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XTT assay of ex vivo saliva biofilms to test antimicrobial influences

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Author(s): Ina Koban | Rutger Matthes | Nils-Olaf Hübner | Alexander Welk | Rabea Sietmann | Jürgen Lademann | Axel Kramer | Thomas Kocher

Journal: GMS Krankenhaushygiene Interdisziplinär
ISSN 1863-5245

Volume: 7;
Issue: 1;
Start page: Doc06;
Date: 2012;
Original page

Keywords: biofilm model | saliva | S. mutans | ozone | chlorhexidine | XTT assay

ABSTRACT
Objective: Many dental diseases are attributable to biofilms. The screening of antimicrobial substances, in particular, requires a high sample throughput and a realistic model, the evaluation must be as quick and as simple as possible. For this purpose, a colorimetric assay of the tetrazolium salt XTT (sodium 3'-[1-[(phenylamino)-carbony]-3,4-tetrazolium]-bis(4-methoxy-6-nitro)benzene-sulfonic acid hydrate) converted by saliva biofilms is recommended. Cleavage of XTT by dehydrogenase enzymes of metabolically active cells in biofilms yields a highly colored formazan product which is measured photometrically. Materials and method: The suitability of the XTT assay for detecting the vitality of ex vivo saliva biofilms was tested to determine the efficacy of chlorhexidine and ozone versus saliva biofilms grown on titanium discs. Results: The XTT method lends itself to testing the vitality of microorganisms in saliva biofilms. The sensitivity of the arrays requires a specific minimum number of pathogens, this number being different for planktonic bacteria and those occurring in biofilms. The antibacterial effect after treatment with chlorhexidine or ozone was measured by XTT conversion that was significantly reduced. The antimicrobial efficacy of 60 s 0.5% and 0.1% chlorhexidine treatment was equal and comparable with 60 s ozone treatment. Conclusion: The XTT assay is a suitable method to determine the vitality in saliva biofilms, permitting assessment of the efficacy of antimicrobial substances. Its quick and easy applicability renders it especially suitable for screening.
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