Effect of indomethacin-loaded nanocapsules incorporation in a dentin adhesive resin.

OBJECTIVES: The aim of this study was to produce indomethacin-loaded nanocapsules (IndOH-NCs) and evaluate the influence of their incorporation into an adhesive resin. MATERIALS AND METHODS: Indomethacin was encapsulated by the deposition of preformed polymer. IndOH-NCs were characterized by laser diffractometry, Fourier transformed infrared spectrometry, transmission electron microscopy (TEM), scanning electron microscopy, high-performance liquid chromatography (HPLC), and MTT assay. Nanocapsules (NCs) were incorporated into an adhesive in concentrations of 1, 2, 5, and 10 %. The addition was visualized by TEM and drug release was evaluated by HPLC until 120 h of immersion in simulated body fluid (SBF). Drug diffusion through dentin was tested using a Franz diffusion cell apparatus and quantified by HPLC. The degree of conversion (DC), softening in ethanol, and microtensile bond strength (µTBS) were evaluated to determine whether the nanocapsules influenced the adhesive. Data were analyzed using one-way ANOVA and Tukey's post hoc test for DC, softening in ethanol, µTBS, and cytotoxicity, and paired t test for comparison between the initial and final Knoop microhardness. RESULTS: IndOH-NCs, with a spherical shape and a mean diameter of 165 nm, were incorporated into an adhesive. Indomethacin content was 7 mg drug/g powder. IndOH-NCs maintained high cell viability. At 120 h, an amount of 13.83 % of indomethacin was released, and after 7 days, 7.07 % of this drug was diffused through dentin for an adhesive containing 10 % of nanocapsules. No alteration in the DC, softening in ethanol, and µTBS resulted from NC addition. CONCLUSIONS: IndOH-NCs may be incorporated into adhesive systems, without compromising properties, to add an anti-inflammatory drug controlled release for restorative procedures in deep cavities. CLINICAL SIGNIFICANCE: Here is the first step toward the goal of providing agents to act at an inflammatory process of pulp tissue through dental adhesives via encapsulation of drug.

Fluorosilicic acid induces DNA damage and oxidative stress in bone marrow mesenchymal stem cells.

Excess fluoride in water can produce changes in tooth enamel mineralization and lead to diseases such as dental or skeletal fluorosis. The present study aimed to assess the genotoxic effects, oxidative stress, and osteoblastic mineralization induced by fluorosilicic acid (FA) in murine bone marrow-derived mesenchymal stem cells (BM-MSCs). BM-MSCs were isolated from the femurs and tibias of rats and cultured under standard conditions. Cells exposure occurred for 3, 7, 14, and 21 days to different concentrations of FA (0.6-9.6 mg/L). Cytotoxicity was observed in 14 and 21 days of exposure for all concentrations of FA (cell proliferation below 60%), and for 3 and 7 days, in which the proliferation was above 80%. Alkaline comet assay results demonstrated significant increased damage at concentrations of 0.3-2.4 mg/L, and the micronucleus test showed increased rates for micronucleus (1.2-2.4 mg/L) and nuclear buds (NBUDs) (0.3-2.4 mg/L) (P < 0.05/Dunnett's test). An alkaline comet assay modified by repair endonuclease (FPG) was used to detect oxidized nucleobases, which occurred at 0.6 mg/L. The oxidative stress was evaluated by lipid peroxidation (TBARS) and antioxidant activity (TAC). Only lipid peroxidation was increased at concentrations of 0.6 mg/L and 1.2 mg/L (P < 0.001/Tukey's test). The osteogenesis process determined the level of extracellular matrix mineralization. The mean concentration of Alizarin red increased significantly in 14 days at the 0.6 mg/L concentration group (P < 0.05/Tukey's test) compared to the control group, and a significant difference between the groups regarding the activity of alkaline phosphatase (ALP) was observed. Unlike other studies, our results indicated that FA in BM-MSCs at concentrations used in drinking water induced genotoxicity, oxidative stress, and acceleration of bone mineralization.

Nanoneedle-like zinc oxide as a filler particle for an experimental adhesive resin.

AIM: The aim of this study was to develop an experimental adhesive resin with nanoneedle-like zinc oxide (N-ZnO), an inorganic filler, that could avoid particle agglomeration and lead to a homogeneous stress distribution within the material and characterize it. MATERIALS AND METHODS: N-ZnO particles obtained by a thermal evaporation technique were characterized regarding size and surface area and added at 0 (control), 1, 2, 5, and 10 wt%, to an experimental adhesive resin. The following experimental adhesive resins' properties were assessed: radiopacity, contact angle to conditioned enamel and dentin, color, degree of conversion, flexural strength, resistance to degradation, and cytotoxicity. Statistical analysis was performed using one-way ANOVA and Tukey's post hoc test and paired Student's t-test. RESULTS: Particles presented a mean particle size of 40 nm and a specific surface area of 16 m2/g. N-ZnO10%showed an increased radiopacity when compared to N-ZnO0%. Contact angles were significantly higher for N-ZnO10%at enamel and N-ZnO2%, N-ZnO5%, and N-ZnO10%at dentin. All groups showed color change when compared to N-ZnO0%. Higher the N-ZnO concentration, lower the degree of conversion. There were no significant differences between the groups for flexural strength and resistance to degradation. The addition of N-ZnO showed no difference in cytotoxicity when compared to positive control, N-ZnO0%, and all groups showed higher values than negative control. CONCLUSIONS: N-ZnO possibly exceeded potential limitations due to particles' agglomeration and improved the transference and distribution of stress within the material. It could be effectively used as a filler for adhesive resins.

Higher maternal age is associated with higher occurrence of cleft lip/palate in neonates under intensive care

Aim: To assess the prevalence of cleft lip and/or cleft palate (CL/P) and associated variables in neonates admitted to neonatal intensive care units (ICU). Methods: Medical charts for neonates born and admitted to the ICU between 2012 and 2018 were reviewed. Obstetric and neonatal variables were collected by a trained researcher. In the case group, all neonates with CL/P were included. The control group was formed by matching sex, prematurity and month of birth using random number generation. Neonates with congenital malformations were excluded from the control group. Adjusted logistic regression was used (p<0.05). Results: The prevalence of CL/P was 0.43% (n=15). Five cases were excluded, as pairing was not possible. Twenty neonates were included in the control group. In the final multivariate model, CL/P was only associated with increased maternal age. For each year of increase in maternal age, neonates had a 35.2% higher chance of presenting CL/P (95% confidence interval: 1.021­1.792). Conclusions: Higher maternal age was associated with higher occurrence of CL/P in neonates admitted to the ICU. No other neonatal or maternal independent variables were associated with CL/P. Due to missing data, interpretation of study results must be approached with caution

Antimicrobial effect and physicochemical properties of an adhesive system containing nanocapsules.

OBJECTIVE: To incorporate indomethacin and triclosan-loaded nanocapsules into primer and adhesive, and evaluate its properties. METHODS: Indomethacin and triclosan were encapsulated by deposition of preformed polymer and subsequently characterized regarding morphology, particle size, drug content and cytotoxicity. Nanocapsules (NCs) were incorporated into primer at 2% and into adhesive at 1, 2, 5, and 10% concentrations. Degree of conversion (DC) and softening in ethanol of the adhesive were evaluated. Drug release and drug diffusion through dentin was quantified by high performance liquid chromatography. Antimicrobial test was performed until 96h. RESULTS: Spherical and biocompatible NCs presented mean size of 159nm. Drugs content was 3mg indomethacin/g powder and 2mg triclosan/g powder. Incorporating NCs in adhesive showed no influence in DC (p=0.335). The addition of 2% of NCs showed no influence in softening in ethanol (p>0.05). After 120h, 93% of indomethacin and 80% of triclosan were released from primer, 20% of indomethacin and 17% of triclosan were released from adhesive with 10% of NCs. Indomethacin showed diffusion through dentin. In 24h, adhesive containing 2 and 5% of NCs using primer with NCs showed antimicrobial effect. In 96h, adhesives containing different concentration of NCs promoted antimicrobial effect. CONCLUSIONS: Indomethacin and triclosan-loaded nanocapsules were successfully incorporated into primer and adhesive, promoting controlled drugs release, indomethacin diffusion through dentin and antimicrobial effect without compromising its physicochemical properties. SIGNIFICANCE: Indomethacin and triclosan-loaded nanocapsules have potential to prevent recurrent caries and to be used in deep cavities controlling pulpar inflammatory process.

Repair of bone defects using adipose-derived stem cells combined with alpha-tricalcium phosphate and gelatin sponge scaffolds in a rat model.

OBJECTIVES: This study aimed to evaluate the potential of adipose-derived stem cells (ASCs) combined with a modified α-tricalcium phosphate (α-TCP) or gelatin sponge (GS) scaffolds for bone healing in a rat model. MATERIAL AND METHODS: Bone defects were surgically created in the femur of adult SHR rats and filled with the scaffolds, empty or combined with ASCs. The results were analyzed by histology and histomorphometry on days seven, 14, 30, and 60. RESULTS: Significantly increased bone repair was observed on days seven and 60 in animals treated with α-TCP/ASCs, and on day 14 in the group treated with GS/ASCs, when compared with the groups treated with the biomaterials alone. Intense fibroplasia was observed in the group treated with GS alone, on days 14 and 30. CONCLUSIONS: Our results showed that the use of ASCs combined with α-TCP or GS scaffolds resulted in increased bone repair. The higher efficacy of the α-TCP scaffold suggests osteoconductive property that results in a biological support to the cells, whereas the GS scaffold functions just as a carrier. These results confirm the potential of ASCs in accelerating bone repair in in vivo experimental rat models. These results suggest a new alternative for treating bone defects.

Gelatin and galactomannan-based scaffolds: Characterization and potential for tissue engineering applications.

In this work, we produced gelatin films containing different concentrations of galactomannan by casting solutions. The films were crosslinked by immersion in 30mM solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC). The crosslinking of gelatin-containing films was confirmed by the reduction of free amine band intensity (3400-3200cm(-1)) in the GEL IR, as well as by the evaluation of its behavior when immersed in phosphate-buffer solution. The crosslinking of galactomannan film was confirmed by the formation of new ether bonds, as observed by increasing intensity of the band at 1148cm(-1), and the reduction of OH band intensity (3600-3200cm(-1)). The presence of galactomannan and the crosslinking mediated by EDC were responsible to improve elasticity in the gelatin-based films. The samples did not show cytotoxicity during 24h or 48h. In addition, rat mesenchymal stem cells adhered to the films regardless of galactomannan concentration. The results indicated that the gelatin/galactomannan films are potential biomaterials for use as scaffolds for tissue engineering.

Glycerol salicylate-based containing α-tricalcium phosphate as a bioactive root canal sealer.

The use of bioactive materials instead of inert materials to fill the root canal space could be an effective approach to achieve a hermetic seal and stimulate the healing of periapical tissues. The purpose of this study was to develop and characterize an endodontic sealer based on a glycerol salicylate resin and α-tricalcium phosphate (αTCP) at physical and chemical properties. Different sealers were formulated using 70% of a glycerol salicylate resin and 30% of a mixture of calcium hydroxide and αTCP (0, 5, 10, or 15%, in weight). Sealers formulated were characterized based on setting time, in vitro degradation over time, pH, cytotoxicity, and mineral deposition. Sealers presented setting time ranging from 240 to 405 min, and basic pH over 8.21 after 28 days. Higher αTCP concentration leads to sealers with low solubility. Cell viability after 48 h in direct contact with sealers was similar to a commercial sealer used as reference. The 10% and 15% αTCP sealers exhibited a calcium-phosphate layer on the surface after immersion in water and SBF for 7 days. Glycerol salicylate sealers with 10% and 15% α-tricalcium phosphate showed reliable physical-chemical properties and apatite-forming ability.

Mesenchymal stem cell adherence on poly(D, L-lactide-co-glycolide) nanofibers scaffold is integrin-beta 1 receptor dependent.

Tissue engineering is a potential approach to regenerate damaged tissue by the combination and synergism among the scaffolding material, cell source and signaling factors. In the present study, mesenchymal stem cells (MSCs) were isolated from C57BL/6 mice, cultured on poly(D, L-lactide-co-glycolide) (PLGA) scaffold produced by electrospinning technique and differentiated into chondrogenic lineage. After seeding, MSCs were responsive and became flattened with fibroblast-like morphology demonstrated by the presence of actin stress fibers. Integrin-beta1 receptor blockage reduced significantly cell adhesion with loss of actin stress fibers, demonstrating the ability of PLGA nanofiber to trigger integrin receptor-mediated cell adhesion. Present data contribute to the understanding of MSCs' behavior on these biodegradable and biocompatible scaffolds that can be used as carriers in treatments involving cell transplantation.

Repair of bone defects using adipose-derived stem cells combined with alpha-tricalcium phosphate and gelatin sponge scaffolds in a rat model

Abstract Objectives This study aimed to evaluate the potential of adipose-derived stem cells (ASCs) combined with a modified α-tricalcium phosphate (α-TCP) or gelatin sponge (GS) scaffolds for bone healing in a rat model. Material and Methods Bone defects were surgically created in the femur of adult SHR rats and filled with the scaffolds, empty or combined with ASCs. The results were analyzed by histology and histomorphometry on days seven, 14, 30, and 60. Results Significantly increased bone repair was observed on days seven and 60 in animals treated with α-TCP/ASCs, and on day 14 in the group treated with GS/ASCs, when compared with the groups treated with the biomaterials alone. Intense fibroplasia was observed in the group treated with GS alone, on days 14 and 30. Conclusions Our results showed that the use of ASCs combined with α-TCP or GS scaffolds resulted in increased bone repair. The higher efficacy of the α-TCP scaffold suggests osteoconductive property that results in a biological support to the cells, whereas the GS scaffold functions just as a carrier. These results confirm the potential of ASCs in accelerating bone repair in in vivo experimental rat models. These results suggest a new alternative for treating bone defects.