Zinc- and Fluoride-Releasing Bioactive Glass as a Novel Bone Substitute.

Bioglass 45S5, a silica-based glass, has pioneered a new field of biomaterials. Bioglass 45S5 promotes mineralization through calcium ion release and is widely used in the dental field, including toothpaste formulations. However, the use of Bioglass 45S5 for bone grafting is limited owing to the induction of inflammation, as well as reduced degradation and ion release. Phosphate-based glasses exhibit higher solubility and ion release than silica-based glass. Given that these glasses can be synthesized at low temperatures (approximately 1,000°C), they can easily be doped with various metal oxides to confer therapeutic properties. Herein, we fabricated zinc- and fluoride-doped phosphate-based glass (multicomponent phosphate [MP] bioactive glass) and further doped aluminum oxide into the MP glass (4% Al-MP glass) to overcome the striking solubility of phosphate-based glass. Increased amounts of zinc and fluoride ions were detected in water containing the MP glass. Doping of aluminum oxide into the MP glass suppressed the striking dissolution in water, with 4% Al-MP glass exhibiting the highest stability in water. Compared with Bioglass 45S5, 4% Al-MP glass in water had a notably reduced particle size, supporting the abundant ion release of 4% Al-MP glass. Compared with Bioglass 45S5, 4% Al-MP glass enhanced the osteogenesis of mouse bone marrow-derived mesenchymal stem cells. Mouse macrophages cultured with 4% Al-MP glass displayed enhanced induction of anti-inflammatory M2 macrophages and reduced proinflammatory M1 macrophages, indicating M2 polarization. Upon implanting 4% Al-MP glass or Bioglass 45S5 in a mouse calvarial defect, 4% Al-MP glass promoted significant bone regeneration when compared with Bioglass 45S5. Hence, we successfully fabricated zinc- and fluoride-releasing bioactive glasses with improved osteogenic and anti-inflammatory properties, which could serve as a promising biomaterial for bone regeneration.

High-Performance Dental Adhesives Containing an Ether-Based Monomer.

Dental adhesives are vital for the success of dental restorations. The objective of this study is to make strong and durable dental adhesives that are free from 2 symbolic methacrylate-based dental resins-2-bis[4-(2-hydroxy-3-methacryl-oxypropoxy)-phenyl]-propane (Bis-GMA) and 2-hydroxyethyl-methacrylate (HEMA)-and have equivalent/improved bonding strength and durability. We formulated, prepared, and evaluated 2 dental adhesives using mixtures of a hydrolytically stable ether-based monomer, triethylene glycol divinylbenzyl ether (TEG-DVBE), with urethane dimethacrylate (UDMA) or pyromellitic glycerol dimethacrylate. These adhesives were composed of equimolar ester-/ether-based vinyl functional groups. They were compared with Bis-GMA/HEMA-based commercial and experimental dental adhesives in terms of shear bond strength and microtensile bond strength (µTBS) to human dentin and the µTBS bond stability under extended thermocycling challenges. In addition, the resins' infiltration to dentin tubules, mechanical performance, and chemical properties were assessed by scanning electron microscopy, ISO standard flexural strength and modulus measurements, contact angle measurements, and water sorption/solubility measurements. The hybrid TEG-DVBE-containing dental adhesives generated equivalent shear bond strength and µTBS in comparison with the controls. Significantly, these adhesives outperformed the controls after being challenged by 10,000 thermocycles between 5 °C and 55 °C. Water contact angle measurements suggested that the hybrid dental adhesives were relatively more hydrophobic than the Bis-GMA/HEMA controls. However, both TEG-DVBE-containing adhesives developed more and deeper resin tags in dentin tubules and formed thicker hybrid layers at the composite-dentin interface. Furthermore, the water solubility of UDMA/TEG-DVBE resins was reduced approximately 89% in comparison with the Bis-GMA/HEMA controls. The relatively hydrophobic adhesives that achieved equivalent/enhanced bonding performance suggest great potentials in developing dental restoration with extended service life. Furthermore, the TEG-DVBE-containing materials may find wider dental applications and broader utility in medical device development.

Gingival Fibroblasts as Autologous Feeders for Induced Pluripotent Stem Cells.

Human gingival fibroblasts (hGFs) present an attractive source of induced pluripotent stem cells (iPSCs), which are expected to be a powerful tool for regenerative dentistry. However, problems to be addressed prior to clinical application include the use of animal-derived feeder cells for cultures. The aim of this study was to establish an autologous hGF-derived iPSC (hGF-iPSC) culture system by evaluating the feeder ability of hGFs. In both serum-containing and serum-free media, hGFs showed higher proliferation than human dermal fibroblasts (hDFs). Three hGF strains were isolated under serum-free conditions, although 2 showed impaired proliferation. When hGF-iPSCs were transferred onto mitomycin C-inactivated hGFs, hDFs, or mouse-derived SNL feeders, hGF and SNL feeders were clearly hGF-iPSC supportive for more than 50 passages, whereas hDF feeders were only able to maintain undifferentiated hGF-iPSC growth for a few passages. After 20 passages on hGF feeders, embryonic stem cell marker expression and CpG methylation at the NANOG and OCT3/4 promoters were similar for hGF-iPSCs cultured on hGF and SNL feeder cells. Long-term cultures of hGF-iPSCs on hGF feeders sustained their normal karyotype and pluripotency. On hGF feeders, hGF-iPSC colonies were surrounded by many colony-derived fibroblast-like cells, and the size of intact colonies at 7 d after passage was significantly larger than that on SNL feeders. Allogeneic hGF strains also maintained hGF-iPSCs for 10 passages. Compared with hDFs, hGFs showed a higher production of laminin-332, laminin α5 chain, and insulin-like growth factor-II, which have been reported to sustain the long-term self-renewal of pluripotent stem cells. These results suggest that hGFs possess an excellent feeder capability and thus can be used as alternatives to conventional mouse-derived SNL and hDF feeders. In addition, our findings suggest that hGF feeders are promising candidates for animal component-free ex vivo expansion of autologous hGF-iPSCs, thus providing an important step toward the future therapeutic application of hGF-iPSCs.

Cyclic mechanical pressure-loading alters epithelial homeostasis in a three-dimensional in vitro oral mucosa model: clinical implications for denture-wearers.

Denture-wearing affects the quality and quantity of epithelial cells in the underlying healthy oral mucosa. The physiologic mechanisms, however, are poorly understood. This study aimed to compare histologic changes and cellular responses of an epithelial cell layer to cyclic mechanical pressure-loading mimicking denture-wearing using an organotypic culture system to develop a three-dimensional in vitro oral mucosa model (3DOMM). Primary human oral keratinocytes and fibroblasts were serially grown in a monolayer culture, and cell viability was measured under continuous cyclic mechanical pressure (50 kPa) for 7 days (cycles of 60 min on, 20 s off to degas and inject air). Upon initiation of an air-liquid interface culture for epithelial stratification, the cyclic pressure, set to the mode above mentioned, was applied to the 3DOMMs for 7 days. Paraffin-embedded 3DOMMs were examined histologically and immunohistochemically. In the monolayer culture, the pressure did not affect the viability of oral keratinocytes or fibroblasts. Few histologic changes were observed in the epithelial layer of the control and pressure-loaded 3DOMMs. Immunohistochemical examination, however, revealed a significant decrease in Ki-67 labelling and an increase in filaggrin and involucrin expression in the suprabasal layer of the pressure-loaded 3DOMMs. Pressure-loading attenuated integrin ß1 expression and increased matrix metalloproteinase-9 activity. Incomplete deposition of laminin and type IV collagen beneath the basal cells was observed only in the pressure-loaded 3DOMM. Cyclic pressure-loading appeared to disrupt multiple functions of the basal cells in the 3DOMM, resulting in a predisposition towards terminal differentiation. Thus, denture-wearing could compromise oral epithelial homeostasis.

Pharmacokinetic evaluation of intraperitoneal doxorubicin in rats.

The intraperitoneal (ip) administration of doxorubicin (DOX) is considered to be an important approach for the treatment of peritoneal tumors, because the prognosis of peritoneal cancer is generally poor due to its refractoriness to conventional chemotherapy. In the present study, we examined the disposition behavior of DOX after ip administration in rats to evaluate the adequacy of the ip administration of DOX on the basis of pharmacokinetic aspects. By comparing the area under the serum concentration-time curve (AUC) after ip and intravenous (iv) dosing of 5 mg/kg DOX, the bioavailability of intraperitoneally administered DOX was estimated as 43.8%. This finding suggests that the majority of DOX remained in the abdominal cavity without being incorporated into the systemic circulation. The mean residence time (MRT) of DOX after its ip administration was about 80% longer than that after its iv administration, which indicated the slow absorption process associated with ip application. No significant difference was observed in the elimination rates of systemically absorbed DOX. These results indicate that the ip administration of DOX likely provided an adequate opportunity for it to interact with peritoneal tumors by maintaining sufficient DOX levels while reducing its systemic exposure

Transcriptional regulation of drug-resistance genes in Candida albicans biofilms in response to antifungals.

Biofilm formation is a major virulence attribute of Candida albicans and is directly associated with therapeutic failure. One method by which Candida acquires antifungal resistance is the expression of drug-resistance genes. This study aimed to evaluate the transcriptional regulation of several genes associated with antifungal resistance of C. albicans under planktonic, recently adhered and biofilm growth modes and in C. albicans biofilms in response to antifungal agents. Initially, the antifungal susceptibility of C. albicans cultures in different growth modes was evaluated by standard antifungal susceptibility testing. Next, to assess CDR1, CDR2, MDR1, ERG11, FKS1 and PIL1 expression, RNA was harvested from cells in each growth mode, and from biofilms after drug treatment, and subjected to quantitative real-time RT-PCR (qRT-PCR). Biofilm C. albicans was more resistant to antifungals than recently adhered cells and stationary-phase planktonic cultures. Transcriptional expression of CDR1, CDR2, MDR1, ERG11 and FKS1 was lower in recently adhered C. albicans than in the stationary-phase planktonic cultures. In contrast, PIL1 levels were significantly increased in recently adhered and biofilm modes of growth. The expression of MDR1 in biofilms greatly increased on challenge with amphotericin B but not with the other drugs tested (P<0.01). ERG11 was significantly upregulated by ketoconazole (P<0.01). Caspofungin and amphotericin B significantly upregulated FKS1 expression, whereas they significantly downregulated PIL1 expression (P<0.01). These results indicate that the expression of drug-resistance genes is associated with higher drug resistance of Candida biofilms, and lay a foundation for future large-scale genome-wide expression analysis.

Effect of filamentation and mode of growth on antifungal susceptibility of Candida albicans.

Biofilm formation involving profuse hyphal growth is a major characteristic of Candida spp. and confers higher antifungal resistance than its planktonic mode of growth. We investigated the antifungal susceptibility of Candida albicans and its hyphal mutants (Delta efg1/efg1, Delta cph1/cph1 and DeltaDelta cph1/cph1 efg1/efg1) to commonly used antifungals during planktonic, adhesion and biofilm modes of growth. The minimum inhibitory concentration (MIC) of each antifungal agent was determined for a lower inoculum (1x10(3) cells/mL) and higher inoculum (1x10(7) cells/mL) of planktonic Candida. Furthermore, MICs of C. albicans biofilms and adhesion modes of growth were determined with a standard XTT assay. Candida albicans in adhesion and biofilm modes of growth, but not in planktonic mode, were resistant to all five antifungal agents tested. Although Delta efg1/efg1 and DeltaDelta cph1/cph1 efg1/efg1 mutants formed less biofilm than wild-type C. albicans SC5314, they were similarly resistant to caspofungin. However, these mutants were more sensitive to amphotericin B and nystatin than the wild-type. Adhesion per se confers increased resistance to antifungal agents, which is further pronounced in the biofilm mode of Candida. Filamentation does not appear to be a major determinant of the antifungal resistance in Candida biofilms.

The influence of single molar crown placement on the smoothness of masticatory movement.

The jerk-cost is an index that can quantify the smoothness of various movements including human body movements. A previous study reported the usefulness of jerk-cost in the evaluation of masticatory movement, and proposed that the masticatory movement of subjects with good occlusion could be explained as a maximum smooth movement. The purpose of this study was to investigate the influence of a single prosthetic molar restoration on the smoothness of masticatory movement. Fourteen adults who visited this hospital seeking a single prosthetic restoration on a molar were selected. Each subject chewed a piece of chewing gum on the molars of the treated side before and after crown placement. Movement trajectory was recorded using the Sirognathograph Analyzing System. Normalized jerk-cost (NJC) was calculated on the closing phase of each chewing cycle and was compared before and after the crown placement. After the prosthetic restoration, NJC significantly decreased (P < 0.05) in seven subjects with a crown placed on a lower molar, whereas significant changes were not observed in seven subjects with a crown placed on an upper molar. These results suggest that restoring a crown on a lower molar could significantly improve the smoothness of masticatory movement on the same side.

Modification of apatite materials for bone tissue engineering and drug delivery carriers.

Apatite-related calcium phosphate, the main component of biological hard tissue, has good biocompatibility and is an economical material. Methods for the synthesis of apatite materials including hydroxyapatite (HAp) have previously been established. Therefore, for many years, apatite materials have been utilized as substitute materials for bone in orthopedic and dental fields. Such types of conventional substitute materials, which are implanted in the human body, should ostensibly be chemically stable to maintain their quality over time. However, recent advances in tissue engineering have altered this concept. Physicians and researchers now seek to identify materials that alter their properties temporally and spatially to achieve ideal tissue regeneration. In order to use apatite materials for tissue engineering and as drug delivery systems, the materials require both a high affinity for cells, tissues and/or functional molecules (e.g. growth factors and genes) and controllable bioabsorbability. To achieve these properties, various physicochemical modifications of apatite materials have been attempted. In addition, fabrication desiring three-dimensional structures (e.g. size, morphology and porosity) of apatite materials for implant sites could be one of the crucial techniques used to obtain ideal prognoses. In this review, the latest research trends relating to the techniques for the fabrication and modification of apatite materials are introduced.

The effect of saliva or serum on bacterial and Candida albicans colonization on type I collagen.

Colonization of Candida albicans on oral surfaces can serve as a reservoir for disseminated infections, such as aspiration pneumonia and gastrointestinal infection, particularly in the immunocompromised host. Therefore, the aim of this study was to investigate the effects of salivary and serum pellicles on C. albicans, Streptococcus mutans, S. sanguis, Lactobacillus and Actinomyces colonization on type I collagen, a major organic component of periodontal ligaments. The colonization potential of two isolates each of C. albicans, S. mutans and S. sanguis, and a single isolate each of Lactobacillus and Actinomyces to uncoated (control), saliva-coated or serum-coated type I collagen plates (surface area 143 mm(2), Cell Disk; Sumitomo, Tokyo, Japan) was examined using a bioluminescent adenosine triphosphate assay based on firefly luciferase-luciferin system. The results revealed that with mutans streptococci, a saliva pellicle was significantly more effective in promoting bacterial colonization compared with the pellicle-free collagen disc, and the serum-coated sample significantly inhibited the colonization of streptococci (anova; P < 0b01). In contrast, in the case of C. albicans, Lactobacillus and Actinomyces isolates, a serum pellicle was significantly more effective in promoting the colonization, followed by saliva pellicle and uncoated specimen (anova; P < 0b01). These results suggested that crevicular fluid rich in seruminous components would promote the colonization of Candida, Lactobacillus and Actinomyces on type I collagen as opposed to streptococci which showed greater avidity to saliva-coated collagen.

An in vitro evaluation of the adhesion of Candida species to oral and lung tissue cells.

The analysis of the adherence capacity of fungi to surfaces of both oral tissue and different tissues would be of interest in the fungal dissemination as an oral and systemic pathogen. We developed an in vitro adenosine triphosphate (ATP)-based assay technique to extract the cellular and fungal ATP separately, which allowed the quantitative evaluation of the adhesion of the yeast to monolayers of human gingival epithelial cells (GEC), gingival fibroblasts (GF) and pulmonary fibroblasts (PF). Seven oral isolates of Candida species (three of Candida albicans, three of Candida tropicalis and one of Candida glabrata) were used in the study. The adherent level of the Candida species varied depending on both the isolates and the cell origins, although all the Candida isolates had a significantly higher level of adherence to GEC than to GF except the single isolate of C. tropicalis. Whereas the adherent level of the five isolates to GEC was significantly higher than that to PF, the adherent level of the remaining two isolates of C. tropicalis to GEC was significantly lower than that to PF. These results suggest that candidal adherence to host tissue cells should be regulated in an isolate-dependent and cell-origin-dependent manner, and that the phenomena may be involved in the colonisation and/or dissemination of the fungi.

In vitro cariogenic potential of Candida albicans.

The adherence and dissociation of Candida albicans, C. tropicalis, Streptococcus mutans and S. sanguis to six substrates including hydroxylapatite (HAP) which exhibit various hydrophobicity, was examined by the use of a bioluminescent adenosine triphosphate (ATP) assay. Dissolution of HAP by C. albicans or S. mutans was determined spectrophotometrically by the use of o-cresolphthalein complexone. In the adherence of C. tropicalis, S. mutans and S. sanguis, the amount of adherent cells correlated with the hydrophobicity of the substrates. In contrast, the adherence of C. albicans to HAP was extraordinary high, although the adherence of the fungi also correlated with the hydrophobicity of the substrates, except for HAP. The yeasts attached to HAP was effectively removed by high concentration of either phosphate or calcium ions. The amount of calcium-release from HAP caused by C. albicans and S. mutans was 113 microg ml(-1) (final pH = 3.45), and 5.4 microg ml(-1) (final pH 4.81), respectively and the maximum growth of C. albicans and S. mutans was 10(7) cfu ml(-1) and 7.4 x 10(12) cfu ml(-1), respectively. The results, taken together, suggest that C. albicans adhere to HAP specifically through electrostatic interaction, and that, in a much smaller number (1.0/7.4 x 10(5)), C. albicans possesses the ability to dissolve HAP to a greater extent (approximately 20-fold) when compared with S. mutans.

A novel technique to evaluate the adhesion of Candida species to gingival epithelial cells.

We developed an in vitro ATP assay technique to extract cellular and fungal ATP separately, which allowed to evaluate quantitatively the adhesion of the yeasts to monolayers of human gingival epithelial cells. Thirteen isolates of Candida spp. representing three species (i.e. Candida albicans, C. tropicalis and C. glabrata) were used in the present study. When the adherent capacity of the Candida species was compared, C. albicans exhibited highest capacity of adherence to gingival epithelial cells, followed by C. tropicalis, and C. glabrata was the lowest [analysis of variance (ANOVA), P < 0.01]. The germ tubes of C. albicans exhibited significantly higher adherence capacity than their blastoconidia cells (ANOVA, P < 0.01), which was not observed with a C. albicans isolate, defect of germ tube formation. Our results suggested that the adherence of C. albicans is promoted by germ tube formation and may play an important role in the pathogenesis of the fungus.

Biofilm formation of Candida albicans on the surfaces of deteriorated soft denture lining materials caused by denture cleansers in vitro.

Candidal colonization and subsequent biofilm formation on denture materials are important in the development of pathogenesis, such as denture stomatitis. Routine use of denture cleansers is one of the most effective methods of denture plaque control, although the incompatibility of soft liners and denture cleansers cause damage to the materials. The present study, biofilm formation of Candida albicans on the surfaces of soft denture lining materials, immersed in denture cleansers for 180 days were studied. Seven commercially available soft denture lining materials, were artificially deteriorated by immersion into three commercially available denture cleansers for 180 days, and subsequent fungal growth and biofilm formation were studied by measuring pH of the media and by the use of adenosine triphosphate (ATP) analysis. Fungal biofilm formation on the deteriorated soft liners varied depending upon the combination of the soft liners and denture cleansers. Several combinations of soft liners with denture cleansers exhibited the significantly high colonization capacity as compared with each sample immersed in distilled water, used as individual controls. The relationship between the biofilm formation on the samples of each material and the surface roughness of the soft lining materials was analyzed. However, no significant correlation was observed. The results, taken together, suggested that fungal colonization could be predominantly regulated by the combination of lining material with denture cleansers. In clinical terms, our findings suggests that daily cleansing of soft lining materials with mismatched denture cleansers promoted the subsequent biofilm formation of fungi on the materials.

Impact of components of denture acrylic resin on gingival cell growth and sensitivity to Candida albicans adhesion.

The effects of four liquid components of denture acrylic resin on host cell activity and fungal adhesion were investigated in this study. The low concentration (1 micromol l(-1)) of the liquid components caused no change in the activities and morphologies of the gingival fibroblast cells, compared with control and dimethylsulphoxide-exposed cells. However, when the cells were exposed to high concentrations (1 mmol l(-1)) of benzqyl peroxide, morphological change was observed, implying that the exposure of the cells to high concentrations of the liquid components of denture acrylic causes the loss of adhesion proteins from the cells. Thus the amount of Candida adhesion to human gingival cells was analysed, and the adherence of fungi to the cell was significantly reduced when the cells were pre-exposed to methyl methacrylate, hydroquinone and benzoyl peroxide at a concentration of 1 micromol l(-1) (P < 0.01), which did not affect either the cell viability or the cell morphology. These results, taken together, suggested that the renewal of dentures could be a possible therapeutic and/or preventive aid for oral candidosis in denture-wearing patients.

Alteration of the coadherence of Candida albicans with oral bacteria by dietary sugars.

Interactions between bacterial oral flora and Candida albicans are important in denture plaque formation. This study therefore first aimed to quantify the coadherence of C. albicans and bacteria by the use of a bioluminescent adenosine triphosphate (ATP) assay based on the firefly luciferase-luciferin system. The second aim was to examine the effect of i) dietary sugars (used for preculture) and ii) enzymatic digestion of fungi on the coadherence. When yeast was preincubated in yeast nitrogen base medium (YNB) supplemented with 250 mM glucose, the yeast coadhered with all isolates of Streptoccus mutans and Streptococcus sanguis, and no significant coadhesion was observed with the isolates of Streptococcus sobrinus, Streptococcus salivarius, Lactobacillus and Actinomyces. However, when the yeast was precultured in YNB supplemented with 500 mM galactose, the yeast coadhered with S. salivarius and Actinomyces, which was not observed when the yeast was grown in YNB with glucose. In addition, the coadherence of the yeast with the isolates of S. sanguis was significantly reduced. Enzymatic digestion of yeast and a reverse transcription polymerase chain reaction assay revealed that expression of at least two types of proteinaceous adhesins are involved in these phenomena.

Body image as a visuomotor transformation device revealed in adaptation to reversed vision.

People adapt with remarkable flexibility to reversal of the visual field caused by prism spectacles. With sufficient time, this adaptation restores visually guided behaviour and perceptual harmony between the visible and tactile worlds. Although it has been suggested that seeing one's own body is crucial for adaptation, the underlying mechanisms are unclear. Here we show that a new representation of visuomotor mapping with respect to the hands emerges in a month during adaptation to reversed vision. The subjects become bi-perceptual, or able to use both new and old representations. In a visual task designed to assess the new hand representation, subjects identified visually presented hands as left or right by matching the picture to the representation of their own hands. Functional magnetic resonance imaging showed brain activity in the left posterior frontal cortex (Broca's area) that was unique to the new hand representations of both hands, together with activation in the intraparietal sulcus and prefrontal cortex. The emergence of the new hand representation coincided with the adaptation of perceived location of visible objects in space. These results suggest that the hand representation operates as a visuomotor transformation device that provides an arm-centred frame of reference for space perception.

Exposure to subtherapeutic concentrations of polyene antifungals suppresses the adherence of Candida species to denture acrylic.

BACKGROUND: The adherence of Candida species to denture acrylic is the initial event leading to Candida-associated denture stomatitis, with Candida albicans being the main aetiological agent. However, the increased incidence of immunocompromised patients in the community has resulted in the emergence of a number of non-albicans Candida species as causative agents of this disease, which is commonly managed by topically delivered polyene antifungals. Hence, we investigated the effect of the exposure of denture acrylic surfaces to nystatin and amphotericin B on the subsequent adhesion of six different Candida species. METHODS: Acrylic strips were exposed to subtherapeutic concentrations of the two polyenes for 30 min, and the adhesion of 4 isolates each of C. albicans, Candida glabrata, Candida guilliermondii, Candida krusei, Candida parapsilosis and Candida tropicalis was assessed using a previously described in vitro method with slight modifications. RESULTS: Overall, the results indicated a 35.9% (p < 0.01) and 63.1% (p < 0. 01) reduction, respectively, in yeast adhesion to denture acrylic following exposure to nystatin and amphotericin B, although this effect was not uniform for all the tested isolates. Thus, all C. glabrata, 3 C. guilliermondii and a single isolate each of C. krusei, C. parapsilosis and C. tropicalis were not significantly affected by nystatin exposure, and a single isolate each of C. glabrata and C. guilliermondii were not significantly affected by amphotericin B. CONCLUSIONS: The present data, the first on the effect of polyenes on a wide range of Candida species, indicate that the in vitro exposure of denture acrylic to subtherapeutic concentrations of nystatin and amphotericin B suppresses the adherence of pathogenic Candida species in general.

Sub-therapeutic exposure to polyene antimycotics elicits a post-antifungal effect (PAFE) and depresses the cell surface hydrophobicity of oral Candida albicans isolates.

Post-antifungal effect (PAFE) is defined as the suppression of growth that persists following limited exposure of fungi to antimycotics and subsequent removal of the drug. The fungal pathogen Candida albicans is the major aetiologic agent of oral candidosis, and the cell surface hydrophobicity (CSH) of this yeast is considered a critical factor contributing to its colonisation potential. As the concentration of topically prescribed antifungals reach sub-therapeutic levels at dosage intervals, the study of the polyene-induced PAFE and its impact on the CSH of oral C. albicans should be of clinical relevance. Hence the aims of this investigation were to measure the PAFE and CSH of 12 isolates of C. albicans following limited exposure (1 h) to nystatin and amphotericin B and also to investigate the ultrastructural features of yeast cells following such antifungal exposure. The yeasts were exposed to sub-lethal concentrations of nystatin (x2 MIC) and amphotericin B (x2 MIC) for a period of 1 h. Following subsequent removal of the drug, the PAFE and the CSH of the isolates were assessed by a turbidometric measurement of growth and a biphasic aqueous-hydrocarbon assay, respectively. The mean duration of PAFE of nystatin and amphotericin B were 5.99 (+/-0.49) h and 8.73 (+/-0.93) h, respectively, while the reduction in CSH following exposure to these drugs were 17.32% (P<0.05 for 83% of the isolates) and 14.26% (P<0.05 for 66% of the isolates), respectively. On scanning electron microscopy the exposed cells were seen to undergo collapse of the internal cell membrane, leaving an intact cell wall, while a proportion of cells were deflated. Some cells showed intense puckering of the cell wall, resulting in a mulberry appearance. Taken together, these data elucidate additional mechanisms by which polyene antimycotics may operate in vivo to suppress candidal pathogenicity.