Action of food preservatives on 14-days dental biofilm formation, biofilm vitality and biofilm-derived enamel demineralisation in situ

Arweiler, Nicole Birgit; Netuschil, L; Beier, Daniel; Grunert, Susanne C.; Heumann, Christian; Altenburger, Markus Jörg; Sculean, Anton; Nagy, Katalin; Al‐Ahmad, Ali; Auschill, Thorsten Mathias

doi:10.1007/s00784-013-1053-9

Cited by 9 publications

(7 citation statements)

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“…The most commonly used techniques in these studies are those based on visualizing the oral biofilm with fluorescence microscopes [both epifluorescence and Confocal Laser Scanning Microscope (CLSM)] (Benelli et al, 1993 ; Netuschil et al, 1998 ; Wood et al, 1999 , 2000 , 2002 ; Giertsen et al, 2000 ; Auschill et al, 2001 , 2002 , 2004 , 2005 ; Palmer et al, 2003 ; Arweiler et al, 2004 ; Al-Ahmad et al, 2007 , 2009 , 2010a , c , 2013 ; Chalmers et al, 2007 ; Dige et al, 2007 , 2009a , b ; Arweiler et al, 2008 ; Hannig et al, 2013a , b , c ; Beyth et al, 2010 ; Burgers et al, 2010 ; Dong et al, 2010 ; Gosau et al, 2010 ; Jung et al, 2010 ; von Ohle et al, 2010 ; Bremer et al, 2011 ; Gu et al, 2012 ; Rupf et al, 2012 ; García-Caballero et al, 2013 ; He et al, 2013 ; Kensche et al, 2013 ; Tomás et al, 2013 ; Arweiler et al, 2014 ; Grychtol et al, 2014 ; Padovani et al, 2015 ; Prada-López et al, 2015a , b ; Quintas et al, 2015a , b ) (45 studies), mainly combined with fluorescence in situ hybridization (FISH) and 4′,6-diamidino-2-phenylindole (DAPI) for bacterial identification. In the case of studies aiming for the analysis of bacterial viability, fluorescence microscopes have usually been combined with staining dyes for live/dead bacterial identification such as SYTO 9/Propidium Iodide and Fluorescein Diacetate/Etidium Bromide.…”

Section: Resultsmentioning

confidence: 99%

“…On the other hand, buccal devices have been used extensively, allowing the definition of six different groups: AcD (Figures 4A,B ) (Nyvad and Fejerskov, 1987a , b , 1989 ; Nyvad and Kilian, 1987 ; Hannig, 1997 ; Netuschil et al, 1998 ; Hannig, 1999a , b ; Fine et al, 2000 ; Auschill et al, 2001 , 2002 ; Jentsch et al, 2002 ; Palmer et al, 2003 ; Scarano et al, 2003 ; Groessner-Schreiber et al, 2004 ; Diaz et al, 2006 ; Chalmers et al, 2007 ; Hannig et al, 2007b ; Scotti et al, 2007 ; Grossner-Schreiber et al, 2009 ; Sreenivasan et al, 2009 ; Azevedo et al, 2012 ; Rehman et al, 2012 ; Do Nascimento et al, 2013 ; Nascimento et al, 2014 ), LiD (Figures 5A,B ; Strassler et al, 1986 ; Robinson et al, 1997 ; Wood et al, 1999 , 2000 , 2002 ; Arai et al, 2000 ; Shore et al, 2001 ; Kato et al, 2004 ; Robinson et al, 2006 ; Dong et al, 2010 ; Kato et al, 2012 ; He et al, 2013 ), AcMD (Figures 6A,B ; Rimondini et al, 1997 ; Zucchelli et al, 1998 ; Giertsen et al, 2000 ; Zucchelli et al, 2000 ; Arweiler et al, 2004 ; Auschill et al, 2004 , 2005 ; Al-Ahmad et al, 2007 , 2010a , c , 2013 ; Dige et al, 2007 ; Arweiler et al, 2008 , 2014 ; Dige et al, 2009a , b ; von...…”

Section: Resultsunclassified

“…Despite the importance of these parameters, only one study which discussed the volunteer's experience with the device was found after this review (Prada-López et al, 2015b ). On the other hand, some papers have registered dropouts by the volunteers during the experiment, because of discomfort (Brambilla et al, 2012 ) or unclear reasons (Arweiler et al, 2014 ).…”

Section: Discussionmentioning

confidence: 99%

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Devices for In situ Development of Non-disturbed Oral Biofilm. A Systematic Review

et al. 2016

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Objective: The aim of this review was to assess the types of devices used for in situ development of oral biofilm analyzed microbiologically.Materials and Methods: A systematic search of the literature was conducted to identify all in situ studies of oral biofilm which used an oral device; the Ovid MEDLINE and EMBASE databases complemented with manual search were used. Specific devices used to microbiologically analyze oral biofilm in adults were included. After reading of the selected full texts, devices were identified and classified according to the oral cavity zone and manufacturing material. The “ideal” characteristics were analyzed in every group.Results: The search provided 787 abstracts, of which 111 papers were included. The devices used in these studies were classified as palatal, lingual or buccal. The last group was sub-classified in six groups based on the material of the device. Considering the analyzed characteristics, the thermoplastic devices and the Intraoral Device of Overlaid Disk-holding Splints (IDODS) presented more advantages than limitations.Conclusions: Buccal devices were the most commonly used for the study of in situ biofilm. The majority of buccal devices seemed to slightly affect the volunteer's comfort, the IDODS being the closest to the “ideal” model.Clinical Relevance: New devices for in situ oral biofilm microbiological studies should take into account the possible effect of their design on the volunteer's comfort and biofilm formation.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsunclassified

Section: Discussionmentioning

confidence: 99%

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Devices for In situ Development of Non-disturbed Oral Biofilm. A Systematic Review

et al. 2016

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“…It is to question why the “traditional” VF stains of FDA and EB used in oral biofilm research should be replaced with other substances that exert a similar health risk (see next paragraph) and are not proven to be suitable and reliable in biofilm studies. Regarding FDA/EB, an assortment of existing publications (apart from numerous cell culture and cytotoxicity investigations) can be cited from research groups Netuschil [ 108 - 110 , 112 , 114 , 115 , 118 - 124 ], Brecx [ 125 - 128 ], Arweiler/Auschill [ 113 , 129 - 143 ] and others [ 144 - 155 ]. In this context the FDA/EB vital fluorescence staining was routinely used together with Confocal Laser Scanning Microscopy (CLSM) [ 119 , 124 , 131 , 134 - 136 , 139 , 140 , 143 , 153 ] to establish the three-dimensional vitality pattern of the (early) dental biofilm or to document the antibacterial effects of dental materials, mouthrinse solutions as well as food preservatives.…”

Section: Discussionmentioning

confidence: 99%

Confusion over live/dead stainings for the detection of vital microorganisms in oral biofilms - which stain is suitable?

Auschill²,

et al. 2014

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BackgroundThere is confusion over the definition of the term “viability state(s)” of microorganisms. “Viability staining” or “vital staining techniques” are used to distinguish live from dead bacteria. These stainings, first established on planctonic bacteria, may have serious shortcomings when applied to multispecies biofilms. Results of staining techniques should be compared with appropriate microbiological data.DiscussionMany terms describe “vitality states” of microorganisms, however, several of them are misleading. Authors define “viable” as “capable to grow”. Accordingly, staining methods are substitutes, since no staining can prove viability.The reliability of a commercial “viability” staining assay (Molecular Probes) is discussed based on the corresponding product information sheet: (I) Staining principle; (II) Concentrations of bacteria; (III) Calculation of live/dead proportions in vitro. Results of the “viability” kit are dependent on the stains’ concentration and on their relation to the number of bacteria in the test. Generally this staining system is not suitable for multispecies biofilms, thus incorrect statements have been published by users of this technique.To compare the results of the staining with bacterial parameters appropriate techniques should be selected. The assessment of Colony Forming Units is insufficient, rather the calculation of Plating Efficiency is necessary. Vital fluorescence staining with Fluorescein Diacetate and Ethidium Bromide seems to be the best proven and suitable method in biofilm research.Regarding the mutagenicity of staining components users should be aware that not only Ethidium Bromide might be harmful, but also a variety of other substances of which the toxicity and mutagenicity is not reported.Summary– The nomenclature regarding “viability” and “vitality” should be used carefully.– The manual of the commercial “viability” kit itself points out that the kit is not suitable for natural multispecies biofilm research, as supported by an array of literature.– Results obtained with various stains are influenced by the relationship between bacterial counts and the amount of stain used in the test. Corresponding vitality data are prone to artificial shifting.– As microbiological parameter the Plating Efficiency should be used for comparison.– Ethidium Bromide is mutagenic. Researchers should be aware that alternative staining compounds may also be or even are mutagenic.

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“…All volunteers received individually customized acrylic appliances for the upper jaw into which six plasma sterilized bovine enamel discs (diameter 3 mm, height 2 mm) were inserted. The three discs on each side were positioned towards the interdental area of two teeth in order to generate and mimic approximal biofilms (for details see [ 31 ] and Figure 1 ). A total of 1872 enamel discs were prepared for the eight wearing cycles.…”

Section: Methodsmentioning

confidence: 99%

Influence of Probiotics on the Salivary Microflora Oral Streptococci and Their Integration into Oral Biofilm

et al. 2020

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Probiotics’ ability to integrate into dental biofilms is not yet clarified. The aim of this trial was to detect probiotic bacteria from probiotic products in dental biofilm and saliva during and after intake. In this parallel, randomized clinical trial, 39 subjects wore customized appliances to build up intra-oral biofilms (72-h periods). The trial was divided into screening (S) to determine baseline biofilm flora, intervention (I), and wash out (WO). During I (28 days), subjects consumed a product containing (a) Enterococcus faecalis (b) Lactobacilluscasei, or (c) Lactobacillus rhamnosus GG. Probiotic bacteria and Streptococci spp. were detected in the biofilms and saliva of the 35 subjects that were included in the analysis. During I and WO, the ratio of probiotics in the biofilm was very low compared to total bacterial load, while saliva had slightly but not significantly higher values. No significant changes of probiotic bacteria (p > 0.05) were found at any visit during I or WO. The proportion of streptococci was significantly reduced (p < 0.05) during I and even lower in WO, compared to S. Probiotic bacteria could neither integrate nor persist in dental biofilm and saliva but did influence the growth of streptococci in biofilm and saliva.

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Action of food preservatives on 14-days dental biofilm formation, biofilm vitality and biofilm-derived enamel demineralisation in situ

Cited by 9 publications

References 53 publications

Devices for In situ Development of Non-disturbed Oral Biofilm. A Systematic Review

Devices for In situ Development of Non-disturbed Oral Biofilm. A Systematic Review

Confusion over live/dead stainings for the detection of vital microorganisms in oral biofilms - which stain is suitable?

Influence of Probiotics on the Salivary Microflora Oral Streptococci and Their Integration into Oral Biofilm

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