Development of an in vitro biofilm model of the human supra-gingival microbiome for Oral microbiome transplantation

IF 1.7 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS
Don H.K. Ketagoda , Peter Varga , Tracy R. Fitzsimmons , Nicole E. Moore , Laura S. Weyrich , Peter S. Zilm
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Abstract

The high prevalence of dental caries and periodontal disease place a significant burden on society, both socially and economically. Recent advances in genomic technologies have linked both diseases to shifts in the oral microbiota – a community of >700 bacterial species that live within the mouth. The development of oral microbiome transplantation draws on the success of fecal microbiome transplantation for the treatment of gut pathologies associated with disease. Many current in vitro oral biofilm models have been developed but do not fully capture the complexity of the oral microbiome which is required for successful OMT. To address this, we developed an in vitro biofilm system that maintained an oral microbiome with 252 species on average over 14 days. Six human plaque samples were grown in 3D printed flow cells on hydroxyapatite discs using artificial saliva medium (ASM). Biofilm composition and growth were monitored by high throughput sequencing and confocal microscopy/SEM, respectively. While a significant drop in bacterial diversity occurred, up to 291 species were maintained in some flow cells over 14 days with 70% viability grown with ASM. This novel in vitro biofilm model represents a marked improvement on existing oral biofilm systems and provides new opportunities to develop oral microbiome transplant therapies.

开发用于口腔微生物组移植的人类龈上微生物组体外生物膜模型。
龋齿和牙周病的高发病率给社会带来了沉重的社会和经济负担。基因组学技术的最新进展将这两种疾病与口腔微生物群的变化联系了起来,口腔微生物群是由生活在口腔中的超过 700 种细菌组成的群落。口腔微生物组移植的发展借鉴了粪便微生物组移植治疗与疾病相关的肠道病变的成功经验。目前已开发出许多体外口腔生物膜模型,但并不能完全捕捉口腔微生物组的复杂性,而口腔微生物组是成功进行口腔微生物组移植所必需的。为了解决这个问题,我们开发了一种体外生物膜系统,该系统能在 14 天内平均维持 252 种口腔微生物群。使用人工唾液培养基(ASM)在羟基磷灰石盘上的 3D 打印流动细胞中培养了六种人体牙菌斑样本。生物膜的组成和生长分别通过高通量测序和共聚焦显微镜/SEM 进行监测。虽然细菌多样性出现了明显下降,但在使用人工唾液培养基生长的 14 天内,一些流动细胞中仍有多达 291 种细菌存活,存活率达 70%。这种新型体外生物膜模型明显改善了现有的口腔生物膜系统,为开发口腔微生物组移植疗法提供了新的机遇。
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来源期刊
Journal of microbiological methods
Journal of microbiological methods 生物-生化研究方法
CiteScore
4.30
自引率
4.50%
发文量
151
审稿时长
29 days
期刊介绍: The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.
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