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Effects of tyrosol and farnesol on Candida albicans biofilm

机译:酪醇和法尼醇对白色念珠菌生物膜的影响

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摘要

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.
机译:目前的体外研究检查了群体感应分子法尼醇和酪醇对白色念珠菌生物膜发育的影响,以阐明它们在口腔卫生中作为新型佐剂的作用。该研究在假牙念珠菌ATCC 10231和假牙念珠菌分离物中进行,并在平底96孔聚苯乙烯平板中进行。在补充了0.001–3 mM法尼醇和/或1–20 mM酪醇的Sabouraud肉汤中于37°C孵育24和48 h后,评估了酵母的生长及其形成生物膜的能力。通过比浊法评估酵母的生长,并在需氧和厌氧条件下通过结晶紫染色对生物膜进行定量。真菌细胞的生存能力通过浮游细胞的培养以及荧光素双乙酸盐和溴化乙锭染色后使用荧光显微镜检查生物膜来控制。当在生物膜形成开始时添加(> 50%抑制)时,3 mM的法尼醇发挥出比在预先形成的生物膜中添加(<10%抑制)时更强的作用。类似地,20 mM的酪醇对生物膜形成的影响(> 80%抑制)比对预先形成的生物膜的影响(<40%抑制)大。尽管所附着的生物量显着减少,但是在所研究的分子存在下,酵母的生长变化很小,这通过比浊法得到了证实,在固体培养基上培养了上清液,随后进行了菌落形成单位的计数和使用荧光显微镜的生存力测试。在最高测试浓度下,分子在生物膜形成的初始阶段具有更大的作用。法尼醇在厌氧期间的作用与在需氧期间观察到的作用没有显着差异,这与在厌氧期间的酪醇表现出略微降低的酵母生物膜抑制作用不同。总之,本研究证明了法尼醇和酪醇的特定抗生物膜作用,独立于杀真菌或抑真菌作用,如在白色念珠菌ATCC 10231和从假牙分离的6个菌株中测试的那样。在建议将这些分子用于口腔卫生的预防目的之前,需要进行进一步的研究,以弄清与其抗生物膜作用有关的代谢途径和细胞机制,以及对口腔微生物组的影响。

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