Low concentration zinc oxide nanoparticles enrichment enhances bacterial and pro-inflammatory resistance of calcium silicate-based cements

Calcium silicate-based cement (CSC) is a commonly used material in endodontic treatment. However, it has limited antibacterial activity, especially for cases involving primary infections. Zinc oxide nanoparticles (ZnONPs) are recognized for their potential in biomedical applications due to their antibacterial properties and ability to reduce inflammation. This study aims to optimize CSC by incorporating ZnO-NPs to maintain its physical properties, enhance its antibacterial activity, and reduce the production of pro-inflammatory cytokines. ZnO-NPs were integrated into a commercial CSC (Endocem MTA) at 1 wt% (CSZ1) or 3 wt% (CSZ3). Setting time, compressive strength, and X-ray diffraction were then measured. In addition, pH, calcium ion release, and zinc ion release were measured for 7 days. Antibacterial activity against Enterococcus faecalis and viability of murine macrophages (RAW264.7) were determined using colorimetric assays. Gene expression levels of proinflammatory cytokines in lipopolysaccharide induced RAW264.7 were evaluated using quantitative polymerase chain reaction. Results were compared to an unmodified CSC group. In the CSZ3 group, there was a significant increase of approximately 12% in setting time and a reduction of about 36.4% in compressive strength compared to the control and CSZ1 groups. The presence of ZnO-NPs was detected in both CSZ1 and CSZ3. Both CSC and CSZ1 groups maintained an alkaline pH and released calcium ions, while zinc ions were significantly released in the CSZ1 group. Additionally, CSZ1 showed a 1.8-fold reduction of bacterial activity and exhibited around 85% reduction in colony-forming units compared to the CSC group. Furthermore, the CSZ1 group showed a more than 39% reduction in pro-inflammatory cytokine levels compared to the CSC group. Thus, enriching CSC with 1 wt% ZnO-NPs can enhance its antibacterial activity and reduce pro-inflammatory cytokines without showing any tangible adverse effects on its physical properties.