Grapefruit seed extract effectively inhibits the Candida albicans biofilms development on polymethyl methacrylate denture-base resin
Grapefruit seed extract effectively inhibits the Candida albicans biofilms development on polymethyl methacrylate denture-base resin
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DOI：
10.1371/journal.pone.0217496
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发表时间：
2019-05
期刊：
影响因子：
3.7
通讯作者：
Chiaki Tsutsumi-Arai;Kensuke Takakusaki;Y. Arai;Chika Terada-Ito;Yusuke Takebe;T. Imamura;Shinji Ide;S. Tatehara;Reiko Tokuyama-Toda;N. Wakabayashi;K. Satomura
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文献类型：
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作者：
Chiaki Tsutsumi-Arai;Kensuke Takakusaki;Y. Arai;Chika Terada-Ito;Yusuke Takebe;T. Imamura;Shinji Ide;S. Tatehara;Reiko Tokuyama-Toda;N. Wakabayashi;K. Satomura
本研究旨在探讨西柚籽提取物(GSE)对义齿基托树脂上白色念珠菌生物膜的净化作用及其对树脂力学性能和表面性能的影响。用蒸馏水稀释成0.1%(0.1%GSE)和1%(1%GSE)的GSE溶液和Polident®义齿清洁片溶于蒸馏水(Polident)或0.1%GSE溶液(0.1%G+P)的溶液作为清洁液。制备了丙烯酸树脂膜片，并在其上形成了白念珠菌生物膜。将含有生物膜的圆盘在25℃下分别用每种溶液处理5min，处理后，用菌落形成单位(CFU)试验、荧光显微镜和扫描电子显微镜(SEM)分析圆盘上的生物膜。为了评估持续清洁效果，将每种溶液处理5min的圆盘在酵母氮基质中有氧孵育24 h，孵育后用CFU法评估持续清洁效果。将一些丙烯酸树脂试件在两种溶液中浸泡7天，观察表面粗糙度(Ra)、维氏硬度(VH)、弯曲强度(FS)和弯曲弹性模量(Fm)的变化。结果表明，1%GSE处理5min几乎完全消除了树脂上形成的生物膜，而0.1%GSE、Polident和0.1%G+P处理5min对生物膜有显著的抑制作用。0.1%的GSE和0.1%的G+P对生物膜有持续的抑制作用。荧光显微镜显示，Polident主要诱导酵母菌死亡，而至少含0.1%GSE的洗涤液除诱导酵母菌死亡外，还诱导菌丝死亡。扫描电子显微镜还显示，Polident主要在酵母细胞表面引起皱纹、收缩和一些深深的凹坑，而至少含有0.1%GSE的溶液不仅在酵母细胞表面引起皱纹、收缩，还对菌丝细胞表面造成一些损伤。浸泡在任何一种溶液中后，Ra、VH、FS或Fm均未观察到显著变化。综上所述，GSE溶液能够清洁义齿基托树脂上的白色念珠菌生物膜，并对生物膜的形成具有持久的抑制作用，而不会对树脂表面造成任何损害。
This study aimed to investigate the cleansing effects of grapefruit seed extract (GSE) on biofilms of Candida albicans (C. albicans) formed on denture-base resin and the influence of GSE on the mechanical and surface characteristics of the resin. GSE solution diluted with distilled water to 0.1% (0.1% GSE) and 1% (1% GSE) and solutions with Polident® denture cleansing tablet dissolved in distilled water (Polident) or in 0.1% GSE solution (0.1% G+P) were prepared as cleansing solutions. Discs of acrylic resin were prepared, and the biofilm of C. albicans was formed on the discs. The discs with the biofilm were treated with each solution for 5 min at 25°C. After the treatment, the biofilm on the discs was analyzed using a colony forming unit (CFU) assay, fluorescence microscopy, and scanning electron microscopy (SEM). In order to assess the persistent cleansing effect, the discs treated with each solution for 5 min were aerobically incubated in Yeast Nitrogen Base medium for another 24 h. After incubation, the persistent effect was assessed by CFU assay. Some specimens of acrylic resin were immersed in each solution for 7 days, and changes in surface roughness (Ra), Vickers hardness (VH), flexural strength (FS), and flexural modulus (FM) were evaluated. As a result, the treatment with 1% GSE for 5 min almost completely eliminated the biofilm formed on the resin; whereas, the treatment with 0.1% GSE, Polident, and 0.1% G+P for 5 min showed a statistically significant inhibitory effect on biofilms. In addition, 0.1% GSE and 0.1% G+P exerted a persistent inhibitory effect on biofilms. Fluorescence microscopy indicated that Polident mainly induced the death of yeast, while the cleansing solutions containing at least 0.1% GSE induced the death of hyphae as well as yeast. SEM also revealed that Polident caused wrinkles, shrinkage, and some deep craters predominantly on the cell surfaces of yeast, while the solutions containing at least 0.1% GSE induced wrinkles, shrinkage, and some damage on cell surfaces of not only yeasts but also hyphae. No significant changes in Ra, VH, FS, or FM were observed after immersion in any of the solutions. Taken together, GSE solution is capable of cleansing C. albicans biofilms on denture-base resin and has a persistent inhibitory effect on biofilm development, without any deteriorations of resin surface.