Biomedical subjects

Matthias Hannig

Publications and source records attributed to Matthias Hannig.

At least 19 recordsLinked to original sources

PubMed2026

Proteomic profile of the dentine pellicle modified with plant polyphenols and fluoride.

OBJECTIVES: Despite its protective role, the dentine pellicle has rarely been studied, therefore we aimed to map out the proteomic profile of in vitro dentine pellicles before and after modification. MATERIALS AND METHODS: A total of 135 human dentine specimens were prepared. After initial pellicle formation with 150 µl pooled human saliva (37 °C, 30 min), the dentine specimens were immersed in one of 9 pellicle modification solutions (2 ml/specimen): deionized water (non-modified pellicle), SnCl2/NaF/AmF (commercial solution containing 800 ppm Sn2+ and 500 ppm F-), NaF solution (500 ppm F- ), and six polyphenol solutions (2 mg / ml) with or without 500 ppm F-: blueberry extract (BBE and BBE + F-), green tea extract (GTE and GTE + F-) and grape seed extract (GSE and GSE + F-). After another aliquot of saliva (150 µl, 37 °C, 60 min), the pellicles were harvested with sodium dodecyl sulphate by rubbing with cotton balls, and taken to proteomic analyses by Liquid Chromatography-Tandem Mass Spectrometry after tryptic digestion. RESULTS: A total of 382 proteins were identified in all the proteomic analyses for all groups. Pellicle modification with fluoride, either NaF or SnCl2/NaF/AmF, led to the presence of 12 or 14 exclusive proteins, respectively, whereas modification with the solutions containing polyphenols presented less exclusive proteins (4-6 proteins). The number of exclusive proteins was even lower for when polyphenols and fluoride (GTE + F- and GSE + F-) were used, with lower abundance of proteases. CONCLUSIONS: We conclude that NaF and SnCl2/NaF/AmF significantly modify the proteome of the dentine pellicle. The combination of fluoride with polyphenols further lowers the abundance of proteins and proteases, which explains the positive effect of these solutions on the dentine pellicles. CLINICAL RELEVANCE: Plant extract solutions with fluoride can significantly modify the proteomic structure of the dentine pellicle, which clarifies the mechanism of action of polyphenols on the protection of dentine demineralization.

Humans↗

PubMed2006

Protective effect of the in situ pellicle on dentin erosion - an ex vivo pilot study.

AIM: The acquired pellicle is well known as an anti-erosive proteinaceous layer on enamel, but its protective properties on dentin have not been investigated in detail until now. The aim of the present ex vivo study was to evaluate the erosive effects on pellicle coated dentin. METHODS: Bovine dentin slabs were exposed to the oral cavity of one subject for 120 min for in situ pellicle formation. Subsequently, the slabs were incubated with HCl (pH 2.3) in vitro for 5 min and erosive calcium-release was measured photometrically. In addition, the acid treated specimens were evaluated by transmission electron microscopy (TEM). Pellicle free samples served as controls. RESULTS: Calcium erosion from the pellicle coated dentin slabs amounted to 23.5+/-2.9 microg Ca/min (pellicle free samples: 32.2+/-4.2 microg Ca/min). The difference was statistically significant (p < or = 0.05). In pellicle coated as well as in uncoated dentin samples, TEM-evaluation showed a demineralised dentinal surface layer which thickness ranged between 3 and 6 microm. The pellicle itself was partially dissolved but not removed by hydrochloric acid treatment. CONCLUSION: The protective properties of the acquired pellicle against an erosive challenge of the dentinal surface are limited. The dentinal pellicle functions like an ion permeable network rather than a barrier.

Adult↗

PubMed2006

Influence of different restorative materials on lysozyme and amylase activity of the salivary pellicle in situ.

Lysozyme and amylase are the most abundant enzymatic components in the salivary pellicle. The purpose of the present study was to determine the influence of different substrata on amylase and lysozyme activity in salivary pellicles formed in situ. Slabs (5 mm diameter) of bovine dentine and enamel, of titanium, gold alloy, resin composite, PMMA, amalgam, and feldspar ceramic were fixed on the buccal sites of individual splints worn by six subjects for 30 min to allow pellicle formation. Thereafter, slabs were removed from the trays and rinsed with running water. Lysozyme activity was determined via lysis of Micrococcus lysodeicticus. Amylase activity was measured with a photometric method using 2-chloro-4-nitrophenyl-4-O-beta-D-galactopyranosylmaltotriosid (GalG2CNP) as substrate. Both pellicle enzymes were evaluated in the immobilized as well as in the desorbed state. Salivary enzyme activities were also measured. All investigated pellicles exhibited lysozyme and amylase activity. Great intraindividual and interindividual differences were observed. Over all samples, immobilized amylase activity amounted to 0.65 +/- 0.64 mU/cm2. Immobilized lysozyme activity was 5.04 +/- 1.55 U/cm2. There were no major effects of the substratum on pellicle-bound amylase and lysozyme activity. Immobilized and desorbed enzyme activities revealed a strong correlation (lysozyme: r = 0.700; amylase: r = 0.990). Salivary enzyme activities had only little impact on pellicle-bound enzyme activities. Amylase and lysozyme are incorporated in the acquired in situ pellicle on different solid surfaces in an active conformation. Dental material and enzyme activity in the saliva have only little impact on enzymatic activity in the pellicle in situ.

Adult↗

PubMed2006

Focal adhesion contact formation by fibroblasts cultured on surface-modified dental implants: an in vitro study.

A major consideration in designing dental implants is to create a surface that provides strong attachment of the implant to bone, connective tissue and epithelium. The aim of the present study was to examine the influence of different treatments of titanium (Ti) implant surfaces on focal adhesion contact (FAC) formation in fibroblast cultures. Human gingival fibroblasts were cultured on glass sheets and polished Ti discs with different surface coatings (applied by physical vapor deposition (PVD): Ti, titanium nitride (TiN), zirconium nitride (ZrN)) or on Ti discs with different surface topographies. For characterization of all surfaces, modified estimation of surface roughness and spacing parameter was carried out using a contact stylus profilometer. Contact angle measurements were carried out to calculate surface energy. Fibroblasts were prepared for transmission electron microscopy at day 3 after seeding, and the number of FACs and the ratio FAC/cellular cross-sections was determined at a length of 300 microm in ultrathin sections. To visualize the extracellular fibronectin and vitronectin molecules and the intracellular actin and vinculin in FAC areas, immunogold labeling was performed. The results revealed a strong correlation between the number of FACs and the surface roughness. The highest number of FACs and the majority of the immunogold-labeled intra- and extracellular matrix molecules were counted on surfaces with the lowest surface roughness: glass sheets coated with either Ti, TiN or ZrN (roughness average=0.03-0.1 microm). These surfaces appear to favor cellular attachment of human gingival fibroblasts and moreover in previous studies the hard coatings have been shown to reduce bacterial adhesion.

Analysis of Variance↗

PubMed2006

Restorative therapy of primary teeth severely affected by amelogenesis imperfecta.

OBJECTIVE: Primary teeth severely affected by amelogenesis imperfecta (AI) often show an extensive loss of enamel. Such defects are difficult to restore with resin composites, since neither the correct anatomic form nor the marginal fit can be guaranteed. METHODS AND MATERIALS: After clinical and scanning electron microscopic examinations were performed on replica models of 5 patients with primary teeth affected by AI, impressions were made without previous preparation by rotary instruments. Composite crowns and veneers were manufactured and luted adhesively using the total bonding technique and low-viscosity resin composite. RESULTS: The pre-restorative scanning electron microscopic analysis showed that the dentinal tubules were exposed and that the border of the residual enamel was in the process of splitting. The preoperative oral examination had revealed tooth discoloration, masticatory disturbances, hypersensitivity, and speech problems. After placement of the restorations, patients reported improvements in tooth sensitivity, articulation, and mastication. CONCLUSIONS: A new protocol for restoration of primary teeth with an extensive loss of enamel is offered. It is quick and easy to perform, highly esthetic, and can be applied in children younger than 4 years old.

Acrylic Resins↗

PubMed2005

Lysozyme activity in the initially formed in situ pellicle.

UNLABELLED: Lysozyme is one of the most abundant enzymatic components in the salivary pellicle. The purpose of the present in situ study was to determine if and to which extent lysozyme immobilised in pellicles exposes enzymatic activity. Influence of different oral sites and pellicle formation time on enzyme activity was also evaluated. Bovine enamel slabs (5mm diameter) were fixed on buccal and oral sites of individual trays worn by six subjects for 3 and 30 min on different days. After pellicle formation, slabs were removed from the trays and rinsed with running water. Afterwards, pellicle-bound lysozyme activity was determined via lysis of Micrococcus lysodeicticus photometrically in two steps. In a first step, lysozyme was desorbed in phosphate buffer and dissolved activity was measured. In a second step, slabs were incubated in phosphate buffer with the substrate and remaining immobilised activity was determined. All investigated pellicles exhibited lysozyme activity. Great intra- and inter-individual differences were observed. Mean desorbed activity of 3 min-pellicles amounted to 26.06+/-17.81 U/cm(2) (30 min; 26.79+/-17.48). The remaining immobilised activity was 13.54+/-11.42 for 3 min-pellicles and 16.08+/-12.81 for 30 min-pellicles. Pellicle derived lysozyme showed a Michaelis type kinetic. CONCLUSION: In situ pellicle exposes lysozyme activity even after a 3 min formation period. Exposed enzyme activity is neither influenced by pellicle formation time nor by the site of pellicle formation. It shows great inter- and intra-individual differences.

Adsorption↗

PubMed2005

Evidence of chemical bonding to hydroxyapatite by phosphoric acid esters.

Phosphoric acid esters (PAEs) have been used as a self-etching primer for composite-to-enamel bonding in adhesive dentistry. However, the chemical mechanism of their interactions with hydroxyapatite (HA) is not clear. In the present study, HA particles were mixed with Resulcin AquaPrime (Merz Co.) priming agent that contains a mixture of PAEs, and dried. The primer, HA and the mixture of both were analyzed by FTIR. Primed and untreated HA discs were analyzed with attenuated total reflectance (ATR). After AquaPrime+D(2)O (1:1) was mixed with HA or Ca(OD)(2) for 48 h, the primer and both mixtures were analyzed with (31)P NMR in D(2)O. The solid and the liquid separated from both mixtures were analyzed with (31)P NMR in CDCl(3). The primer's characteristic bands (nu(CO), nu(CC)) were found on the primer-subtracted mixture's spectrum and the primed HA disc. (31)P NMR data indicated that the reactions of PAEs with HA produced PAEs-HA complexes, and were not a simple acid-base reaction like those with Ca(OD)(2), either in liquid or in solid. It is concluded that phosphoric acid esters can decalcify and adhere to hydroxyapatite simultaneously.

Acid Etching, Dental↗

PubMed2005

Cytochemical differences in bacterial glycocalyx.

To examine new cytochemical aspects of the bacterial adhesion, a strain 41452/01 of the oral commensal Streptococcus sanguis and a wild strain of Staphylococcus aureus were grown with and without sucrose supplementation for 6 days. Osmiumtetraoxyde (OsO4), uranyl acetate (UA), ruthenium red (RR), cupromeronic blue (CB) staining with critical electrolytic concentrations (CECs), and the tannic acid-metal salt technique (TAMST) were applied for electron microscopy. Cytochemically, only RR-positive fimbriae in S. sanguis were visualized. By contrast, some types of fimbriae staining were observed in S. aureus glycocalyx: RR-positive, OsO4-positive, tannophilic and CB-positive with ceasing point at 0.3 M MgCl2. The CB staining with CEC, used for the first time for visualization of glycoproteins of bacterial glycocalyx, also reveals intacellular CB-positive substances-probably the monomeric molecules, that is, subunits forming the fimbriae via extracellular assembly. Thus, glycosylated components of the biofilm matrix can be reliably related to single cells. The visualization of intracellular components by CB with CEC enables clear distinction between S. aureus and other bacteria, which do not produce CB-positive substances. The small quantities of tannophilic substances found in S. aureus makes the use of TAMST for the same purpose difficult. The present work protocol enables, for the first time, a partial cytochemical differentiation of the bacterial glycocalyx.

Coloring Agents↗

PubMed2005

Sinus floor elevation via hydraulic detachment and elevation of the Schneiderian membrane.

OBJECTIVES: Minor sinus floor elevation is a method with relatively high predictability but is technically demanding. Improvement of the technique and increase in the predictability are desirable. MATERIAL AND METHODS: A clinical protocol for minor sinus floor elevation with SLA-ITI (large grit acid-etched implants with diameter of 4.8 mm) is described. Using trephine instead of spiral burrs enables the harvesting of autogenous grafts from the implant socket and guarantees a perfect implant socket. The latter is necessary for optimal implant anchoring and for the hydraulic seal between socket and the osteotome. The whole allows a hydraulic detachment of the Schneiderian membrane, where the blood cushion gradually detaches and elevates the membrane, preventing its contact with the graft. RESULTS: Eight patients were successfully treated with the method described above. No membrane perforation occurred and an uneventful healing was observed in all patients. All implants were loaded prosthodontically 3 months after the implantation. CONCLUSIONS: The clinical protocol presented provides high predictability in clinical outcome, together with extremely low morbidity and shortened surgery.

Adult↗

PubMed2005

Enzymes in the acquired enamel pellicle.

The acquired pellicle is a biofilm, free of bacteria, covering oral hard and soft tissues. It is composed of mucins, glycoproteins and proteins, among which are several enzymes. This review summarizes the present state of research on enzymes and their functions in the dental pellicle. Theoretically, all enzymes present in the oral cavity could be incorporated into the pellicle, but apparently enzymes are adsorbed selectively onto dental surfaces. There is clear evidence that enzymes are structural elements of the pellicle. Thereby they exhibit antibacterial properties but also facilitate bacterial colonization of dental hard tissues. Moreover, the immobilized enzymes are involved in modification and in homeostasis of the salivary pellicle. It has been demonstrated that amylase, lysozyme, carbonic anhydrases, glucosyltransferases and fructosyltransferase are immobilized in an active conformation in the pellicle layer formed in vivo. Other enzymes, such as peroxidase or transglutaminase, have been investigated in experimental pellicles. Despite the depicted impact of enzymes on the formation and function of pellicle, broader knowledge on their properties in the in vivo-formed pellicle is required. This might be beneficial in the development of new preventive and diagnostic strategies.

Adsorption↗

PubMed2004

Immobilisation and activity of human alpha-amylase in the acquired enamel pellicle.

UNLABELLED: Amylase is an important salivary component and structural element of the acquired enamel pellicle. Aim of the study was to establish a method for precise and direct determination of pellicle bound amylase activity in order to analyse kinetics and activity of the immobilised enzyme. Six bovine enamel slabs (5mm diameter) were fixed on individual maxillary trays and worn by five subjects for different times (3, 30 and 120 min) on buccal and palatal sites on different days. Slabs were removed from the trays and rinsed with aqua dest. Afterwards, pellicle bound amylase activity was determined directly with a photometric method using 2-chloro-4-nitrophenyl-4-O-beta-D-galactopyranosylmaltotriosid (GalG2CNP) as substrate yielding the coloured product chloronitrophenolate (CNP). All investigated pellicles exhibited immobilised amylase activity. Mean activity was 1.39 +/- 187 mU/cm(2) (n=87, range 0.14-11.5 mU/cm(2)). Product formation of CNP by immobilised amylase was linear over time. Pellicle bound amylase showed a Michaelis type kinetic (Km = 3.3 x 10(-3) M). Immobilised activity on buccal surfaces ranged between 0.25 and 11.1 mU/cm(2) (palatal slabs: 0.14-3.06 mU/cm(2)). Thirty minutes pellicles formed on buccal sites exhibited significantly higher immobilised amylase activity (2.85 +/- 3.65 mU/cm(2)) than palatal ones (0.63 +/- 0.32 mU/cm(2)). Amylase activity showed great intraindividual variability when comparing same positions on different days. CONCLUSION: Pellicle bound amylase activity can be determined directly with GalG2CNP and shows a Michaelis Menten kinetic. Enzyme activity of the amylase immobilised in the in situ pellicle reveals great intra- and interindividual differences.

Analysis of Variance↗

PubMed2004

Protective effect of the in situ formed short-term salivary pellicle.

Salivary pellicle, as previously investigated, protects the enamel surface after certain processes of maturation against the influence of acidic agents. The aim of the present study was to investigate the protective effect of the short-term salivary pellicle formed in situ over periods of 3, 60 and 120 min. Six human volunteers used intraoral acrylic splints with bovine enamel samples fixed at the buccal and palatal sites of the maxillary first molars and second premolars. Enamel specimens (n = 252) with and without pellicle were immersed for 60 s in 1.0% citric acid solution under agitation. Knoop surface hardness (KHN) of uneroded polished enamel was measured as a baseline and estimated immediately after erosive treatment reflecting the microhardness loss (DeltaKHN). The amounts of calcium dissolved from the eroded enamel surface were analysed by atomic absorption spectroscopy and scored in mg/l per 10 mm2 of enamel surface area. In addition, the scanning electron microscope was used for the micromorphological examination of the erosive alterations of the enamel surface. The average microhardness loss values after erosion of the enamel samples with buccally/palatally formed pellicle layers were measured as 139.1/144.9 DeltaKHN for 3 min pellicle, 145.9/146.9 DeltaKHN for 60 min pellicle and 141.7/138.6 DeltaKHN for 120 min pellicle. Calcium release values from the specimens with buccal/palatal pellicles were amounted to 15.0/14.9, 16.5/15.9 and 15.3/17.4 mg/l per 10 mm2 for 3, 60 and 120 min-old pellicles, respectively. No significant differences were related to the pellicle formation time and intraoral site (buccal or palatal) in all tested series (ANOVA, P < 0.05). However, significant protection of the enamel surface provided by the pellicle layer was observed on all pellicle-covered surfaces if compared to the non-covered enamel samples (calcium release: 25.6 mg/l per 10 mm2; microhardness loss 187.0 DeltaKHN). These data were in accordance with the morphologic alterations caused by citric acid on the pellicle-covered and pellicle non-covered specimens. It could be concluded that salivary pellicle formed in situ within a period of 3 min offers protection of enamel against citric acid. However, pellicle does not completely inhibit the erosive action of citric acid under the conditions of the present study.

Adult↗

PubMed2004

Supramolecular pellicle precursors.

Saliva contacting with solid surfaces in the oral cavity forms a coat termed the pellicle. However, its formation is not fully understood. Although indications for the existence of supramolecular pellicle precursors have been reported, the possible relationship between them and pellicle formation is unclear. This study investigates the ability of supramolecular precursors to form the pellicle via interaction with a solid surface. Fixed and unfixed salivary globes were spread onto a microscopic grid and examined by transmission electron microscopy. Biochemical pretreatment of saliva revealed that neither disulphide links nor transglutaminase-mediated crosslinking are responsible for maintaining the salivary globes, i.e. supramolecular pellicle precursors. However, the detergent, sodium dodecyl sulphate, caused dissociation of the salivary globes, indicating their micellar nature. Saliva contacting a formvar film for 10 s did not form a complete surface coating, but single supramolecular pellicle precursors were observed attached to the surface. After extension of the contact time to 60 s, a surface layer was formed by clustering and fusion of the supramolecular pellicle precursors. The supramolecular pellicle precursors are unstable and attain a thermodynamically more favourable state by adhesion to a solid surface. As a result, a layer of fused precursors covering the solid surface is formed -- the salivary pellicle.

Dental Pellicle↗

PubMed2004

Evidence of chemisorption of maleic acid to enamel and hydroxyapatite.

Maleic acid has been used as an etchant or non-rinse conditioner in adhesive dentistry. However, the inherent mechanisms of the interaction of maleic acid with hydroxyapatite/enamel have never been fully elucidated. The purpose of this study was to provide evidence for the chemisorption of maleic acid onto hydroxyapatite/enamel, and to identify the reaction products obtained following the interaction of maleic acid with hydroxyapatite. Hydroxyapatite particles were dissolved in a 15% (w/v) aqueous solution of maleic acid (pH = 0.98). Half of the solution was dried to obtain a desiccated mixture. This mixture, hydroxyapatite, maleic acid and self-prepared calcium maleate were analysed by X-ray diffraction (XRD). Acetone was added to the other half of the solution to obtain a precipitate. This precipitate, hydroxyapatite, maleic acid, unetched enamel and maleic acid-etched enamel were analysed by X-ray photoelectron spectroscopy (XPS). The precipitate was also analysed by (1)H NMR. A new binding energy, indicating carboxylate groups, was detected by XPS on the precipitate and maleic acid-etched enamel surface. XRD data indicated the formation of calcium maleate and calcium hydrogen phosphate after the reaction. NMR data revealed that one carboxylic group of maleic acid reacted with hydroxyapatite. Hence, maleic acid can chemisorb to hydroxyapatite and enamel via ionic interactions.

Acid Etching, Dental↗

PubMed2004

Electron microscopic detection of salivary alpha-amylase in the pellicle formed in situ.

Immunological and biochemical analyses have shown that alpha-amylase is an essential component of the acquired pellicle. After adsorption, this enzyme might act as a receptor for bacterial adherence. However, data indicating that amylase is bound to the pellicle surface in vivo and thus available for adhering bacteria are rare. Therefore, the present study focused on alpha-amylase within the pellicle formed in situ, using gold-immunolabeling electron microscopic techniques. Pellicles were formed by intra-oral exposure of enamel specimens for 30 and 120 min in six subjects. The results obtained by transmission electron microscopy indicate that amylase was randomly distributed in the pellicle layer without any preferential localization within the pellicle. Thus, salivary alpha-amylase might be considered as an important structural component that is even involved in the early stages of pellicle formation. The findings of field emission in-lens scanning electron microscopy provided evidence that the enzyme is located on the pellicle surface. It could be concluded that alpha-amylase might act as a receptor for bacterial adherence to the pellicle in vivo.

Adsorption↗

PubMed2004

Do different implant surfaces exposed in the oral cavity of humans show different biofilm compositions and activities?

Osseointegrated dental implants play an important role in restorative dentistry. However, plaque accumulation may cause inflammatory reactions around the implants, sometimes leading to implant failure. In this in vivo study the influence of two physical hard coatings on bacterial adhesion was examined in comparison with a pure titanium surface. Thin glass sheets coated with titanium nitride (TiN), zirconium nitride (ZrN) or pure titanium were mounted on removable intraoral splints in two adults. After 60 h of intraoral exposure, the biofilms were analyzed to determine the number of bacteria, the types of bacteria [by applying single-strand conformation polymorphism (SSCP analysis) of 16S rRNA genes], and whether or not the bacteria were active (by SSCP analysis of 16S rRNA). The results showed that bacterial cell counts were higher on the pure titanium-coated glass sheets than on the glass sheets coated with TiN or ZrN. The lowest number of bacterial cells was present on theZrN-coated glass. However, the metabolic activity (RNA fingerprints) of bacteria on TiN- and ZrN-coated glass sheets seemed to be lower than the activity of bacteria on the titanium-coated surfaces, whereas SSCP fingerprints based on 16S rDNA revealed that the major 16S bands are common to all of the fingerprints, independently of the surface coating.

Adult↗

PubMed2004

Assessment of enamel erosion and protective effect of salivary pellicle by surface roughness analysis and scanning electron microscopy.

PURPOSE: To assess dental erosion caused by 0.1% and 1.0% citric acid in vitro and to estimate the protective influence of experimentally formed salivary pellicle. MATERIALS AND METHODS: Bovine enamel slabs (n = 80) were polished and embedded in epoxy resin. For the formation of pellicle layer 40 specimens were immersed for 24 h in pooled human saliva. Erosion was caused by immersion in citric acid solution for 1, 5, 10 and 30 min. Erosive alterations on the pellicle-covered and non-covered enamel specimens were scored as a change (delta) of surface roughness parameters Ra, Rt and RzDIN using contact profilometer and observed in scanning electron microscope. RESULTS: Profilometric analysis of eroded enamel specimens emphasized the aggressiveness of even low concentrated citric acid with a short period of challenge. The change of roughness parameters after 1-min immersion in 0.1% citric acid were 16.4, 182.6 and 132.2 nm for deltaRa, deltaRt and deltaRzDIN, respectively, and 54.8, 516.6 and 258.2 nm after 1-min immersion in 1.0% citric acid. Changes of the surface roughness were dependent on the exposure time and concentration of acidic solution. Pellicle layer significantly reduced the extent of erosive destruction, which was additionally documented on SEM-micrographs. Residual pellicle-like structures were detected after 5 min of immersion in 0.1% citric acid. However, there were no significant differences in pellicle-covered and non-covered enamel slabs measured profilometrically for 1.0% citric acid with 10 min and 30 min exposure time. CONCLUSION: The findings confirm the property of pellicle layer to resist against erosive influence of organic acids, which is, however, limited by duration of acidic treatment and concentration of erosive agent.

Animals↗

PubMed2003

Glycocalyx morphology of Candida albicans.

In this study, we examine new cytochemical aspects of the fimbria-mediated adhesion of the oral facultative pathogen Candida albicans. A wild-type strain of the yeast was grown with and without sucrose supplementation for 8 days. Osmium tetroxide, uranyl acetate (UA), ruthenium red (RR), and cupromeronic blue (CB) staining with critical electrolytic concentrations (CECs) and tannic acid-metal salt technique (TAMST) were applied to specimens separately or in combination for transmission electron microscopy (TEM) examination. Cytochemically, two types of fimbriae of C. albicans were distinguished: RR-positive fimbriae of polyanionic glycoconjugates and CB-positive fimbriae with a ceasing point of 0.3 M MgCl2 where no staining of sulfated carboxyl-rich and/or phospho-glycoconjugates occurred. Additionally, CB-positive intercellular fibers were observed, which seemed to be involved in intercellular adhesion. The present protocol enables, for the first time, a partial cytochemical differentiation between at least two kinds of yeast fimbriae.

Animals↗