Construction of a versatile in vitro cultivation screening platform using human oral microbiota

Author:

Sasaki Kengo1ORCID,Takeshima Yasunobu1,Fujino Ayami1,Yamashita Junya2,Kimoto Akira2,Sasaki Daisuke3,Kondo Akihiko3,Akashi Masaya2,Okumura Ryo1

Affiliation:

1. Bio Palette Co., Ltd Kobe Japan

2. Department of Oral and Maxillofacial Surgery Kobe University Graduate School of Medicine Kobe Hyogo Japan

3. Graduate School of Science, Technology and Innovation, Kobe University Kobe Hyogo Japan

Abstract

AbstractWe developed a simulation model of human oral microbiota using Bio Palette oral medium (BPOM) containing 0.02% glucose and lower bacterial nitrogen sources, derived from saliva and dental plaque. By decreasing the concentration of Gifu anaerobic medium (GAM) from 30 to 10 g L−1, we observed increased ratios of target pathogenic genera, Porphyromonas and Fusobacterium from 0.5% and 1.7% to 1.2% and 3.5%, respectively, in the biofilm on hydroxyapatite (HA) discs. BPOM exhibited the higher ratios of Porphyromonas and Fusobacterium, and amplicon sequence variant number on HA, compared with GAM, modified GAM and basal medium mucin. Mixing glycerol stocks of BPOM culture solutions from four human subjects resulted in comparable ratios of these bacteria to the original saliva. In this simulation model, sitafloxacin showed higher inhibitory effects on P. gingivalis than minocycline hydrochloride at a low dosage of 0.1 μg mL−1. Probiotics such as Streptococcus salivarius and Limosilactobacillus fermentum also showed significant decreases in Porphyromonas and Fusobacterium ratios on HA, respectively. Overall, the study suggests that BPOM with low carbon and nutrients could be a versatile platform for assessing the efficacy of antibiotics and live biotherapeutics in treating oral diseases caused by Porphyromonas and Fusobacterium.

Publisher

Wiley

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