Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro
| Autor: | Archita Gadkari, Thao T. To, Kristopher A. Kerns, Xuesong He, Georgios A. Kotsakis, Diane M. Daubert, Jeffrey S. McLean, Batbileg Bor, Eleanor I. Lamont |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Microbiology (medical) Fastidious organism Firmicutes Immunology Dental Plaque Microbiology SHI‐media 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine RNA Ribosomal 16S Prevotella Humans Periodontal Pocket Periodontitis General Dentistry Growth medium Treponema biology Biofilm subgingival microbiota 030206 dentistry Original Articles 16S ribosomal RNA biology.organism_classification In vitro biofilm model 030104 developmental biology chemistry Biofilms Original Article Bacteria |
| Zdroj: | Molecular Oral Microbiology |
| ISSN: | 2041-1014 2041-1006 |
| Popis: | Developing a laboratory model of oral polymicrobial communities is essential for in vitro studies of the transition from healthy to diseased oral plaque. SHI medium is an enriched growth medium capable of supporting in vitro biofilms with similar diversity to healthy supragingival inocula; however, this medium does not maintain the diversity of gram‐negative bacteria more associated with subgingival plaque. Here, we systematically modified SHI medium components to investigate the impacts of varying nutrients and develop a medium capable of supporting a specific disease‐state subgingival community. A diseased subgingival plaque sample was inoculated in SHI medium with increasing concentrations of sucrose (0%, 0.1%, 0.5%), fetal bovine serum (FBS) (0%, 10%, 20%, 30%, 50%), and mucin (0.1, 2.5, 8.0 g/L) and grown for 48 hrs, then the 16S rRNA profiles of the resulting biofilms were examined. In total, these conditions were able to capture 89 of the 119 species and 43 of the 51 genera found in the subgingival inoculum. Interestingly, biofilms grown in high sucrose media, although dominated by acidogenic Firmicutes with a low final pH, contained several uncultured taxa from the genus Treponema, information that may aid culturing these periodontitis‐associated fastidious organisms. Biofilms grown in a modified medium (here named subSHI‐v1 medium) with 0.1% sucrose and 10% FBS had a high diversity closest to the inoculum and maintained greater proportions of many gram‐negative species of interest from the subgingival periodontal pocket (including members of the genera Prevotella and Treponema, and the Candidate Phyla Radiation phylum Saccharibacteria), and therefore best represented the disease community. |
| Databáze: | OpenAIRE |
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