Biocompatibility and Cytotoxicity of Pulp-Capping Materials on DPSCs, With Marker mRNA Expressions

Banu Çiçek Tez; Bahar Başak Kızıltan Eliaçık; Pakize Neslihan Taşlı; Hazal Yılmaz; Fikrettin Şahin

doi:10.1016/j.identj.2024.04.006

Biocompatibility and Cytotoxicity of Pulp-Capping Materials on DPSCs, With Marker mRNA Expressions

Int Dent J. 2024 Apr 30:S0020-6539(24)00113-8. doi: 10.1016/j.identj.2024.04.006. Online ahead of print.

Authors

Banu Çiçek Tez¹, Bahar Başak Kızıltan Eliaçık², Pakize Neslihan Taşlı³, Hazal Yılmaz³, Fikrettin Şahin³

Affiliations

¹ Department of Pediatric Dentistry, Faculty of Dentistry, Ankara Medipol University, Ankara, Türkiye.
² Department of Pediatric Dentistry, Faculty of Dentistry, University of Health Sciences, Istanbul, Türkiye.
³ Department of Genetics and Bioengineering, Faculty of Engineering and Architecture, Yeditepe University, Istanbul, Türkiye.

PMID: 38692961
DOI: 10.1016/j.identj.2024.04.006

Abstract

Objectives: The present study aimed to (1) investigate biocompatibility and cytotoxicity of pulp-capping materials on viability of human dental pulp stem cells (hDPSCs); (2) determine angiogenic, odontogenic, and osteogenic marker mRNA expressions; and (3) observe changes in surface morphology of the hDPSCs using scanning electron microscopy (SEM).

Methods: Impacted third molars were used to isolate the hDPSCs, which were treated with extract-release fluids of the pulp-capping materials (Harvard BioCal-Cap, NeoPUTTY MTA, TheraCal LC, and Dycal). Effects of the capping materials on cell viability were assessed using 3-(4,5-di-methyl-thiazol-2-yl)-5-(3-carboxy-methoxy-phenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium (MTS) assay and the apoptotic/necrotic cell ratios and reactive oxygen species (ROS) levels from flow cytometry. Marker expressions (alkaline phosphatase [ALP], osteocalcin [OCN], collagen type I alpha 1 [Col1A], secreted protein acidic and rich in cysteine [SPARC], osteonectin [ON], and vascular endothelial growth factor [VEGF]) were determined by quantitative reverse-transcription polymerase chain reaction. Changes in surface morphology of the hDPSCs were visualised by SEM.

Results: The MTS assay results at days 1, 3, 5, and 7 indicated that Harvard BioCal-Cap, NeoPUTTY MTA, and TheraCal LC did not adversely affect cell viability when compared with the control group. According to the MTS assay results at day 14, no significant difference was found amongst Dycal, Harvard BioCal-Cap, NeoPUTTY MTA, and TheraCal LC affecting cell viability. Dycal was the only capping material that increased ROS level. High levels of VEGF expression were observed with Harvard BioCal-Cap, TheraCal LC, and NeoPUTTY MTA. NeoPUTTY MTA, and Dycal upregulated OCN expression, whereas TheraCal LC upregulated Col1A and SPARC expression. Only Dycal increased ALP expression. HDSCs were visualized in characteristic spindle morphology on SEM when treated with TheraCal LC and Harvard BioCal-Cap.

Conclusions: NeoPUTTY MTA and Harvard BioCal-Cap showed suitable biocompatibility values; in particular, these pulp-capping materials were observed to support the angiogenic marker.

Keywords: Biocompatibility; Capping materials; Cytotoxicity; Dental pulp stem cells; mRNA expression.