Using Antimicrobial Photodynamic Therapy with Ultrasound Devices and Bioactive Glasses as a Combined Approach for Treating Dentin Caries Lesions.

Novel approaches for caries lesion removal and treatment have been proposed. This study evaluates the combined use of an experimental ultrasound, aPDT (antimicrobial photodynamic therapy) and bioactive glasses on the removal, decontamination and remineralization of dentin caries lesions. A biological model created with a duo species biofilm (Streptococcus mutans and Lactobacillus acidophilus) was used for the development of a caries-like lesion over the dentin for 7 days. Bovine dentin specimens (4 × 4 × 2 mm) were randomized according to the following caries removal techniques: bur (BUR) or ultrasound (ULT), decontamination (with or without aPDT) and remineralization materials (45S5 or F18 bioactive glasses). The following different groups were investigated: caries lesion (control); sound dentin (control); BUR; BUR + aPDT; ULT; ULT + aPDT; BUR + 45S5, BUR + F18; ULT + 45S5; ULT + F18; BUR + aPDT + 45S5; BUR + aPDT + F18; ULT + aPDT + 45S5; and ULT + aPDT + F18. Transverse microradiography (TMR), cross-sectional microhardness (CSH), FT-Raman spectroscopy and confocal microscopy (CLSM) were performed. A two-way ANOVA and Tukey's test were used (α = 0.05). (3) Results: The TMR revealed a lesion depth of 213.9 ± 49.5 µm and a mineral loss of 4929.3% vol.µm. The CSH increases as a function of depth, regardless of the group (p < 0.05). Removal with BUR (24.40-63.03 KHN) has a greater CSH than ULT (20.01-47.53 KHN; p < 0.05). aPDT did not affect the CSH (p > 0.05). No difference was observed between 45S5 or F18 (p > 0.05), but a change was observed for ULT (p > 0.05). The FT-Raman shows no differences for the phosphate (p > 0.05), but a difference is observed for the carbonate and C-H bonds. The CLSM images show that aPDT effectively inactivates residual bacteria. A combination of ULT, aPDT and bioactive glasses can be a promising minimally invasive treatment.

Effects of the Incorporation of Bioactive Particles on Physical Properties, Bioactivity and Penetration of Resin Enamel Infiltrant.

Purpose: The resinous infiltrant lacks remineralizing activity. This research aimed to develop and evaluate bioactivity, physico-mechanical properties and penetration of resin infiltrants containing Biosilicate or nanohydroxyapatite. Methods: Experimental resin infiltrant (ERI; 75/25 wt.% TEGDMA/BisEMA) was divided among the groups Pure Experimental (PE); ERI + Biosilicate 5 or 10% (Bio5; Bio10), ERI + 10% nanohydroxyapatite (Hap10), and Icon (DMG, Germany). Bioactivity was analyzed by SEM, EDS and FT-IR/ATR after soaking in SBF. Degree of conversion (DC), sorption and solubility (SO; SOL), flexural strength, modulus of elasticity (FS; E-modulus), contact angle (CA) and penetration were characterized. Extent of penetration was analyzed by treating white spot lesions (WSL) in human dental enamel samples with the infiltrants and subsequently analyzing specimens by confocal laser scanning microscopy. Data from each test were submitted to ANOVA and Tukey's tests (p < 0.01). Results: SEM, EDS and FT-IR showed the formation of precipitates and increase in the rates of Ca and P in the groups with bioactive particles, after storage in SBF. Hap10 showed higher DC and CA values than all the other groups. Groups Bio5 and Bio10 showed CA values similar to those of Icon, higher SO and SOL values, and reduction in other properties. All infiltrants were capable of penetrating into the WSLs. Conclusion: The incorporation of Biosilicate (5 or 10%) or nanohydroxyapatite (10%) into ERI induced mineral deposition on the surface and did not compromise infiltration and penetration into WSLs, however, compromising their physico-mechanical properties.

Impact of silanization of different bioactive glasses in simplified adhesives on degree of conversion, dentin bonding and collagen remineralization.

OBJECTIVE: To analyze simplified adhesive containing pure or silanized bioglass 45S5 (with calcium) or Sr-45S5 (strontium-substituted) fillers applied on dentin and to evaluate the microtensile bond strength (µTBS), interface nanoleakage, degree of conversion of adhesive, collagen degradation and remineralization. METHODS: Ambar Universal adhesive (FGM) was doped with 10 wt% bioactive glasses to form following groups: Control (no bioglass), 45S5 (conventional bioglass 45S5), Sr-45S5 (Sr-substituted bioglass 45S5), Sil-45S5 (silanized bioglass 45S5) and Sil-Sr-45S5 (silanized bioglass Sr-45S5). Adhesives were applied after dentin acid-etching using phosphoric acid at extracted human molars. Resin-dentin sticks were obtained and tested for µTBS, nanoleakage at 24 h or 6 months. Degree of conversion was measured using micro-Raman spectroscopy. Dentin remineralization was assessed by FTIR after 6-month storage in PBS. Hydroxyproline (HYP) release was surveyed by UV-Vis spectroscopy. Statistical analysis was performed using ANOVA and Tukey's test (p < 0.05). RESULTS: Regarding µTBS, Sr-45S5 and 45S5 presented higher and stable results (p > 0.05). Control (p = 0.018) and Sil-Sr-45S5 (p < 0.001) showed µTBS reduction after 6-month aging. Sil-Sr-45S5 showed higher HYP release than that obtained in the 45S5 group. Sil-45S5 showed mineral deposition and increase in µTBS (p = 0.028) after 6-months. All experimental adhesives exhibited higher degree of conversion compared to Control group, except for 45S5. All adhesives created gap-free interfaces, with very low silver impregnation, except for Sil-Sr-45S5. SIGNIFICANCE: The incorporation of silanized 45S5 bioglass into the universal adhesive was advantageous in terms of dentin remineralization, bonding performance and adhesive polymerization. Conversely, Sil-Sr-45S5 compromised the µTBS, interface nanoleakage and had a negative impact on HYP outcomes.

In vitro dentin permeability and tubule occlusion of experimental in-office desensitizing materials.

OBJECTIVES: This study investigates the dentin permeability (by hydraulic conductance) and tubule occlusion (by confocal and scanning electron microscopies) of in-office desensitizing materials. MATERIALS AND METHODS: Bovine dentin blocks were immersed in EDTA to open dentinal tubules. Placebo varnish (PLA), fluoride varnish (FLU), NaF 5% + 5% nanoparticulate sodium trimetaphosphate varnish (TMP), universal adhesive system (SBU), S-PRG filler varnish (SPRG), Biosilicate (BIOS), and amelotin (AMTN) solution were the materials tested. After application, the specimens underwent an erosive-abrasive challenge. Dentin permeability was evaluated at T0 (initial), T1 (after treatment), and T2 (after challenge). Confocal and scanning electron microscopy (SEM) were used to evaluate, respectively, length and number of dentinal tubule occlusions and opened dentinal tubules, after challenge. Permeability and SEM data were analyzed by two-way repeated measures ANOVA and Tukey's tests. Confocal data were analyzed by one-way ANOVA, Tukey's test, and Kruskal-Wallis and Dunn's tests. Spearman and Pearson's correlation tests were also used. Significance level was set at 5%. RESULTS: At T1, the AMTN group showed the lowest permeability value, following the increasing order at T2: AMTN = SBU < BIOS = SPRG < TMP < FLU < PLA. The SBU group had the highest value of occluded dentinal tubule length. The AMTN group presented more occluded dentinal tubules compared to PLA and FLU. AMTN and SBU had the lowest values of opened dentin tubules. Results showed a negative correlation between the analyses. CONCLUSION: The SBU and AMTN solution were more effective in reducing dentin permeability by occluding dentin tubules. CLINICAL RELEVANCE: All materials reduced permeability after challenge, except fluoride varnish.

Analysis of permeability and biological properties of dentin treated with experimental bioactive glasses.

OBJECTIVES: To evaluate obliterating capability and biological performance of desensitizing agents. METHODS: 50 dentin blocks were distributed according to the desensitizing agent used (n = 10): Control (Artificial saliva); Ultra EZ (Ultradent); Desensibilize Nano P (FGM); T5-OH Bioactive Glass (Experimental solution); F18 Bioactive Glass (Experimental solution). Desensitizing treatments were performed for 15 days. In addition, specimens were subjected to acid challenge to simulate oral environment demineralizing conditions. Samples were subjected to permeability analysis before and after desensitizing procedures and acid challenge. Cytotoxicity analysis was performed by using Alamar Blue assay and complemented by total protein quantification by Pierce Bicinchoninic Acid assay at 15 min, 24-h and 48-h time points. Scanning electron microscopy and energy dispersion X-ray spectroscopy were performed for qualitative analysis. Data of dentin permeability was analyzed by two-way repeated measures ANOVA and Tukey's test. For cytotoxicity, Kruskal-Wallis and Newman-Keuls tests. RESULTS: for dentin permeability there was no significant difference among desensitizing agents after treatment, but control group presented highest values (0.131 ± 0.076 Lp). After acid challenge, control group maintained highest values (0.044 ± 0.014 Lp) with significant difference to other groups, except for Desensibilize Nano P (0.037 ± 0.019 Lp). For cytotoxicity, there were no significant differences among groups. CONCLUSION: Bioglass-based desensitizers caused similar effects to commercially available products, regarding permeability and dentin biological properties. CLINICAL SIGNIFICANCE: There is no gold standard protocol for dentin sensitivity. The study of novel desensitizing agents that can obliterate dentinal tubules in a faster-acting and long-lasting way may help meet this clinical need.

Bactericidal activity and biofilm inhibition of F18 bioactive glass against Staphylococcus aureus.

Antimicrobial treatment failure has been increasing at alarming rates. In this context, the bactericidal properties of biocompatible antimicrobial agents have been widely studied. F18 is a recently developed bioactive glass that presents a much wider working range when compared to other bioactive glasses, a feature that allows it to be used for coating metallic implants, sintering scaffolds or manufacturing fibers for wound healing applications. The aim of this study was to investigate the in vitro bactericidal and anti-biofilm activity of F18 glass as a powder and as a coating on steel samples, and to explore the effects of its dissolution products at concentrations from 3 mg/mL to 50 mg/mL against the Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) biofilms. Furthermore, we intend to verify whether changes in the medium pH could influence the bactericidal activity of F18. The results indicated that F18 presented bactericidal activity in preformed S. aureus and MRSA biofilms, reducing more than 6 logs of the viable cells that remained in contact with 50 mg/mL for 24 h. Moreover, an anti-biofilm activity was observed after 12 h of direct contact, with a drop of more than 6 logs of the viable bacterial population. Neutralization of the F18 solution pH decreased its bactericidal efficacy. These results indicate that the F18 glass could be considered as an alternative material for controlling and treating infections by S. aureus.

Biocompatibility, Biomineralization, and Maturation of Collagen by RTR®, Bioglass and DM Bone® Materials.

This study evaluated the biocompatibility, biomineralization, and collagen fiber maturation induced by Resorbable Tissue Replacement (RTR®; ß-tricalcium phosphate [TCP]), Bioglass (BIOG; bioactive glass), and DM Bone® (DMB; hydroxyapatite and ß-TCP) in vivo. Sixty-four polyethylene tubes with or without (control group; CG) materials (n=8/group/period) were randomly implanted in the subcutaneous tissue of 16 male Wistar rats (four per rat), weighting 250 to 280 g. The rats were killed after 7 and 30 days (n=8), and the specimens were removed for analysis of inflammation using hematoxylin-eosin; biomineralization assay using von Kossa (VK) staining and polarized light (PL); and collagen fiber maturation using picrosirius red (PSR). Nonparametric data were statistically analyzed by Kruskal-Wallis and Dunn tests, and parametric data by one-way ANOVA test (p<0.05). At 7 days, all groups induced moderate inflammation (p>0.05). At 30 days, there was mild inflammation in the BIOG and CG, and moderate inflammation in the RTR and DMB groups, with a significant difference between the CG and RTR (p<0.05). The fibrous capsule was thick at 7 days and predominantly thin at 30 days in all groups. All materials exhibited structures that stained positively for VK and PL. Immature collagen fibers were predominant at 7 and 30 days in all groups (p>0.05), although DMB exhibited more mature fibers than BIOG at 30 days (p<0.05). RTR, BIOG, and DMB were biocompatible, inducing inflammation that reduced over time and biomineralization in the subcutaneous tissue of rats. DMB exhibited more mature collagen fibers than BIOG over a longer period.

Biofilm Formation and Expression of Virulence Genes of Microorganisms Grown in Contact with a New Bioactive Glass.

Bioactive glass F18 (BGF18), a glass containing SiO2-Na2O-K2O-MgO-CaO-P2O5, is highly effective as an osseointegration buster agent when applied as a coating in titanium implants. Biocompatibility tests using this biomaterial exhibited positive results; however, its antimicrobial activity is still under investigation. In this study we evaluated biofilm formation and expression of virulence-factor-related genes in Candida albicans, Staphylococcus epidermidis, and Pseudomonas aeruginosa grown on surfaces of titanium and titanium coated with BGF18. C. albicans, S. epidermidis, and P. aeruginosa biofilms were grown on specimens for 8, 24, and 48 h. After each interval, the pH was measured and the colony-forming units were counted for the biofilm recovery rates. In parallel, quantitative real-time polymerase chain reactions were carried out to verify the expression of virulence-factor-related genes. Our results showed that pH changes of the culture in contact with the bioactive glass were merely observed. Reduction in biofilm formation was not observed at any of the studied time. However, changes in the expression level of genes related to virulence factors were observed after 8 and 48 h of culture in BGF18. BGF18 coating did not have a clear inhibitory effect on biofilm growth but promoted the modulation of virulence factors.

Biocompatibility, biomineralization, and maturation of collagen by rtr®, bioglass and dm bone® materials

Abstract This study evaluated the biocompatibility, biomineralization, and collagen fiber maturation induced by Resorbable Tissue Replacement (RTR®; β-tricalcium phosphate [TCP]), Bioglass (BIOG; bioactive glass), and DM Bone® (DMB; hydroxyapatite and β-TCP) in vivo. Sixty-four polyethylene tubes with or without (control group; CG) materials (n=8/group/period) were randomly implanted in the subcutaneous tissue of 16 male Wistar rats (four per rat), weighting 250 to 280 g. The rats were killed after 7 and 30 days (n=8), and the specimens were removed for analysis of inflammation using hematoxylin-eosin; biomineralization assay using von Kossa (VK) staining and polarized light (PL); and collagen fiber maturation using picrosirius red (PSR). Nonparametric data were statistically analyzed by Kruskal-Wallis and Dunn tests, and parametric data by one-way ANOVA test (p<0.05). At 7 days, all groups induced moderate inflammation (p>0.05). At 30 days, there was mild inflammation in the BIOG and CG, and moderate inflammation in the RTR and DMB groups, with a significant difference between the CG and RTR (p<0.05). The fibrous capsule was thick at 7 days and predominantly thin at 30 days in all groups. All materials exhibited structures that stained positively for VK and PL. Immature collagen fibers were predominant at 7 and 30 days in all groups (p>0.05), although DMB exhibited more mature fibers than BIOG at 30 days (p<0.05). RTR, BIOG, and DMB were biocompatible, inducing inflammation that reduced over time and biomineralization in the subcutaneous tissue of rats. DMB exhibited more mature collagen fibers than BIOG over a longer period.

Resumo Este estudo avaliou a biocompatibilidade, biomineralização e maturação das fibras de colágeno induzidas por Resorbable Tissue Replacement (RTR®; fosfato β-tricálcico [TCP]), Bioglass (BIOG; vidro bioativo) e DM Bone® (DMB; hidroxiapatita e β-TCP) in vivo. Sessenta e quatro tubos de polietileno com ou sem (grupo controle; GC) os materiais (n=8/grupo/período) foram implantados aleatoriamente em tecido subcutâneo de 16 ratos machos Wistar (quatro por rato), pesando entre 250 a 280g. Os ratos foram mortos após 7 e 30 dias (n=8), e as amostras foram removidas para análise da inflamação utilizando hematoxilina-eosina; avaliação da biomineralização utilizando coloração de von Kossa (VK) e luz polarizada (LP); e maturação das fibras colágenas, utilizando picrosirius red (PSR). Os dados não-paramétricos foram analisados pelos testes de Kruskal-Wallis e Dunn, e os paramétricos pelo teste de one-way ANOVA (p<0.05). Aos 7 dias, todos os grupos induziram inflamação moderada (p>0,05). Aos 30 dias, houve inflamação leve nos grupos BIOG e GC, e inflamação moderada nos grupos RTR e DMB, com diferença significativa entre os GC e RTR (p<0,05). A cápsula fibrosa foi espessa aos 7 dias, e predominantemente fina aos 30 dias em todos os grupos. Todos os materiais exibiram estruturas positivas para VK e LP. Fibras colágenas imaturas foram predominantes aos 7 e 30 dias em todos os grupos (p>0,05), embora o DMB exibiu fibras mais maduras do que o BIOG aos 30 dias (p<0,05). RTR, BIOG e DMB foram biocompatíveis, induzindo inflamação que reduziu com o tempo, e biomineralização no tecido subcutâneo de ratos. O DMB exibiu mais fibras colágenas maduras do que o BIOG em período mais longo.

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.

Bioactive Glass Fiber-Reinforced PGS Matrix Composites for Cartilage Regeneration.

Poly(glycerol sebacate) (PGS) is an elastomeric polymer which is attracting increasing interest for biomedical applications, including cartilage regeneration. However, its limited mechanical properties and possible negative effects of its degradation byproducts restrict PGS for in vivo application. In this study, a novel PGS-bioactive glass fiber (F18)-reinforced composite was developed and characterized. PGS-based reinforced scaffolds were fabricated via salt leaching and characterized regarding their mechanical properties, degradation, and bioactivity in contact with simulated body fluid. Results indicated that the incorporation of silicate-based bioactive glass fibers could double the composite tensile strength, tailor the polymer degradability, and improve the scaffold bioactivity.