Chitosan-Calcium Aluminate as a Cell-homing Scaffold: Its Bioactivity Testing in a Microphysiological Dental Pulp Platform.

In vitro models of the dental pulp microenvironment have been proposed for the assessment of biomaterials, to minimise animal use in operative dentistry. In this study, a scaffold/3-D dental pulp cell culture interface was created in a microchip, under simulated dental pulp pressure, to evaluate the cell-homing potential of a chitosan (CH) scaffold functionalised with calcium aluminate (the 'CHAlCa scaffold'). This microphysiological platform was cultured at a pressure of 15 cm H2O for up to 14 days; cell viability, migration and odontoblastic differentiation were then assessed. The CHAlCa scaffold exhibited intense chemotactic potential, causing cells to migrate from the 3-D culture to its surface, followed by infiltration into the macroporous structure of the scaffold. By contrast, the cells in the presence of the non-functionalised chitosan scaffold showed low cell migration and no cell infiltration. CHAlCa scaffold bioactivity was confirmed in dentin sialophosphoprotein-positive migrating cells, and odontoblastic markers were upregulated in 3-D culture. Finally, in situ mineralised matrix deposition by the cells was confirmed in an Alizarin Red-based assay, in which the CHAlCa and CH scaffolds were adapted to fit within dentin discs. More intense deposition of matrix was observed with the CHAlCa scaffold, as compared to the CH scaffold. In summary, we present an in vitro platform that provides a simple and reproducible model for selecting and developing innovative biomaterials through the assessment of their cell-homing potential. By using this platform, it was shown that the combination of calcium aluminate and chitosan has potential as an inductive biomaterial that can mediate dentin tissue regeneration during cell-homing therapies.

Surface and intra-pulpal temperature variation during tooth whitening photoactivated with LED/laser.

This study evaluated the variation of surface and intra-pulpal temperature, during bleaching protocol, using LED/laser. The 35% (HP35), 15% (HP15) and 6% (HP6) gels were used associated with LED/laser applied every 1 min for 30 min in a human canine. The evaluation of surface temperature variation (∆Ts) was performed using a pHmeter and the intra-pulpal temperature variation (∆Ti) was performed using a digital thermometer, at times of 1-, 5-, 10- 15- and 30-min. Statistical analysis was performed using the two-way repeated measures ANOVA test and Bonferroni post-hoc test was used at a significance level of 5%. HP35 and HP15 showed greater temperature variation than HP6 up to 10 min of surface evaluation, showing no differences between them. In the intra-pulpal evaluation, no group showed differences throughout the procedure.

Assessment of the regenerative potential of macro-porous chitosan-calcium simvastatin scaffolds on bone cells.

This study evaluated the bioactive potential of a macro-porous chitosan scaffold incorporated with calcium hydroxide (CH-Ca) and functionalized with bioactive doses of simvastatin (SV) for bone tissue regeneration. Initially, the bioactive dose of SV in osteoblastic cells (SAOS-2) was determined. For the direct contact experiment, SAOS-2 cells were plated on scaffolds to assess cell viability and osteogenic differentiation. The second assay was performed at a distance using extracts from scaffolds incubated in culture medium to assess the effect of conditioned medium on viability and osteogenic differentiation. The initial screening showed that 1 µM SV presented the best biostimulating effects, and this dose was selected for incorporation into the CH-Ca and pure chitosan (CH) scaffolds. The cells remained viable throughout the direct contact experiment, with the greatest cell density in the CH-Ca and CH-Ca-SV scaffolds because of their higher porosity. The CH-Ca-SV scaffold showed the most intense bio-stimulating effect in assays in the presence and absence of osteogenic medium, leading to an increased deposition of mineralized matrix. There was an increase in the viability of cells exposed to the extracts for CH-Ca, CH-SV, and CH-Ca-SV during the one-day period. There was an increase in ALP activity in the CH-Ca and CH-Ca-SV; however, the CH-Ca-SV scaffold resulted in an intense increase in the deposition of mineralized nodules, approximately 56.4% at 7 days and 117% at 14 days, compared with CH (control). In conclusion, functionalization of the CH-Ca scaffold with SV promoted an increase in bioactivity, presenting a promising option for bone tissue regeneration.

Assessment of the regenerative potential of macro-porous chitosan-calcium simvastatin scaffolds on bone cells

Abstract This study evaluated the bioactive potential of a macro-porous chitosan scaffold incorporated with calcium hydroxide (CH-Ca) and functionalized with bioactive doses of simvastatin (SV) for bone tissue regeneration. Initially, the bioactive dose of SV in osteoblastic cells (SAOS-2) was determined. For the direct contact experiment, SAOS-2 cells were plated on scaffolds to assess cell viability and osteogenic differentiation. The second assay was performed at a distance using extracts from scaffolds incubated in culture medium to assess the effect of conditioned medium on viability and osteogenic differentiation. The initial screening showed that 1 μM SV presented the best biostimulating effects, and this dose was selected for incorporation into the CH-Ca and pure chitosan (CH) scaffolds. The cells remained viable throughout the direct contact experiment, with the greatest cell density in the CH-Ca and CH-Ca-SV scaffolds because of their higher porosity. The CH-Ca-SV scaffold showed the most intense bio-stimulating effect in assays in the presence and absence of osteogenic medium, leading to an increased deposition of mineralized matrix. There was an increase in the viability of cells exposed to the extracts for CH-Ca, CH-SV, and CH-Ca-SV during the one-day period. There was an increase in ALP activity in the CH-Ca and CH-Ca-SV; however, the CH-Ca-SV scaffold resulted in an intense increase in the deposition of mineralized nodules, approximately 56.4% at 7 days and 117% at 14 days, compared with CH (control). In conclusion, functionalization of the CH-Ca scaffold with SV promoted an increase in bioactivity, presenting a promising option for bone tissue regeneration.

Violet LED associated with high concentration hydrogen peroxide: Effects on bleaching efficacy, pH, and temperature.

BACKGROUND: This study aimed to evaluate the bleaching efficacy, pH, and temperature of 35% hydrogen peroxide (HP) gel used alone or associated with violet LED. METHODS: Sixty bovine crowns were sectioned (5 × 5 × 2mm). After staining with black tea, the specimens were randomized into four groups (n = 10) according to the bleaching protocol: HP35R: 3 × 15 min 35% HP; HP35: 1 × 45 min 35% HP; HP35VR: 3 × 8min 35% HP + Violet LED; HP35V: 1 × 24 min + Violet LED. Two bleaching sessions were performed for all the groups. Color change was evaluated before, 24h after each session, 7 days and 15 days after the last session. The variables ∆E00 [CIEDE2000] and WID were used for color analysis. The pH variation (initial and final) and the temperature of the gel were recorded (n = 5). ANOVA two-way for repeated measures and Bonferroni post-test was used at a significance level of 5%. RESULTS: HP35VR and HP35V the most noticeable color change(p < 0.05). The final values of pH were lower than the initial ones, but with no difference between the groups (p > 0.05). Groups HP35VR and HP35V showed an increase in temperature in relation to HP35R (p < 0.05). CONCLUSIONS: Violet LED improved the bleaching efficacy of 35% HP in a time-saving manner without negatively affecting the pH and temperature of 35% HP. The renewal of HP did not influence the outcomes.

TiO2 nanoparticles added to dense bovine hydroxyapatite bioceramics increase human osteoblast mineralization activity.

OBJECTIVES: This study evaluated the effect of TiO2 nanoparticles + dense hydroxyapatite (HA) on human osteoblast cells (SAOS-2). METHODS: Particulate bovine HA powder with or without the addition of either 5 or 8 % TiO2 (HA, HA/TiO2Np5 % or HA/TiO2Np8 %) were pressed into disks (Ø = 12.5 mm; thickness = 1.3 mm) uniaxially (100 MPa) and isostatically (200 MPa/1 min) and sintered at 1300 °C. Y-TZP disks were used as control. The following tests were performed: Scanning Electron Microscopy and Dispersive Energy Spectroscopy (SEM/EDS), Atomic Force Microscopy (AFM), cell viability assay (Alamar Blue-AB) and mineralized matrix deposition (Alizarin Red-AR). AB and AR data were submitted to 2-way ANOVA/Tukey tests and ANOVA/Tukey tests, respectively. RESULTS: SEM revealed that the surface of HA/TiO2Np5% resembles DPBHA surface, but also contains smaller granules. HA/TiO2Np8% characteristics resembles HA/TiO2Np5% surface, but with irregular topography. Y-TZP showed a typical oxide ceramic surface pattern. EDS revealed Ca, O, and P in all samples. C, O, and Zr appeared in Y-TZP samples. AFM data corroborates SEM analysis. AB test revealed excellent cellular viability for HA/TiO2Np5% group. AR test showed that all groups containing TiO2np had more mineralized matrix deposition than all other groups, with statistically differences between HA/TiO2Np8% and HA cultivated in non-osteogenic medium. Culture in osteogenic medium exhibited much more mineralized matrix deposition by TiO2np groups. SIGNIFICANCE: In conclusion, the addition of TiO2np showed chemical, superficial, and biological changes in the reinforced materials. HA/TiO2Np5% showed the best results for cell viability and HA/TiO2Np8% for mineralized matrix deposition.

An organotypic model of oral mucosa cells for the biological assessment of 3D-printed resins for interim restorations.

STATEMENT OF PROBLEM: Three-dimensionally (3D) printed resins have become popular as a new class of materials for making interim restorations. However, little is known about how the fabrication parameters can influence biological compatibility with oral tissues. PURPOSE: The purpose of this in vitro study was to evaluate the effect of the postpolymerization time on the cytotoxicity of resins for printing interim restorations by using a 3D organotypic model of the oral mucosa. MATERIAL AND METHODS: Cylindrical specimens were prepared with conventional acrylic resin (AR), computer-aided design and computer-aided manufacture (CAD-CAM) resin (CC), composite resin (CR), and 2 resins for 3D printing (3DP) marketed as being biocompatible. The 3DPs were submitted to postpolymerization in an ultraviolet (UV) light chamber for 1, 10, or 20 minutes (90 W, 405 nm). Standard specimens of the materials were incubated for 1, 3, and 7 days in close contact with an organotypic model of keratinocytes (NOK-Si) in coculture with gingival fibroblasts (HGF) in a 3D collagen matrix, or directly with 3D HGF cultures. Then, the viability (Live/Dead n=2) and metabolism (Alamar Blue n=6) of the cells were assessed. Spectral scanning of the culture medium was performed to detect released components (n=6) and assessed statistically with ANOVA and the Tukey post hoc test (α=.05). RESULTS: Severe reduction of metabolism (>70%) and viability of keratinocytes occurred for 3DP resin postpolymerized for 1 minute in all periods of analysis in a time-dependent manner. The decrease in cell metabolism and viability was moderate for the 3D culture of HGFs in both experimental models, correlated with the intense presence of resin components in the culture medium. The resins postpolymerized for 10 and 20 minutes promoted a mild-moderate cytotoxic effect in the period of 1 day, similar to AR. However, recovery of cell viability occurred at the 7-day incubation period. The 3DP resins submitted to postpolymerization for 20 minutes showed a pattern similar to that of CR and CC at the end of the experiment. CONCLUSIONS: The cytotoxic potential of the tested 3DP resins on oral mucosa cells was influenced by postprinting processing, which seemed to have been related with the quantity of residual components leached.

Mineral-induced bubbling effect and biomineralization as strategies to create highly porous and bioactive scaffolds for dentin tissue engineering.

The objective of the study was to assess the biological and mechanical characteristics of chitosan-based scaffolds enriched by mineral phases and biomineralized in simulated body fluid (SBF) as a possible biomaterial for dentin regeneration. Thus, porous chitosan scaffolds were prepared by the mineral-induced bubbling-effect technique and subjected to biomineralization to create biomimetic scaffolds for dentin tissue engineering. Suspensions containing calcium hydroxide, nanohydroxyapatite, or ß-tricalcium phosphate were added to the chitosan (CH) solution and subjected to gradual freezing and freeze-drying to obtain CHCa, CHnHA, and CHßTCP porous scaffolds, respectively, by the bubbling effect. Then, scaffolds were incubated in SBF for 5 days at 37°C, under constant stirring, to promote calcium-phosphate (CaP) biomineralization. Scanning electron microscopy revealed increased pore size and porosity degree on mineral-containing scaffolds, with CHCa and CHnHA presenting as round, well-distributed, and with an interconnected pore network. Nevertheless, incubation in SBF disrupted the porous architecture, except for CHCaSBF , leading to the deposition of CaP coverage, confirmed by Fourier Transform Infrared Spectroscopy analyses. All mineral-containing and SBF-treated formulations presented controlled degradation profiles and released calcium throughout 28 days. When human dental pulp cells (HDPCs) were seeded onto scaffold structures, the porous and interconnected architecture of CHCa, CHnHA, and CHCaSBF allowed cells to infiltrate and spread throughout the scaffold structure, whereas in other formulations cells were dispersed or agglomerated. It was possible to determine a positive effect on cell proliferation and odontogenic differentiation for mineral-containing formulations, intensely improved by biomineralization. A significant increase in mineralized matrix deposition (by 8.4 to 18.9 times) was observed for CHCaSBF , CHnHASBF , and CHßTCPSBF in comparison with plain CH. The bioactive effect on odontoblastic marker expression (ALP activity and mineralized matrix) was also observed for HDPCs continuously cultivated with conditioned medium obtained from scaffolds. Therefore, biomineralization of chitosan scaffolds containing different mineral phases was responsible for increasing the capacity for mineralized matrix deposition by pulpal cells, with potential for use in dentin tissue engineering.

Novel cinnamon-laden nanofibers as a potential antifungal coating for poly(methyl methacrylate) denture base materials.

OBJECTIVES: To modify the surface of denture base material by coating it with cinnamon-laden nanofibers to reduce Candida albicans (C. albicans) adhesion and/or proliferation. MATERIALS AND METHODS: Heat-cured poly(methyl methacrylate) (PMMA) specimens were processed and coated, or not, with cinnamon-laden polymeric nanofibers (20 or 40 wt.% of cinnamon relative to the total polymer weight). Scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR) analyses of the nanofibers were performed. Antifungal activity was assessed through agar diffusion and colony-forming unit (CFU/mL) assays. Representative SEM morphological analysis was carried out to observe the presence/absence of C. albicans on the fibers. Alamar blue assay was used to determine cell toxicity. Analysis of variance and the Tukey's test were used to analyze the data (α = 0.05). RESULTS: SEM imaging revealed nanofibers with adequate (i.e., bead-free) morphological characteristics and uniform microstructure. FTIR confirmed cinnamon incorporation. The cinnamon-laden nanofibers led to growth inhibition of C. albicans. Viable fungal counts support a significant reduction on CFU/mL also directly related to cinnamon concentration (40 wt.%: mean log 6.17 CFU/mL < 20 wt.%: mean log 7.12 CFU/mL), which agrees with the SEM images. Cinnamon-laden nanofibers at 40 wt.% led to increased cell death. CONCLUSIONS: The deposition of 20 wt.% cinnamon-laden nanofibers onto PMMA surfaces led to a significant reduction of the adhesive and/or proliferative ability of C. albicans, while maintaining epithelial cells' viability. CLINICAL RELEVANCE: The high recurrence rates of denture stomatitis are associated with patient non-adherence to treatments and contaminated prostheses use. Here, we provide the non-patients' cooperation sensible method, which possesses antifungal action, hence improving treatment effectiveness.

Bioactivity effects of extracellular matrix proteins on apical papilla cells.

BACKGROUND: Potent signaling agents stimulate and guide pulp tissue regeneration, especially in endodontic treatment of teeth with incomplete root formation. OBJECTIVE: This study evaluated the bioactive properties of low concentrations of extracellular matrix proteins on human apical papilla cells (hAPCs). METHODOLOGY: Different concentrations (1, 5, and 10 µg/mL) of fibronectin (FN), laminin (LM), and type I collagen (COL) were applied to the bottom of non-treated wells of sterilized 96-well plates. Non-treated and pre-treated wells were used as negative (NC) and positive (PC) controls. After seeding the hAPCs (5×103 cells/well) on the different substrates, we assessed the following parameters: adhesion, proliferation, spreading, total collagen/type I collagen synthesis and gene expression (ITGA5, ITGAV, COL1A1, COL3A1) (ANOVA/Tukey; α=0.05). RESULTS: We observed greater attachment potential for cells on the FN substrate, with the effect depending on concentration. Concentrations of 5 and 10 µg/mL of FN yielded the highest cell proliferation, spreading and collagen synthesis values with 10 µg/mL concentration increasing the ITGA5, ITGAV, and COL1A1 expression compared with PC. LM (5 and 10 µg/mL) showed higher bioactivity values than NC, but those were lower than PC, and COL showed no bioactivity at all. CONCLUSION: We conclude that FN at 10 µg/mL concentration exerted the most intense bioactive effects on hAPCs.

Management of Tooth Discoloration after Orthognathic Surgery Complications: Report of Two Cases

ABSTRACT: Orthognathic surgery is performed for the correction of craniofacial discrepancies. However, complications, such as tooth discoloration are possible. This case report presents two patients who underwent bilateral sagittal split ramus osteotomy associated with segmental Le Fort I osteotomy and genioplasty. During surgeries, the apical region of anterior teeth was accidentally injured in both cases. After three-week surgery follow-up, the injured teeth showed a change in color to dark pink. In both teeth, the root canal treatment was performed followed by the non vital tooth bleaching. Three sessions were necessary to achieve a significant color change of the teeth. The two-year follow-up showed that both teeth preserved an acceptable color. It was concluded that tooth discoloration after orthognathic surgery is a possible complication, which could be overcome following a conservative approach. Additionally, patients should be informed preoperatively.

RESUMEN: La cirugía ortognática es comúnmente realizada para corregir las discrepancias cráneo-faciales. Sin embargo, se pueden producir complicaciones tales como la pigmentación dentaria. Este reporte de casos presenta a dos pacientes que fueron sometidos osteotomía sagital bilateral de la rama mandibular asociada a osteotomía segmentaria Le Fort I y genioplastía. Durante la fase quirúrgica, la región apical de dientes anteriores fueron accidentalmente dañados en ambos casos. Después de tres semanas de seguimiento de la cirugía, los dientes afectados mostraron un cambio de color a rosado oscuro. Se realizó el tratamiento de conductos seguido de aclaramiento interno en ambas piezas dentarias. Fueron necesarias tres sesiones para lograr un cambio de color significativo. El seguimiento de dos años mostró que ambos dientess conservaron un color aceptable. En conclusión, la pigmentación dentaria después de la cirugía ortognática es una posible complicación, sin embargo, puede ser tratada por medio de un enfoque conservador. Además, los pacientes deben ser informados antes de la cirugía.

Bioactivity effects of extracellular matrix proteins on apical papilla cells

Abstract Potent signaling agents stimulate and guide pulp tissue regeneration, especially in endodontic treatment of teeth with incomplete root formation. Objective This study evaluated the bioactive properties of low concentrations of extracellular matrix proteins on human apical papilla cells (hAPCs). Methodology Different concentrations (1, 5, and 10 µg/mL) of fibronectin (FN), laminin (LM), and type I collagen (COL) were applied to the bottom of non-treated wells of sterilized 96-well plates. Non-treated and pre-treated wells were used as negative (NC) and positive (PC) controls. After seeding the hAPCs (5×103 cells/well) on the different substrates, we assessed the following parameters: adhesion, proliferation, spreading, total collagen/type I collagen synthesis and gene expression (ITGA5, ITGAV, COL1A1, COL3A1) (ANOVA/Tukey; α=0.05). Results We observed greater attachment potential for cells on the FN substrate, with the effect depending on concentration. Concentrations of 5 and 10 µg/mL of FN yielded the highest cell proliferation, spreading and collagen synthesis values with 10 µg/mL concentration increasing the ITGA5, ITGAV, and COL1A1 expression compared with PC. LM (5 and 10 µg/mL) showed higher bioactivity values than NC, but those were lower than PC, and COL showed no bioactivity at all. Conclusion We conclude that FN at 10 µg/mL concentration exerted the most intense bioactive effects on hAPCs.

Simvastatin-Enriched Macro-Porous Chitosan-Calcium-Aluminate Scaffold for Mineralized Tissue Regeneration.

The present study evaluated the odontogenic potential of human dental pulp cells (HDPCs) exposed to chitosan scaffolds containing calcium aluminate (CHAlCa) associated or not with low doses of simvastatin (SV). Chitosan scaffolds received a suspension of calcium aluminate (AlCa) and were then immersed into solutions containing SV. The following groups were established: chitosan-calcium-aluminate scaffolds (CHAlCa - Control), chitosan calcium-aluminate with 0.5 µM SV (CHAlCa-SV0.5), and chitosan calcium-aluminate with 1.0 µM SV (CHAlCa-SV1.0). The morphology and composition of the scaffolds were evaluated by SEM and EDS, respectively. After 14 days of HDPCs culture on scaffolds, cell viability, adhesion and spread, mineralized matrix deposition as well as gene expression of odontogenic markers were assessed. Calcium aluminate particles were incorporated into the chitosan matrix, which exhibited regular pores homogeneously distributed throughout its structure. The selected SV dosages were biocompatible with HDPCs. Chitosan-calcium-aluminate scaffolds with 1 µM SV induced the odontoblastic phenotype in the HDPCs, which showed enhanced mineralized matrix deposition and up-regulated ALP, Col1A1, and DMP-1 expression. Therefore, one can conclude that the incorporation of calcium aluminate and simvastatin in chitosan scaffolds had a synergistic effect on HDPCs, favoring odontogenic cell differentiation and mineralized matrix deposition.

Antibiofilm efficacy of tea tree oil and of its main component terpinen-4-ol against Candida albicans.

Candida infection is an important cause of morbidity and mortality in immunocompromised patients. The increase in its incidence has been associated with resistance to antimicrobial therapy and biofilm formation. The aim of this study was to evaluate the efficacy of tea tree oil (TTO) and its main component - terpinen-4-ol - against resistant Candida albicans strains (genotypes A and B) identified by molecular typing and against C. albicans ATCC 90028 and SC 5314 reference strains in planktonic and biofilm cultures. The minimum inhibitory concentration, minimum fungicidal concentration, and rate of biofilm development were used to evaluate antifungal activity. Results were obtained from analysis of the biofilm using the cell proliferation assay 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and confocal laser scanning microscopy (CLSM). Terpinen-4-ol and TTO inhibited C. albicans growth. CLSM confirmed that 17.92 mg/mL of TTO and 8.86 mg/mL of terpinen-4-ol applied for 60 s (rinse simulation) interfered with biofilm formation. Hence, this in vitro study revealed that natural substances such as TTO and terpinen-4-ol present promising results for the treatment of oral candidiasis.

Characterization of novel calcium hydroxide-mediated highly porous chitosan-calcium scaffolds for potential application in dentin tissue engineering.

The aim of this study was to develop a highly porous calcium-containing chitosan scaffold suitable for dentin regeneration. A calcium hydroxide (Ca[OH]2 ) suspension was used to modulate the degree of porosity and chemical composition of chitosan scaffolds. The chitosan solution concentration and freezing protocol were adjusted to optimize the porous architecture using the phase-separation technique. Scanning electron microscopy/energy-dispersive spectroscopy demonstrated the fabrication of a highly porous calcium-linked chitosan scaffold (CH-Ca), with a well-organized and interconnected porous network. Scaffolds were cross-linked on glutaraldehyde (GA) vapor. Following a 28-day incubation in water, cross-linked CH scaffold had no changes on humid mass, and CH-Ca featured a controlled degradability profile since the significant humid mass loss was observed only after 21 (26.0%) and 28 days (42.2%). Fourier-transform infrared spectroscopy indicated the establishment of Schiff base on cross-linked scaffolds, along with calcium complexation for CH-Ca. Cross-linked CH-Ca scaffold featured a sustained Ca2+ release up to 21 days in a humid environment. This porous and stable architecture allowed for human dental pulp cells (HDPCs) to spread throughout the scaffold, with cells exhibiting a widely stretched cytoplasm; whereas, the cells seeded onto CH scaffold were organized in clusters. HDPCs seeded onto CH-Ca featured significantly higher ALP activity, and gene expressions for ALP, Col1, DMP-1, and DSPP in comparison to CH, leading to a significant 3.5 times increase in calcium-rich matrix deposition. In sum, our findings suggest that CH-Ca scaffolds are attractive candidates for creating a highly porous and bioactive substrate for dentin tissue engineering.

Human pulp response to conventional and resin-modified glass ionomer cements applied in very deep cavities.

OBJECTIVES: This study assessed the human pulp response after adhesive restoration of cavities by indirect pulp capping with a conventional or a resin-modified glass ionomer cement. MATERIALS AND METHODS: Deep cavities prepared in 26 human premolars were lined with Riva Light Cure (Riva LC), Riva Self Cure (Riva SC), or Dycal, and then restored with composite resin. Four teeth were used as intact control. After time intervals of 7 or 30 days, the teeth were extracted, processed for histological evaluation of the pulp, and the remaining dentin thickness (RDT) between the cavity floor and the pulp was measured. RESULTS: At 7 days, a slight pulp inflammation associated with discrete tissue disorganization was observed in most of t the teeth lined with Riva LC and Riva SC. Moderate pulp inflammation occurred in one tooth lined with Riva LC. Bacteria were identified in one specimen of the same group that exhibited no pulp damage. At 30 days, slight pulp inflammation and discrete tissue disorganization persisted in two specimens treated with Riva LC, in which a thin layer of tertiary dentin was deposited. Mean RDTs ranged from 383.0 to 447.8 µm. CONCLUSIONS: Riva LC produced more damage to the pulp than Riva SC. However, the initial pulp damage decreased over time and after 30 days both GICs were labeled as biocompatible. CLINICAL RELEVANCE: In this study conducted with human teeth, the conventional and the resin-modified glass ionomer cements investigated were shown not to cause post-operative sensitivity or persistent pulp damage when applied as liners in very deep cavities, thereby indicating their biocompatibility.

Synergistic potential of 1α,25-dihydroxyvitamin D3 and calcium-aluminate-chitosan scaffolds with dental pulp cells.

OBJECTIVES: This study aimed to develop a porous chitosan-calcium-aluminate scaffold (CH-AlCa) in combination with a bioactive dosage of 1α,25-dihydroxyvitamin D3 (1α,25VD), to be used as a bioactive substrate capable to increase the odontogenic potential of human dental pulp cells (HDPCs). MATERIALS AND METHODS: The porous CH-AlCa was developed by the incorporation of an AlCa suspension into a CH solution under vigorous agitation, followed by phase separation at low temperature. Scaffold architecture, porosity, and calcium release were evaluated. Thereafter, the synergistic potential of CH-AlCa and 1 nM 1α,25VD, selected by a dose-response assay, for HDPCs seeded onto the materials was assessed. RESULTS: The CH-AlCa featured an organized and interconnected pore network, with increased porosity in comparison with that of plain chitosan scaffolds (CH). Increased odontoblastic phenotype expression on the human dental pulp cell (HDPC)/CH and HDPC/CH-AlCa constructs in the presence of 1 nM 1α,25VD was detected, since alkaline phosphatase activity, mineralized matrix deposition, dentin sialophosphoprotein/dentin matrix acidic phosphoprotein 1 mRNA expression, and cell migration were overstimulated. This drug featured a synergistic effect with CH-AlCa, since the highest values of cell migration and odontoblastic markers expression were observed in this experimental condition. CONCLUSIONS: The experimental CH-AlCa scaffold increases the chemotaxis and regenerative potential of HDPCs, and the addition of low-dosage 1α,25VD to this scaffold enhances the potential of these cells to express an odontoblastic phenotype. CLINICAL RELEVANCE: Chitosan scaffolds enriched with calcium-aluminate in association with low dosages of 1α,25-dihydroxyvitamin D3 provide a highly bioactive microenvironment for dental pulp cells prone to dentin regeneration, thus providing potential as a cell-free tissue engineering system for direct pulp capping.

Antibiofilm efficacy of tea tree oil and of its main component terpinen-4-ol against Candida albicans

Abstract Candida infection is an important cause of morbidity and mortality in immunocompromised patients. The increase in its incidence has been associated with resistance to antimicrobial therapy and biofilm formation. The aim of this study was to evaluate the efficacy of tea tree oil (TTO) and its main component - terpinen-4-ol - against resistant Candida albicans strains (genotypes A and B) identified by molecular typing and against C. albicans ATCC 90028 and SC 5314 reference strains in planktonic and biofilm cultures. The minimum inhibitory concentration, minimum fungicidal concentration, and rate of biofilm development were used to evaluate antifungal activity. Results were obtained from analysis of the biofilm using the cell proliferation assay 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) and confocal laser scanning microscopy (CLSM). Terpinen-4-ol and TTO inhibited C. albicans growth. CLSM confirmed that 17.92 mg/mL of TTO and 8.86 mg/mL of terpinen-4-ol applied for 60 s (rinse simulation) interfered with biofilm formation. Hence, this in vitro study revealed that natural substances such as TTO and terpinen-4-ol present promising results for the treatment of oral candidiasis.

Increased whitening efficacy and reduced cytotoxicity are achieved by the chemical activation of a highly concentrated hydrogen peroxide bleaching gel.

OBJECTIVE: This study was designed for the chemical activation of a 35% hydrogen peroxide (H2O2) bleaching gel to increase its whitening effectiveness and reduce its toxicity. METHODOLOGY: First, the bleaching gel - associated or not with ferrous sulfate (FS), manganese chloride (MC), peroxidase (PR), or catalase (CT) - was applied (3x 15 min) to enamel/dentin discs adapted to artificial pulp chambers. Then, odontoblast-like MDPC-23 cells were exposed for 1 h to the extracts (culture medium + components released from the product), for the assessment of viability (MTT assay) and oxidative stress (H2DCFDA). Residual H2O2 and bleaching effectiveness (DE) were also evaluated. Data were analyzed with one-way ANOVA complemented with Tukey's test (n=8. p<0.05). RESULTS: All chemically activated groups minimized MDPC-23 oxidative stress generation; however, significantly higher cell viability was detected for MC, PR, and CT than for plain 35% H2O2 gel. Nevertheless, FS, MC, PR, and CT reduced the amount of residual H2O2 and increased bleaching effectiveness. CONCLUSION: Chemical activation of 35% H2O2 gel with MC, PR, and CT minimized residual H2O2 and pulp cell toxicity; but PR duplicated the whitening potential of the bleaching gel after a single 45-minute session.

Positive influence of simvastatin used as adjuvant agent for cavity lining.

OBJECTIVES: To assess the biological, antimicrobial, and mechanical effects of the treatment of deep dentin with simvastatin (SV) before application of a glass-ionomer cement (GIC). MATERIALS AND METHODS: Dentin discs were adapted to artificial pulp chambers and SV (2.5 or 1.0 mg/mL) was applied to the occlusal surface, either previously conditioned or not with EDTA (±EDTA). The extracts (culture medium + SV that diffused through dentin) was obtained and then applied to cultured odontoblast-like MDPC-23 cells. Cell viability, alkaline phosphatase (ALP) activity, and mineralization nodule (MN) deposition were evaluated. Untreated discs were used as control. The antibacterial activity of SV (2.5 or 1.0 mg/mL) against Streptococcus mutans and Lactobacillus acidophilus, as well as the bond strength of GIC to dentin in the presence of SV 2.5 mg/mL (±EDTA) were also assessed. The data were analyzed by ANOVA/Tukey tests (α = 5%). RESULTS: EDTA + SV 2.5 mg/mL significantly enhanced the ALP activity and MN deposition in comparison with the control, without changing in the cell viability (p < 0.05). The association EDTA + SV 2.5 mg/mL + GIC determined the highest ALP and MN values (p < 0.05). SV presented intense antimicrobial activity, and the EDTA dentin conditioning followed by SV application increased bond strength values compared with SV treatment alone (p < 0.05). CONCLUSION: SV presents antimicrobial activity and diffuses across conditioned dentin to biostimulate odontoblast-like pulp cells. CLINICAL SIGNIFICANCE: The use of SV as adjuvant agent for indirect pulp capping may biostimulate pulp cells thus preserving vitality and function of the pulp-dentin complex.