Antimicrobial and anti-biofilm properties of polypropylene meshes coated with metal-containing DLC thin films.

A promising strategy to reduce nosocomial infections related to prosthetic meshes is the prevention of microbial colonization. To this aim, prosthetic meshes coated with antimicrobial thin films are proposed. Commercial polypropylene meshes were coated with metal-containing diamond-like carbon (Me-DLC) thin films by the magnetron sputtering technique. Several dissimilar metals (silver, cobalt, indium, tungsten, tin, aluminum, chromium, zinc, manganese, tantalum, and titanium) were tested and compositional analyses of each Me-DLC were performed by Rutherford backscattering spectrometry. Antimicrobial activities of the films against five microbial species (Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Enterococcus faecalis) were also investigated by a modified Kirby-Bauer test. Results showed that films containing silver and cobalt have inhibited the growth of all microbial species. Tungsten-DLC, tin-DLC, aluminum-DLC, zinc-DLC, manganese-DLC, and tantalum-DLC inhibited the growth of some strains, while chromium- and titanium-DLC weakly inhibited the growth of only one tested strain. In-DLC film showed no antimicrobial activity. The effects of tungsten-DLC and cobalt-DLC on Pseudomonas aeruginosa biofilm formation were also assessed. Tungsten-DLC was able to significantly reduce biofilm formation. Overall, the experimental results in the present study have shown new approaches to coating polymeric biomaterials aiming antimicrobial effect.

Effect of green tea extract and mouthwash without alcohol on Candida albicans biofilm on acrylic resin.

OBJECTIVE: To evaluate the effect of aqueous extract of green tea and the oral antiseptic without alcohol, on Candida albicans biofilm formation to heat-curing acrylic resin plates. BACKGROUND: Candida is associated with oral candidiasis in poorly cleaned dentures. MATERIAL AND METHODS: Standardised specimens of heat-cured (Conv; n = 30) or microwave-cured acrylic resin (Mw; n = 30) were obtained and divided into six groups (n = 10): G1 = Conv resin and green tea aqueous extract, G2 = Conv resin and mouthwash, G3 = control of Conv resin, G4 = Mw resin and green tea aqueous extract, G5 = Mw resin and mouthwash and G6 = control of Mw-cured resin. The specimens were contaminated with 10 ml of Sabouraud dextrose broth inoculated with 0.1 ml of standard suspension containing 1 × 10(6) cells/ml of C. albicans and incubated for 24 h at 37°C. After this period, they were immersed in the aqueous extract or in mouthwash for 15 min. The control groups were treated with sterile distilled water. Aliquots of 0.1 ml were plated on Sabouraud dextrose agar and incubated at 37°C for 24 h. The numbers of colony-forming units per test specimen (CFU/TS) were calculated, and the results statistically analysed by two-way anova and Tukey's tests (5%). RESULTS: Statistically significant difference was observed for the aqueous extract groups (G1; 33.65%) and mouthwash (G2; 17.06%), when compared to control (G3; 100%), for Conv resin. For the Mw resin, there was significant difference between mouthwash (G5; 43.16%) and control (G6; 100%). CONCLUSION: The aqueous extract of green tea and mouthwash led to a reduction in the number of viable fungal cells in biofilm formed on acrylic resin.

Influence of TNF-α blockers on the oral prevalence of opportunistic microorganisms in ankylosing spondylitis patients.

OBJECTIVES: To compare the oral prevalence and antimicrobial susceptibility of candida spp., staphylococci, enterobacteriaceae, and pseudomonas spp. from ankylosing spondylitis (AS) patients receiving conventional and anti-TNF-α therapy. METHODS: The study included 70 AS patients, diagnosed according to the modified New York criteria (1984). The volunteers were divided into 2 groups: a biological group (AS BioG) (n=35) (on anti-TNF-α therapy) and a conventional group (AS ConvG) (n=35). The control group (ContG) (n=70) was made up of healthy individuals matched for age, gender, and oral conditions. After clinical examination, oral rinse samples were collected and plated in specific culture media. The number of colony-forming units per milliliter (cfu/ml) was obtained, and isolates were identified using the API system. Antimicrobial susceptibility tests were performed according to the NCCLS guidelines. Prevalence and counts of microorganisms were statistically compared between the 3 groups, using the Mann-Whitney and Chi-square tests. Significance level was set at 5%. RESULTS: In both the AS BioG and the AS ConvG, staphylococci counts were higher than that in the ContG (p<0.0001). Candida albicans and staphylococcus epidermidis were the most commonly found species in all the groups. Serratia marcescens and klebsiella oxytoca were more prevalent in the AS BioG and the AS ConvG, respectively. Two candida isolates (2.8%) from the AS BioG and 5 (10.8%) from the AS ConvG were resistant to amphotericin B and 5-fluorocytosine. A low percentage of staphylococci isolates was resistant to amoxicillin, ciprofloxacin, and doxycycline. CONCLUSIONS: Higher counts of staphylococci were observed in both AS groups, regardless of the current therapy, age, sex, and oral conditions. Anti-TNF-α therapy could not be correlated with increased counts of microorganisms.

Antibiofilm effects of Thymus vulgaris and Hyptis spicigera essential oils on cariogenic bacteria.

Aim: The inhibitory and antibiofilm effects of Thymus vulgaris (EOTv) and Hyptis spicigera essential oils (EOHs) on cariogenic microorganisms were evaluated. Materials & methods: The chemical characterization of EOTv was performed by gas chromatography/mass spectrometry. Streptococcus mutans, Streptococcus gordonii, Streptococcus sanguinis, Streptococcus mitis, Streptococcus sobrinus, Lactobacillus acidophilus and Actinomyces naeslundii were used for agar diffusion assays and determination of minimal inhibitory and minimal bactericide concentrations. In addition, 20 streptococci and lactobacilli clinical isolates were also tested. The effects of essential oil on microbial initial biofilm formation and on preformed microcosm biofilm formed from human saliva were studied. Results & conclusion: Both essential oils had inhibitory effects on the cariogenic species and reduced the bacterial adherence to dental enamel. Essential oils were able to disrupt preformed microcosm biofilms. Thymus vulgaris and Hyptis spicigera essential oils have potential to be used in the development of formulations to the control of cariogenic biofilms.

Non-steroidal anti-inflammatory drugs may modulate the protease activity of Candida albicans.

The phenotypic pressure exerted by non-steroidal anti-inflammatory drugs (NSAIDs) on autochthonous and pathogenic microbiota remains sparsely known. In this study, we investigated if some NSAIDs increment or diminish the secretion of aspartyl-proteases (Sap) by Candida albicans grown under different phenotypes and oxygen availability using a set of SAP knock-out mutants and other set for genes (EFG1 and CPH1) that codify transcription factors involved in filamentation and protease secretion. Pre-conditioned cells were grown under planktonic and biofilm phenotypes, in normoxia and anoxia, in the presence of plasma concentrations of acetylsalicylic acid, diclofenac, indomethacin, nimesulide, piroxicam, ibuprofen, and acetaminophen. For diclofenac, indomethacin, nimesulide, and piroxicam the secretion rates of Sap by SAP1-6, EFG1, and CPH1 mutants were similar or, even, inferior to parental wild-type strain. This suggests that neither Sap 1-6 isoenzymes nor Efg1/Cph1 pathways may be entirely responsible for protease release when exposed to these NSAIDs. Ibuprofen and acetaminophen enhanced Sap secretion rates in three environmental conditions (normoxic biofilm, normoxic planktonic and anoxic planktonic). In other hand, aspirin seems to reduce the Sap-related pathogenic behavior of candidal biofilms. Modulation of Sap activity may occur according to candidal phenotypic state, oxygen availability, and type of NSAID to which the cells are exposed.

Biopharmaceutical and antifungal properties of ellagic acid-cyclodextrin using an in vitro model of invasive candidiasis.

Aim: To investigate biopharmaceutical and antifungal properties of pure and complexed ellagic acid. Materials & methods: Caco-2 cells cultured in a Transwell® inserts were infected with Candida albicans to develop an in vitro model. Ellagic acid was complexed with cyclodextrins. Microbial compositions, ellagic acid concentration as function of time and characterization studies of complexes were evaluated. Results: Ellagic acid presented ability to reduce C. albicans invasion, although this was not statistically significant. Its poor water solubility and absorption probably limited this ability. Water solubility was increased after complexation with hydroxypropyl-ß-CD; however, ellagic acid/hydroxypropyl-ß-CD did not improve the antifungal activity. Conclusion: Although ellagic acid presented a promising antifungal activity, its biopharmaceutical properties limit such activity and should be improved.

In vitro antifungal susceptibility of clinical isolates of Candida spp. obtained from patients with different predisposing factors to candidosis.

The increase in the number of infections caused by Candida species and the consequent use of antifungal agents favours an increase of resistant isolates. The aim of this study was to evaluate the antifungal susceptibility of Candida spp. isolates from patients with different systemic predisposing factors to candidosis. Seventy-nine Candida spp. isolates were assayed for in vitro susceptibility to amphotericin B, fluconazole, 5-flucytosine and itraconazole using the technique proposed by the Clinical and Laboratory Standards Institute (CLSI). Four C. albicans, one C. guilliermondii, four C. parapsilosis and two C. tropicalis isolates were resistant to amphotericin B. Only two isolate was resistant to itraconazole. All the isolates tested were susceptible to fluconazole and flucytosine. It could be concluded that the most efficient drugs against the Candida isolates studied were fluconazole and flucytosine and that all of the antifungal agents used in this study were effective against the Candida spp. isolates tested.

Effects of Acetone Fraction From Buchenavia tomentosa Aqueous Extract and Gallic Acid on Candida albicans Biofilms and Virulence Factors.

A promising anti-Candida activity of Buchenavia tomentosa extracts was recently described. In the present work, experiments were carried out to determine the fraction with higher antifungal activity from a B. tomentosa extract. Acetone fraction (AF) was obtained from the aqueous extract from dried leaves (5 min/100°C) and it was the most effective one. Gallic acid (GA) was identified by electrospray ionization mass spectrometry (ESI-MS) and also chosen to perform antifungal tests due to its promising activity on Candida albicans. Minimal inhibitory and fungicidal concentrations (MIC and MFC) were determined by broth microdilution technique. The effect on virulence factors of C. albicans was evaluated, and the cytotoxicity was determined. MIC50 and MIC90 values were both equal to 0.625 mg ml-1 for AF and 2.5 and 5 mg ml-1, respectively, for GA. AF and GA showed ability to inhibit C. albicans adherence and to disrupt 48 h-biofilm. AF and GA were effective in reducing the formation of hyphae of C. albicans SC5314. AF and GA decreased adherence of C. albicans to oral epithelial cells. AF and GA showed slight to moderate toxicity to Vero cells. This result suggests further studies for topic use of these compounds. AF, which contains a combination of several molecules, presented greater potential of antimicrobial activity than GA, with lower values of MIC and lower cytoxicity.

Potential Use of Phenolic Acids as Anti-Candida Agents: A Review.

There has been a sharp rise in the occurrence of Candida infections and associated mortality over the last few years, due to the growing body of immunocompromised population. Limited number of currently available antifungal agents, undesirable side effects and toxicity, as well as emergence of resistant strains pose a considerable clinical challenge for the treatment of candidiasis. Therefore, molecules that derived from natural sources exhibiting considerable antifungal properties are a promising source for the development of novel anti-candidal therapy. Phenolic compounds isolated from natural sources possess antifungal properties of interest. Particularly, phenolic acids have shown promising in vitro and in vivo activity against Candida species. However, studies on their mechanism of action alone or in synergism with known antifungals are still scarce. This review attempts to discuss the potential use, proposed mechanisms of action and limitations of the phenolic acids in anti-candidal therapy.

Antifungal activity of extracts and isolated compounds from Buchenavia tomentosa on Candida albicans and non-albicans.

AIM: This study aimed to evaluate the antifungal activity of Buchenavia tomentosa extract and bioactive compounds on six Candida species. MATERIALS & METHODS: The antimicrobial activity of extract was evaluated using standard strains and clinical isolates. Cytotoxicity was tested in order to evaluate cell damage caused by the extract. Extract was chemically characterized and the antifungal activity of its compounds was evaluated. RESULTS: Extract showed antifungal activity on Candida species. Candida non-albicans were more susceptible than Candida albicans. Low cytotoxicity for extract was observed. The isolated compounds presented antifungal activity at least against one Candida spp. and all compounds presented antifungal effect on Candida glabrata. CONCLUSION: Extracts from Buchenavia tomentosa showed promising antifungal activity on Candida species with low cytotoxicity. Gallic acid, corilagin and ellagic acid showed promising inhibitory activity on Candida glabrata.

Genetic variability analysis among clinical Candida spp. isolates using random amplified polymorphic DNA.

The patterns of genetic variation of samples of Candida spp. isolated from patients infected with human immunodeficiency virus in Vitória, state of Espírito Santo, Brazil, were examined. Thirty-seven strains were isolated from different anatomical sites obtained from different infection episodes of 11 patients infected with the human immunodeficiency virus (HIV). These samples were subjected to randomly amplified polymorphic DNA (RAPD) analysis using 9 different primers. Reproducible and complex DNA banding patterns were obtained. The experiments indicated evidence of dynamic process of yeast colonization in HIV-infected patients, and also that certain primers are efficient in the identification of species of the Candida genus. Thus, we conclude that RAPD analysis may be useful in providing genotypic characters for Candida species typing in epidemiological investigations, and also for the rapid identification of pathogenic fungi.

rDNA-RFLP identification of Candida species in immunocompromised and seriously diseased patients.

PCR was used to amplify a targeted region of the ribosomal DNA of 76 Candida spp. isolates from immunocompromised and seriously diseased patients. Thirty-seven strains isolated from different anatomical sites of 11 patients infected with HIV (Vitória, ES, Brazil), 26 isolates from patients under treatment at Odilon Behrens Hospital and 13 isolates from skin and urine samples from São Marcos Clinical Analysis Laboratory (Belo Horizonte, Brazil) were scored. Fragments of rDNA were amplified using primer pairs ITS1-ITS4, for the amplification of ITS1 and ITS2 regions, including the gene for the 5.8 s subunit. Amplification resulted in fragments ranging in size from 350 to 950 bp. Amplicons were digested with eight restriction enzymes. A pattern of species-specificity among the different medically important Candida species could be identified following restriction digestion of the PCR products. Candida albicans was the species most frequently observed, except for the group of newborns under treatment at the Odilon Behrens Hospital and for the isolates from the clinical analysis laboratory. C. parapsilosis was the species most frequently observed in these two groups.

Genetic variability analysis among clinical Candida spp. isolates using random amplified polymorphic DNA

The patterns of genetic variation of samples of Candida spp. isolated from patients infected with human immunodeficiency virus in Vitória, state of Espírito Santo, Brazil, were examined. Thirty-seven strains were isolated from different anatomical sites obtained from different infection episodes of 11 patients infected with the human immunodeficiency virus (HIV). These samples were subjected to randomly amplified polymorphic DNA (RAPD) analysis using 9 different primers. Reproducible and complex DNA banding patterns were obtained. The experiments indicated evidence of dynamic process of yeast colonization in HIV-infected patients, and also that certain primers are efficient in the identification of species of the Candida genus. Thus, we conclude that RAPD analysis may be useful in providing genotypic characters for Candida species typing in epidemiological investigations, and also for the rapid identification of pathogenic fungi.