National Engineering Laboratory for Druggable Gene and Protein Screening, School of Life Sciences, Northeast Normal University, Changchun, China.
College of Ecology and Environment, Hainan Tropical Ocean University, Sanya, China.
Front Cell Infect Microbiol. 2023 Aug 28;13:1140689. doi: 10.3389/fcimb.2023.1140689. eCollection 2023.
Dental caries is one of the most common and costly biofilm-dependent oral diseases in the world. is the major cariogenic pathogen of dental caries. synthesizes extracellular polysaccharides by autologous glucosyltransferases, which then promotes bacterial adhesion and cariogenic biofilm formation. The biofilm is the principal target for caries treatment. This study was designed to explore the antibacterial activity and mechanisms of areca nut essential oil (ANEO) against .
The ANEOs were separated by negative pressure hydro-distillation. The Kirby-Bauer method and broth microdilution method were carried out to evaluate the antibacterial activity of different ANEOs. The antibacterial mechanism was revealed by crystal violet staining, XTT reduction, microbial adhesion to hydrocarbon test, extracellular polysaccharide production assay, glucosyltransferase activity assay, lactate dehydrogenase leaking, propidium iodide staining and scanning electron microscopy (SEM). The cytotoxicity of ANEOs was determine by MTT assay.
The ANEOs separated at different temperatures exhibited different levels of antibacterial activity against , and the ANEO separated at 70°C showed the most prominent bacteriostatic activity. Anti-biofilm experiments showed that the ANEOs attenuated the adhesion ability of by decreasing the surface hydrophobicity of the bacteria, prevented biofilm formation by inhibiting glucosyltransferase activity, reducing extracellular polysaccharide synthesis, and reducing the total biofilm biomass and activity. SEM further demonstrated the destructive effects of the ANEOs on the biofilm. Cell membrane-related experiments indicated that the ANEOs destroyed the integrity of the cell membrane, resulting in the leakage of lactic dehydrogenase and nucleic acids. SEM imaging of cell showed the disruption of the cellular morphology by the ANEOs. The cytotoxicity assay suggested that ANEO was non-toxic towards normal oral epithelial cells.
This study displayed that ANEOs exerted antibacterial activity against primarily by affecting the biofilm and disrupting the integrity of the cell membrane. ANEOs has the potential to be developed as an antibacterial agent for preventing dental caries. Additionally, a new method for the separation of essential oil components is presented.
龋齿是世界上最常见和最昂贵的生物膜依赖型口腔疾病之一。 是龋齿的主要致龋病原体。通过自身的葡糖基转移酶合成细胞外多糖，从而促进细菌黏附和致龋生物膜的形成。 生物膜是龋齿治疗的主要靶标。本研究旨在探索槟榔籽油（ANEO）对 的抗菌活性及其机制。
采用减压水蒸馏法分离 ANEO。采用 Kirby-Bauer 法和肉汤微量稀释法评价不同 ANEO 的抗菌活性。通过结晶紫染色、XTT 还原、微生物附着烃试验、细胞外多糖产生测定、葡糖基转移酶活性测定、乳酸脱氢酶渗漏、碘化丙啶染色和扫描电子显微镜（SEM）揭示抗菌机制。通过 MTT 测定法测定 ANEO 的细胞毒性。
在不同温度下分离的 ANEO 对 表现出不同水平的抗菌活性，70°C 下分离的 ANEO 表现出最强的抑菌活性。抗生物膜实验表明，ANEO 通过降低细菌表面疏水性来减弱 黏附能力，通过抑制葡糖基转移酶活性、减少细胞外多糖合成、降低总生物膜生物量和活性来阻止 生物膜形成。SEM 进一步证明了 ANEO 对 生物膜的破坏作用。细胞膜相关实验表明，ANEO 破坏了细胞膜的完整性，导致乳酸脱氢酶和核酸漏出。SEM 对 细胞的成像显示，ANEO 破坏了细胞形态。细胞毒性测定表明，ANEO 对正常口腔上皮细胞无毒。
本研究表明，ANEO 主要通过影响生物膜和破坏细胞膜完整性来发挥对 的抗菌活性。ANEO 有可能被开发为预防龋齿的抗菌剂。此外，还提出了一种分离精油成分的新方法。
Zotero 插件
