond' and 'Clearfil SE Bond' primers/adhesives increased DNA damage in human peripheral lymphocytes in high doses.
