Intranuclear cell uptake and toxicity of titanium dioxide and zirconia particles as well as bacterial adhesion on dental titanium- and zirconia-implants

Abstract

Objective: Previous studies have shown that particles can be released from dental titanium (Ti)- and zirconia (ZrO₂)-implants. Titanium dioxide (TiO₂)- and ZrO₂-particles were compared regarding their toxicity and intranuclear cell uptake as well as the adhesion of various anaerobic bacteria on Ti- and ZrO₂-implants.

Methods: Cyto- and genotoxicity of TiO₂-microparticles (TiO₂-MPs) and TiO₂-nanoparticles (TiO₂-NPs) in periodontal ligament (PDL)-hTERT cells were determined with XTT test and DNA damage with comet assay. Particle sizes of TiO₂- and ZrO₂-particles were measured with scanning electron microscope. Intranuclear uptake in PDL-hTERT cells was determined with laser scanning confocal microscopy. Adhesions of relevant anaerobic mouth bacteria Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans on Ti- and ZrO₂-implants were investigated by cultivation and counting bacterial colonies.

Results: Particle size measurements revealed that 99% of the TiO₂-NPs had a size below 100 nm and 88% of the TiO₂-MPs sizes were between 50 and 200 nm. Following EC₅₀ values were found for particles (mg/l): 92 (TiO₂-MPs) and 15 (TiO₂-NPs). A significant increase in olive tail moment (OTM) was found for TiO₂-NPs at a concentration of 1/10 EC₅₀. TiO₂- and ZrO₂-NPs had a higher intranuclear cell uptake efficiency, compared to corresponding TiO₂- and ZrO₂-MPs. All investigated particles could be detected in cell nucleus. Adhesion of all investigated bacterial species was significantly higher on Ti-implants, compared to ZrO₂-implants.

Conclusion: Ti usually develops an oxide layer (TiO₂). Particles released from Ti-implants should be TiO₂-particles or Ti-particles coated with a TiO₂-layer. Toxicity of released Ti-particles depends on their oxidation state and on their size (NP or MP). Particularly, NPs were more cyto- and genotoxic compared to the corresponding MPs. TiO₂- and ZrO₂-NPs showed a significant increase in the intranuclear cell uptake ratio at higher exposure concentration, compared to lower concentrations and consequently might lead to a higher potential of DNA damage. Adhesion of bacteria to ZrO₂-implants is reduced, compared to Ti-implants. Therefore, ZrO₂-implants might contribute to reduced biological complications (e.g. periimplantitis).

Keywords: Bacterial adhesion; DNA damage; Genotoxicity; Nanoparticles; Nuclear uptake; PDL cells; SEM; SLA; Titaniumoxide, Zirconia, Cytotoxicity; XTT.