Division of Periodontology, Faculty of Dentistry & Graduate School of Medical and Dental Sciences, Niigata University, 2-5274 Gakkocho-dori, Chuo-ku, 951-8514 Niigata, Japan; Department of Periodontology, University of Dental Medicine, 582, Than Thu Mar Road, Thingangyun Township, 11071 Yangon, Myanmar.
Division of Periodontology, Faculty of Dentistry & Graduate School of Medical and Dental Sciences, Niigata University, 2-5274 Gakkocho-dori, Chuo-ku, 951-8514 Niigata, Japan; Division of Periodontics, Niigata University Medical and Dental Hospital, 2-5274 Gakkocho-dori, Chuo-ku, 951-8514 Niigata, Japan.
Arch Oral Biol. 2022 Oct;142:105497. doi: 10.1016/j.archoralbio.2022.105497. Epub 2022 Jul 9.
This study aimed to clarify the antibacterial mechanism and antibiofilm effect of soybean-derived peptide BCBS-11 against periodontopathic bacteria.
The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of BCBS-11 against Porphyromonas gingivalis (P. gingivalis), Fusobacterium nucleatum (F. nucleatum), and Streptococcus mitis (S. mitis) were determined for the antibacterial mechanism. The effect of BCBS-11 on membrane permeability and depolarization activity were investigated using propidium iodide (PI) staining and 3, 3'-dipropylthiadicarbocyanine iodide (DiSC-(5)) analysis. Monospecies and multispecies biofilms were cultured on 96-well plates. The amount of biofilm was determined using crystal violet staining to determine the inhibition of biofilm formation and the eradication of established biofilm using BCBS-11. The cytotoxicity of BCBS-11 was evaluated using 3-(4, 5-Dimethylthiazol-2-yl)- 2, 5-diphenyltetrazolium bromide (MTT) assay.
The MIC and MBC indicated the bactericidal activity of BCBS-11 against P. gingivalis and F. nucleatum. The PI staining revealed that BCBS-11 disrupted the bacterial membrane integrity. The DiSC-(5) analysis indicated that BCBS-11 depolarized the bacterial cytoplasmic membrane. These results indicate the antimicrobial action of BCBS-11 through membrane disruption and the collapse of membrane electrochemical gradient. BCBS-11 significantly inhibited the monospecies biofilm formation of P. gingivalis and F. nucleatum and also inhibited dual-species biofilm. BCBS-11 was not cytotoxic toward human oral epithelial cells.
BCBS-11 inhibits the monospecies and multispecies biofilm formation of P. gingivalis and F. nucleatum, and their bactericidal activity results from membrane disruption.
本研究旨在阐明大豆衍生肽 BCBS-11 对牙周病原菌的抗菌机制和抗生物膜作用。
测定 BCBS-11 对牙龈卟啉单胞菌（P. gingivalis）、核梭杆菌（F. nucleatum）和缓症链球菌（S. mitis）的最小抑菌浓度（MIC）和最小杀菌浓度（MBC），以了解其抗菌机制。通过碘化丙啶（PI）染色和 3,3'-二丙基噻二碳菁碘化物（DiSC-(5))分析研究 BCBS-11 对膜通透性和去极化活性的影响。在 96 孔板上培养单种和多种生物膜。用结晶紫染色法测定生物膜量，以确定 BCBS-11 对生物膜形成的抑制作用和对已建立的生物膜的清除作用。用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法评估 BCBS-11 的细胞毒性。
MIC 和 MBC 表明 BCBS-11 对 P. gingivalis 和 F. nucleatum 具有杀菌活性。PI 染色显示 BCBS-11 破坏了细菌膜的完整性。DiSC-(5)分析表明 BCBS-11 使细菌细胞质膜去极化。这些结果表明，BCBS-11 通过破坏细胞膜和膜电化学梯度的崩溃发挥抗菌作用。BCBS-11 显著抑制 P. gingivalis 和 F. nucleatum 的单种生物膜形成，也抑制双种生物膜形成。BCBS-11 对人口腔上皮细胞无细胞毒性。
BCBS-11 抑制 P. gingivalis 和 F. nucleatum 的单种和多种生物膜形成，其杀菌活性源于膜破坏。
