Effect of disinfectants on multispecies biofilm, the physical and mechanical properties of polymethyl methacrylate, and the corrosion of cobalt chromium alloy.

STATEMENT OF PROBLEM: The optimal disinfection protocol that controls adverse effects and promotes effective antimicrobial action on removable prostheses is unclear. PURPOSE: This in vitro study investigated the effect of disinfectant solutions on the biological, physical, mechanical, and chemical properties of removable prosthesis materials. MATERIAL AND METHODS: Specimens of polymethyl methacrylate (PMMA) and cobalt chromium (Co-Cr) alloy were immersed in distilled water (PMMA) or artificial saliva (Co-Cr) as the control and in 0.25% sodium hypochlorite (NaOCl0.25%), 0.5% chloramine T (CT0.5%), and 0.15% Triclosan (TR0.15%). The antibiofilm activity was evaluated by microbial load and cell metabolisms of the mixed biofilm. Physical (color change, sorption, solubility, and surface roughness), mechanical (hardness, flexural, and impact strength), and chemical (corrosion) properties were analyzed before and after simulating a 5-year immersion. Laser confocal microscopy, scanning electron microscopy (SEM), and dispersive energy spectroscopy (EDS) complemented the analyses. The data were analyzed by using the Mann-Whitney U test, Kruskal-Wallis with Dunn posttests, 1-way ANOVA, and repeated measures ANOVA (α=.05). RESULTS: All solutions were effective against bacteria, but only NaOCl0.25% eliminated Candida spp. TR0.15%, and CT0.5% increased cell metabolisms. For interaction (time and solution), there was a reduction in PMMA hardness in the control and TR0.15%. Color, sorption, solubility, and flexural strength did not change. CT0.5% and TR0.15% were similar for impact resistance. CT0.5% caused the lowest roughness. NaOCl0.25% showed the greatest corrosive potential. Dark spots were seen under SEM in Co-Cr stored with NaOCl0.25% and TR0.15%. EDS indicated different proportions of oxygen, cobalt, chromium, and molybdenum. CONCLUSIONS: NaOCl0.25% had the best antimicrobial action. CT0.5% and TR0.15% have potential. Hardness and roughness changes were clinically acceptable, and the other properties remained unchanged. All the solutions caused color changes. NaOCl0.25% was unsatisfactory for use with Co-Cr, CT0.5% was intermediate, and TR0.15% was suitable.

In vitro antibacterial activity of green tea-loaded chitosan nanoparticles on caries-related microorganisms and dentin after Er:YAG laser caries removal.

This study aimed to determine the inhibitory effects of green tea (Gt), EGCG, and nanoformulations containing chitosan (Nchi) and chitosan+green tea (Nchi+Gt) against Streptococcus mutans and Lactobacillus casei. In addition, the antibacterial effect of nanoformulations was evaluated directly on dentin after the selective removal of carious lesion. At first, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against S. mutans and L. casei isolates were investigated. In parallel, dentin specimens were exposed to S. mutans to induce carious lesions. Soft dentin was selectively removed by Er:YAG laser (n=33) or bur (n=33). Remaining dentin was biomodified with Nchi (n=11) or Gt+Nchi (n=11). Control group (n=11) did not receive any treatment. Dentin scraps were collected at three time points. Microbiological analyses were conducted and evaluated by agar plate counts. Gt at 1:32 dilution inhibited S. mutans growth while 1:16 was efficient against L. casei. EGCG at 1:4 dilution completely inhibited S. mutans and L. casei growth. Independently of the association with Gt, Nchi completely inhibited S. mutans at 1:4 dilution. For L. casei, different concentrations of Nchi (1:32) and Nchi+Gt (1:8) were required to inhibit cell growth. After selective carious removal, viability of S. mutans decreased (p<0.001), without difference between bur and Er:YAG laser (p>0.05). Treatment with Nchi and Nchi+Gt did not influence the microbial load of S. mutans on dentin (p>0.05). Although variations in concentrations were noticed, all compounds showed antibacterial activity against S. mutans and L. casei. Both bur and Er:YAG laser have effectively removed soft dentin and reduced S. mutans counts. Nanoformulations did not promote any additional antibacterial effect in the remaining dentin.

Comparative analysis of Candida spp. isolated from complete denture wearers with and without an HIV diagnosis.

STATEMENT OF PROBLEM: Denture-related stomatitis (DRS), an inflammation frequently present in human immunodeficiency virus-positive (HIV+) individuals, can be attributable to colonization by Candida spp., which is considered a main factor. The virulence factors of these species are often modulated by the systemic condition of their hosts. PURPOSE: The purpose of this clinical study was to evaluate the incidence, virulence, and morphology of Candida spp. isolated from biofilms of complete denture wearers with DRS, with and without an HIV diagnosis. In addition, the interaction of the systemic condition with the ability of Candida spp. to colonize was evaluated. MATERIAL AND METHODS: Fifty-five complete denture wearers diagnosed with DRS were divided into 2 groups: experimental (HIV+) and control (human immunodeficiency virus-noninfected participants [HIV-]). Biofilm was collected by a standardized method of ultrasonification of prostheses. The incidence was evaluated by a chromogenic method and polymerase chain reaction (PCR). The virulence factors were assessed by using the capacity for biofilm formation by counting colony-forming units (CFUs/mL), biofilm metabolism by tetrazolium salt metabolization, and proteinase and phospholipase production by using a fluorimetric kit. Morphology was verified by using the hyphae-inducing test, and participants' health data were collected with a form. Data were analyzed by using the Student t, Mann-Whitney U, Spearman, and Fisher tests (α=.05). RESULTS: The results of incidence were related to 55 participants (22 experimental and 33 control); in total, 63 Candida spp. samples were isolated, showing 28 Candida albicans and 36 nonalbicans strains. No significant difference was found between groups in baseline CFU/mL counts, biofilm formation capacity, cell metabolism, and phospholipase production. Proteinase production was higher for C. albicans in the control (P=.031) and for nonalbicans in the experimental (P=.016) groups. Relative to health data, the experimental group showed a moderate negative correlation between the CFU count/mL at baseline for nonalbicans and DRS classification (P=.020). CONCLUSIONS: C. albicans was the most prevalent species. No difference was found in the Candida spp. of complete denture wearers with DRS, with and without an HIV diagnosis, with regard to virulence factors (except for proteinase production) and morphology.

Action of chitosan-based solutions against a model four-species biofilm formed on cobalt-chromium and acrylic resin surfaces.

OBJECTIVE: To evaluate the anti-biofilm action of chitosan, nanoparticulate chitosan, and denture cleanser Nitradine™ against biofilms comprising Candida albicans, Candida glabrata, Staphylococcus aureus, and Streptococcus mutans. BACKGROUND: Biofilm removal from removable partial dentures (RPD) is important for success in prosthetic rehabilitation. MATERIALS AND METHODS: The anti-biofilm action of the experimental chitosan-based solutions and Nitradine™ was evaluated on acrylic resin and cobalt-chromium alloy through assessing cell viability, cell metabolism, residual aggregated biofilm, and extracellular polymeric substance and biofilm morphology. RESULTS: Only chitosan reduced the viability of C. albicans on cobalt-chromium alloy surface, by 98% (a 1.7 log10 reduction in cfu). Chitosan-based solutions neither promoted substantial alteration of the metabolic activity of the four-species biofilm nor reduced the amount of the aggregated biofilm. After immersion in chitosan and nanoparticulate chitosan, viable microorganisms and extracellular polymeric substances distributed over the entire specimens' surfaces were observed. Nitradine™ reduced the viability and metabolic activity of biofilm grown on both surfaces, but it did not remove all aggregated biofilm and extracellular polymeric substances. After immersion in Nitradine™, approximately 35% of the specimens' surfaces remained covered by aggregated biofilm, mainly composed of dead cells. CONCLUSION: Although chitosan and Nitradine™ promoted changes in the viability of microorganisms, neither solution completely removed the four-species biofilm from the Co-Cr and acrylic resin surfaces. Thus, isolated use of hygiene solutions is not indicated for biofilm control on RPDs; this requires complementary mechanical removal.

Protocol with non-toxic chemicals to control biofilm in dental unit waterlines: physical, chemical, mechanical and biological perspective.

Biosafety in dentistry aims to combat cross-contamination and biofilm in dental unit waterlines. The aim was to investigate from a physical, chemical, mechanical and biological perspective, a protocol for using chemical products (citric acid, sodium bicarbonate and sodium chloride) to improve and maintain water quality in dental unit waterlines. Change in microhardness and corrosion tendency were observed in stainless steel samples. On the polyurethane surfaces, there were changes in color, microhardness and roughness. Anti-biofilm evaluations revealed a significant reduction in the biofilm biomass, metabolic activity and residual biofilm. These findings suggest that the protocol analyzed in this study showed an innovative potential against biofilm in dental unit waterlines, preserving the physical, chemical and mechanical properties of the materials.

Clinical study of the biofilm of implant-supported complete dentures in healthy patients.

OBJECTIVE: The purpose of this study was to quantify the area covered by biofilm and identify bacteria and yeasts present in mandibular acrylic resin full-arch implant-supported fixed prostheses. BACKGROUND: Biofilm control of implant-supported fixed prosthesis is hampered by their design, and it can cause oral and systemic problems, mainly in immunocompromised patients like the elder. Knowledge about microbiota reinforces the awareness about the need for periodic professional cleaning maintenance. MATERIALS AND METHODS: Twenty prostheses were unscrewed, washed in 0.89% sodium chloride, stained with eosin 1% and photographed. The area covered by biofilm was digitally delimited and quantified. Biofilm samples were collected, diluted up to 1:107 , seeded in chromogenic agar media and incubated for 48 hours, at 37°C, for counting of colony-forming units (CFU/mL). DNA hybridization was performed to complement the identification and quantification of microorganisms. Data were analyzed using Mann-Whitney test, Spearman correlation and Fisher's exact test (α = .05). RESULTS: An average of 62% of the gingival surface of the prostheses was covered by biofilm. Enterococcus spp. (5.82 ± 1.38 log10 CFU/mL) and Staphylococcus aureus (5.75 ± 2.02 log10 CFU/mL) showed higher prevalence in cultures. Patients with five implants had less biofilm compared to those with four implants (P = .031) but had higher Escherichia coli counts (P = .039). In DNA hybridization, Streptococcus pneumoniae, Veillonella parvula and Fusobacterium nucleatum presented higher quantification and were present in all the samples; patients over 65 years old contained more Candida tropicalis (P = .049); prostheses on five implants presented lower quantification for several species. CONCLUSION: Biofilm was present on all prostheses, containing potentially pathogenic microorganisms. The number of implants may play a role in quantification of biofilm and in microorganism counts.

Increased diversity, fungal burden, and virulence of oral Candida spp. in patients undergoing anti-tuberculosis treatment.

Some studies have demonstrated a high prevalence of Candida species in patients with tuberculosis (TB). This is most likely due to long-term antimicrobial therapy. To date, no longitudinal studies addressed the effects of anti-TB treatment on the fungal burden and virulence of Candida spp. This study investigated the prevalence and virulence of Candida spp. in the oral cavity of 30 TB patients at different stages of treatment through a cohort study. These results were compared with those of 60 systemically healthy individuals in a cross-sectional study. Oral rinse samples from TB patients were collected before 45 and after 120 days of treatment. In the control group, the biological samples were collected only once. Candida spp. were identified by restriction fragment length polymorphism (RFLP) assays, and the following virulence factors were studied: phospholipase C and proteinase production, as well as Candida spp. biofilm and hyphae formation. The clinical diagnosis of TB and its treatment time were associated with the greater fungal burden (p < 0.0001), presence of non-albicans Candida (NAC) species (p = 0.0003), and increased virulence factors when compared with the Candida spp. isolated from systemically healthy individuals. The results showed that anti-TB treatment time was responsible for the increased fungal burden and isolation of NAC in TB patients (p = 0.0233). The increased prevalence, quantification, and virulence of Candida spp. isolated from the oral cavity of TB patients highlight the greater risk of oral lesions and cases of systemic dissemination in these patients.

Expression of virulence factors by Pseudomonas aeruginosa biofilm after bacteriophage infection.

The use of bacteriophages for the treatment of bacterial infections has been extensively studied. Nonetheless, the stress response regarding bacteriophage infection and the expression of virulence factors of Pseudomonas aeruginosa after phage infection is poorly discussed. In this study, we evaluated biofilm formation capacity and expression of virulence factors of P. aeruginosa after bacteriophage infection. Biofilm growth rates, biofilm morphology, pyocyanin production and elastase activity were evaluated after 2, 8, 24 and 48 h of co-cultivation with bacteriophages that was recently characterized and showed to be infective towards clinical isolates. In parallel, quantitative real-time polymerase chain reactions were carried out to verify the expression of virulence-related genes. Bacteriophages promoted substantial changes in P. aeruginosa biofilm growth at early co-culture time. In addition, at 8 h, we observed that some cultures developed filaments. Although bacteriophages did not alter both pyocyanin and protease activity, changes on the expression level of genes related to virulence factors were detected. Usually, lasI, pslA, lasB and phzH genes were upregulated after 2 and 48 h of co-culture. These results highlight the need for extensive investigation of pathways and molecules involved in phage infection, since the transcriptional changes would suggest a response activation by P. aeruginosa.

Comparative effects on acrylic resin properties of short- and long-term immersion in alkaline peroxide solution.

PURPOSE: To evaluate the color stability (ΔE), surface roughness (ΔRa), and flexural strength of denture base acrylic resin after short (20 minutes) and long-term (8 hours) immersions simulating a period of 5 years of use. METHODS: 120 disk-shaped (16 mm x 4 mm) and 120 rectangular (65 mm x 10 mm x 3.3 mm) specimens were prepared (Lucitone 550) and distributed into three groups (n=20): distilled water (Control); Corega Tabs (CT) and Polident (Po), for immersions of 20 minutes and 8 hours. ΔE were determined by a colorimeter, ΔRa by a profilometer and the flexural strength by a universal testing machine. Data were evaluated by two-way ANOVA followed by Tukey's test and Bonferroni correction (α= 0.05). RESULTS: Short-term immersions for 20 minutes resulted in significant differences in ΔE, as CT values (1.46 ± 1.41) were higher than Po (0.55 ± 0.25) and Control (0.47 ± 0.21). Specimens immersed for 8 hours showed more extensive alterations overall than the control (ΔE [Po: 7.27 ± 0.53; CT: 5.58 ± 0.49; Control: 1.26 ± 0.23]; 916;Ra [Po: 0.16 ± 0.08; CT: 0.12 ± 0.11; Control: 0.07 ± 0.06]; flexural strength [Po: 42.99 ± 9.34; CT: 63.96 ± 12.98; Control: 64.59 ± 12.87]). The tested alkaline peroxide-based solutions promoted significant alteration on acrylic resin properties after overnight immersion. Therefore, short-term immersions seem to be more suitable for daily denture hygiene. CLINICAL SIGNIFICANCE: Alkaline peroxides were able to alter color stability, surface roughness and flexural strength of acrylic resin after overnight immersions, in a 5-year-simulated period of use.

Brushing associated with oral irrigation in maintaining implants and overdentures hygiene - a randomized clinical trial.

Evaluate, through a randomized clinical trial, the efficacy of brushing associated with oral irrigation in maintaining implant and overdenture hygiene. Thirty-eight participants, who had a clinically acceptable conventional maxillary complete denture and mandibular overdenture retained by either implants or mini-implants using an O-ring-retained system, were enrolled to participate in the study. They were instructed to use two different hygiene methods, in a random sequence for a period of 14 days, with a 7-day wash-out interposed period: (I) mechanical brushing (MB); (II) association of mechanical brushing with oral irrigation (WP). Biofilms from both subgingival sulci and overdentures were collected and processed by Checkerboard DNA-DNA hybridization method at baseline and after using the proposed hygiene protocols. Comparisons were performed using Wilcoxon test and Friedman test with Benjamini-Hochberg false discovery rate, followed by Conover post-hoc test (α = 0.05). In the subgingival sulci-related biofilm, a lower number of microbial cells were detected, after WP compared to the MB method (P < 0.001). The findings of overdenture-related biofilm suggest that both methods were similar (P = 0.607) being the identified microbiota qualitatively coincident after each method. Despite the number of microbial counts, it was concluded that the association of mechanical brushing with oral irrigation was more effective in reducing microorganisms in the subgingival sulci biofilm; however, the same outcome was not observed in the overdentures.

Biomaterials for orthopedics: anti-biofilm activity of a new bioactive glass coating on titanium implants.

This study evaluated adhesion and biofilm formation by Candida albicans, Pseudomonas aeruginosa and Staphylococcus epidermidis on surfaces of titanium (Ti) and titanium coated with F18 Bioactive Glass (BGF18). Biofilms were grown and the areas coated with biofilm were determined after 2, 4 and 8 h. Microscopy techniques were applied in order to visualize the structure of the mature biofilm and the extracellular matrix. On the BGF18 specimens, there was less biofilm formation by C. albicans and S. epidermidis after incubation for 8 h. For P. aeruginosa biofilm, a reduction was observed after incubation for 4 h, and it remained reduced after 8 h on BGF18 specimens. All biofilm matrices seemed to be thicker on BGF18 surface than on titanium surfaces. BGF18 showed significant anti-biofilm activity in comparison with Ti in the initial periods of biofilm formation; however, there was extensive biofilm after incubation for 48 h.

The effects of three disinfection protocols on Candida spp., denture stomatitis, and biofilm: A parallel group randomized controlled trial.

STATEMENT OF PROBLEM: Antifungals are used to treat Candida infections. However, because of increased antifungal resistance and the length of antifungal therapy, Candida spp. infections can be prevented using the prosthesis hygiene method. Therefore, establishing efficient, safe, and low-cost hygiene protocols for complete denture wearers is necessary. PURPOSE: The purpose of this clinical trial was to compare 10% Ricinus communis (RC10%) and 0.5% chloramine-T (CT0.5%) with negative (water) and positive (0.25% sodium hypochlorite [SH0.25%]) controls to establish a protocol to treat denture stomatitis (DS), remove denture biofilm, reduce overall microbiota, and decrease Candida spp. on the palate and denture bases. MATERIAL AND METHODS: This randomized, double blind, controlled clinical trial allocated 60 DS-positive participants in parallel groups: RC10%, CT0.5%, negative control, and SH0.25%. All participants brushed their palate and dentures and applied 1 of the solutions only to the denture. The following outcomes were assessed at baseline and after 7 and 37 days: Candida spp. counts, frequency of species by presumptive identification, DS severity, and photographic quantification of biofilm. The Kruskal-Wallis and Friedman tests with stepwise step-down post hoc test compared the anticandidal effect and the DS score (between groups and time). ANOVA and the Tukey post hoc test were used for biofilm removal comparison (α=.05). RESULTS: Microbial counts were solution- and time-dependent for dentures, with C. albicans, C. tropicalis, and C. glabrata being the most prevalent species. RC10% presented similar results to baseline and control after 7 and 37 days. CT0.5% reduced the CFU/mL compared with the baseline. SH0.25% was the most effective. DS reduced in all groups, independent of the solution. SH0.25% reduced biofilm the most, followed by RC10%. CT0.5% was similar to the control. CONCLUSIONS: SH0.25% demonstrated potential for Candida spp. control in denture wearers with DS. The other protocols showed intermediate activity and might be more suitable for longer immersion periods.

In Vitro Evaluation of Resilient Liner after Brushing with Conventional and Experimental Ricinus communis-Based Dentifrices.

PURPOSE: To evaluate the effect of experimental (Ricinus communis) and commercial dentifrices used for denture cleaning on abrasiveness (gravimetric method; roughness), hardness, and color stability of a resilient relining material. MATERIALS AND METHODS: Sixty circular (15 × 3 mm) specimens were distributed into four groups: C (control; brushing with water); CO (brushing with Colgate - for natural teeth); CB (brushing with Corega Brite - for complete dentures); RC (brushing with experimental dentifrice). Brushing was performed in a toothbrushing machine with a soft brush and a dentifrice suspension for 50 minutes, calculated to correspond to 1 year of regular brushing. Variables were measured initially and after the trial period. For the gravimetric method, the difference in mass was considered. The surface roughness was measured by a rugosimeter, and the hardness test was performed by a Shore A durometer. Color changes (ΔE; CIE L*a*b* and NBS systems) were measured by a portable spectrophotometer. Results were analyzed by ANOVA/Tukey's test (α = 0.05). RESULTS: The largest mass variation (µg; p < 0.0001) occurred in C (-6.21 ± 3.18). Concerning roughness, CB (0.26 ± 0.04) showed the lowest value, followed by RC (0.29 ± 0.08) and CO (0.34 ± 0.24) (p < 0.0001). Group C produced the greatest surface roughness (0.72 ± 0.25). Hardness values decreased after brushing with water (p = 0.014). No significant differences were found among RC (50.31 ± 1.03), CO (49.11 ± 1.31), CB (49.17 ± 1.23), and C (48.02 ± 1.26). Color stability was similar in all groups (p = 0.135; C: 2.3 ± 0.77; CO: 2.6 ± 0.54; CB: 2.2 ± 0.44; RC: 2.9 ± 1.56). CONCLUSIONS: The use of experimental dentifrice could be indicated, as it showed similar results to the specific dentifrice, keeping the resilient material properties within acceptable values.

Preclinical study of a cost-effective photodynamic therapy protocol for treating oral candidoses.

Photodynamic therapy (PDT) is a promising treatment for oral candidoses. Its use as an alternative to antifungals prevents several adverse effects, including microbial resistance. However, most PDT protocols do not employ devices and consumables commonly available in dental practice, thus influencing treatment affordability. This study aimed to determine the efficacy of a PDT method based on light curing units' blue LEDs combined to a plaque-disclosing composition (5% erythrosine) against C. albicans in culture and in a murine model of oral candidosis. Standard and resistant fungal strains were tested in vitro in planktonic and biofilm forms. PDT (pre-irradiation time periods: 30 and 60 s; irradiation time: 3 min) was compared to control conditions without light and/or erythrosine. Mice with induced oral candidosis (n = 40) randomly received PDT or similar control conditions with subsequent C. albicans count. These mice underwent histological analysis, as well as 12 healthy mice submitted to experimental treatments. PDT completely inactivated C. albicans planktonic cells and biofilm. Control conditions presented minor differences (ANOVA, p < 0.05), with mean values ranging from 5.2 to 6.8 log10 (UFC/mL). Infected mice presented no significant difference in C. albicans counts consequent to treatments (ANOVA, p = 0.721), although the PDT protocol was able to enhance the inflammatory infiltrate in healthy mice. It can be concluded that the tested PDT protocol can inactivate C. albicans but still needs further investigation in order to achieve efficacy and safety.

In Vitro Analysis of Surface Roughness of Acrylic Resin Exposed to the Combined Hygiene Method of Brushing and Immersion in Ricinus communis and Sodium Hypochlorite.

PURPOSE: To evaluate a solution based on Ricinus communis (Castor oil) for denture cleansing, comparing it to sodium hypochlorite (NaOCl) for the surface roughness of heat-polymerized acrylic resin. MATERIALS AND METHODS: Forty polished and unpolished resin specimens (90 × 30 × 4 mm) were evaluated before and after their exposure to protocol hygiene: brushing the specimens with a specific denture brush and mild soap for 3 minutes, three times a day, and immersing them in hygiene solutions (0.25% NaOCl-S1 and 0.5% NaOCl-S2; 10% R. communis-S3; saline-S4: control) for 20 minutes. Surface roughness was evaluated by rugosimeter and scanning electron microscopy (SEM) before and after the protocol. For evaluation of surface roughness, polished and unpolished surfaces were used. RESULTS: The roughness of the polished surface was not affected by time (p = 0.062), but was affected by solutions (p < 0.0001) and the interaction between factors (p = 0.005). For S1 and S4, the period did not influence the roughness. For S2, there was a change after 7 days, remaining stable after 14 days. For S3, there were changes, and stabilization occurred after 14 days. After 7 and 14 days, S2 and S3 promoted major changes, but after 21 days, there were no differences among solutions, except saline. The unpolished surface was not influenced by factors: period (p = 0.115), solution (p = 0.120), and their interaction (p = 0.382). SEM analysis showed similar results on the evaluation of surface roughness. CONCLUSIONS: The polished surface of the prosthesis was more susceptible to changes when exposed to hygiene solutions, and although the 0.5% NaOCl solution promoted an increase in the surface roughness compared with the same solution at 0.25% and R. communis at 10%, the values are clinically acceptable.

Efficient recovery of undifferentiated human embryonic stem cell cryopreserved with hydroxyethyl starch, dimethyl sulphoxide and serum replacement.

BACKGROUND: The therapeutic use of human embryonic stem cells (hESCs) is dependent on an efficient cryopreservation protocol for long-term storage. The aim of this study was to determine whether the combination of three cryoprotecting reagents using two freezing systems might improve hESC recovery rates with maintenance of hESC pluripotency properties for potential cell therapy application. METHODS: Recovery rates of hESC colonies which were frozen in three cryoprotective solutions: Me2SO/HES/SR medium, Defined-medium® and Me2SO/SFB in medium solution were evaluated in ultra-slow programmable freezing system (USPF) and a slow-rate freezing system (SRF). The hESC pluripotency properties after freezing-thawing were evaluated. RESULTS: We estimated the distribution frequency of survival colonies and observed that independent of the freezing system used (USPF or SRF) the best results were obtained with Me2SO/HES/SR as cryopreservation medium. We showed a significant hESC recovery colonies rate after thawing in Me2SO/HES/SR medium were 3.88 and 2.9 in USPF and SRF, respectively. The recovery colonies rate with Defined-medium® were 1.05 and 1.07 however in classical Me2SO medium were 0.5 and 0.86 in USPF and SRF, respectively. We showed significant difference between Me2SO/HES/SR medium×Defined-medium® and between Me2SO/HES/SR medium×Me2SO medium, for two cryopreservation systems (P<0.05). CONCLUSION: We developed an in house protocol using the combination of Me2SO/HES/SR medium and ultra-slow programmable freezing system which resulted in hESC colonies that remain undifferentiated, maintain their in vitro and in vivo pluripotency properties and genetic stability. This approach may be suitable for cell therapy studies.

Evaluation of surface roughness, wettability and adhesion of multispecies biofilm on 3D-printed resins for the base and teeth of complete dentures.

OBJECTIVE: This study evaluated the surface roughness, wettability and adhesion of multispecies biofilms (Candida albicans, Staphylococcus aureus and Streptococcus mutans) on 3D-printed resins for complete denture bases and teeth compared to conventional resins (heat-polymerized acrylic resin; artificial pre-fabricated teeth). METHODOLOGY: Circular specimens (n=39; 6.0 mm Ø × 2.0 mm) of each group were subjected to roughness (n=30), wettability (n=30) and biofilm adhesion (n=9) tests. Three roughness measurements were taken by laser confocal microscopy and a mean value was calculated. Wettability was evaluated by the contact angle of sessile drop method, considering the mean of the three evaluations per specimen. In parallel, microorganism adhesion to resin surfaces was evaluated using a multispecies biofilm model. Microbial load was evaluated by determining the number of Colony Forming Units (CFU/mL) and by scanning electron microscopy (SEM). Data were subjected to the Wald test in a generalized linear model with multiple comparisons and Bonferroni adjustment, as well as two-way ANOVA (α=5%). RESULTS: The roughness of the conventional base resin (0.01±0.04) was lower than that of the conventional tooth (0.14±0.04) (p=0.023) and 3D-printed base (0.18±0.08) (p<0.001). For wettability, conventional resin (84.20±5.57) showed a higher contact angle than the 3D-printed resin (60.58±6.18) (p<0.001). Higher microbial loads of S. mutans (p=0.023) and S. aureus (p=0.010) were observed on the surface of the conventional resin (S. mutans: 5.48±1.55; S. aureus: 7.01±0.57) compared to the 3D-printed resin (S. mutans: 4.11±1.96; S. aureus: 6.42±0.78). The adhesion of C. albicans was not affected by surface characteristics. The conventional base resin showed less roughness than the conventional dental resin and the printed base resin. CONCLUSION: The 3D-printed resins for base and tooth showed less hydrophobicity and less adhesion of S. mutans and S. aureus than conventional resins.

Assessment of peripheral venous catheters microbiota and its association with phlebitis.

BACKGROUND: Peripheral venous catheters (PVCs) remain the primary mode of short-term venous access for managing intravenous fluid, obtaining blood samples, and peripheral parenteral nutrition. They may get contaminated and require regular monitoring to prevent complications. This study evaluated the occurrence of phlebitis and its associated-clinical and microbiological indicators. METHODS: The frequency of phlebitis was evaluated in hospitalized patients of both medical and surgical fields. Subsequently, the dichotomous association between the presence of phlebitis and the clinical aspects was investigated. In parallel, the bacterial contamination of PVCs was assessed through culture-based methods, microscopy observation, and 16S rRNA gene sequencing. RESULTS: Approximately one in four patients presented phlebitis (28.4%). The most frequent symptom was erythema at access site, with or without pain, corresponding to Score 1 on the phlebitis scale (17.9%). Colonization of both lumen and external surface of PVC was observed in 31.3% of the samples. Staphylococcus and Pseudomonas were the most isolated bacterial genera on the PVC surface. No significant association was observed between the presence of phlebitis and the clinical aspects, as well as the presence of microorganisms. CONCLUSION: Microorganism were present on both internal and external PVC surface, without being associated to phlebitis.

The implications of healthcare professionals wearing jewelry on patient care biosafety: observational insights and experimental approaches.

The use of jewelry among healthcare professionals poses a risk of cross contamination due to potential bacterial accumulation and spread. Through a mixed-method design, this study first analyzed the implications of healthcare professionals wearing jewelry on patient care biosafety as well as on the residual bacterial load of hands and rings after hand hygiene. Firstly, an observational prevalence study to verify whether nursing professionals wear personal accessories during healthcare assistance was carried out. Second, an experimental design involving intentional contamination and hygiene of the hands, with and without a ring, was conducted. The bacterial load of both hands and rings was measured by counting colony forming units. The observational study showed that nursing workers frequently wear jewelry during healthcare assistance. Nonetheless, the experimental study did not indicate differences in bacterial contamination between hands with and without a ring, despite the hand hygiene procedure applied. In conclusion, many nursing workers wear jewelry in the workplace. Although hands with and without a ring exhibited similar microbial load, rings appeared as a potential source of bacterial contamination, reinforcing the need to remove jewelry during working hours. Hand hygiene using alcohol, or soap and water significantly decreased the bacterial load on the participants' hands, with handwashing proving to be the most efficient method for removing intentional contamination.

Unveiling the efficacy of latex gloves in preventing viral infections during needlestick injuries: An in vitro mixed-methods study.

BACKGROUND: Gloves are personal protective equipment designed to prevent contamination and reduce the spread of microorganisms. This study aimed to assess in vitro the physical integrity of latex gloves and the retention of biological contamination in healthcare simulation. METHOD: Three different batches of latex procedure gloves from five different brands and specific batches were evaluated before use for physical integrity by the standard protocols of the Society for Testing and Materials (ASTM) and of the American Food and Drug Administration (FDA). Moreover, the retention of biological contamination by latex procedure gloves in needlestick injury simulation with crystal violet and bacteriophages were applied in order to mimic human blood and virus presence. RESULTS: Brands D and C showed the best and worst results in the immediate inspections and after 2 min, respectively. For Brand C, damage occurred in one finger/region in a total of 12 gloves, while seven gloves were damaged/unable to be worn. Brand D presented only two gloves with tears and/or holes in one finger/region. Regarding the viral contamination, in a simulated needlestick injury, data showed no significant difference among the groups. CONCLUSION: All glove brands presented physical damage that might affect the spread of microorganisms. The gloves did not exert an additional protective effect during a needlestick injury simulation in accordance with the two techniques used in this study.