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Time-lapse confocal microscopy to study in vitro Streptococcus mutans surface colonization.

Jason T F Wing1, Michael A L Hayashi1, Aneesa F Redissi1

  • 1Department of Epidemiology, University of Michigan School of Public Health, Ann Arbor, MI 48109, United States.

Letters in Applied Microbiology
|February 8, 2024
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Sucrose promotes dense, immobile Streptococcus mutans biofilms, while sucralose and no sweetener result in loose, mobile aggregates. This study highlights a new flowcell method for observing early biofilm development.

Keywords:
in vitro modelsbiofilmsmethod developmentmicrobial-cell interactionstreptococci

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Area of Science:

  • Microbiology
  • Biomaterials Science
  • Dental Research

Background:

  • Streptococcus mutans is a key cariogenic bacterium responsible for dental caries.
  • Biofilm formation by S. mutans on tooth surfaces is crucial for its virulence.
  • Understanding initial adhesion and accumulation is vital for developing preventive strategies.

Purpose of the Study:

  • To develop a flowcell system for time-lapse confocal microscopy.
  • To compare S. mutans adhesion and accumulation in different media conditions.
  • To analyze the impact of sucrose and sucralose on early biofilm formation.

Main Methods:

  • A 3D-printed flowcell system was utilized.
  • Fluorescent S. mutans was cultured in unsupplemented, sucrose-, or sucralose-supplemented media.
  • Time-lapse confocal microscopy and a novel object-movement-based image analysis were employed.

Main Results:

  • Sucrose-supplemented media led to small, dense, immobile S. mutans clumps.
  • Unsupplemented and sucralose-supplemented media resulted in large, loose, mobile aggregates.
  • Significant differences (P < 0.05) in architectural metrics and movement were observed with sucrose.

Conclusions:

  • The developed flowcell system is effective for studying initial biofilm formation.
  • Sucrose significantly enhances the adhesion and reduces the mobility of S. mutans biofilms.
  • Sucralose does not promote the same dense biofilm structure as sucrose.