在存在或不存在蔗糖的情况下原位生长的牙生物膜中碳水化合物成分的荧光凝集素结合分析。

IF 2.8 3区 医学 Q1 DENTISTRY, ORAL SURGERY & MEDICINE
Irene Dige, Pune N Paqué, Yumi Chokyu Del Rey, Marie Braad Lund, Andreas Schramm, Sebastian Schlafer
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引用次数: 1

摘要

碳水化合物成分,如糖缀合物和多糖,是牙齿生物膜基质的组成部分,在生物膜的稳定性和毒力中起着重要作用。变形链球菌生物膜中的外多糖已经被广泛表征,但对于复杂的原位生长的牙齿生物膜中的基质碳水化合物知之甚少。本研究采用荧光凝集素结合分析(FLBA)研究了10个参与者的生物膜(n = 306)中糖缀合物/多糖的丰度和空间分布,这些生物膜分别在(SUC)和(H2O)蔗糖环境下原位生长。用不同碳水化合物特异性的10种荧光标记凝集素(AAL、ABA、ASA、HPA、LEA、MNA-G、MPA、PSA、VGA和WGA)对生物膜进行染色,并通过共聚焦显微镜和数字图像分析进行分析。采用16S rRNA基因测序法测定微生物组成。除ABA外,所有凝集素靶向的基质生物体积都相当大,占生物膜微生物生物体积的19.3%至194.0%,这表明原位生长的牙生物膜中碳水化合物种类繁多。MNA-G、AAL和ASA分别是半乳糖、焦糖和甘露糖特异性的,染色的生物量最大。在SUC生物膜中,AAL和ASA生物量均有所增加,但由于存在较大的生物学变异,差异不显著。SUC生物膜中链球菌丰富,奈瑟菌和嗜血杆菌丰度降低,但凝集素染色的生物量与细菌丰度之间没有显著相关性。总之,FLBA证明了在复杂的原位生长的牙齿生物膜中存在由各种不同碳水化合物成分组成的大量生物膜基质。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Fluorescence lectin binding analysis of carbohydrate components in dental biofilms grown in situ in the presence or absence of sucrose.

Fluorescence lectin binding analysis of carbohydrate components in dental biofilms grown in situ in the presence or absence of sucrose.

Carbohydrate components, such as glycoconjugates and polysaccharides, are constituents of the dental biofilm matrix that play an important role in biofilm stability and virulence. Exopolysaccharides in Streptococcus mutans biofilms have been characterized extensively, but comparably little is known about the matrix carbohydrates in complex, in situ-grown dental biofilms. The present study employed fluorescence lectin binding analysis (FLBA) to investigate the abundance and spatial distribution of glycoconjugates/polysaccharides in biofilms (n = 306) from 10 participants, grown in situ with (SUC) and without (H2O) exposure to sucrose. Biofilms were stained with 10 fluorescently labeled lectins with different carbohydrate specificities (AAL, ABA, ASA, HPA, LEA, MNA-G, MPA, PSA, VGA and WGA) and analyzed by confocal microscopy and digital image analysis. Microbial composition was determined by 16S rRNA gene sequencing. With the exception of ABA, all lectins targeted considerable matrix biovolumes, ranging from 19.3% to 194.0% of the microbial biovolume in the biofilms, which illustrates a remarkable variety of carbohydrate compounds in in situ-grown dental biofilms. MNA-G, AAL, and ASA, specific for galactose, fucose, and mannose, respectively, stained the largest biovolumes. AAL and ASA biovolumes were increased in SUC biofilms, but the difference was not significant due to considerable biological variation. SUC biofilms were enriched in streptococci and showed reduced abundances of Neisseria and Haemophilus spp., but no significant correlations between lectin-stained biovolumes and bacterial abundance were observed. In conclusion, FLBA demonstrates the presence of a voluminous biofilm matrix comprising a variety of different carbohydrate components in complex, in situ-grown dental biofilms.

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来源期刊
Molecular Oral Microbiology
Molecular Oral Microbiology DENTISTRY, ORAL SURGERY & MEDICINE-MICROBIOLOGY
CiteScore
6.50
自引率
5.40%
发文量
46
审稿时长
>12 weeks
期刊介绍: Molecular Oral Microbiology publishes high quality research papers and reviews on fundamental or applied molecular studies of microorganisms of the oral cavity and respiratory tract, host-microbe interactions, cellular microbiology, molecular ecology, and immunological studies of oral and respiratory tract infections. Papers describing work in virology, or in immunology unrelated to microbial colonization or infection, will not be acceptable. Studies of the prevalence of organisms or of antimicrobials agents also are not within the scope of the journal. The journal does not publish Short Communications or Letters to the Editor. Molecular Oral Microbiology is published bimonthly.
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