Multiple-enzyme treatment reduces biofilm formation in a highly acidogenic in vitro model

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Background: Biofilm matrix-degrading enzymes are a promising approach to caries control, as they target determinants of biofilm virulence without compromising the microbial balance of the oral cavity. This study aimed to test the effect of treatment with a multiple-enzyme combination on the prevention and removal of a highly acidogenic in vitro-grown biofilm model dominated by Streptococcus spp.
Materials and methods: Biofilms were grown in 96-well plates under aerobic conditions at 37°C for 24 hours using pooled saliva as inoculum and brain-heart infusion broth supplied with 5% sucrose and sterile saliva as growth medium. The effect of treatment with a combination of mutanase, glucanase and DNase on biofilm prevention and removal was investigated. Treatment was performed either during (prevention) or after biofilm growth (removal) and the remaining biofilm was quantified by crystal violet staining. Additionally, the treatment effect on biofilm matrix compounds was analyzed using confocal microscopy. Extracellular DNA was stained with TOTO-1, microbial cells with SYTO41 and matrix polysaccharides were visualized by including fluorescently labeled dextran in the growth medium. Treatment effects were analyzed using unpaired t-tests (p<0.05 was considered statistically significant).
Results: Enzyme treatment reduced mean biofilm formation with 94%±1% SD (p=0.0001) and 69%±2% SD (p<0.0001) when performed during and after growth, respectively. Extracellular polysaccharides were on average reduced by 28%±6% SD compared to control treatment (p=0.02), but the mean ratio between polysaccharides and microbial cells remained unchanged (0.70±0.37 SD vs. 0.77±0.22 SD; p=0.74). No significant reduction in the amount of eDNA (p=0.74) was observed.
Conclusion: Multiple-enzyme treatment is a promising non-biocidal approach to biofilm control.
Original languageEnglish
Article number52
JournalCaries Research
Pages (from-to)333
Number of pages1
Publication statusPublished - Oct 2023
EventPoster session presented at the 70th Annual ORCA Congress - Egmond Aan Zee, Netherlands
Duration: 5 Jul 20238 Sept 2023


ConferencePoster session presented at the 70th Annual ORCA Congress
CityEgmond Aan Zee

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