ABSTRACT
The study was conducted to inform risk assessments concerning microbial exposure to quaternary ammonium biocides (QUATs) by investigating their effects on 10 microbial strains of hygiene relevance. Biocides were divided into three categories: simple aqueous solutions, biocide mixtures, and formulated biocides. Organisms were grown in the presence of biocides for 10 generations and then subsequently for another 10 generations in biocide-free media. Control organisms were passaged 20 times in biocide-free media. Strains were then assessed for biocide and antibiotic susceptibility, changes in growth dynamics, and single nucleotide polymorphisms (SNPs). Biocide mixtures demonstrated greater antimicrobial potency than singular and formulated biocides. Susceptibility changes of under twofold were observed for all biocides tested. Susceptibility decreased significantly for organisms passaged with singular biocides (1.29- to 4.35-fold) and biocide mixtures (1.4- to 1.5-fold), but not for formulated biocides (1.3- to 1.84-fold) compared to controls. Antibiotic susceptibility both increased and decreased in passaged organisms, with heightened susceptibility occurring more frequently in the singular biocide group. Changes in susceptibility and growth dynamics were similar in the passaged and unexposed controls for fitness measures of adapted bacteria; there were no significant differences between biocide groups, but significantly lower generation and doubling times in organisms exposed to singular biocides. Similar frequencies in SNPs occurred for the three biocide groups and unexposed controls. While some adaptations occurred, particularly with singular biocides, the impact on antibiotic resistance and genomic mutations was limited. These findings suggest that the use of formulated QUATs may pose a comparatively lower risk for antimicrobial resistance.IMPORTANCEBiocides are used globally to control microbial growth and effective assessment of the risks and benefits of their use is therefore a high priority. Much of the data used to assess risk has been based on sub-lethal exposure of bacteria to singular biocides in simple aqueous solutions. This work builds on limited prior realism-based studies to demonstrate enhanced potency in biocidal mixtures; the mitigation of resistance selection by formulations and inconsistent cross-resistance effects with both increases and decreases in susceptibility for a wide range of antibiotics. These data can be used to better inform risk assessments of biocide deployment.
ABSTRACT
PURPOSE: To develop a biocompatible denture base resin/TiO2 nanocomposite material with antifungal characteristics that is suitable for 3D-printing denture bases. MATERIALS AND METHODS: TiO2 nanoparticles (NPs) with a 0.10, 0.25, 0.50, and 0.75 weight percent (wt.%) were incorporated into a commercially available 3D-printed resin material. The resulting nanocomposite material was analyzed using Lactate dehydrogenase (LDH) and AlamarBlue (AB) assays for biocompatibility testing with human gingival fibroblasts (HGF). The composite material was also tested for its antifungal efficacy against Candida albicans. Fourier transform infrared (FTIR) and Energy Dispersive X-ray Spectroscopy (EDX) mapping were conducted to assess the surface coating and the dispersion of the NPs. RESULTS: LDH and AB assays confirmed the biocompatibility of the material showing cell proliferation at a rate of nearly 100% at day 10, with a cytotoxicity of less than 13% of the cells at day 10. The concentrations of 0.10, 0.25, and 0.50 wt.% caused a significant reduction (p < 0.05) in the number of candida cells attached to the surface of the specimens (p < 0.05), while 0.75 wt.% did not show any significant difference compared to the control (no TiO2 NPs) (p > 0.05). FTIR and EDX analysis confirmed the presence of TiO2 NPs within the nanocomposite material with a homogenous dispersion for 0.10 and 0.25 wt.% groups and an aggregation of the NPs within the material at higher concentrations. CONCLUSION: The addition of TiO2 NPs into 3D-printed denture base resin proved to have an antifungal effect against Candida albicans. The resultant nanocomposite material was a biocompatible material with HGFs and was successfully used for 3D printing.