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洛神花萼提取物对口腔病原菌体外活力和生物膜形成能力的影响。

Effect of roselle calyx extract on in vitro viability and biofilm formation ability of oral pathogenic bacteria.

机构信息

Department of Oral Microbiology, School of Dentistry, Health Sciences University of Hokkaido, Hokkaido 061-0293, Japan.

Department of Oral Microbiology, School of Dentistry, Health Sciences University of Hokkaido, Hokkaido 061-0293, Japan.

出版信息

Asian Pac J Trop Med. 2016 Feb;9(2):119-24. doi: 10.1016/j.apjtm.2016.01.020. Epub 2016 Jan 13.

Abstract

OBJECTIVE

To investigate the effect of the roselle calyx extract (RCE) (Hibiscus sabdariffa L.) on the in vitro viability and biofilm formation ability of oral pathogenic bacteria.

METHODS

RCE was prepared by soaking roselle calyx powder with ethyl alcohol for 24 h at room temperature. After centrifugation, the extract was lyophilized. Then, the extract was dissolved in phosphate-buffered saline, the pH was adjusted, and the extract was aseptically filtered. We used Streptococcus mutans, Streptococcus sanguinis, Lactobacillus casei, Actinomyces naeslundii, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis and Prevotella intermedia in this study. The antibacterial activity of the RCE was determined by treating the cells of these bacteria with the extract for 10 or 20 min at room temperature. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration was determined using the microdilution method, and the effect of the RCE on the ability to form biofilm was determined using a polystyrene micro plate assay. In addition, we used the WST-1 assay to determine the cytotoxicity of the RCE on HGF, Ca9-22 and KB cells.

RESULTS

The RCE had antibacterial activity against oral bacteria used in this study. In particular, most significant antibacterial activity was observed against Fusobacterium nucleatum, Prevotella intermedia and Porphyromonas gingivalis. The MIC and minimum bactericidal concentration were 7.2 mg/mL-28.8 mg/mL and 14.4 to >57.6 mg/mL. The RCE had an inhibitory effect on biofilm formation at the MIC and sub-MIC levels. In addition, the RCE had low cytotoxic effects on HGF, Ca9-22 and KB cells.

CONCLUSIONS

Thus, our results indicate that the RCE may be used for preventing oral diseases.

摘要

目的

研究洛神花萼提取物(Hibiscus sabdariffa L.)对口腔病原菌体外活力和生物膜形成能力的影响。

方法

将洛神花萼粉用乙醇室温浸泡 24 小时制备罗勒花萼提取物(RCE)。离心后,将提取物冻干。然后,将提取物溶解于磷酸盐缓冲盐水(PBS)中,调整 pH 值,无菌过滤提取物。我们在本研究中使用了变形链球菌、血链球菌、干酪乳杆菌、奈瑟放线菌、伴放线放线杆菌、核梭杆菌、牙龈卟啉单胞菌和中间普氏菌。将这些细菌的细胞在室温下用提取物处理 10 或 20 分钟,以确定 RCE 的抗菌活性。采用微量稀释法测定最小抑菌浓度(MIC)和最小杀菌浓度,采用聚苯乙烯微量板法测定 RCE 对生物膜形成能力的影响。此外,我们还使用 WST-1 测定 RCE 对 HGF、Ca9-22 和 KB 细胞的细胞毒性。

结果

RCE 对本研究中使用的口腔细菌具有抗菌活性。特别是,对核梭杆菌、中间普氏菌和牙龈卟啉单胞菌表现出最强的抗菌活性。MIC 和最小杀菌浓度分别为 7.2mg/mL-28.8mg/mL 和 14.4 至>57.6mg/mL。RCE 在 MIC 和亚 MIC 水平上对生物膜形成具有抑制作用。此外,RCE 对 HGF、Ca9-22 和 KB 细胞的细胞毒性较低。

结论

因此,我们的结果表明,RCE 可能用于预防口腔疾病。

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