Yang Sirui, Li Na, Wu Haoze, Zhang Mai, Wang Luoqianhui, Xiao Min, Cheng Xiaogang, Yu Qing
State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, National Clinical Research Center for Oral Diseases, Shaanxi Key Laboratory of Stomatology, Department of Operative Dentistry and Endodontics, School of Stomatology, The Fourth Military Medical University, Xi'an, Shaanxi, China.
Int Endod J. 2025 Apr;58(4):613-626. doi: 10.1111/iej.14199. Epub 2025 Jan 17.
Previous studies have shown the important relationships between Enterococcus faecalis and Candida albicans in post-treatment endodontic disease (PTED). However, the fungal-bacterial interactions and their possible functional routes are less understood. In this study, we investigated the effect of extracellular vesicles (EVs) derived from C. albicans on E. faecalis growth and pathogenicity.
Candida albicans EVs were isolated from a yeast nitrogen base (YNB) medium, and their morphology, size distribution, and protein concentration were observed and identified. The effects of EVs on planktonic E. faecalis were evaluated using growth curves and colony-forming unit counts, whereas the effects on E. faecalis biofilms were determined using scanning electron and confocal laser scanning microscopes. The ability of E. faecalis to resist a detrimental environment, infect dentinal tubules, and biofilm formation on gutta percha was examined. Additionally, the effect of EVs on cell invasion and cytotoxicity of E. faecalis were assessed. Statistical analysis was performed using one-way analysis of variance, and p-values of <.05 were considered significantly different.
Candida albicans EVs were nanoparticles with bilayer membranes and with peak sizes of 111.9 and 230 nm. EVs exhibited a complex effect on E. faecalis and its biofilms; 5 μg/mL of EVs showed inhibitory effects whereas 0.156 μg/mL of EVs facilitated their growth. The EVs showed consistent effects on E. faecalis virulence. Notably, 5 μg/mL of EVs reduced the damage to RAW264.7 cells caused by E. faecalis, as well as the invasion ability of E. faecalis to macrophages and the intracellular survival ability of E. faecalis after macrophage phagocytosis, whereas 0.156 μg/mL of EVs had completely opposite effects.
Candida albicans EVs showed dual effects on E. faecalis growth and virulence in vitro, suggesting C. albicans EVs are involved in fungal-bacterial communication. Moreover, the inhibitory effects exhibited by 5 μg/mL of EVs in vitro may suggest a new agent for the control of E. faecalis.
以往研究表明粪肠球菌与白色念珠菌在根管治疗后疾病(PTED)中存在重要关联。然而,真菌与细菌之间的相互作用及其可能的功能途径尚不清楚。在本研究中,我们调查了白色念珠菌衍生的细胞外囊泡(EVs)对粪肠球菌生长和致病性的影响。
从酵母氮源(YNB)培养基中分离白色念珠菌EVs,并观察和鉴定其形态、大小分布及蛋白质浓度。使用生长曲线和菌落形成单位计数评估EVs对浮游粪肠球菌的影响,而使用扫描电子显微镜和共聚焦激光扫描显微镜确定其对粪肠球菌生物膜的影响。检测粪肠球菌抵抗有害环境、感染牙本质小管以及在牙胶上形成生物膜的能力。此外,评估EVs对粪肠球菌细胞侵袭和细胞毒性的影响。采用单因素方差分析进行统计分析,p值<0.05被认为具有显著差异。
白色念珠菌EVs是具有双层膜的纳米颗粒,峰值大小分别为111.9和230nm。EVs对粪肠球菌及其生物膜表现出复杂的影响;5μg/mL的EVs显示出抑制作用,而0.156μg/mL的EVs促进其生长。EVs对粪肠球菌毒力表现出一致的影响。值得注意的是,5μg/mL的EVs减少了粪肠球菌对RAW264.7细胞的损伤,以及粪肠球菌对巨噬细胞 的侵袭能力和巨噬细胞吞噬后粪肠球菌的细胞内存活能力,而0.156μg/mL的EVs则产生完全相反的效果。
白色念珠菌EVs在体外对粪肠球菌的生长和毒力表现出双重作用,表明白色念珠菌EVs参与了真菌与细菌的通讯。此外,5μg/mL的EVs在体外表现出的抑制作用可能提示其为控制粪肠球菌的新型药物。