Fluorescence microscopic visualization of non cellular components during initial bioadhesion in situ

Kensche, Anna; Basche, Sabine; Bowen, William H.; Hannig, Matthias; Hannig, Christian

doi:10.1016/j.archoralbio.2013.07.006

Cited by 28 publications

(33 citation statements)

References 46 publications

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“…Aside from pellicle ultrastructure and GTF distribution, the initial bioadhesion patterns were visualized additionally. Thereby, simultaneous immunofluorescencebased visualization of bacteria, glucans and enzymes took place [Kensche et al, 2013]. DAPI staining indicates adhering bacteria at the pellicle layer as well as bacteria-glucan agglomerates in combination with the glucan staining (concanavalin A) .…”

Section: Discussionmentioning

confidence: 99%

“…In the present study lysozyme, peroxidase, α-amylase and GTF B, C, and D were visualized in the in situ formed pellicle by immunofluorescence microscopy [Kensche et al, 2013]. For this purpose, in situ pellicles on enamel slabs were labeled with specific fluorophore-conjugated markers [primary antibodies: mouse monoclonal immunoglobulins against GTF B and rabbit polyclonal immunoglobulins against GTF C and GTF D; secondary antibodies: anti-mouse GTF B, Promocell, Heidelberg, and antirabbit immunoglobulin G (GTF C and D), Biotrend, Cologne, Germany].…”

Section: Immunofluorescence Microscopic Visualization Of α-Amylase Gmentioning

confidence: 99%

“…The specific secondary antibodies were conjugated with Dylight 488 (green), Texas red (red) or FITC (green) and incubated with the enamel slabs for 2 h at room temperature or overnight at 4 ° C, each in a dark chamber. Controls were simultaneously conducted to validate the correctness of the labeling [Kensche et al, 2013]. The glucan visualization was performed with Alexa Fluor 574-conjugated concanavalin A .…”

Section: Immunofluorescence Microscopic Visualization Of α-Amylase Gmentioning

confidence: 99%

“…For in situ pellicle formation, individual upper jaw splints were customized from stainless-steel clamps and polymethylmethacrylate [Kensche et al, 2013;Weber et al, 2015]. Cavities for the insertion of the enamel slabs were prepared on the buccal sides of the premolars and the 1st molar on both sides ( n = 8/splint).…”

Section: Pellicle Formationmentioning

confidence: 99%

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Enzymology and Ultrastructure of the in situ Pellicle in Caries-Active and Caries-Inactive Patients

et al. 2017

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Aim: The present study aimed to evaluate the impact of caries activity on the key enzymes and the ultrastructure of the in situ pellicle. Methods: Pellicle formation was performed on bovine enamel slabs. Intraoral exposure (3, 30, and 120 min) was accomplished by 14 caries-active (DMFS: 22.7 ± 12.1) and 13 caries-inactive (DMFS: 1.5 ± 1.8) individuals. The enzyme activities (lysozyme, peroxidase, α-amylase, glycosyltransferase [GTF]) in the in situ pellicle and resting saliva of all participants were analyzed directly after oral exposure. In addition, a simultaneous visualization of these enzymes, extracellular glucans, and adherent bacteria was carried out. Fluorescent patterns were analyzed with fluorescence labeling and 4′,6-diamidino-2-phenylindole/concanavalin A staining. In addition, the distribution of GTF B, C, and D and the ultrastructure of the pellicle were examined by gold immunolabeling and transmission electron microscopy with selected samples. Results: Enzyme activities of amylase, peroxidase, lysozyme, and GTF were detected on all enamel slabs in an active conformation. Neither exposure time nor caries activity had an impact on the enzyme activities. Gold immunolabeling indicated that the pellicle of caries-active subjects tends to more GTF D molecules. The pellicles of caries-inactive and -active individuals revealed a similar ultrastructural pattern. Conclusion: The enzyme activities as well as the pellicle's ultrastructure are of high similarity in caries-active and -inactive subjects. Thereby, oral exposure time has no significant influence. This reflects a high uniformity during the initial phase of bioadhesion (3-120 min) concerning enzymatic functions. However, there is a tendency towards more GTF D in caries-active individuals.

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Section: Discussionmentioning

confidence: 99%

Section: Immunofluorescence Microscopic Visualization Of α-Amylase Gmentioning

confidence: 99%

Section: Immunofluorescence Microscopic Visualization Of α-Amylase Gmentioning

confidence: 99%

Section: Pellicle Formationmentioning

confidence: 99%

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Enzymology and Ultrastructure of the in situ Pellicle in Caries-Active and Caries-Inactive Patients

et al. 2017

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“…To generate in situ pellicles, individual maxillary wire/polymethyl methacrylate splints were customized for the volunteers by a dental technician [Hannig et al, 2013;Kensche et al, 2013a]. At the buccal sides of the splints, cavities were prepared in the regions 14-16 and 24-26.…”

Section: Formation Of In Situ Pelliclementioning

confidence: 99%

Application of Plant Extracts for the Prevention of Dental Erosion: An in situ/in vitro Study

et al. 2015

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Objectives: Antiadherent and antibacterial effects of certain plant extracts have been proven to be beneficial in preventive dentistry. In the present in situ/in vitro crossover study, the impact of plant extracts rich in polyphenols on the erosion-protective properties of the in situ pellicle was evaluated. Methods: Individual splints were prepared for 12 subjects for intraoral exposure of bovine enamel specimens. Following formation of a 1-min pellicle, watery plant extracts (leaves of the wild form of Ribes nigrum, the wild form of Origanum as well as a combination of both) were administered for 10 min in situ. Alternatively, a mouth rinse with fluorides (Elmex Kariesschutz) was performed for 1 min. After further oral exposure for 19/28 min, respectively, slabs were removed and incubated with HCl in vitro over 120 s (pH 2, 2.3, 3). The resulting calcium and phosphate release was quantified photometrically. Slabs with and without a 30-min in situ pellicle served as controls. The modification of pellicle ultrastructure was evaluated by transmission electron microscopy (TEM). Results: Plant extracts modulated the erosion-protective properties of the native in situ pellicle in all test groups in a pH-dependent manner. The combination of R. nigrum leaves and Origanum enhanced the protective properties of the pellicle at all pH values; the administration of this preparation was comparable, yet superior, to the effect of the fluoridated mouth rinse. TEM images indicated that rinsing with R. nigrum leaves/Origanum yielded a distinctly thicker and more electron-dense pellicle. Conclusion: The combination of certain plant extracts offers a novel approach to the complementary prevention of dental erosion.

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Bioadhesion and oral fluids—perireceptor modulators of taste perception?

Hannig

Morzel

2016

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Fluorescence microscopic visualization of non cellular components during initial bioadhesion in situ

Cited by 28 publications

References 46 publications

Enzymology and Ultrastructure of the in situ Pellicle in Caries-Active and Caries-Inactive Patients

Enzymology and Ultrastructure of the in situ Pellicle in Caries-Active and Caries-Inactive Patients

Application of Plant Extracts for the Prevention of Dental Erosion: An in situ/in vitro Study

Bioadhesion and oral fluids—perireceptor modulators of taste perception?

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