Drug release kinetics and biological properties of a novel local drug carrier system.

BACKGROUND: The purpose of this in vitro study was to investigate drug release kinetics and cytotoxicity of a novel drug delivery system for treatment of periodontitis. MATERIALS AND METHODS: This in vitro study addresses the fabrication of a polycaprolactone/alginic acid-based polymeric film loaded with metronidazole, as a basic drug in the treatment of periodontal diseases. Films were prepared by solvent casting technique. Four formulations with different percentages of drug by weight (3%, 5%, 9%, and 13%) were prepared. Drug release kinetics were investigated using ultraviolet-visible spectroscopy during (one week). Data were analyzed using repeated measures ANOVA. Cytotoxicity of drug-loaded system extracts was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay using L929 cells after 24-h incubation. The results were evaluated according to ISO standard 10993-5 and assessed using ANOVA and Tukey's tests at a significance level of P < 0.05. RESULTS: All polymeric films showed a burst drug release followed by a gradual release. Drug release data were fitted well with the first-order kinetic model in all drug-containing formulations indicating that drug release is a fraction of remaining drug in the matrix. Drug release is mainly driven by diffusion of medium into the composite matrix. 3%wt metronidazole-containing formulation exhibited the best MTT result. CONCLUSION: The findings of this study supported the synthesis of drug-loaded periodontal films with 3% metronidazole due to better biological properties along with the ability of acceptable drug release to eradicate anaerobic periodontal bacteria.

The effect of heavy metals on thiocyanate biodegradation by an autotrophic microbial consortium enriched from mine tailings.

Bioremediation systems represent an environmentally sustainable approach to degrading industrially generated thiocyanate (SCN-), with low energy demand and operational costs and high efficiency and substrate specificity. However, heavy metals present in mine tailings effluent may hamper process efficiency by poisoning thiocyanate-degrading microbial consortia. Here, we experimentally tested the tolerance of an autotrophic SCN--degrading bacterial consortium enriched from gold mine tailings for Zn, Cu, Ni, Cr, and As. All of the selected metals inhibited SCN- biodegradation to different extents, depending on concentration. At pH of 7.8 and 30 °C, complete inhibition of SCN- biodegradation by Zn, Cu, Ni, and Cr occurred at 20, 5, 10, and 6 mg L-1, respectively. Lower concentrations of these metals decreased the rate of SCN- biodegradation, with relatively long lag times. Interestingly, the microbial consortium tolerated As even at 500 mg L-1, although both the rate and extent of SCN- biodegradation were affected. Potentially, the observed As tolerance could be explained by the origin of our microbial consortium in tailings derived from As-enriched gold ore (arsenopyrite). This study highlights the importance of considering metal co-contamination in bioreactor design and operation for SCN- bioremediation at mine sites. KEY POINTS: • Both the efficiency and rate of SCN- biodegradation were inhibited by heavy metals, to different degrees depending on type and concentration of metal. • The autotrophic microbial consortium was capable of tolerating high concentrations of As, potential having adapted to higher As levels derived from the tailings source.

In-vitro bioactivity evaluation and physical properties of an epoxy-based dental sealer reinforced with synthesized fluorine-substituted hydroxyapatite, hydroxyapatite and bioactive glass nanofillers.

The purpose of this study was to evaluate the physical properties and bioactivity potential of epoxy-based dental sealers modified with synthesized bioactive glass (BAG), hydroxyapatite (HA) and fluorine substituted hydroxyapatite (FHA) nanoparticles. The synthesized powders were incorporated at 10% and 20% into the epoxy-based dental sealer. The setting time, flow and solubility and microhardness of the modified and unmodified samples were examined. The bioactivity was evaluated using FESEM-EDX and elemental mapping, ATR-FTIR and XRD. The flow value of all of the experimental groups except the FHA modified samples, was greater than 20 mm. Concerning solubility, no specimens exhibited more than 1% weight loss. The solubility value of the FHA groups was statistically significant lower than other groups (p ≤ 0.001). The mean hardness values of all of the modified samples were significantly higher than the unmodified group (p ≤ 0.001). Regarding bioactivity, in vitro study revealed that after 3 days immersion in SBF a compact and continuous calcium phosphate layer formed on the surface of epoxy sealers containing BAG and HA nanoparticles. Based on these results, the addition of BAG and HA nanoparticles did not adversely alter the physical properties of epoxy sealers. Additionally, they improved the in vitro bioactivity of the epoxy sealer.

Bond strength of self-adhesive resin cement to base metal alloys having different surface treatments.

BACKGROUND: This study aimed to assess and compare the shear bond strength of self-etch and self-adhesive resin cement to nickel-chromium-cobalt alloy with different surface treatments. MATERIALS AND METHODS: In this in vitro study, a total of 120 disks were fabricated of VeraBond II base metal alloy. Specimens were divided into 15 groups of 8 based on the type of cement and surface treatment. The five surface treatments studied included sandblasting alone, application of Alloy Primer with and without sandblasting, and application of Metal Primer II with and without sandblasting. The three cement tested included Panavia F2.0, RelyX Unicem (RU), and G-Cem (GC). After receiving the respective surface treatments, the specimens were thermocycled for 1500 cycles and underwent shear bond strength testing. Data were analyzed using SPSS 20.0 and three-way analysis of variance. P values of the significant level of 0.05 were reported. RESULTS: The results exhibited that the mean bond strengths in sandblasted groups were higher than nonsandblasted one. These differences were significantly higher in the sandblasted groups of Panavia F2.0 and RU cement (P < 0.05). The mean bond strength values between GC and Panavia F2.0 were not statistically significant (P > 0.05). The highest bond strength was recorded for Panavia F2.0 with the surface treatment of both sandblasting and Metal Primer II. CONCLUSION: Based on the results, sandblasting improves the shear bond strength of self-etch and self-adhesive resin cement to base metal alloys. The best results can be achieved with a combination of sandblasting and metal primers. The performance of resin cement depends on to their chemical composition, not to the type of system.

Study on the influence of leucine-rich amelogenin peptide (LRAP) on the remineralization of enamel defects via micro-focus x-ray computed tomography and nanoindentation.

Regeneration of severely damaged enamel (e.g. deep demineralized lesions) is currently not possible, because the structural units of enamel crystal construction are removed after its maturation. The aim of this in vitro study was to evaluate the effect of surface impregnation by leucine-rich amelogenin peptide (LRAP) on the remineralization of eroded enamel using micro-focus x-ray computed tomography (µCT). Fifteen bovine enamel blocks were embedded in resin and three zones (sound, demineralization, and remineralization) were defined on each specimen. Lesions were prepared by immersing the samples in demineralization solution for 7 d. The samples were soaked in distilled water or 60 or 120 µg mL(-1) solution of LRAP in water for 30 min. After the surface treatment, specimens were incubated in artificial saliva for either 5 or 10 d at 37 °C. The amount of mineral gain (dΔZ%) and the relative changes in the lesion depth (dLD%), obtained from µCT, were used to evaluate the effect of LRAP on the remineralization of lesions. The effects of LRAP on cross-sectional integrated hardness ΔINH were studied after 10 d using nanoindentation. ANOVA test was used to determine the effect of time and/or LRAP concentration on dΔZ%, dLD% and ΔINH mean values. Tukey's analysis was used for multiple comparison testing (α = 0.05). Analysis of µCT data showed significant effect of time and LRAP concentration on the dΔZ% (p = 0.013, p = 0.003) and the dLD% (p < 0.001, p = 0.002) mean values. The nanoindentation hardness was significantly improved by 120 µg mL(-1) LRAP (p = 0.02). Also, the peptide treatment affected the mineral distribution throughout the lesion by inhibiting of superficial deposition. This study showed that the treatment of eroded lesions in enamel by LRAP can improve and regulate the pattern of remineralization in vitro.

Leucine-rich amelogenin peptide (LRAP) as a surface primer for biomimetic remineralization of superficial enamel defects: An in vitro study.

This study was carried out to obtain more information about the assembly of hydroxyapatite bundles formed in the presence of Leucine-Rich Amelogenin Peptide (LRAP) and to evaluate its effect on the remineralization of enamel defects through a biomimetic approach. One or 2 mg/mL LRAP solutions containing 2.5 mM of Ca(+2) and 1.5 mM phosphate were prepared (pH = 7.2) and stored at 37 °C for 24 h. The products of the reaction were studied using atomic force microscopy (AFM), transmission electron microscopy (TEM), and selected area electron diffraction (SAED). Vickers surface microhardness recovery (SMR%) of acid-etched bovine enamel, with or without LRAP surface treatment, were calculated to evaluate the influence of peptide on the lesion remineralization. Distilled water and 1 or 2 mg/mL LRAP solution (pH = 7.2) were applied on the lesions and the specimens were incubated in mineralization solution (2.5mM Ca(+2) , 1.5mM PO4 (-3) , pH = 7.2) for 24 h. One-way ANOVA and Tukey's multi-comparison tests were used for statistical analysis. The pattern of enamel surface repair was studied using FE-SEM. AFM showed the formation of highly organized hierarchical structures, composed of hydroxyapatite (HA) crystals, similar to the dental enamel microstructure. ANOVA procedure showed significant effect of peptide treatment on the calculated SMR% (p < 0.001). Tukey's test revealed that peptide treated groups had significantly higher values of SMR%. In conclusion, LRAP is able to regulate the formation of HA and enhances the remineralization of acid-etched enamel as a surface treatment agent.

Push-out bond strength of Resilon/Epiphany self-etch to intraradicular dentin after retreatment: a preliminary study.

INTRODUCTION: Retreatment procedures might affect the adhesion capability of refilling materials to dentinal walls. The purpose of this study was to compare the effect of different retreatment techniques on bond strength of Resilon (Resilon Research LLC, Madison, CT)/Epiphany (Pentron Clinical Technologies, LLC, Wallingford, CT) self-etch (SE) with radicular dentin after canal reobturation. METHODS: Sixty extracted single-rooted human teeth were prepared using Mtwo rotary files (VDW, Munich, Germany) and obturated with Resilon/Epiphany SE. The roots were randomly divided into four groups; in group 1, no retreatment procedure was done; group 2 was retreated using Mtwo R/Mtwo files; group 3 was retreated using Mtwo R/Mtwo combined with chloroform; and group 4 was retreated using Mtwo R/Mtwo combined with Endosolv R (Septodont, Paris, France). The root canals were then reobturated with Resilon/Epiphany SE. One-millimeter slices of midroot dentin were prepared for the push-out test (n=30 slices per group). Failure modes after the push-out test were examined under microscopy. The data were analyzed by using a one-way analysis of variance and the Dunnett post hoc test. RESULTS: Group 3 showed significantly the lowest mean bond strength (p<0.02). No significant difference was found between the values of groups 1, 2, and 4 (p>0.26). The mode of bond failure was predominantly adhesive for all groups. CONCLUSIONS: This study showed that the bond strength of Resilon/Epiphany SE after root canal retreatment using Mtwo files, either alone or combined with Endosolv R, was not significantly different from that of nonretreated specimens. Chloroform used for retreatment had an adverse effect on the bond strength of Resilon/Epiphany SE after root canal reobturation.

Epigenetic regulation of E-cadherin controls endometrial receptivity.

Key to the success of human reproduction is the capacity of an embryo to attach and implant into the endometrial wall after which a nutrient supply is established through placentation. Herein, we have examined the potential epigenetic regulation of uterine receptivity by use of the receptive RL95-2 and nonreceptive AN3-CA endometrial epithelial carcinoma cell lines. Using an in vitro model of embryo implantation, we demonstrate that inhibition of DNA methylation by 5'-aza-2'-deoxycytidine (AZA), resulted in the nonreceptive AN3-CA cell line becoming receptive to BeWo cell spheroid attachment. Examination of components of the adherens junction complex revealed that AZA specifically increased the expression of E-cadherin and plakoglobin at the mRNA and protein levels in AN3-CA cells, and E-cadherin protein expression was found to localize to sites of intercellular contact. Forced expression of E-cadherin in AN3-CA cells significantly enhanced receptivity. Small interfering RNA (siRNA)-mediated depletion of the individual DNA methyltransferase (DNMT) molecules did not induce E-cadherin expression in AN3-CA cells; however, concomitant siRNA-mediated depletion of both DNMT3A and DNMT3B induced the expression of E-cadherin. Furthermore, E-cadherin expression was significantly increased after the concomitant siRNA-mediated depletion of DNMT-1, -3A, and -3B in AN3-CA cells. Therefore, we have provided evidence that E-cadherin plays an important role in uterine receptivity and that E-cadherin expression is epigenetically regulated in AN3-CA cells, suppressed by the combined actions of DNMT-1, -3A, and -3B.

Compressive fatigue behavior of dental restorative composites.

The purpose of this study was to investigate the compressive fatigue behavior of five dental composites. Cylindrical specimens of 8 mm length and 4 mm diameter were made according to manufacturers' recommendations and stored for two weeks in distilled water at 37 degrees C. Compressive fracture strength was measured, and subsequently fatigue tests at 10 Hz frequency were carried out in distilled water. Compressive fatigue strength was thereby obtained using the staircase method for 10(5) cycles (n = 17) under sinusoidal loading. Acquired data for compressive fracture strength were analyzed using ANOVA and Weibull statistics. Among the dental composites examined, Filtek Z250 exhibited the highest fatigue strength. This seemed to be due to the superior matrix properties coupled with a specific filler type at the highest weight%/volume% ratio. In addition, fracture mechanisms of the composites were examined.

Epigenetic regulation of human trophoblastic cell migration and invasion.

Pivotal to successful mammalian reproduction is the ability of a developing embryo to implant to the uterine wall and establish a nutrient supply via placentation. Herein, we have examined the potential epigenetic regulation of human trophoblastic cell migration and invasion by use of the choriocarcinoma cell line, BeWo. Treatment of BeWo cells with a DNA methyltransferase inhibitor, 5'-aza-2'-deoxycytidine (AZA), resulted in conversion of cell morphology to a nonmigratory phenotype. This was exemplified by the ability of AZA to prevent BeWo cell migration in wound healing and transwell migration assays. AZA consequently inhibited BeWo cell invasion through reconstituted basement membrane. Examination of components of the adherens junction complex pivotal for determination of cell phenotype revealed that AZA specifically increased the mRNA level of E-cadherin and plakoglobin (gamma-catenin), but not alpha-catenin and beta-catenin. AZA also increased the gene promoter activity of both plakoglobin and E-cadherin. Protein levels of both plakoglobin and E-cadherin were increased by AZA, and AZA enhanced their localization to sites of intercellular contact. Forced expression of plakoglobin and E-cadherin abrogated BeWo cell migration, indicative that repression of these genes was required for BeWo cell migration. Small interfering RNA-mediated depletion of the individual DNA methyltransferase (DNMT) molecules did not affect plakoglobin and E-cadherin promoter activity or BeWo cell migration. However, increases in plakoglobin and E-cadherin promoter activity and inhibition of BeWo cell migration was achieved with small interfering RNA-mediated depletion of both DNMT-3a and DNMT-3b. Epigenetic regulation of plakoglobin and E-cadherin is therefore pivotal for appropriate trophoblastic invasion in vitro.