Monitoring of extracellular pH in young dental biofilms grown in vivo in the presence and absence of sucrose
Background and objective: pH in dental biofilms is of central importance for the development of caries. We used the ratiometric pH-sensitive dye C-SNARF-4 in combination with digital image analysis to monitor extracellular pH in dental biofilms grown in situ with and without sucrose supply. Design:...
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2016-02-01
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doaj-c0c93735b3c14dfd9773b8341bb047aa2020-11-24T21:01:40ZengTaylor & Francis GroupJournal of Oral Microbiology2000-22972016-02-01801810.3402/jom.v8.3039030390Monitoring of extracellular pH in young dental biofilms grown in vivo in the presence and absence of sucroseIrene Dige0Vibeke Baelum1Bente Nyvad2Sebastian Schlafer3Department of Dentistry, HEALTH, Aarhus University, Aarhus, DenmarkDepartment of Dentistry, HEALTH, Aarhus University, Aarhus, DenmarkDepartment of Dentistry, HEALTH, Aarhus University, Aarhus, DenmarkDepartment of Dentistry, HEALTH, Aarhus University, Aarhus, DenmarkBackground and objective: pH in dental biofilms is of central importance for the development of caries. We used the ratiometric pH-sensitive dye C-SNARF-4 in combination with digital image analysis to monitor extracellular pH in dental biofilms grown in situ with and without sucrose supply. Design: Dental biofilms (48 h) from 10 individuals were collected on glass slabs mounted on intra-oral appliances. During growth, appliances were immersed extra-orally in either physiological saline or 4% sucrose for 2 min, eight times per day. Fluorescence emissions of C-SNARF-4 in deep layers of the biofilms were recorded ex vivo with confocal microscopy for 15 min or for 1 h after exposure to 0.4% glucose. Extracellular pH was determined ratiometrically using digital image analysis. Results: Extracellular pH dropped rapidly in most examined sites after addition of glucose. Distinct pH microenvironments were observed within single biofilms. The variation in pH was similar between sites within the same biofilm and sites from different individuals. pH drop patterns did not differ between biofilms exposed to sucrose-free and sucrose-rich environments. Conclusion: The present study is the first of its kind to apply the combination of pH ratiometry and digital image analysis to systematically record extracellular pH in intact dental biofilms from several individuals for up to 1 h. We observed highly heterogeneous pH landscapes and the presence of acidogenic microenvironments – ‘acidogenic hotspots’ – within the biofilms. The data suggest that pH drops in young (48 h) dental biofilms are independent of the sucrose supply during growth.http://www.journaloforalmicrobiology.net/index.php/jom/article/view/30390/45004pHdental biofilmsC-SNARF-4imagingsucroseratiometric imagingdigital image analysis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Irene Dige Vibeke Baelum Bente Nyvad Sebastian Schlafer |
spellingShingle |
Irene Dige Vibeke Baelum Bente Nyvad Sebastian Schlafer Monitoring of extracellular pH in young dental biofilms grown in vivo in the presence and absence of sucrose Journal of Oral Microbiology pH dental biofilms C-SNARF-4 imaging sucrose ratiometric imaging digital image analysis |
author_facet |
Irene Dige Vibeke Baelum Bente Nyvad Sebastian Schlafer |
author_sort |
Irene Dige |
title |
Monitoring of extracellular pH in young dental biofilms grown in vivo in the presence and absence of sucrose |
title_short |
Monitoring of extracellular pH in young dental biofilms grown in vivo in the presence and absence of sucrose |
title_full |
Monitoring of extracellular pH in young dental biofilms grown in vivo in the presence and absence of sucrose |
title_fullStr |
Monitoring of extracellular pH in young dental biofilms grown in vivo in the presence and absence of sucrose |
title_full_unstemmed |
Monitoring of extracellular pH in young dental biofilms grown in vivo in the presence and absence of sucrose |
title_sort |
monitoring of extracellular ph in young dental biofilms grown in vivo in the presence and absence of sucrose |
publisher |
Taylor & Francis Group |
series |
Journal of Oral Microbiology |
issn |
2000-2297 |
publishDate |
2016-02-01 |
description |
Background and objective: pH in dental biofilms is of central importance for the development of caries. We used the ratiometric pH-sensitive dye C-SNARF-4 in combination with digital image analysis to monitor extracellular pH in dental biofilms grown in situ with and without sucrose supply. Design: Dental biofilms (48 h) from 10 individuals were collected on glass slabs mounted on intra-oral appliances. During growth, appliances were immersed extra-orally in either physiological saline or 4% sucrose for 2 min, eight times per day. Fluorescence emissions of C-SNARF-4 in deep layers of the biofilms were recorded ex vivo with confocal microscopy for 15 min or for 1 h after exposure to 0.4% glucose. Extracellular pH was determined ratiometrically using digital image analysis. Results: Extracellular pH dropped rapidly in most examined sites after addition of glucose. Distinct pH microenvironments were observed within single biofilms. The variation in pH was similar between sites within the same biofilm and sites from different individuals. pH drop patterns did not differ between biofilms exposed to sucrose-free and sucrose-rich environments. Conclusion: The present study is the first of its kind to apply the combination of pH ratiometry and digital image analysis to systematically record extracellular pH in intact dental biofilms from several individuals for up to 1 h. We observed highly heterogeneous pH landscapes and the presence of acidogenic microenvironments – ‘acidogenic hotspots’ – within the biofilms. The data suggest that pH drops in young (48 h) dental biofilms are independent of the sucrose supply during growth. |
topic |
pH dental biofilms C-SNARF-4 imaging sucrose ratiometric imaging digital image analysis |
url |
http://www.journaloforalmicrobiology.net/index.php/jom/article/view/30390/45004 |
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