Candida Albicans Biofilm Formation Research Articles

Sodium New Houttuyfonate Inhibits Candida albicans Biofilm Formation by Inhibiting the Ras1-cAMP-Efg1 Pathway Revealed by RNA-seq.

Here, we aim to investigate the antifungal effect and mechanism of action of sodium new houttuyfonate (SNH) against Candida albicans. Microdilution analysis results showed that SNH possesses potent inhibitory activity against C. albicans SC5314, with a MIC80 of 256 μg/mL. Furthermore, we found that SNH can effectively inhibit the initial adhesion of C. albicans. Inverted microscopy, crystal violet staining, scanning electron microscopy and confocal laser scanning microscopy results showed that morphological changes during the transition from yeast to hypha and the biofilm formation of C. albicans are repressed by SNH treatment. We also found that SNH can effectively inhibit the biofilm formation of clinical C. albicans strains (Z103, Z3044, Z1402, and Z1407) and SNH in combination with fluconazole, berberine chloride, caspofungin and itraconazole antifungal agents can synergistically inhibit the biofilm formation of C. albicans. Eukaryotic transcriptome sequencing and qRT-PCR results showed that SNH treatment resulted in significantly down-regulated expression in several biofilm formation related genes in the Ras1-cAMP-Efg1 pathway (ALS1, ALA1, ALS3, EAP1, RAS1, EFG1, HWP1, and TEC1) and significantly up-regulated expression in yeast form-associated genes (YWP1 and RHD1). We also found that SNH can effectively reduce the production of key messenger cAMP in the Ras1-cAMP-Efg1 pathway. Furthermore, using Galleria mellonella as an in vivo model we found that SNH can effectively treat C. albicans infection in vivo. Our presented results suggest that SNH exhibits potential antibiofilm effects related to inhibiting the Ras1-cAMP-Efg1 pathway in the biofilm formation of C. albicans.

Biocompatible silver nanoparticles incorporated in acrylic resin for dental application inhibit Candida albicans biofilm.

Although silver nanoparticles (SNP) have proven antimicrobial activity against different types of microorganisms, the effect of SNP incorporation into acrylic resin to control Candida albicans biofilm formation aiming at the prevention of Candida-associated denture stomatitis has not yet been fully elucidated. This study aimed to evaluate the antimicrobial effect of an acrylic resin containing SNP on C. albicans biofilm growth, the flexural strength of this material and tissue reaction in the subcutaneous connective tissue of rats to SNP. SNP were synthesized through silver nitrate reduction by sodium citrate. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were used to verify the size and colloidal stability. SNP were added to acrylic resin monomer (Lucitone 550) at 0.05, 0.5 and 5vol%. The antimicrobial effect against C. albicans (ATCC 10231) was investigated by the enumeration of colony-forming units (CFUs) and SEM. The three-point bending test was performed to analyze the flexural strength. Tissue reaction was evaluated after 7 and 60days of implantation in the connective tissue of Wistar rats. Spherical particles of 5 and 10nm were obtained. SNP at 0.05 and 0.5% incorporated into acrylic resin was effective in reducing C. albicans biofilm growth (p<.001). SEM revealed that the material was able to disrupt C. albicans biofilm formation and did not reduce the flexural strength compared to control (p>.05). The inflammatory response observed 60days after implantation SNP in the subcutaneous tissue was similar to control. It was concluded that SNP addition at 0.05 and 0.5% into acrylic resin exhibited antimicrobial effects against C. albicans biofilm, did not interfere in the flexural strength and may be considered biocompatible.

The effects of clioquinol in morphogenesis, cell membrane and ion homeostasis in Candida albicans

BackgroundCandida albicans is the most prevalent opportunistic fungal pathogen. Development of antifungals with novel targets is necessary for limitations of current antifungal agents and the emergence of drug resistance. The antifungal activity of clioquinol was widely accepted while the precise mechanism was poorly understood. Hence, we aimed to seek for the possible mechanism of clioquinol against Candida albicans in the present study.ResultsClioquinol could inhibit hyphae formation in a concentration-dependent manner in multiple liquid and solid media. The concentration and time-dependent anti-biofilm activities were observed in different incubation periods quantitatively and qualitatively. Further investigation found that clioquinol disrupted cell membrane directly in high concentration and induced depolarization of the membrane in low concentration. As for the influence on ion homeostasis, the antifungal effects of clioquinol could be reversed by exogenous addition of metal ions. Meanwhile, the minimum inhibitory concentration of clioquinol was increased in media supplemented with exogenous metal ions and decreased in media supplemented with exogenous metal chelators. We also found that the cellular labile ferrous iron level decreased when fungal cells were treated with clioquinol.ConclusionThese results indicated that clioquinol could inhibit yeast-hyphae transition and biofilm formation in Candida albicans. The effect on the cell membrane was different depending on different concentrations of clioquinol. Meanwhile, clioquinol could interfere with ion homeostasis as metal chelators for zinc, copper and iron, which was quite different with current common antifungal agents. All in all, clioquinol can be a new promising antifungal agent with novel target though more studies are needed to better understand the precise antifungal mechanism.

Biofilm Formation of Candida albicans Facilitates Fungal Infiltration and Persister Cell Formation in Vaginal Candidiasis.

BackgroundVaginal candidiasis is an important medical condition awaiting more effective treatment. How Candida albicans causes this disease and survives antifungal treatment is not yet fully understood. This study aimed to establish a comprehensive understanding of biofilm-related defensive strategies that C. albicans uses to establish vaginal candidiasis and to survive antifungal treatment.MethodsA mouse model of vaginal candidiasis was adopted to examine the formation of biotic biofilms on the vaginal epithelium and fungal infiltration by laboratory and clinical strains of C. albicans. Histopathological changes and local inflammation in the vaginal epithelium caused by C. albicans of different biofilm phenotypes were compared. Antifungal susceptibility testing was carried out for C. albicans grown as planktonic cells, microplate-based abiotic biofilms, and epithelium-based biotic biofilms. Formation of persister cells by C. albicans in different growth modes was also quantified and compared.ResultsC. albicans wild-type reference strains and clinical isolates, but not the biofilm-defective mutants, formed a significant number of biotic biofilms on the vaginal epithelium of mice and infiltrated the epithelium. Biofilm formation and epithelial invasion induced local inflammatory responses and histopathological changes in the vaginal epithelium including neutrophil infiltration and subcorneal microabscesses. Biofilm growth on the vaginal epithelium also led to high resistance to antifungal treatments and promoted the formation of antifungal-tolerant persister cells.ConclusionThis study comprehensively assessed biofilm-related microbial strategies that C. albicans uses in vaginal candidiasis and provided experimental evidence to support the important role of biofilm formation in the histopathogenesis of vaginal candidiasis and the recalcitrance of the infection to antifungal treatment.

Antioxidant and antimicrobial activity of two Asplenium species

Extracts of the aerial (fronds) and underground parts (rhizome and roots) of Asplenium adiantum-nigrum L. and Asplenium ruta-muraria L. were analyzed for their phenolic composition and antioxidant properties (scavenging capacities against DPPH• and ABTS•+), and were comparatively assayed for antimicrobial and antibiofilm activity. Total of the 17 phenolic compounds, belonging to the classes of phenolic acids (hydroxybenzoic and hydroxycinnamic acids), flavonoids (flavan-3-ols, flavonols), and xanthones were identified and quantified using UHPLC/DAD/(–)HESI-MS/MS analysis. The present study shows that two analyzed ferns are significantly diverse in terms of their phenolic profile. Flavonoids considerably contribute to the total phenolic content of A. ruta-muraria, ranging from 47% in roots to around 75% in rhizome and fronds, while A. adiantum-nigrum preferably accumulates xanthones. Antioxidant activity of fern methanol extracts was ascribed to their phenolics content. Extracts of rhizomes of both Asplenium ferns exhibited the highest antimicrobial activity, while frond extracts were more efficient in inhibition of Candida albicans biofilm formation. The present study provides experimental evidence that methanol extracts from two analyzed Asplenium fern species exhibit strong antibacterial effect against Gram-negative bacteria which candidate them as a potentially useful natural antimicrobial agents.

FDA Approved Drug Library Screening Identifies Robenidine as a Repositionable Antifungal.

Due to the increasing prevalence of pathogenic fungal infections, the emergence of antifungal resistant clinical isolates worldwide, and the limited arsenal of available antifungals, developing new antifungal strategies is imperative. In this study, we screened a library of 1068 FDA-approved drugs to identify hits that exhibit broad-spectrum antifungal activity. Robenidine, an anticoccidial agent which has been widely used to treat coccidian infections of poultry and rabbits, was identified in this screen. Physiological concentration of robenidine (8 μM) was able to significantly inhibit yeast cell growth, filamentation and biofilm formation of Candida albicans – the most extensively studied human fungal pathogen. Moreover, we observed a broad-spectrum antifungal activity of this compound against fluconazole resistant clinical isolates of C. albicans, as well as a wide range of other clinically relevant fungal pathogens. Intriguingly, robenidine-treated C. albicans cells were hypersensitive to diverse cell wall stressors, and analysis of the cell wall structure by transmission electron microscopy (TEM) showed that the cell wall was severely damaged by robenidine, implying that this compound may target the cell wall integrity signaling pathway. Indeed, upon robenidine treatment, we found a dose dependent increase in the phosphorylation of the cell wall integrity marker Mkc1, which was decreased after prolonged exposure. Finally, we provide evidence by RNA-seq and qPCR that Rlm1, the downstream transcription factor of Mkc1, may represent a potential target of robenidine. Therefore, our data suggest that robenidine, a FDA approved anti-coccidiosis drug, displays a promising and broadly effective antifungal strategy, and represents a potentially repositionable candidate for the treatment of fungal infections.

Terpenoids from the Liverwort Plagiochila fruticosa and Their Antivirulence Activity against Candida albicans.

Fourteen new terpenoids plagicosins A-N (1-14), including seven sesquiterpenoids (1-7) consisting of six ent-bicyclogermacrenes and one ent-2,3-seco-aromadendrane, as well as seven diterpenoids (8-14) comprising five fusicoccanes, a eunicellane, and a rare gersemiane, were isolated from the Chinese liverwort Plagiochila fruticosa Mitt. The structures of these terpenoids were determined on the basis of comprehensive analysis of MS and NMR spectroscopic data coupled with electronic circular dichroism (ECD) and coupling constant calculations. Plagicosin F (6) displayed potent antivirulence activity through inhibiting the hyphal morphogenesis, adhesion, and biofilm formation of Candida albicans. The genes related to hyphal formation were regulated by 6.

Environmental pH modulates biofilm formation and matrix composition in Candida albicans and Candida glabrata

Candida species are fungal opportunistic pathogens capable of colonizing and infecting various human anatomical sites, where they have to adapt to distinct niche-specific pH conditions. The aim of this study was to analyse the features of Candida albicans and Candida glabrata biofilms developed under neutral and vaginal acidic (pH 4) conditions. C. albicans produced thicker and more filamentous biofilms under neutral than under acidic conditions. On the other hand, the formation of biofilms by C. glabrata was potentiated by the acidic conditions suggesting the high adaptability of this species to the vaginal environment. In general, both species developed biofilms containing higher amounts of matrix components (protein and carbohydrate) under neutral than acidic conditions, although the opposite result was found for one C. glabrata strain. Overall, this study contributes to a better understanding of the modulation of C. albicans and C. glabrata virulence by specific pH conditions.

Combination of Antifungal Drugs and Protease Inhibitors Prevent Candida albicans Biofilm Formation and Disrupt Mature Biofilms.

Biofilms formed by the fungal pathogen Candida albicans are resistant to many of the antifungal agents commonly used in the clinic. Previous reports suggest that protease inhibitors, specifically inhibitors of aspartyl proteases, could be effective antibiofilm agents. We screened three protease inhibitor libraries, containing a total of 80 compounds for the abilities to prevent C. albicans biofilm formation and to disrupt mature biofilms. The compounds were screened individually and in the presence of subinhibitory concentrations of the most commonly prescribed antifungal agents for Candida infections: fluconazole, amphotericin B, or caspofungin. Although few of the compounds affected biofilms on their own, seven aspartyl protease inhibitors inhibited biofilm formation when combined with amphotericin B or caspofungin. Furthermore, nine aspartyl protease inhibitors disrupted mature biofilms when combined with caspofungin. These results suggest that the combination of standard antifungal agents together with specific protease inhibitors may be useful in the prevention and treatment of C. albicans biofilm infections.

Ras activates the first step of GPI anchor biosynthesis in Candida albicans

Glycosylphosphatidylinositol (GPI) anchor biosynthesis is a multi‐step pathway found in all eukaryotes. GPI anchored proteins are involved in a variety of functions like adhesion, host cell attachment, invasion and biofilm formation in Candida albicans, a human pathogenic fungus. The first and committed step of the pathway i.e., transfer of GlcNAc from UDP‐GlcNAc to phosphatidylinositol (PI) to form GlcNAc‐PI, involves an enzyme complex (GPI‐GnT) of six subunits viz. CaGpi1, CaGpi2, CaGpi3, CaGpi15, CaGpi19 and CaEri1. A cross‐talk exists between GPI anchor biosynthesis, Ras signaling and ergosterol biosynthesis in Candida albicans, in which CaGPI2 controls hyphal morphogenesis through CaRAS1 while CaGPI19 seems to regulate azole drug response via co‐regulation of CaERG11. Interestingly, both CaGPI2 and CaGPI19 also negatively regulate each other. Further investigations on how Ras signaling cross‐talks with GPI biosynthesis in C. albicans suggest that Ras activates GPI‐GnT activity, which runs counter to what is reported in Saccharomyces cerevisiae. Out of the two Ras proteins of Candida albicans, CaRas1 alone activates the GPI‐GnT activity. Different forms of CaRas1 too can activate the GPI‐GnT in vivo. This evidence is further strengthened by the fact that CaRas1 physically interacts with CaGpi2, as evidenced by FRET. Furthermore, CaGpi2 appears to be important for virulence since a heterozygous mutant of CaGPI2 is unable to kill macrophage cells and the mutant is more susceptible for macrophage‐mediated killing. Current work is focused on identifying the domains of interaction between CaGpi2 and CaRas1. Understanding this interaction will enable us to target any or both of these molecules to control the virulence of this pathogen.Support or Funding InformationSSK received funding from Department of Science and Technology (DST) India Grant SB/SO/BB‐011/2014 and an umbrella grant to the School of Life Sciences from the DST under Promotion of University research and Scientific Excellence Grant DST PURSE No. SR/PURSE/PHASE 2/11 (C), and Department of Biotechnology (DBT) India Grant BT/PR3749/BRB/10/986/2011. SCS received Junior and Senior Research Fellowships from University Grants Commission (UGC), India.

Global proteomic analysis deciphers the mechanism of action of plant derived oleic acid against Candida albicans virulence and biofilm formation

Candida albicans is a commensal fungus in humans, mostly found on the mucosal surfaces of the mouth, gut, vagina and skin. Incidence of ever increasing invasive candidiasis in immunocompromised patients, alarming occurrence of antifungal resistance and insufficient diagnostic methods demand more focused research into C. albicans pathogenicity. Consequently, in the present study, oleic acid from Murraya koenigii was shown to have the efficacy to inhibit biofilm formation and virulence of Candida spp. Results of in vitro virulence assays and gene expression analysis, impelled to study the protein targets which are involved in the molecular pathways of C. albicans pathogenicity. Proteomic studies of differentially expressed proteins reveals that oleic acid induces oxidative stress responses and mainly targets the proteins involved in glucose metabolism, ergosterol biosynthesis, lipase production, iron homeostasis and amino acid biosynthesis. The current study emphasizes anti-virulent potential of oleic acid which can be used as a therapeutic agent to treat Candida infections.

Effect of progesterone on Candida albicans biofilm formation under acidic conditions: A transcriptomic analysis

Vulvovaginal candidiasis (VVC) caused by Candida albicans is a common disease worldwide. A very important C. albicans virulence factor is its ability to form biofilms on epithelium and/or on intrauterine devices promoting VVC. It has been shown that VVC has a hormonal dependency and that progesterone affects virulence traits of C. albicans cells. To understand how the acidic environment (pH 4) and progesterone (either alone and in combination) modulate C. albicans response during formation of biofilm, a transcriptomic analysis was performed together with characterization of the biofilm properties. Compared to planktonic cells, acidic biofilm-cells exhibited major changes in their transcriptome, including modifications in the expression of 286 genes that were not previously associated with biofilm formation in C. albicans. The vast majority of the genes up-regulated in the acidic biofilm cells (including those uniquely identified in our study) are known targets of Sfl1, and consistently, Sfl1 deletion is herein shown to impair the formation of acidic biofilms (pH 4). Under the acidic conditions used, the presence of progesterone reduced C. albicans biofilm biomass and structural cohesion. Transcriptomic analysis of biofilms developed in the presence of progesterone led to the identification of 65 down-regulated genes including, among others, the regulator Tec1 and several of its target genes, suggesting that the function of this transcription factor is inhibited by the presence of the hormone. Additionally, progesterone reduced the susceptibility of biofilm cells to fluconazole, consistent with an up-regulation of efflux pumps. Overall, the results of this study show that progesterone modulates C. albicans biofilm formation and genomic expression under acidic conditions, which may have implications for C. albicans pathogenicity in the vaginal environment.

In vitro efficacy of the lipopeptide biosurfactant surfactin-C15 and its complexes with divalent counterions to inhibit Candida albicans biofilm and hyphal formation

Surfactin is a type of cyclic lipopeptide biosurfactant implicated in a wide range of applications. Although its antimicrobial activity has been characterized, its effect on Candida albicans physiology remains to be elucidated. The present study evaluated the influence of surfactin-C15 (SF) and its complexes with divalent counterions on C. albicans biofilm formation and preformed biofilms. The SF and metal(II)-SF complexes inhibited biofilm formation and reduced the metabolic activity of mature biofilms in a concentration-dependent manner. The same concentrations of the compounds studied dislodged preexisting biofilms grown on polystyrene plates. Moreover, SF and its metal(II) complexes reduced the mRNA expression of hypha-specific genes HWP1, ALS1, ALS3, ECE1 and SAP4 without exhibiting significant growth inhibition. Further research showed that the compounds tested reduced cellular surface hydrophobicity (CSH). These results suggest that SF and metal(II)-SF complexes could be used as anti-biofilm agents against C. albicans hypha-related infections in clinical practice.

The Effects of Ellagic Acid on Growth and Biofilm Formation of Candida albicans

The Effects of Ellagic Acid on Growth and Biofilm Formation of Candida albicans

How to Use a Mutant Library to Identify Genes Required for Biofilm Formation in the Pathogenic Fungus Candida albicans.

With over 1 billion infections and the causative agents showing critical diseases such as pancreatic cancer, the study of pathogenic fungi has never been more critical. In 2017, the United States spent $7.2 billion on fungal diseases. $4.5 billion was allocated to 75,055 hospitalizations, while $2.6 billion went to 8,993,230 outpatient visits. For Candida infections specifically, the cost was $1.4 billion. Currently, there are few classes of antifungals available, and resistance is growing. The identification of genes required for biofilm formation is essential for new antifungal development. This review details how to identify, verify, and characterize defective biofilm formation mutants in C. albicans. This includes how to run an in vitro biofilm formation assay, how to create clean deletions using the modified CRISPR-Cas9 system, how to assay to identify the potential causes of the defect, and how to create complementation strains to confirm the mutant defect.

In vitro Candida albicans biofilm formation on different titanium surface topographies

Objectives To investigate if differences in titanium implant surface topography influence Candida albicans biofilm formation. Materials and Methods Titanium discs were prepared and characterized using a profilometer: Group A (R a 0.15 µm, smooth), Group B (R a 0.64 µm, minimally rough) and Group C (R a 1.3 µm, moderately rough). Contact angle and surface free energy (SFE) were determined for each group. Non-preconditioned titanium discs were incubated with C. albicans for 24 h. In additional experiments, the titanium discs were initially coated with human saliva, bovine serum albumin or phosphate-buffered saline for 2 h before incubation with C. albicans for 24 h. The amount of fungal biofilm formation was quantified using a colorimetric assay. Results C. albicans biofilm formation was significantly lower (p < 0.05) on the minimally rough titanium surface compared to smooth and moderately rough surfaces. The titanium surface displaying the lowest SFE (Group B) was associated with significantly lower (p < 0.05) C. albicans biofilm formation than the other two groups. Salivary coating resulted in greater adherence of C. albicans with increased surface roughness. Conclusions The minimally rough titanium discs displayed lowest SFE compared to smooth and moderately rough surfaces and showed the least C. albicans biofilm formation. This study demonstrated that C. albicans biofilm formation increased in a SFE-dependent manner. These findings suggest that SFE might be a more explanatory factor for C. albicans biofilm formation on titanium surfaces than roughness. The presence of a pellicle coating may negate the impact of SFE on C. albicans biofilm formation on titanium surfaces.

Novel mixture of chloroxylenol and copper alters Candida albicans biofilm formation, biochemical characteristics, and morphological features

ABSTRACT The use of disinfectants has drastically increased in response to the growing public awareness of risks of microbial infection. However, this practice has a negative impact on the environment as it results in dumping large volumes of antimicrobial and microbicidal compounds in aquatic and terrestrial ecosystems. Chloroxylenol (PCMX) is a phenolic compound that poses public health and environmental risks art high concentrations. Here, a novel disinfectant was proposed to increase the lethality of PCMX by adding copper (Cu) to it. However, it was necessary to demonstrate that this mixture is fungicidal and not merely fungistatic, and that the addition of Cu does, in fact, increase PCMX lethality. We evaluated the fungicidal efficacy of PCMX/Cu on Candida albicans. PCMX/Cu synergy was explored on biofilm formation, biochemistry, and morphology of C. albicans. Inhibition increased with PCMX increased concentration. The combination of Cu and PCMX disrupted fungal ultrastructure, biofilm formation, and biochemical characteristic.

Candida albicans biofilms on different materials for manufacturing implant abutments and prostheses.

Background Morphological, physical and chemical properties of both implants and prostheses can determine the biofilm formation on their surface and increase the risk of biological complications. The aim of this study was to evaluate the capacity of biofilm formation of Candida albicans on different materials used to manufacture abutments and prostheses. Material and Methods Biofilm formation was analyzed on cp grade II titanium, cobalt-chromium alloy and zirconia, silicone, acrylic resin (polymethylmethacrylate) and nano-hybrid composite. Some samples were partially covered with lithium disilicate glass ceramic to study specifically the junction areas. C. albicans was incubated in a biofilm reactor at 37 °C with agitation. The biofilm formation was evaluated at 24 and 48 hours. In addition, the morphology of the biofilm was evaluated by scanning electron microscopy. Results C. albicans developed biofilms on the surface of all materials tested. Cobalt-chromium alloy showed the lowest density of adhered biofilm, followed by zirconia and titanium. Silicone and resin showed up to 20 times higher density of biofilm. A higher biofilm formation was observed when junctions of materials presented micropores or imperfections. Conclusions The biofilm formed in the three materials used in the manufacture of abutments and prostheses showed no major differences, being far less dense than in the resins. Two clinical recommendations can be made: to avoid the presence of resins in the subgingival area of implant prostheses and to design prostheses placing cobalt-chromium alloy/ceramic or titanium/ceramic junctions as far as possible from implants. Key words:Dental implants, cobalt-chromium alloy, titanium, zirconia.

Reduction of Candida albicans biofilm formation by coating polymethyl methacrylate denture bases with a photopolymerized film

Statement of problemAs Candida albicans biofilm formation is associated with severe local and systemic infections in denture-wearing patients, its prevention or reduction becomes an essential factor in the health of this population. PurposeThe purpose of this in vitro study was to investigate whether 2 photopolymerized coatings of poly(acrylic acid) (PAA) and poly(itaconic acid) (PIA) can effectively reduce the adhesion of C albicans on denture base acrylic resin surfaces. Material and methodsThe surface of the polymethyl methacrylate (PMMA) denture base was modified through photopolymerization of a thin film of PAA or PIA. The polymeric coatings were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), contact angle goniometry (CA), and surface roughness measurement (Ra). For biological evaluation, the coated PMMA surfaces were tested in a C albicans biofilm dynamic formation model, observed by confocal laser scanning microscopy (CLSM), and quantified by the number of colony-forming units (CFUs). The cytotoxicity of the polymeric coatings was also evaluated by using a lactic dehydrogenase-based (LDH) test. For statistical analysis, ANOVA and the nonparametric Kruskal–Wallis test were used (α=.05). ResultsThe PMMA resin base surfaces coated with PAA and PIA had an inhibitory effect on C albicans growth, the wettability of the coated surface, and the average roughness. The PAA and PIA coatings had no statistically significant cytotoxic effect on periodontal ligament fibroblasts. ConclusionsPMMA acrylic resin base material was superficially modified through the incorporation of carboxylic acid groups by using PAA and PIA coatings that reduced the adherence of C albicans biofilm by 90%.

Impact of caffeine on metabolic activity and biofilm formation of Candida albicans on acrylic denture resin in the presence of nicotine

Statement of problemCandida albicans has been implicated in denture stomatitis, and this effect is exacerbated by nicotine exposure. However, studies have also suggested that caffeine exposure inhibits the growth of C. albicans. The interaction effects of nicotine and caffeine are not yet clear on the growth of C. albicans. PurposeThe purpose of this in vitro study was to determine the effect of caffeine on metabolic activity and biofilm formation of C. albicans growing on acrylic denture resin while simultaneously exposed to nicotine and, if an effect were to be identified, whether this effect would vary depending on the caffeine concentration. Material and methodsA total of 240 acrylic resin specimens were divided into 2 equal groups (120 each). Specimens in one group were processed to measure C. albicans metabolic activity, and those in the other group were processed to measure C. albicans biofilm attachment. Ten subgroups (n=12) were established within each group with different concentration combinations of nicotine and caffeine to test the interaction effect. The first subgroup was designed as a negative control, containing 0 mg/mL of nicotine and caffeine. The following subgroups all contained 8.00 mg/mL of nicotine, and the caffeine concentrations were prepared at the following 9 levels: 0, 0.25, 0.50, 1.00, 2.00, 4.00, 8.00, 16.00, and 32.00 mg/mL. Metabolic activity was measured by using a 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-carboxanilide (XTT) assay. Biofilm attachment was measured by using spiral plating and calculated in terms of the number of colony-forming units (CFUs)/mL. Descriptive statistics and a 2-way ANOVA were conducted to determine whether the concentrations of nicotine and caffeine used affected the biofilm attachment and metabolic activity of C. albicans (α=.05). ResultsThe presence of 8 mg/mL of nicotine increased the metabolic activity and biofilm formation of C. albicans. When compared with the 0 mg/mL of caffeine and 8.00 mg/mL of nicotine group, caffeine from 1.00 to 4.00 mg/mL significantly increased C. albicans biofilm metabolic activity. Caffeine at 16.00 and 32.00 mg/mL significantly decreased C. albicans biofilm metabolic activity in the presence of 8 mg/mL of nicotine. Caffeine from 1.00 to 32.00 mg/mL significantly decreased the biofilm formation of C. albicans in the presence of 8 mg/mL of nicotine. ConclusionsThe presence of 8 mg/mL of nicotine alone increased the metabolic activity and biofilm formation of C. albicans. In the presence of 8 mg/mL of nicotine with different caffeine concentrations, the results suggest that, overall, caffeine at higher concentrations (16 and 32 mg/mL) inhibited the metabolic activity and biofilm formation of C. albicans on acrylic denture resin most.