PubMed Health⌕ Search

Biomedical subjects

M Hannig

Publications and source records attributed to M Hannig.

At least 19 recordsLinked to original sources

Does irradiation affect the protein composition of saliva?

The purpose of this study was to compare the relative amount of low molecular weight salivary proteins in patients with head and neck tumours treated with radiotherapy and healthy subjects. Reverse-phase high-pressure liquid chromatography was used for protein separation. Nine protein fractions (including acidic and basic proline-rich proteins (PRPs), cystatins, histatins and statherin) were identified in saliva from irradiated patients as well as healthy subjects. However, compared with non-irradiated healthy subjects, the fraction of acidic PRPs was significantly reduced in irradiated patients. These data indicate an alteration of the relative amount of low molecular weight salivary proteins in irradiated patients besides the reduction of salivary flow.

Acute-Phase Proteins↗

Advances in in vitro and in vivo models for studying the staphylococcal factors involved in implant infections.

Implant infections due to staphylococci are one of the greatest threats facing patients receiving implant devices. For many years researchers have sought to understand the mechanisms involved in the adherence of the bacterium to the implanted device and the formation of the unique structure, the biofilm, which protects the indwelling bacteria from the host defence and renders them resistant to antibiotic treatment. A major goal has been to develop in vitro and in vivo models that adequately reflect the real-life situation. From the simple microtiter plate assay and scanning electron microscopy, tools for studying adherence and biofilm formation have since evolved to include specialised equipment for studying adherence, flow cell systems, real-time analysis of biofilm formation using reporter gene assays both in vitro and in vivo, and a wide variety of animal models. In this article, we discuss advances in the last few years in selected in vitro and in vivo models as well as future developments in the study of adherence and biofilm formation by the staphylococci.

Animals↗

Chlorhexidine-induced ultrastructural alterations in oral biofilm.

Chlorhexidine, the most used biocide in periodontology, alters the permeability of the bacterial cell membrane. However, the chlorhexidine-induced morphological alterations in the oral biofilm have not been studied. To examine the effects of chlorhexidine on oral biofilm on an electron microscopic level, gingival epithelial cells with attached biofilm were collected from 10 volunteers, subjected to 0.1% chlorhexidine for 1 or 5 min, stained with ruthenium red-tetroxide, and analyzed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). SEM visualized the bacterial glycocalyces and the biofilm matrix on the biofilm surface; however, no chlorhexidine-induced alterations were observed. TEM revealed loss of bacterial membrane integrity and fimbrial disintegration in a few bacteria. In the proximity of these alterations, a restricted matrix disintegration was also observed. However, the chlorhexidine-induced alterations only effected a minor part of the oral biofilm and did not cause its disintegration. These findings suggest the insufficient efficiency of chlorhexidine against oral biofilm.

Adolescent↗

Interfacial interaction of tartaric acid with hydroxyapatite and enamel.

The purpose of this study was to investigate the interfacial interaction of tartaric acid with hydroxyapatite and enamel. Hydroxyapatite particles were mixed with 15% (w/v) alcohol-aqueous (1:1) solution of tartaric acid for 72 h. After the mixture, the filtrate was separated from the solid. The filtrate, the solid, hydroxyapatite, and tartaric acid were dried for FTIR and XRD analysis. Enamel disks etched with tartaric acid for 60 s were analyzed by attenuated total reflectance (ATR). Enamel disks etched with 15% tartaric acid for 15, 30, 60, 120 s were analyzed by SEM. FTIR spectra showed carboxylate in the solid and in subtraction spectra. ATR spectra revealed carboxylate on the enamel surfaces. XRD data indicated the formation of calcium tartrate and calcium hydrogen phosphate after the reaction of tartaric acid with hydroxyapatite. SEM observations revealed typical decalcification of the periphery of the enamel rods due tartaric acid etching. It is concluded that tartaric acid can decalcify and chemisorb onto HA simultaneously, hence, tartaric acid could be used as etchant and ingredient of self-etching primers in adhesive dentistry.

Acid Etching, Dental↗

Bacterial internalization in periodontitis.

BACKGROUND: Bacterial invasion of host epithelial cells plays an important role in the pathogenesis of periodontitis; however, the exact mechanism of the invasion has not been investigated. METHODS: Pocket epithelium biopsies in periodontitis were analysed via scanning and transmission electron microscopy using ultra-histochemical staining with ruthenium red for glycocalyx visualization. RESULTS: We demonstrated that oral bacteria adhered via fimbriae-mediated adhesion only. The bacterial internalization in periodontitis was marked by the hallmark of the fimbriae-induced zipper mechanism--the phagocytic cup formation--but we found no sign of the trigger mechanism of internalization. In addition, we frequently observed apoptosis in the phagocytizing epithelial cells. CONCLUSION: Fimbriae-mediated adhesion is a prerequisite for bacterial invasion in periodontitis. This occurs by the fimbriae-induced zipper mechanism of internalization. As internalization induces apoptosis, the subsequent exfoliation might play a significant role in the clearance of periodontal pathogens.

Apoptosis↗

Ex vivo gingival-biofilm consortia.

AIMS: To develop a protocol for harvesting ex vivo samples of gingival-biofilm consortia and to investigate their basic characteristics. METHODS AND RESULTS: Gingival epithelial cells with attached biofilm were collected from healthy subjects by taking a smear. The bacterial viability was estimated via the alteration of the membrane permeability and metabolic activity via the double/single-stranded nucleic acid ratio using a confocal laser-scanning microscope. Morphological analysis was performed by scanning and transmission electron microscopy. Additionally, microbiological estimations were made. The electron microscopy revealed fimbriae-mediated adhesion and the formation of a biofilm matrix. Most bacteria were viable and had a high metabolic activity. CONCLUSIONS: The presented study offers an easy to follow approach for harvesting samples of gingival-biofilm consortia. The latter differs considerably from the supragingival plaque in viability and zonal distribution. Related to free-living and in vitro-grown biofilms, the gingiva-associated biofilm revealed an atypically high metabolic activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Biofilm fragments should possess the basic features of the entire gingiva-associated biofilm; which as yet cannot be simulated in vitro. Thus, samples of ex vivo gingival-biofilm consortia can be used to investigate the resistance of oral biofilms against antibiotics and biocides.

Bacteria↗

Transmission electron microscopy comparison of methods for collecting in situ formed enamel pellicle.

The in vivo formed salivary pellicle is composed of an outer globular and a densely structured basal layer. This study developed a method for selective recovering of these pellicle layers from the enamel surface. Two-hour in situ pellicles were formed by intraoral exposure of enamel specimens in two adults. Pellicle-covered enamel specimens were treated either mechanically (scraping with scaler, curette or razor blade, or rubbing with a sponge) or chemically (phosphate buffer, NaCl, NaOCl, CaCl2, NaSCN, urea, tetrahydrofurane, guanidine, SDS, HCl, or EDTA with or without additional ultrasonication). Specimens were processed for transmission electron microscopic analysis to detect pellicle residues remaining on the enamel surface after the different treatments. Most of the chemical treatments caused partial, incomplete removal of the globular layer. Complete removal of the globular layer without disruption of the basal layer was obtained by sponge rubbing or by CaCl2 combined with ultrasonication, whereas scraping caused partial disruption of the basal layer. Removal of the basal layer was observed after treatment with HCl, EDTA, or NaOCl combined with ultrasonication. Electrophoretical analysis of recovered pellicle fractions indicate that combination of sponge-rubbing followed by EDTA treatment can be recommended for stepwise removal of the globular and basal pellicle layers.

Animals↗

Microleakage and SEM evaluation of fissure sealants placed by use of self-etching priming agents.

OBJECTIVES: This study evaluated the microleakage and internal seal of fissure sealants placed by use of self-etching priming agents in comparison to phosphoric acid etching of enamel. METHODS: Seventy-two caries-free extracted human molars were divided into six groups with 12 teeth each. Occlusal surfaces were cleansed by either pumicing (Groups I, III, V) or by 15-s air-abrasion treatment with 25 microm aluminum oxide particles (Groups II, IV, VI). Fissures were sealed with the self-etching priming systems, Clearfil Liner Bond 2 (Groups I, II) or Resulcin AquaPrime (Groups III, IV). In Groups V and VI, sealants were placed after phosphoric acid etching. Half of the teeth in each group were thermocycled. After staining with 0.5% methylene blue, the teeth were sectioned for evaluation of microleakage. Internal adaptation of the fissure sealants was analyzed by SEM on replicas of cross sections. RESULTS: Independent of the methods used for cleansing of the occlusal surfaces, fissure sealants in Groups I and II showed significantly more microleakage and less sufficient internal seal as compared to sealants placed in Groups III to VI. Sealants placed by Resulcin AquaPrime (Groups III, IV) leaked significantly more than sealants applied after phosphoric acid etching (Groups V, VI) of the enamel. However, statistical analysis (H-test) did not reveal significant differences concerning the internal adaptation of sealants placed in Groups III, IV, V and VI. CONCLUSIONS: Concerning the microleakage data, use of the self-etching bonding systems, Clearfil Liner Bond 2 and Resulcin AquaPrime, cannot be recommended for fissure sealing, since the sealing ability is less effective as compared to the conventional acid-etching technique.

Acid Etching, Dental↗

Initial salivary pellicle formation on solid substrates studied by AFM.

Organic layers of salivary biopolymers adsorbed on soft and hard oral tissues, referred to also as salivary pellicle, play a critical role with respect to all surface phenomena taking place in the oral cavity. The initial stages of pellicle formation are of great interest since they determine the ensuing processes of salivary biopolymer adsorption and subsequent adherence of bacteria. In spite of the important physiological role of the pellicle in protecting the enamel surface against short-term acidic attacks, the composition and ultrastructure of the pellicle layer are not yet understood and resolved in detail. The present study utilized atomic force microscopy (AFM), for the first time, to elucidate the morphogenesis and ultrastructural pattern of initial salivary pellicle formation taking place in situ on solid substrates of mica, silicon wafer and graphite. Using tapping mode AFM, salivary pellicles were found in all intraorally exposed specimens and revealed a globular surface morphology of the adsorbed protein layer. The average diameter and height of the adsorbed salivary proteins were determined to be 15 +/- 3 nm and 2.0 +/- 0.5 nm, respectively. It was also found that the surface energy of the substrates affects the rate of pellicle formation, while the overall size of the adsorbed salivary proteins appears to be identical on all studied substrates.

Adsorption↗

Influence of salivary pellicle formation time on enamel demineralization--an in situ pilot study.

This study assessed the protective potential of salivary pellicles formed in situ over periods ranging from 2 to 24 h. Pellicles were produced on enamel slabs mounted on the palatal aspect of removable acrylic splints and exposed to the oral environment in three subjects for 2, 6, 12 and 24 h. Enamel specimens with and without pellicles were immersed in citric acid (1%) for 60 s, and the amount of dissolved calcium was measured by atomic absorption spectroscopy. In addition, specimens were processed for transmission electron microscopy (TEM). Mean values (standard deviations) for calcium release (mg/l related to the specimen's surface area of 5 x 5 mm(2)) were: 2-h pellicle 6.94 (1.55); 6-h pellicle 6.69 (2.05); 12-h pellicle 6.57 (2.31); 24-h pellicle 5.71 (2.46); enamel without pellicle 8.95 (1.66). There were no significant differences in calcium release that were dependent on pellicle formation time, but in comparison to enamel specimens without pellicle, significantly less (p <0.05) demineralization of the enamel was observed in pellicle-covered specimens. TEM showed that the pellicle was partly, but not completely dissolved following acid exposure. It is concluded that even a 2-h in-situ-formed pellicle layer protects the enamel surface to a certain extent against demineralization.

Analysis of Variance↗

Effect of different luting materials on the marginal adaptation of Class I ceramic inlay restorations in vitro.

OBJECTIVES: The aim of this in vitro study was to assess the marginal adaptation of prefabricated Class I ceramic inlay restorations placed with various luting materials. METHODS: Forty-two standardized occlusal cavities were prepared in extracted human molars with diamond burs exactly corresponding to the dimensions of prefabricated glass ceramic inlays. The prepared teeth were randomly assigned to seven groups of six teeth each and restored using (1). the composite resin Tetric Ceram in increment technique [Group I] or (2). ceramic inlays (Cerana) luted with: the composite based materials Dual Cement [Gr. II] and Panavia 21 [Gr. III], the compomer material Dyract Cem [Gr. IV], Dyract Cem with additional use of Prime & Bond 2.1 [Gr. V], the silicophosphate cement Trans-Lit [Gr. VI], or the ethylcyanoacrylate Cyano-Veneer [Gr. VII]. Marginal adaptation was evaluated by SEM-analyses before and after thermal cycling (2500 cycles; 5-55 degrees C) and mechanical loading (100N; 500000 cycles) using replica models. Kruskal-Wallis H-test and Mann-Whitney U-test were used for statistical analyses. RESULTS: Group I (increment technique) as well as Groups II-V (inlay technique) revealed high percentages of perfect marginal adaptation in over 95% of the analyzed margins, both before and after thermo-mechanical loading. Statistical significant differences could not be detected within these groups. All inlays luted with silicophosphate cement (Group VI) and four of six inlays applied with Cyano-Veneer (Group VII) fractured under occlusal load. SIGNIFICANCE: A stable bonding to the enamel and to the ceramic inlay was achievable with the composite luting resins Dual Cement and Panavia 21 as well as with the compomer based luting material Dyract Cem but not with the use of the silicophosphate cement Trans-Lit or the ethylcyanoacrylate Cyano-Veneer.

Compomers↗

Candida attachment to oral epithelium.

Inflamed oral mucosa biopsies from patients with thrush and high candidal density were observed in a transmission electron microscope (TEM) using ultra-histochemical staining with ruthenium red for glycocalyx visualization. Fimbriae comprising the glycocalyx and enabling yeast adhesion to epithelial cells were clearly visualized by ruthenium red. All internalized portions of the yeast walls were devoid of glycocalyx, indicating that the growing tips of the hyphae mechanically penetrated the host cells. The attachment of Candida occurred in two ways: by fimbria-mediated adhesion enabling colonization of the epithelial surface, and by invasion of the superficial epithelial cells via hyphae. As the interaction between adhesin receptors and adhesins stimulates the production of proinflammatory cytokines, Candida adhesion itself is assumed to induce mucosal inflammation.

Adult↗

Fimbria-mediated bacterial adhesion to human oral epithelium.

Oral mucosa biopsies and saliva samples from 12 individuals were processed for transmission (TEM) and scanning (SEM) electron microscopy with and without ruthenium red staining. Additionally performed microbiological estimations indicated in all bacteriological samples a facultative pathogenic flora. SEM and TEM investigation showed a diverse bacterial flora attached to the mucosal surface. Fimbriae comprising the glycocalyx and enabling bacterial attachment to the epithelial cells could be clearly visualised by ruthenium red. The only mode of bacterial attachment to the oral mucosa detected in the present investigation was fimbria-mediated adhesion and co-adhesion. The fimbria-mediated adhesion enables the bacterial persistence in the oral cavity and is the first step in the bacterial colonisation process.

Adult↗

Effect of air abrasion and resin composite on microleakage of Class V restorations bonded with self-etching primers.

PURPOSE: The purpose of this in vitro study was to determine the effect of air-abrasion pretreatment on microleakage of Class V resin composite restorations bonded with self-etching primers. MATERIALS AND METHODS: A total of 54 Class V cavities prepared either with diamond burs or air abrasion (KCP 1000) was restored with resin composites using adhesive systems with self-etching priming agents (Clearfil Liner Bond 2 or Resulcin AquaPrime + Monobond) and thermocycled 2,500 times. The degree of dye penetration (0.5% methylene blue solution) was scored on an ordinal scale at 25X magnification on longitudinally cut sections of the specimens. Wall adaptation at the tooth-restoration interfaces was evaluated under a SEM using replicas of the sectioned teeth. RESULTS: Severe microleakage occurred at the gingival dentinal margins, and less microleakage was detected at the enamel cavity margins. Nonparametrical statistical analysis (H-test) demonstrated no significant differences in microleakage among the different groups. Comparison of dye penetration data and SEM findings showed that with both self-etching priming agents, microleakage occurred in the absence of gaps. CONCLUSION: Use of air abrasion in combination with self-etching priming agents cannot prevent microleakage at the dentin-resin interface of Class V resin composite restorations.

Air Abrasion, Dental↗

In vitro study of enamel and dentin marginal integrity of composite and compomer restorations placed in primary teeth after diamond or Er:YAG laser cavity preparation.

PURPOSE: The aim of this in vitro study was to evaluate the effect of laser vs diamond instruments on Class II and Class V cavity margins in primary teeth with SEM after restoration with composite and compomer materials and subsequent thermocycling. MATERIALS AND METHODS: Class V cavities with margins completely located in enamel were prepared in 36 extracted primary teeth to examine the restoration-enamel interface. Eighteen cavities were prepared with an Er:YAG laser operated at 3 Hz, 300 mJ/pulse, and 18 with a spherical fine diamond bur. In 24 primary teeth, Class II cavities with cervical proximal margins located in dentin were prepared for examination of the restoration-dentin interface. All Class II cavities were prepared with a conventional cylindrical diamond bur. In 12 cavities, the margins located in dentin were conditioned with an Er:YAG laser (1 Hz, 100 mJ/pulse). All cavities were restored with a fine hybrid composite resin or a compomer material. RESULTS: SEM examination of Class V cavities after thermocycling (between 5 degrees C and 55 degrees C, 2500 cycles) revealed over 90% "perfect" margins in all composite fillings and in those compomer restorations applied after conventional preparation and acid etching. Compomer restorations placed without acid etching revealed a significantly superior marginal integrity in laser-prepared cavities (81% perfect margins) vs conventionally prepared cavities (60% perfect margins). SEM analysis of the dentinal margins of both compomer and composite restorations in Class II cavities showed mean percentages of perfect marginal adaptation ranging from only 13% to 66%. CONCLUSION: Er:YAG laser treatment can be recommended for composite restorations in Class V cavities in primary teeth. Compomer restorations can be placed after conventional preparation of Class V cavities and acid etching or after laser preparation. Dentin bonding in Class II cavities in primary teeth is not sufficient and cannot be improved with Er:YAG laser pretreatment.

Acid Etching, Dental↗

Ultrastructural investigation of pellicle morphogenesis at two different intraoral sites during a 24-h period.

The purpose of the present in vivo study was to examine salivary pellicle formation on enamel surfaces at two different intraoral sites for periods of 1 min up to 24 h by means of transmission electron microscopy. Bovine enamel specimens were attached to the buccal and lingual surfaces of the upper first molars in three subjects using removable intraoral splints. Specimens were carried over period of 1, 10, 30 and 60 min, 2, 6 and 24 h and were processed for transmission electron microscopy. After 1 min, an electron dense pellicle layer, 10-20 nm thick, was observed on the enamel surfaces. The subsequent adsorption of salivary biopolymers was governed by local influences of the oral cavity. Specimens located on the lingual aspect were covered within 2 h by a 20- to 80-nm-thick, homogeneous, predominantly granular-structured pellicle. The thickness of the surface coatings that were adsorbed on lingually carried specimens increased to 100-200 nm after 24 h. In contrast, on the buccally mounted specimen surfaces, a variably structured pellicle with granular and globular components could be detected after intraoral exposure for 2 h. The thickness of the 2-h buccal pellicles ranged between 200 and 700 nm. After 24 h, the buccally positioned specimens were covered by a dense globular pellicle layer varying in thickness from 1000 to 1300 nm. It is suggested that in vivo pellicle formation is initiated by adsorption of an electron-dense layer of salivary proteins. Further adsorption of salivary biopolymers leads to the formation of an outer loosely arranged pellicle layer. Under oral conditions, the locally available salivary biopolymers and the influence of locally effective shearing forces are of significance for the ultrastructural pattern and extent of pellicle formation.

Adult↗

Effect of Carisolv solution on sound, demineralized and denatured dentin--an ultrastructural investigation.

The purpose of this in vitro study was to analyze the micromorphological changes caused by Carisolv gel on sound, demineralized, and denatured dentin. Fractured dentinal surfaces, dentinal surfaces demineralized superficially by phosphoric acid etching and dentinal surfaces denatured due to lactic acid and collagenase pretreatment were exposed to freshly mixed Carisolv gel or 0.25% sodium hypochlorite. No additional mechanical action was exerted during the 20-min exposure of specimens to the Carisolv solution. Specimens were evaluated by transmission electron microscopy. Electron microscopic evaluation did not indicate any ultrastructural changes of the fractured or demineralized dentinal surfaces due to the 20-min Carisolv treatment. Denatured dentin was partially removed within a 20 min period of chemical action of the Carisolv solution leaving only a 1- to 2-micron thick layer of residual denatured dentin on the specimen's surface. In contrast, 0.25% sodium hypochlorite treatment completely dissolved the demineralized as well as denatured dentin layer within 20 min. It is concluded that Carisolv gel (1) does not affect sound fractured dentin, (2) does not dissolve demineralized dentin, and (3) has a limited potential to chemically dissolve denatured dentin.

Cariostatic Agents↗