Candida Albicans Biofilm Research Articles

Cryptocarya moschata fractions decrease planktonic cells and biofilms of Candida albicans and Streptococcus mutans.

Extracts of Cryptocarya species have been shown to reduce biofilms, demonstrating their antimicrobial effects. The extracts can be fractionated to optimize their potential. In this study, we evaluated the antimicrobial activity of Cryptocarya moschata fractions against planktonic cells and biofilms of Candida albicans and Streptococcus mutans. Four fractions were prepared: 100% hexane, acetate/hexane 1:1, 100% ethyl acetate, and water. The effect of the fractions on planktonic cells was assessed by counting the colony-forming units per milliliter (CFU/mL). Biofilm tests included CFU/mL, cell metabolic activity, and qualitative analysis using confocal laser scanning microscopy (CLSM). Results were analyzed by the Mann-Whitney U test (α = 0.05). The fractions contained lipophilic constituents, styrylpyrones, glycosylated flavonoids, and alkaloids. Acetate/hexane (1:1) and 100% ethyl acetate fractions reduced the CFU/mL of planktonic C. albicans. C. moschata fractions did not affect planktonic S. mutans. For biofilms, the fractions reduced the CFU/mL (from 2-5 logs) and cell metabolic activity (approximately 80% reduction in a single-species biofilm). CLSM showed the fractions reduced microorganism viability and damaged the extracellular matrix of biofilms. We conclude that the acetate/hexane 1:1 and 100% ethyl acetate C. moschata fractions exhibit antimicrobial effects against biofilms.

PHOTOINACTIVATION OF CANDIDA ALBICANS BIOFILM WITH GREEN LASER MEDIATED BY THE PAPAYA LEAF EXTRACT CHLOROPHYLL

This study aims to activate the effectiveness of Photodynamic Inactivation (PDI) as an antibacterial agent by using a green laser and papaya leaf chlorophyll extract to prevent Candida albicans cell death. Papaya leaf extract chlorophyll is known to have potential as a photosensitizer (PS) through its antimicrobial properties and ability to absorb optimal light photons at a wavelength range of 405–680 nm. Activation of chlorophyll molecules with appropriate light produces Reactive Oxygen Species (ROS), which are toxic to pathogenic microbes such as Candida albicans. The research method involves using PDI with a green laser light source and chlorophyll extract on Candida albicans biofilms. Four main treatment groups were applied, negative control (C-), positive controls with 10% (C1+) and 15% chlorophyll (C2+), irradiation for 60, 120, 180, 240, and 300 seconds (L1–L5), and combinations of irradiation with chlorophyll (L1F1–L5F2, where F1 for 10% chlorophyll and F2 for 15% chlorophyll), with measurements performed three times for each treatment. Living Candida albicans cells were detected using the XTT assay staining method. The results showed a significant decrease in activity in all treatment groups. Maximum activity was achieved in the L5F1 and L5F2 treatment groups with inactivation of 80% (p<0.05) and 83% (p<0.05), respectively. This study concludes that high papaya leaf extract chlorophyll concentrations combined with a green laser effectively inhibit Candida albicans biofilm.

Influence of nanohydroxyapatite mouthwash on the growth of Candida albicans biofilm on milled denture surfaces: An in vitro study.

There is a need for effective solutions for the persistent prevalence of denture stomatitis (DS) in denture-wearing populations. This study assessed the impact of nanohydroxyapatite (nanoHAP) nanoparticles on Candida albicans biofilm formation on milled polymethylmethacrylate (PMMA) denture bases. Acquired salivary pellicle was formed on disks milled from a prepolymerized PMMA puck. The disks were assigned to two groups, A (n=18) and B (n=18), and each group was further subdivided into three subgroups (6/subgroup), each treated with either Polident solution (antibacterial denture cleanser), nanoHAP mouthwash, or distilled water. Disks in group A subgroups were soaked in their respective treatment solutions for 8h, followed by biofilm formation on thedisk for 24h. Disks in group B subgroups had C. albicans biofilm formed on them for 24h, followed by soaking in their respective treatment solutions for 8h. Biofilm thickness, biomass, and live/dead cell ratio were determined using aconfocal laser scanning microscope. Biofilm morphology was examined with ascanning electron microscope (SEM). Data were analyzed by ANOVA and Tukey-Kramer multiple comparisons (α=0.05). In group A, nanoHAP mouthwash displayed significant anti-adhesive properties. In group B, biofilm biomass and thickness significantly decreased (p<0.05), with the nanoHAP showing the most substantial reduction in existing biofilm compared to other solutions, but it did not significantly affect cell viability. This study demonstrated the efficacy of nanoHAP mouthwash in inhibiting C. albicans biofilm formation when used as a storage medium for acrylic dentures. It suggests its potential clinical application for preventing denture stomatitis in patients.

Effect of Cannabidiol (CBD), a cannabis plant derivative, against Candida albicans growth and biofilm formation.

This study aimed to evaluate the antifungal activities of cannabidiol (CBD) against C. albicans. Yeast cells were treated once or twice with different concentrations (from 0 to 20 mg/ml) of CBD, showing a significant (p < 0.05) decreased the growth of C. albicans, with cell concentrations ranging from 5.1 × 106 cell/mL in the control to 1.8 × 106 cell/mL after one exposure to 20 µg/mL CBD. This growth reduction was greater after two exposures to CBD. After two exposures to 20 µg/mL CBD, the cell concentration was only 1.1 × 106 cell/mL. Such a growth decrease in C. albicans was confirmed by a reduced number of CFUs and a lower MTT value compared to the control. The growth inhibition was supported by a significant (p < 0.001) decrease in the yeast-to-hyphae transition, ranging from 20 ± 0.2% in the control to 2 ± 0.5% after exposure to 20 µg/mL CBD. Biofilm formation was also significantly reduced following CBD exposure. CBD at 10 and 20 µg/mL promoted the death of C. albicans through an apoptosis/necrotic pathway. Altogether, our results suggest the possible use of CBD, a cannabis derivative, to control C. albicans infection, including oral candidiasis.

Antifungal Ability of Novel Silane on Titanium Implant Surface

Titanium and its alloys are commonly used in dentistry for implants due to their strength, lightweight nature, durability, corrosion resistance, and biocompatibility. These implants can osseointegrate after surface treatments such as SLA, plasma-spray, and nanotubes, providing a stable foundation for prostheses. However, Candida albicans, an opportunistic fungal pathogen, can threaten the success of titanium dental implants, causing oral infections in vulnerable individuals. A dual novel silane blend of 3-acryloxypropyltrimethoxysilane (ACPS) and bis-1,2-(triethoxysilyl)ethane (BTSE) has been shown to improve the shear bond strength in resin cement bonds with titanium and ceramics. This study evaluated the effects of Candida albicans colonization on blended silane-coated SLA-Ti surfaces compared to non-coated SLA-Ti (positive control) and flat titanium (negative control). Candida albicans biofilms were cultured on all surfaces, and it was found that silane-coated SLA-Ti had significantly lower CFU counts than non-coated SLA-Ti. However, no significant differences were observed in the RT-PCR results. In conclusion, a combination of 1.0 vol% ACPS and 0.3 vol% BTSE shows promise as a silane coupling agent with potential antifungal properties for inhibiting Candida albicans proliferation.

Antifungal activity’s study of the of new derivatives of pyrrolo[3,4-c]pyrazol-3-ones and pyrazol-3- carboxamides in the biofilm model of Candida spp.

The search for new antifungal drugs is current interest due to the wide spread of fungal infections. To simulate the antifungal effect of new promising compounds with high antifungal activity in planktonic culture in clinical practice, it is necessary to study their effect on the biofilm of micromycetes.Aim: To study the antifungal activity of new promising representatives of silver salts of pyrazoles and their condensed systems under conditions of biofilm formation.Material and Methods. To study the antifungal activity of silver salts I and II, the micromethod of two-fold serial dilutions was used. Activity against a typical and 14 clinical highly virulent isolates of C. albicans was studied. The study of the antimycotic activity of substances under biofilm conditions was carried out using resazurin to quantify the degree of biofilm formation. The minimum inhibitory concentrations for biofilms were calculated (sMIC50 - the concentration of the antifungal substance at which a 50% decrease in fluorescence is observed compared to the positive control).Results and Discussion. It was shown that the antifungal effect of the studied compounds in biofilm culture of clinical strains of Candida spp. is significantly lower than in planktonic culture. High antifungal activity of the silver salt of pyrazole-3- carboxamide in planktonic and biofilm cultures of resistant strains of Candida albicans, exceeding the effect of the reference drug fluconazole by 2.8–11.2 times, was revealed.Conclusion. A promising pyrazole-3-carboxamide derivative has been identified that effectively inhibits the growth of Candida albicans biofilms, which can be recommended for further study.

Efficacy of silver diamine fluoride (SDF) in arresting dentin caries against inter-kingdom biofilms of Streptococcus mutans and Candida albicans.

To compare, in vitro, the efficacy of three proprietary silver diamine fluoride (SDF) products in mitigating progression of dentinal caries induced by an inter-kingdom, dual-species, bacterial-yeast biofilm. Human dentin blocks were demineralized to create artificial caries lesions and randomized into three SDF test groups: Saforide, Topamine, T-SDF, and an aqueous control (n = 26 per group). After application of foregoing SDF variants, the blocks were incubated with Streptococcus mutans and Candida albicans for 24 h for biofilm development, and subsequently subjected to a microbe-induced, pH-cycling process for 7 days, to mimic the oral eco-system. The biofilm cell viability and surface topography were assessed by colony-forming units (CFUs) and scanning electron microscopy respectively. The lesion depth and mineral density were evaluated by micro-computer tomography. SDF precipitate and matrix-to-mineral ratio were evaluated by X-ray diffraction and Fourier transform infrared spectroscopy, respectively. Standard, accepted methodology was used for all these evaluations and procedures. After pH cycling, the SDF groups demonstrated comparable inhibition of the biofilm relative to the control. the log CFU of S. mutans for Saforide, Topamine, T-SDF, and control were 6.69±0.73, 6.48±0.56, 6.63±0.66, and 8.01±0.45, respectively. For C. albicans, the log CFU were 4.86±0.44, 4.72±0.53, 4.92±0.29, and 5.60±0.27, respectively. The log CFU of S. mutans and C. albicans in the SDF groups were significantly lower than the control group (p<0.001). Further, the lesion depth decreased by approximately 14.79±7.00% in the SDF groups, while it increased by 11.07±8.61% in the control (p<0.001), and the mineral density increased by 16.36±4.58% in the SDF group, as opposed to a 5.59±2.64% reduction in the control (p<0.001) implying their caries mitigating effect. These findings were corroborated by SEM images of the lesions. SDF significantly mitigated dentin caries due to an assault by a polymicrobial plaque biofilm whilst arresting mineral loss and lesion growth. There was no difference in the caries-arresting efficacy of the compared SDF variants.

Enhancing Removable Partial Dentures Hygiene: Investigating Mucolytic Agents and Biocides for Disrupting Biofilms and Improving Antimicrobial Efficacy.

This study evaluates the antibiofilm action of 2.5 mg/mL peracetic acid (PA), 0.5 mg/mL cetylpyridinium chloride (CPC), and 160 mg/mL N-Acetylcysteine (NAC) against multispecies biofilm of Streptococcus mutans, Staphylococcus aureus, Candida albicans, and Candida glabrata, developed on surfaces of heat-polymerizing acrylic resin (AR) and cobaltchromium (Co-Cr) alloy. A multispecies biofilm was grown on the surface of AR and Co-Cr specimens (Ø 12×3mm). After biofilm maturation, the specimens were immersed in experimental solutions and evaluated through biofilm viability (CFU) (n&#61;9), biofilm metabolic activity (XTT) (n&#61;9), biofilm-covered areas (Live/Dead) (n&#61;2), effects on the extracellular polymeric substance (EPS) (n&#61;2) and biofilm morphology (n&#61;1). Data were analyzed by ANOVA and the Tukey post-test or Kruskal-Wallis followed by the Dunn post-test (α&#61;.05). Overall, all evaluated solutions impacted biofilm viability. PA presented wider activity by reducing CFU of all microorganisms on both surfaces, XTT (P<.001) and Live/Dead (P<.001). NAC had a notorious effect in reducing the viability of bacteria without affecting the yeasts. NAC reduced XTT on AR (P&#61;.006) and Co-Cr (P&#61;.003) but did not reduce the aggregated biofilm layer. CPC had distinct effect according to the surface, being most effective in reducing CFU on AR than the Co-Cr surface. However, it did not influence XTT, and the amount of residual aggregated biofilm. PA provided the greatest antibiofilm action, while CPC and NAC showed intermediate action. Nonetheless, no solution was able to completely remove the biofilm adhered to the surfaces of heat-polymerizing AR and Co-Cr alloy.

Biofilm formation of Streptococcus mutans, Streptococcus sanguinis, Staphylococcus epidermidis, Staphylococcus aureus, Lactobacillus casei, and Candida Albicans on 5 thermoform and 3D printed orthodontic clear aligner and retainer materials at 3 time points: an in vitro study

IntroductionOrthodontic clear aligners and retainers have numerous advantages that is making them ever increasingly popular. However, they might, similar to any other oral appliance, contribute to biofilm formation and finally dental caries or white spot lesions or gingival inflammations. The literature on biofilm formation on orthodontic clear appliances is very scarce and limited to a few microorganisms and materials. Therefore, this experimental study evaluated the biofilm formation on 5 thermoformed and 3D printed CAD/CAM orthodontic retainers in 3 intervals.MethodsIn this in vitro study, 345 specimens (270 test discs and 45 negative controls) were created from fabricated retainers. Retainers included a 3D printed CAD/CAM material (Detax) and four thermoformed retainers [Erkodent (polyethylene terephthalate glycol [PETG]); EasyVac (polyethylene); DB (polyester based on terephthalic acid); and Clear Tech]. They were all 1 mm thick, and all completely fabricated, i.e., heated or printed. The discs were placed in 96-well plates. Microorganisms were cultured on 270 discs for 24 h (90 discs), 72 h (90 other discs), and 5 days or 120 h (90 other discs). Biofilm formation of the strains and negative controls was measured using the microtiter plate assay by ELISA reading. The microbes’ ability to produce biofilm was categorized based on the comparison of average optical density (OD) of tests versus a cut-off point OD (ODc) calculated as the average of the OD of corresponding negative controls plus 3× its standard deviation: non-biofilm former [OD ≤ ODc], weak biofilm former [ODc < OD ≤ (2 × ODc)], moderate biofilm former [(2 × ODc) < OD ≤ (4 × ODc)], and strong biofilm former [(4 × ODc) < OD]. These were also converted to ranked scores between zero (no biofilm) and 3. The difference between ODs with control ODs were calculated. These were analyzed using 3-way ANOVA, 2-way ANOVA, and Tukey tests (α = 0.05, α = 0.008).ResultsThe 3-way ANOVA showed that the overall difference among the ΔODs of 5 retainers (all microorganisms and all intervals combined, n = 270) was not significant (F = 1.860, P = 0.119). Nevertheless, the difference among 3 intervals (F = 31.607, P = 0.0000) and the difference among the 6 microorganisms (F = 24.044, P = 0.0000) were significant. According to the Tukey test, the differences between the 1st interval with either of the other two intervals was significant (both P values = 0.000). There were significant differences between Candida albicans with all other organisms (all 5 P values = 0.0000). All other pairwise comparisons were insignificant (all 10 P values ≥ 0.1). After taking the averages of the 3 intervals, the order of the biofilm generation for different materials were as follows: Detax (average score: 1.56), Easyvac (1.67), Erkodent (1.78), Clear Tech (1.83), BD (2.28).ConclusionsAs far as these 6 microorganisms are of concern, there might not be a significant overall difference among the clear retainer materials tested in this study. A significant overall increase was observed between the first and third days, which later did not significantly increase more until day 5. The Candida albicans biofilm was more intense than the tested 5 bacteria, which themselves showed rather similar growth patterns to each other.

Mining for antifungal agents to inhibit biofilm formation of Candida albicans: A study on green synthesis, antibiofilm, cytotoxicity, and in silico ADME analysis of 2-amino-4H-pyran-3-carbonitrile derivatives

Candida albicans (C. albicans) biofilm infections are quite difficult to manage due to their resistance against conventional antifungal drugs. To address this issue, there is a desperate need for new therapeutic drugs. In the present study, a green and efficient protocol has been developed for the synthesis of 2-amino-4H-pyran-3-carbonitrile scaffolds 4a-i, 6a-j, and 8a-g by Knoevenagel-Michael-cyclocondensation reaction between aldehydes, malononitrile, and diverse enolizable C-H activated acidic compounds using guanidinium carbonate as a catalyst either under grinding conditions or by stirring at room temperature. This protocol is operationally simple, rapid, inexpensive, has easy workup and column-free purification. A further investigation of the synthesized compounds was conducted to examine their antifungal potential and their ability to inhibit the growth and development of biofilm-forming yeasts like fungus C. albicans. According to our findings, 4b, 4d, 4e, 6e, 6f, 6g, 6i, 8c, 8d, and 8g were found to be active and potential inhibitors for biofilm infection causing C. albicans. The inhibition of biofilm by active compounds were observed using field emission scanning electron microscopy (FESEM). Biofilm inhibiting compounds were also tested for in vitro toxicity by using 3T3-L1 cell line, and 4b, 6e, 6f, 6g, 6i, 8c, and 8d were found to be biocompatible. Furthermore, the in silico ADME descriptors revealed drug-like properties with no violation of Lipinski's rule of five. Hence, the result suggested that synthesized derivatives could serve as a useful aid in the development of novel antifungal compounds for the treatment of fungal infections and virulence in C. albicans.

EFFECT OF MORINGA (Moringa oleifera) LEAF EXTRACT ON IN VITRO INHIBITION OF Candida albicans BIOFILM

Background: Oral candidiasis is an opportunistic infection in the oral cavity. This infection is caused by the fungus Candida albicans. The most important virulent attribute of this fungus is its ability to form biofilms, which can adhere to mucosa, epithelial lining, organs, prostheses or dentures. The formed biofilm is resistant to antifungal drugs. Objectives: The purpose of this study was to determine the effect of Moringa (Moringa oleifera) leaf extract on the inhibition of Candida albicans biofilm formation in vitro. Methods: This type of research is experimental study using post tests with control group design. Fungus used in this study was Candida albicans ATCC 10231. Determination of minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were conducted by microdilution method. Inhibition test of C. albicans biofilm formation was carried out using the polystyrene microplate assay method. Concentration of extracts used in this study were 12.5%, 6.25%, 3.13% and 1.57%. Inhibitory activity of biofilm was measured using a crystal violet (CV) assay. Results: The results showed that MIC of Moringa leaf extract against the fungus C. albicans was at a concentration of 6.25%. Moringa leaf extract starting at a concentration of 3.13% already has had ability to inhibit formation of C. albicans biofilm. Conclusion: It can be concluded that Moringa leaf extract can inhibit formation of Candida albicans biofilm, so it can be developed as an alternative herbal ingredient to prevent oral candidiasis.EFFECT OF MORINGA (Moringa oleifera) LEAF EXTRACT ON IN VITRO INHIBITION OF Candida albicans BIOFILM

Psidium guajav-mediated zinc oxide nanoparticles as a multifunctional, microbicidal, antioxidant and antiproliferative agent against destructive pathogens.

Bio-inspired zinc oxide nanoparticles are gaining immense interest due to their safety, low cost, biocompatibility, and broad biological properties. In recent years, much research has been focused on plant-based nanoparticles, mainly for their eco-friendly, facile, and non-toxic character. Hence, the current study emphasized a bottom-up synthesis of zinc oxide nanoparticles (ZnO NPs) from Psidium guajava aqueous leaf extract and evaluation of its biological properties. The structural characteristic features of biosynthesized ZnO NPs were confirmed using various analytical methods, such as UV-Vis spectroscopy, X-ray diffraction (XRD), energy-dispersive X-ray analysis (EDX), Fourier transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS), Scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (HR-TEM). The synthesized ZnO NPs exhibited ahydrodynamic shape with an average particle size of 11.6-80.2nm. A significant antimicrobial efficiency with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 40 and 27µg/ml for Enterococcus faecalis, followed by 30 and 40µg/ml for Staphylococcus aureus, 20 and 30µg/ml for Staphylococcus mutans, 30µg/ml for Candida albicans was observed by ZnO NPs. Additionally, they showed significant breakdown of biofilms of Streptococcus mutans and Candida albicans indicating their future value in drug-resistance research. Furthermore, an excellent dose-dependent activity of antioxidant property was noticed with an IC50 of 9.89µg/ml. The antiproliferative potential of the ZnO NPs was indicated by the viability of MDA MB 231 cells, which showed adrastic decrease in response to increased concentrations of biosynthesized ZnO NPs. Thus, the present results open up vistas to explore their pharmaceutical potential for the development of targeted anticancer drugs in the future.

Optimizing therapeutic efficacy of antifungal peptides via strategic terminal amino acid modification

IntroductionAntifungal peptides (AFPs) have the potential to treat antifungal-resistant infections; however, their structure–function relationship remains unknown, hindering their rapid development. Therefore, it is imperative to investigate and clarify the structure–function relationships of AFPs. ObjectivesThis study aimed to investigate the impact of end-tagging single hydrophobic amino acids and capping the N-terminus with glycine (Gly) on the antifungal activity of peptide W4. MethodsThe antifungal efficacy of the engineered peptides was initially assessed by determining the minimum inhibitory concentration (MIC) /minimal fungicidal concentration (MFC), killing kinetics, and drug resistance induction, in addition to evaluating the biocompatibility and stability. Subsequently, the antifungal mechanism was investigated using fluorescence labeling, electron microscopy, reactive oxygen species (ROS) detection, and measurement of mitochondrial membrane potential and apoptosis. The impact of the engineered peptides on Candida albicans (C. albicans) biofilm and their potential application in the scratch keratomycosis model were investigated. ResultsThe antifungal activity of W4 was significantly enhanced by capping Gly at the N-terminus, resulting in a decrease in average activity from 11.86 μM to 6.25 μM (GW4) and an increase in TI values by 1.9-fold (TIGW4 = 40.99). Mechanistically, GW4 exerted its antifungal effect by disrupting the cellular membrane structure in C. albicans, forming pores and subsequent leakage of intracellular contents. Concurrently, it facilitated intracellular ROS accumulation while decreasing the mitochondrial membrane potential. Additionally, GW4 demonstrated an excellent ability to inhibit and eliminate biofilms of C. albicans. Notably, GW4 demonstrated significant therapeutic potential in a C. albicans-associated keratitis model. ConclusionCapping Gly at the N-terminus increased residue length while significantly enhancing the helical propensity of W4, thereby augmenting its antifungal activity. Our exploratory study demonstrated the potential strategies and avenues for optimizing the structure–function relationships of AFPs and developing highly effective antifungal drugs.

Interkingdom biofilm of Streptococcus pyogenes and Candida albicans: establishment of an in vitro model and dose-response validation of antimicrobials

Although Streptococcus pyogenes and Candida albicans may colonize tonsillar tissues, the interaction between them in mixed biofilms has been poorly explored. This study established an interkingdom biofilm model of S. pyogenes and C. albicans and verified the dose-response validation of antimicrobials. Biofilms were formed on microplates, in the presence or absence of a conditioning layer of human saliva, using Brain Heart Infusion (BHI) broth or artificial saliva (AS) as a culture medium, and with variations in the microorganism inoculation sequence. Biofilms grown in AS showed higher mass than those grown in BHI broth, and an opposite trend was observed for metabolism. The number of S. pyogenes colonies was lower in AS. Amoxicillin and nystatin showed dose-dependent effects. The inoculation of the two species at the same time, without prior exposure to saliva, and using BHI broth would be the model of choice for future studies assessing the effects of antimicrobials on dual S. pyogenes/C. albicans biofilms.

Preventing proximal enamel caries in neighboring tooth with glass ionomer cement restoration and silver diamine fluoride pretreatment

ObjectiveTo investigate caries preventive effects of 38 % silver diamine fluoride (SDF) pretreatment on neighboring tooth proximal to glass ionomer cement (GIC), including conventional GIC (CGIC) and resin-modified GIC (RMGIC) restorations in an in vitro model. Methods: Human tooth blocks were restored withSDF+CGIC (Group 1), CGIC (Group 2), SDF+RMGIC (Group 3) or RMGIC (Group 4). Enamel specimen simulating proximal surface of neighboring tooth was placed in proximity to the restorations. The specimen underwent cariogenic challenge with cross-kingdom biofilm of Streptococcus mutans, Lacticaseibacillus casei and Candida albicans. After cariogenic challenge, the biofilm's growth kinetics, viability, and morphology were evaluated by propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR), confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM), respectively. The enamel lesion depth, surface morphology and crystal characteristics were determined by micro-computed tomography (micro-CT), SEM and X-ray diffraction (XRD), respectively. ResultsPMA-qPCR demonstrated lower microbial growth in Group 1 and 3 compared with Group 2 and 4 (p < 0.05). CLSM showed the dead−to-live ratio in Groups 1–4 were 1.15±0.12, 0.53±0.13, 1.10±0.24 and 0.63±0.10, respectively (Group 1,3 > 2,4, p < 0.05). SEM revealed Groups 1 and 3 had scattered biofilm whereas Group 2 and 4 had confluent biofilm. Micro-CT showed the enamel lesion depths (µm) were 98±9, 126±7, 103±6 and 128±7 for Group 1 to 4, respectively (Group 1,3 < 2,4, p < 0.05). SEM revealed oriented and ordered enamel prismatic patterns in Group 1 and 3, not in Group 2 and 4. XRD showed the reflections of hydroxyapatite in Groups 1 and 3 were sharper than Groups 2 and 4. ConclusionSDF pretreatment enhances the preventive effect of GIC on proximal enamel surface on neighboring tooth through inhibiting cariogenic biofilm, reducing enamel demineralization and promoting enamel remineralization. Clinical significanceSDF pretreatment of GIC restorations can help prevent caries on neighboring teeth, particular for patients with high caries risk.

Inhibition of Development and Metabolism of Dual-Species Biofilms of Candida albicans and Candida krusei (Pichia kudriavzevii) by Organoselenium Compounds.

Although Candida albicans is the most frequently identified Candida species in clinical settings, a significant number of infections related to the non-albicans Candida (NAC) species, Candida krusei, has been reported. Both species are able to produce biofilms and have been an important resistance-related factor to antimicrobial resistance. In addition, the microbial relationship is common in the human body, contributing to the formation of polymicrobial biofilms. Considering the great number of reports showing the increase in cases of resistance to the available antifungal drugs, the development of new and effective antifungal agents is critical. The inhibitory effect of Organoselenium Compounds (OCs) on the development of Candida albicans and Candida krusei was recently demonstrated, supporting the potential of these compounds as efficient antifungal drugs. In addition, OCs were able to reduce the viability and the development of biofilms, a very important step in colonization and infection caused by fungi. Thus, the objective of this study was to investigate the effect of the Organoselenium Compounds (p-MeOPhSe)2, (PhSe)2, and (p-Cl-PhSe)2 on the development of dual-species biofilms of Candida albicans and Candida krusei produced using either RPMI-1640 or Sabouraud Dextrose Broth (SDB) media. The development of dual-species biofilms was evaluated by the determination of both metabolic activity, using a metabolic assay based on the reduction of XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide sodium salt) assay and identification of either Candida albicans and Candida krusei on CHROMagar Candida medium. Biofilm formation using RPMI-1640 was inhibited in 90, 55, and 20% by 30 µM (p-MeOPhSe)2, (PhSe)2, and (p-Cl-PhSe)2, respectively. However, biofilms produced using SDB presented an inhibition of 62, 30 and 15% in the presence of 30 µM (p-MeOPhSe)2, (PhSe)2, and (p-Cl-PhSe)2, respectively. The metabolic activity of 24 h biofilms was inhibited by 35, 30 and 20% by 30 µM (p-MeOPhSe)2, (PhSe)2, and (p-Cl-PhSe)2, respectively, with RPMI-1640; however, 24 h biofilms formed using SDB were not modified by the OCs. In addition, a great reduction in the number of CFUs of Candida albicans (93%) in biofilms produced using RPMI-1640 in the presence of 30 µM (p-MeOPhSe)2 was observed. However, biofilms formed using SDB and treated with 30 µM (p-MeOPhSe)2 presented a reduction of 97 and 69% in the number of CFUs of Candida albicans and Candida krusei, respectively. These results demonstrated that Organoselenium Compounds, mainly (p-MeOPhSe)2, are able to decrease the metabolic activity of dual-species biofilms by reducing both Candida albicans and Candida krusei cell number during biofilm formation using either RPMI-1640 or SDB. Taken together, these results demonstrated the potential of the OCs to inhibit the development of dual-species biofilms of Candida albicans and Candida krusei.

Antifungal Effect of Poly(methyl methacrylate) with Farnesol and Undecylenic Acid against Candida albicans Biofilm Formation.

The control of Candida albicans biofilm formation on dentures made of poly(methyl methacrylate) (PMMA) is an important challenge due to the high resistance to antifungal drugs. Interestingly, the natural compounds undecylenic acid (UDA) and farnesol (FAR) both prevent C. albicans biofilm formation and could have a synergetic effect. We therefore modified PMMA with a combination of UDA and FAR (UDA+FAR), aiming to obtain the antifungal PMMA_UDA+FAR composites. Equal concentrations of FAR and UDA were added to PMMA to reach 3%, 6%, and 9% in total of both compounds in composites. The physico-chemical properties of the composites were characterized by Fourier-transform infrared spectroscopy and water contact angle measurement. The antifungal activity of the composites was tested on both biofilm and planktonic cells with an XTT test 0 and 6 days after the composites' preparation. The effect of the UDA+FAR combination on C. albicans filamentation was studied in agar containing 0.0125% and 0.4% UDA+FAR after 24 h and 48 h of incubation. The results showed the presence of UDA and FAR on the composite and decreases in the water contact angle and metabolic activity of both the biofilm and planktonic cells at both time points at non-toxic UDA+FAR concentrations. Thus, the modification of PMMA with a combination of UDA+FAR reduces C. albicans biofilm formation on dentures and could be a promising anti-Candida strategy.

The arrangement of dual-species biofilms of Candida albicans and Issatchenkia orientalis can be modified by the medium: effect of Voriconazole

Both Candida albicans and Issatchenkia orientalis have been isolated from different types of infections over the years. They have the ability to form communities of microorganisms known as biofilms. It has been demonstrated that the medium employed in studies may affect the biofilm development. The aim of this study was to investigate the arrangement of dual-species biofilms of C. albicans and I. orientalis cultivated on either RPMI-1640 or Sabouraud Dextrose Broth (SDB), as well as the inhibitory effect of Voriconazole (VRC). For the experiments performed, ATCC strains were used, and yeast-mixed suspensions were inoculated in 96-well plates with either RPMI-1640 or SDB, in the presence or absence of VRC. The results were observed by counting the number of CFU obtained from scraping off the biofilms produced and plating the content on CHROMagar Candida medium. It was observed that for all conditions tested the medium chosen affected the arrangement of dual-species biofilms: when RPMI-1640 was used, there was a prevalence of C. albicans, while the opposite was noted when SDB was used. It could be suggested that the medium and environment could regulate interactions between both yeast species, including the response to different antifungal drugs.

Green Synthesis of Silver Nanoparticles using Moringa oleifera: Implementation to Photoantimicrobial of Candida albicans with LED Light

Green synthesis of silver nanoparticles using Moringa oleifera (AgNPs/MO) was carried out to produce nanoscale antifungal agents which are used as photosensitizing agents in photoantimicrobial therapy. This study focused on investigating the potential of AgNPs/MO producing some toxic radical compounds that inhibit the growth of Candida albicans biofilms. The inhibition mechanism uses the principle of photoinactivation treatment which combines a photosensitizer with an LED light source at a power of 100 mW for 60 - 300 s of exposure. Quantitative data analyzed were the number of viable cells and radical compounds, formed by malondialdehyde (MDA) level. It is investigated through staining the XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) and TBARS (Thiobarbituric Acid Reactive Substances). The results show that the maximum wavelength of AgNPs is 440 nm while the Moringa oleifera has 2 peaks at 425 and 635 nm. The maximum effect occurred in the group of photosensitizer AgNPs/MO combined LED with percentage inactivation about 76.80 % for blue LEDs and 76.20 % for red LEDs. The group of LED irradiation without photosensitizer obtained about 51.85 % for blue LEDs and 50.04 % for red LEDs. The MDA level group of AgNPs/MO combined LED also produced MDA levels of 1.772 nmol/mL for blue LED and 1.617 nmol/mL for red LED. Meanwhile, the application of photoantimicrobial using only LEDs produced MDA levels of only 1.353 nmol/mL for blue LEDs and 1.347 nmol/mL for red LEDs. This research has shown that the green synthesis of AgNPs/MO has a good potential to inhibit the growth of Candida albicans biofilms as a photosensitizer agent. HIGHLIGHTS Silver nanoparticles (AgNPs) combined with Moringa oleifera (MO) extract have been shown to have potential as a photosensitizer to inhibit the growth of Candida albicans Green synthesis AgNPs/MO have a characteristic spectrum at 440 nm which is close to the blue light spectrum. Green synthesis AgNPs/MO combined with LED light irradiation energy can reduce the cell viability of Candida albicans up to 76.80 % through the XTT assay staining test method and produce 1.772 nmol/mL of Malondialdehyde that reacts with ROS compounds to cause cell damage. GRAPHICAL ABSTRACT

Biofilm Formation on Denture Base Material Reinforced With a Novel Organic Material.

Microcrystalline cellulose (MCC) is a novel organic material developed by one of the authors in this study. When MCC was incorporated with conventionally available denture base resin, it demonstrated increased flexural strength and flexural modulus. However, it was speculated that because the material is organic, it can promote the growth of Candida. The purpose of this study is to evaluate the Candida albicans biofilm formation on polymethyl methacrylate (PMMA) denture base resin incorporated with MCC. MCC is an organic material extracted from the oil palm empty fruit bunch (OPEFB). The growth of C. albicans and biofilm formation in three test groups were compared by biofilm assay and imaging techniques like microscopy (by safranin staining) and scanning electron microscopy. The three test groups were comprised of MCC-reinforced PMMA containing OPEFB fibers of 50-micrometer thickness at 5% weight reinforcement, conventionally and commercially available heat cure PMMA, and an empty well to assess any discrepancies from the environment. The test groups showed increased biofilm formation by C. albicans compared to commercially and conventionally available heat cure PMMA. Reinforcement with MCC showed higher biofilm formation of 1.43 times higher compared to conventional PMMA. Biofilms formed by Candida albicans on MCC-reinforced PMMA appeared heterogeneous in structure, comprised of yeast cells and hyphae, surrounded by a higher density of polysaccharide extracellular matrix material compared to that of conventionally available heat cure PMMA. Biofilm formation is increased in denture base resin incorporated with MCC. More investigation is warranted to study the antifungal efficacy of the addition of antifungal agents to the reinforced denture base resin.