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一种新型抗菌肽对浮游和多微生物生物膜状态下牙周病原体的疗效。

Efficacy of a novel antimicrobial peptide against periodontal pathogens in both planktonic and polymicrobial biofilm states.

作者信息

Wang Hong-Yan, Cheng Jya-Wei, Yu Hui-Yuan, Lin Li, Chih Ya-Han, Pan Ya-Ping

机构信息

Department of Periodontology, School of Stomatology, China Medical University, Shenyang 110002, China.

Institute of Biotechnology and Department of Medical Science, National Tsing Hua University, Hsinchu 300, Taiwan.

出版信息

Acta Biomater. 2015 Oct;25:150-61. doi: 10.1016/j.actbio.2015.07.031. Epub 2015 Jul 22.

Abstract

UNLABELLED

Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis represent the early, middle and late colonizers of the bacterial accretion in dental plaque biofilms. These sessile communities constitute a protected mode of growth that promotes survival in a hostile environment. This study describes a novel and unrecognized role for a synthetic cationic antimicrobial peptide, Nal-P-113, which inhibits and kills periodontal bacteria in planktonic state, inhibits the formation of biofilms and eradicates polymicrobial biofilms. Nal-P-113 is also stable in saliva, serum and saline solution. At a concentration less than 320 μg/mL which is harmless to normal oral cells, Nal-P-113 can kill bacteria in planktonic state. At a concentration of antimicrobial peptide Nal-P-113 (1280 μg/mL) which only causes slight damages to normal oral cells is needed to kill bacteria in biofilm state. It is worth mentioning that this concentration of Nal-P-113 is harmless to rat oral mucosa compared to chlorhexidine. The mechanism of Nal-P-113 inhibiting and killing periodontal bacteria might rely on the abilities to permeabilize and/or to form pores within the cytoplasmic membranes, thus causes the death of bacteria. Here, we provided a novel and stable antimicrobial peptide with very low mammalian cytotoxicity, which can inhibit and kill periodontal bacteria in both planktonic and polymicrobial biofilm states.

STATEMENT OF SIGNIFICANCE

Nal-P-113 is a potent antimicrobial peptide with strong antimicrobial ability, improved deficiency compared with other antibacterial peptides, and remains stable in phosphate buffered saline, saliva, brain-heart infusion medium and bovine calf serum. Nal-P-113 exhibits a broad spectrum of bacteriocidal activity with excellent eradicating capability on oral pathogens and the respective biofilms. In this study, we used propidium iodide staining, scanning electron microscopy and transmission electron microscopy to confirm that Nal-P-113 can perforate plasmalemma thereby resulting in the death of oral pathogens and disintegrate the respective biofilms. Nal-P-113 also showed effective anti-plaque biofilms and cytotoxicity in the rat periodontitis model. No adverse effects can be observed on the gingivomucosa tissue. In short, the antimicrobial peptide Nal-P-113 presented to be an effective yet have low mammalian cytotoxicity agent with potential application in the clinic. This study provides a proof of concept in applying antimicrobial peptides in the clinical perspective.

摘要

未标记

戈登链球菌、具核梭杆菌和牙龈卟啉单胞菌分别代表牙菌斑生物膜中细菌黏附的早期、中期和晚期定植菌。这些固着群落构成了一种受保护的生长模式,促进了在恶劣环境中的生存。本研究描述了一种新型且未被认识的合成阳离子抗菌肽Nal-P-113的作用,它能抑制和杀死浮游状态的牙周细菌,抑制生物膜形成并根除多微生物生物膜。Nal-P-113在唾液、血清和盐溶液中也很稳定。在浓度低于320μg/mL时(对正常口腔细胞无害),Nal-P-113能杀死浮游状态的细菌。要杀死生物膜状态的细菌需要抗菌肽Nal-P-113的浓度为1280μg/mL(此浓度仅对正常口腔细胞造成轻微损伤)。值得一提的是,与洗必泰相比,该浓度的Nal-P-113对大鼠口腔黏膜无害。Nal-P-113抑制和杀死牙周细菌的机制可能依赖于其使细胞质膜通透性增加和/或形成孔的能力,从而导致细菌死亡。在此,我们提供了一种新型且稳定的抗菌肽,其对哺乳动物细胞毒性极低,能抑制和杀死浮游状态及多微生物生物膜状态的牙周细菌。

意义声明

Nal-P-113是一种强效抗菌肽,抗菌能力强,与其他抗菌肽相比有所改进,并且在磷酸盐缓冲盐水、唾液、脑心浸液培养基和小牛血清中保持稳定。Nal-P-113具有广谱杀菌活性,对口腔病原体及其生物膜具有出色的根除能力。在本研究中,我们使用碘化丙啶染色、扫描电子显微镜和透射电子显微镜证实Nal-P-113可穿透质膜,从而导致口腔病原体死亡并分解各自的生物膜。Nal-P-113在大鼠牙周炎模型中也显示出有效的抗菌斑生物膜作用和细胞毒性。在牙龈黏膜组织上未观察到不良反应。简而言之,抗菌肽Nal-P-113是一种有效但对哺乳动物细胞毒性低的药物,具有潜在的临床应用价值。本研究为从临床角度应用抗菌肽提供了概念验证。

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