Viviane de Cássia Oliveira

Color Stability, Surface Roughness and Flexural Strength of an Acrylic Resin Submitted to Simulated Overnight Immersion in Denture Cleansers

This study evaluated color stability, surface roughness and flexural strength of acrylic resin specimens after immersion in alkaline peroxide and alkaline hypochlorite, simulating a period of one and a half year of use of overnight immersion. Sixty disc-shaped (16x4 mm) and 80 rectangular specimens (65x10x3.3 mm) were prepared from heat-polymerized acrylic resin (Lucitone 550) and distributed into 4 groups (n=20): C1: without immersion, C2: 8 h immersion in distilled water; AP: 8 h immersion in alkaline peroxide effervescent tablet; SH: 8 h immersion in 0.5% NaOCl solution. Properties were evaluated at baseline and after the immersion. Color data were also calculated according the National Bureau of Standards (NBS). Results were analyzed statistically by ANOVA and Tukeys HSD test (α=0.05). AP (2.34 ± 0.41) caused color alteration significantly higher than C2 (0.39 ± 0.30) and SH (1.73 ± 0.52). The mean ΔE values were classified as indicial for C2 (0.36 ± 0.29) and noticeable for AP (2.12 ± 0.39) and SH (1.59 ± 0.48). SH (0.0195 ± 0.0150) caused significantly higher ΔRa (p=0.000) than the C2 (0.0005 ± 0.0115) and PA (0.0005 ± 0.0157) groups. There was no statistically significant difference (p=0.063) among the solutions for flexural strength (C1: 105.43 ± 14.93, C2: 100.30 ± 12.43, PA: 97.61 ± 11.09, SH: 95.23 ± 10.18). In conclusion, overnight immersion in denture cleansing solutions simulating a year and a half of use did not alter the flexural strength of acrylic resin but caused noticeable color alterations, higher for alkaline peroxide. The 0.5% NaOCl solution caused increase in surface roughness.

Growth and functional harvesting of human mesenchymal stromal cells cultured on a microcarrier-based system.

Human mesenchymal stromal cells (hMSCs) cells are attractive for applications in tissue engineering and cell therapy. Because of the low availability of hMSCs in tissues and the high doses of hMSCs necessary for infusion, scalable and cost‐effective technologies for in vitro cell expansion are needed to produce MSCs while maintaining their functional, immunophenotypic and cytogenetic characteristics. Microcarrier‐based culture systems are a good alternative to traditional systems for hMSC expansion. The aim of the present study was to develop a scalable bioprocess for the expansion of human bone marrow mesenchymal stromal cells (hBM‐MSCs) on microcarriers to optimize growth and functional harvesting. In general, the results obtained demonstrated the feasibility of expanding hBM‐MSCs using microcarrier technology. The maximum cell concentration (n = 5) was ∼4.82 ± 1.18 × 105 cell mL−1 at day 7, representing a 3.9‐fold increase relative to the amount of inoculated cells. At the end of culture, 87.2% of the cells could be harvested (viability = 95%). Cell metabolism analysis revealed that there was no depletion of important nutrients such as glucose and glutamine during culture, and neither lactate nor ammonia byproducts were formed at inhibitory concentrations. The cells that were recovered after the expansion retained their immunophenotypic and functional characteristics. These results represent an important step toward the implementation of a GMP‐compliant large‐scale production system for hMSCs for cellular therapy.

Antimicrobial activity of complete denture cleanser solutions based on sodium hypochlorite and Ricinus communis – a randomized clinical study

ABSTRACT To preserve oral health and to maintain the prosthetic devices, it is important not only to improve the properties of commonly known hygiene products, but also to investigate new materials with antimicrobial action. Objectives This study evaluated the antimicrobial activity of sodium hypochlorite (0.25% and 0.50%) and 10% Ricinus communis’ solutions against specific microorganisms. Material and Methods Sixty four maxillary complete denture wearers were instructed to brush their dentures three times a day and to soak them (20 min/day) in the solutions: SH1: 0.25% sodium hypochlorite; SH2: 0.5% sodium hypochlorite; RC: 10% R. communis oil; and C: 0.85% saline (control). The solutions were used for 7 days in a randomized sequence. Following each period of use, there was a 1-week washout period. Antimicrobial activity was determined by Colony Forming Units (CFU) counts of Streptococcus mutans, Candida spp., and gram-negative microorganisms. For collecting biofilm, the internal surface of maxillary dentures was brushed with saline solution, and biofilm suspension obtained. After dilutions (100 - 10-3), aliquots were seeded in Mitis salivarius, CHROMagar Candida®, and MacConkey agar for detecting S. mutans, Candida spp., or gram-negative microorganisms, respectively. After incubation, colonies were counted, and CFU/mL values were calculated. Then, transformation - log10 (CFU+1) - data were analyzed using the Friedman test (α=0.05). Results showed significant differences between the solutions (p<0.001). Results All three solutions showed antimicrobial activity against S. mutans. Against Candida spp., RC and SH1 solutions showed similar effect while SH2 showed superior activity. SH1 and SH2 solutions showed antimicrobial action against gram-negative microorganisms. The Candida species most frequently isolated was C. albicans, followed by C. tropicalis and C. glabrata. Conclusions The 0.5% sodium hypochlorite solution was the most effective and might be used to control denture biofilm. C. albicans was the most frequently isolated Candida sp.

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.

Antimicrobial action of sodium hypochlorite and castor oil solutions for denture cleaning - in vitro evaluation.

The objective of this in vitro study was to evaluate the antimicrobial action of sodium hypochlorite (0.25% and 0.50%) and 10% castor oil solutions against specific microorganisms, by counting Colony Forming Units (CFU) of clinically important bacteria and Candida species. Acrylic resin specimens (n = 320; Lucitone 550) were obtained from square metal matrices (10 x 10 x 2 mm), sterilized by microwave (650W, for 6 minutes) and contaminated by Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Bacillus subtilis, Escherichia coli, Streptococcus mutans, Enterococcus faecalis and Candida glabrata. The specimens were immersed for 20 minutes in one of the following hygiene solutions (n = 10/each): A - 0.25% Sodium hypochlorite; B - 0.5% Sodium hypochlorite; C - 10% Castor oil solution; and D (Control) - saline. Adhered cells were suspended and inoculated into a selective solid medium (37ºC for 24 h). The Students t-test (α = 0.05) was performed to compare log10(CFU+1)/mL between Groups C and D. The results showed that sodium hypochlorite (0.25% and 0.5%) completely eliminated all detectable microorganisms. The castor oil solution eliminated B. subtilis and reduced counts for other strains. Differences between C and D were significant (p < 0.05) for all species except for E. faecalis. Both sodium hypochlorite solutions (0.25% and 0.5%) were effective in eliminating all microorganisms evaluated, and may be useful as cleaning solutions for complete dentures. The castor oil solution provided moderate efficacy and performed differently on the tested species, with the strongest effect on B. subtilis and with non-significant action on E. faecalis.

Effect of sodium hypochlorite and Ricinus communis solutions on control of denture biofilm: A randomized crossover clinical trial

Statement of problem The prevalence of complete edentulism remains high in the elderly, and previous data have shown that poor denture hygiene is common among patients with edentulism. Purpose The purpose of this randomized crossover trial was to evaluate the efficacy of denture cleansers in terms of biofilm removal, antimicrobial action, and the remission of denture stomatitis. Material and methods Fifty denture wearers with denture stomatitis were instructed to brush their dentures (brush and soap) and to soak them (20 minutes/14 days) in 4 solutions, as follows: C (control), 0.85% saline; SH1, 0.1% sodium hypochlorite; SH2, 0.2% sodium hypochlorite; and RC, 8% Ricinus communis. The biofilm in the intaglio surface of maxillary dentures was stained, photographed, and quantified by software (Image Tool). It was then collected (brushed with saline solution), and the obtained suspension was diluted (100 to 10‐3) and seeded (50 &mgr;L) in CHROMagar for Candida spp. After incubation, colony‐forming units per milliliter values were calculated. Denture stomatitis remission was classified according to the Newton classification. Data were analyzed by Friedman (&agr;=.05) and Wilcoxon tests and corrected by the Bonferroni test (&agr;=.005). Results SH1 (mean rank [MR]=1.98) and SH2 (MR=1.64) showed lower biofilm coverage than C (MR=3.73) that was similar to RC (MR=2.92). SH1 (MR=2.43) and SH2 (MR=2.10) showed antimicrobial action for Candida spp, and RC (MR=3.36) showed similar results to C (MR=3.51) and baseline (MR=3.50). Clinical signs of denture stomatitis were reduced by SH1 (MR=2.44), while SH2 (MR=2.56) and RC (MR=2.74) showed intermediate results. Conclusions The two sodium hypochlorite solutions were the most effective means of biofilm control. All tested solutions were effective in reducing the signs of denture stomatitis.

Clinical trial for evaluation of Ricinus communis and sodium hypochlorite as denture cleanser

Abstract The development of opportunistic infections due to poor denture hygiene conditions justified the search for effective hygiene protocols for controlling denture biofilm. Objective This study evaluated Ricinus communis and sodium hypochlorite solutions in terms of biofilm removal ability, remission of candidiasis, antimicrobial activity, and participant satisfaction. Material and Methods It was conducted a controlled clinical trial, randomized, double-blind, and crossover. Sixty-four denture wearers with (n=24) and without candidiasis (n=40) were instructed to brush (3 times/day) and immerse their dentures (20 min/day) in different storage solutions (S1 / S2: 0.25% / 0.5% sodium hypochlorite; S3: 10% R. communis; S4: Saline).The trial period for each solution was seven days and a washout period of seven days was used before starting the use of another solution. The variables were analyzed at baseline and after each trial period. The biofilm of inner surfaces of maxillary dentures was disclosed, photographed, and total and dyed areas were measured (Image Tool software). The percentage of biofilm was calculated. Remission of candidiasis was assessed by visual scale and score were attributed. Antimicrobial activity was assessed by the DNA-Checkerboard hybridization method. Patient satisfaction was measured using a questionnaire. Results S1 (4.41±7.98%) and S2 (2.93±5.23%) were more effective then S3 (6.95±10.93%) in biofilm remotion(P<0.0001). All solutions were different from the control (11.07±11.99%). S3 was the most effective solution in remission of candidiasis (50%), followed by S1 (46%). Concerning antimicrobial action, S1/S2 were similar and resulted in the lowest microorganism mean count (P=0.04), followed by S3. No significant differences were found with patient’s satisfaction. Conclusions 10% R. communis and 0.25% sodium hypochlorite were effective in biofilm removal, causing remission of candidiasis and reducing the formation of microbial colonies in denture surfaces. All solutions were approved by patients.

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

Abstract LB-304: Bone marrow-derived endothelial cells migrate to tumor sites and contribute to functional tumor vasculature

Tumor expansion is dependent on neovascularization, a process that requires sustained new vessel formation. Although the critical role of angiogenesis by endothelial sprouting in this process, controversy still prevails on the whether vasculogenesis, involving bone marrow (BM)-derived endothelial cells, does contribute to this process. The objective of this study was to investigate the contributions of BM-derived endothelial cells to tumor vasculature, in a melanoma model using chimeric GFP mice. Methodology : First, wild-type C57BL/6 mice were was exposed to a single dose of 10 Gy X-irradition and rescued by transplantation of BM cells from C57BL/6-GFP mice. At d30 post-transplant, the animals received a subcutaneous infusion of melanoma cells stably transfected with the luciferase gene (B16/F10-Luc + ). After 20 days, the tumors were harvested, and tumor stroma characterized using flow cytometry, bioluminescence-based image processing and confocal microscopy, to define the specific contribution of BM-derived cells. Results : Mice transplanted with donor GFP+ cells showed significant BM chimerism (90.9 ± 0.87%) demonstrating successful engraftment of donor BM stem/progenitor cells. Analyses of tumor specimens showed the presence of donor cells in the tumor sites (3.5±1.7%) in all animals, showing that BM-derived cells were effectively recruited to the developing melanoma. Interestingly, these cells represent endothelial cells (CD31+ cells; mean=X%) and myeloid-lineage cells (CD11b+ cells; mean=80%), but also tumor-infiltrating lymphocytes (CD8+ T cells, mean = 13.31; CD4+ T-cells, mean=2.1). Examination of the tumor endothelium by confocal microscopy clearly showed the presence of donor CD31/GFP cells in the vessel wall, namely in vessels lacking other mural components (as pericytes). Conclusions : Taken together, this study demonstrates that BM-derived cells are actively recruited to tumor sites, contributing to the tumor stroma and the formation of blood vessels in the developing tumor. We are performing studies to elucidate the molecular mechanisms involved in mobilization of of BM-derived precursor cells to the tumor microenvironment, and whether disruption of this vasculogeneic process significant impairs tumor development. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-304. doi:10.1158/1538-7445.AM2011-LB-304