Laboratory of Physiology and Pharmacology, Université Libre de Bruxelles, B‑1070 Brussels, Belgium.
Laboratory of Antibiotics and Antifungals, Physico‑Chemistry, Synthesis and Biological Activity, University of Tlemcen, 13000 Tlemcen, Algeria.
Mol Med Rep. 2019 Apr;19(4):3201-3209. doi: 10.3892/mmr.2019.9981. Epub 2019 Feb 22.
The present in vitro study examined the effects of the quorum‑sensing molecules farnesol and tyrosol on the development of Candida albicans biofilm in order to elucidate their role as novel adjuvants in oral hygiene. The investigation was conducted in C. albicans ATCC 10231 and C. albicans isolates from dentures and was performed in flat‑bottomed 96‑well polystyrene plates. Yeast growth and their capacity to form biofilms were evaluated following 24 and 48 h incubations at 37˚C in Sabouraud broth supplemented with 0.001‑3 mM farnesol and/or 1‑20 mM tyrosol. Yeast growth was assessed by turbidimetry and biofilms were quantitated by crystal violet staining, under aerobic and anaerobic conditions. The viability of the fungal cells was controlled by the culture of planktonic cells and by examination of the biofilms using fluorescence microscopy following staining with fluorescein diacetate and ethidium bromide. Farnesol at 3 mM exerted a stronger action when added at the beginning of biofilm formation (>50% inhibition) than when added to preformed biofilms (<10% inhibition). Similarly, tyrosol at 20 mM had a greater effect on biofilm formation (>80% inhibition) than on preformed biofilms (<40% inhibition). Despite significant reductions in attached biomass, yeast growth varied little in the presence of the investigated molecules, as corroborated by the turbidimetry, culture of supernatants on solid culture medium followed by counting of colony‑forming units and viability tests using fluorescence microscopy. At the highest tested concentration, the molecules had a greater effect during the initial phases of biofilm formation. The effect of farnesol during anaerobiosis was not significantly different from that observed during aerobiosis, unlike that of tyrosol during anaerobiosis, which exhibited slightly reduced yeast biofilm inhibition. In conclusion, the present study demonstrated the specific anti‑biofilm effect, independent of fungicidal or fungistatic action, of farnesol and tyrosol, as tested in C. albicans ATCC 10231 and 6 strains isolated from dentures. Prior to suggesting the use of these molecules for preventive purposes in oral hygiene, further studies are required in order to clarify the metabolic pathways and cellular mechanisms involved in their antibiofilm effect, as well as the repercussions on the oral microbiome.
本体外研究旨在探讨群体感应分子法呢醇和酪醇对白色念珠菌生物膜形成的影响,以阐明它们作为口腔卫生新型佐剂的作用。该研究在白色念珠菌 ATCC 10231 和义齿分离株中进行,并在平底 96 孔聚苯乙烯板中进行。在 37°C 的萨布罗肉汤中孵育 24 和 48 小时后,评估酵母生长及其形成生物膜的能力,该肉汤补充有 0.001-3 mM 法呢醇和/或 1-20 mM 酪醇。通过浊度法评估酵母生长,通过结晶紫染色定量生物膜,在有氧和厌氧条件下进行。通过浮游细胞培养和用荧光二乙酸酯和溴化乙锭染色后用荧光显微镜检查生物膜来控制真菌细胞的活力。法呢醇在生物膜形成开始时添加 3 mM 时的作用比添加到已形成的生物膜时更强(>50%抑制)。同样,20 mM 酪醇对生物膜形成的影响大于对已形成的生物膜的影响(>80%抑制)。尽管附着生物量有显著减少,但由于浊度法、在固体培养基上培养上清液后计数集落形成单位以及使用荧光显微镜进行的活力测试,浮游细胞的酵母生长变化很小。在最高测试浓度下,这些分子在生物膜形成的初始阶段的作用更大。与有氧条件下观察到的情况不同,法呢醇在厌氧条件下的作用与有氧条件下的作用没有显著差异,而在厌氧条件下的酪醇作用则略有降低酵母生物膜抑制。总之,本研究证明了法呢醇和酪醇在白色念珠菌 ATCC 10231 和从义齿分离的 6 株菌株中的特定抗生物膜作用,独立于杀菌或抑菌作用。在建议将这些分子用于口腔卫生的预防性目的之前,需要进一步研究以阐明其抗生物膜作用涉及的代谢途径和细胞机制,以及对口腔微生物组的影响。