Liposomal Doxorubicin Kinetic Study in an In vitro 2D and 3D Tumor Model for Osteosarcoma in a Perfusion Bioreactor.

BACKGROUND: In vivo drug screening in animal models is contrary to ethical values, costly and time-consuming. Traditional static in vitro models do not reflect the basic characteristics of bone tumor microenvironments; therefore, perfusion bioreactors, in particular, would be an applicable choice due to their advantages to regenerate versatile bone tumor models for studying in vitro novel drug delivery systems. METHODS: In this study, an optimal drug formulation of liposomal doxorubicin was prepared, and the release kinetics of the drug and its toxicity effect on MG-63 bone cancer cell line were investigated in two-dimensional, static three-dimensional media on a PLGA/ß-TCP scaffold and also in a dynamic media in a perfusion bioreactor. In this assay, the efficacy of the IC50 of this formulation which had been obtained in two-dimensional cell culture (= 0.1 µg/ml), was studied in static and dynamic threedimensional media after 3 and 7 days. Liposomes with good morphology and encapsulation efficiency of 95% had release kinetics of the Korsmeyer-Peppas model. RESULTS: The results of cell growth before treatment and cell viability after treatment in all three environments were compared. Cell growth in 2D was rapid, while it was slow in static 3D conditions. In the dynamic 3D environment, it was significant compared to the static tumor models. Cell viability after 3 and 7 days from treatment was 54.73% and 13.39% in 2D conditions, 72.27% and 26.78% in the static 3D model, while 100% and 78.92% in the dynamic culture indicating the effect of drug toxicity over time, but drug resistance of 3D models compared to 2D culture. In the bioreactor, the formulation used in the mentioned concentration showed very small cytotoxicity demonstrating the dominance of mechanical stimuli on cell growth over drug toxicity. CONCLUSION: Increasing drug resistance in 3D models compared to 2D models indicates the superiority of liposomal Dox over free form to reduce IC50 concentration.

Potential Biomarkers for Depression Associated with Coronary Artery Disease: A Critical Review.

Depression, the most common mood disorder, is a leading contributor to the global burden of disease affecting more than 120 million individuals worldwide. Various pathophysiological processes underlie depression; this complexity renders it difficult to identify clinically useful diagnostic and prognostic markers, as well as treatment options. The current state of knowledge driving the management and treatment of depression remains incomplete, which underscores the need for further insight into pathways relevant to depression. Exploring co-morbid conditions, such as coronary artery disease, may be useful to further elucidate the etiopathology of depression. The present review therefore systematically identifies and critically evaluates relevant markers of depression as assessed in a high-risk population, namely patients with coronary artery disease. Biomarkers related to hypothalamicpituitary- adrenal axis dysregulation, inflammation, endothelial dysfunction, platelet activation and aggregation, serotonin activity, sympathetic nervous system activation, thyroid function, structural and morphological brain abnormalities, genetic variation, lipid metabolism, one-carbon metabolism, endocannabinoid signalling irregularities, and vitamin D deficiency are reviewed. Markers exhibiting the most consistent associations with depression include tumour necrosis factor-α, flow-mediated dilation, endothelin-1, endothelial progenitor cells, brain-derived neurotrophic factor, and docosahexaenoic acid. Further investigating the mechanisms underlying those markers and exploring novel pathways, such as oxidative stress, will extend the current state of knowledge and potentially lead to the identification of novel therapeutic targets.

Leaching of hydrogen peroxide from bleached bovine enamel.

Accurately weighed bovine enamel slabs were individually immersed in 2 ml of 35% hydrogen peroxide for 1, 3, 5, 30, or 60 min. A control group was obtained by individual immersion of bovine enamel slabs in 2 ml of saline for 60 min. All samples were washed, dried, acid-etched with 37% phosphoric acid for 60 s, then washed and dried again. Two milliliters of double-distilled water were used for individual sample leaching. Leaching was done for 1, 5, 10, 20 min, or 7 days for the experimental groups and for 7 days for the control group. The samples of one of the experimental groups were leached for a second time for 1 min. A total of 112 samples was used in this study. Hydrogen peroxide was spectrophotometrically identified and quantified in all leaching solutions based on the oxidation reaction of leuco-crystal violet buffer solution by hydrogen peroxide, a reaction catalyzed by horseradish peroxidase. The results revealed a significant difference in the quantity of leached peroxide between bleached samples (irrespective of the duration of leaching) and control, saline-treated ones. No difference was observed in the quantity of leached peroxide between releached samples and control, saline-treated ones. However, these were small, random, and numerically insignificant. Statistically significant differences were also noted among some of the experimental groups. They were thought to hold no clinical significance. The results suggested that upon immersion, the complete leaching of peroxide from bleached enamel occurs rapidly.

Effect of water leaching the adhesion of composite resin to bleached and unbleached bovine enamel.

Standardized cylinders of light-cured composite resin were bonded to the ground labial enamel surface of bovine incisor teeth that had been immersed in double-distilled water for 7 days after having been (a) immersed in hydrogen peroxide for 5, 30, or 60 min, then etched for 60 s with 37% phosphoric acid; (b) immersed in saline for 5, 30, or 60 min, then etched for 60 s with 37% phosphoric acid; (c) etched with 37% phosphoric acid for 60 s, then immersed in hydrogen peroxide for 5, 30, or 60 min; or (d) etched with 37% phosphoric acid for 60 s, then immersed in saline for 5, 30, or 60 min. The enamel surface was washed with water for 1 min and dried with compressed air for 30 s prior to applying the resin. The tooth and applied resin were stored in water at 37 degrees C for 1 day prior to shear and tensile testing. A total of 192 specimens was used, 8 for each enamel preparation mode, for each time period, and for each test. Test values were tabulated and statistically analyzed. Analysis of variance revealed significantly higher bond strength values (p less than 0.005) for hydrogen peroxide-treated as compared with saline-treated specimens. A significant interaction was also noted between test solution and etching order. Scanning electron microscopic examination of failed shear- and tensile-tested specimens revealed no significant solution-related differences in the fracture pattern or the resin quality.(ABSTRACT TRUNCATED AT 250 WORDS)

Scanning electron microscopy observations on the penetration and structure of resin tags in bleached and unbleached bovine enamel.

The purpose of this study was to determine the effect of hydrogen peroxide on the ability of composite resin to penetrate bovine enamel etched with phosphoric acid. In a previous investigation, the flattened enamel surfaces of extracted bovine incisors were immersed in either saline (control) or 35% hydrogen peroxide (experimental) for 5 or 30 min before or after acid etching with 37% phosphoric acid gel for 60 s. A standard-sized light-cured resin cylinder was then bonded to the enamel surface. The specimens were stored in a water bath at 37 degrees C for 1 day or 7 days, after which the enamel-resin bond was tensile tested to failure. Sixteen of the failed specimens (eight control and eight experimental) were randomly selected for the present scanning electron microscopic study to evaluate the appearance of the resin tags at the resin-enamel interface. In the control specimens, the tags were well defined and contiguous with resin which was uniformly adherent to the enamel surface. In the experimental specimens, large areas of the enamel surface were free of resin. When tags were present, they were fragmented, poorly defined, and penetrated to a lesser depth than in the saline controls. The changes observed suggest that there may be interaction between resin and residual peroxide at or near the enamel surface.

The influence of time of hydrogen peroxide exposure on the adhesion of composite resin to bleached bovine enamel.

Standard sized cylinders of a small particle light-cured resin were bonded to the flattened labial surface of young bovine incisor teeth which had been previously subjected to four different treatments: (a) immersion in 35% hydrogen peroxide and etched with 37% phosphoric acid gel for 60 s, (b) immersion in saline and etched for 60 s, (c) etched for 60 s and immersion in hydrogen peroxide, and (d) etched for 60 s and immersion in saline. Two hydrogen peroxide and saline immersion periods were used, 5 and 30 min. Specimens were stored in water at 37 degrees C for 1 and 7 days before tensile and shear testing. A total of 256 teeth were used, 8 for each treatment group, each immersion period, and each water storage period for each test. Statistical analysis of the results indicated that there was a highly significant reduction in the adhesive bond strength of the resin when the enamel was exposed to hydrogen peroxide and that the reduction was, within the limits of this study, time dependent. The bond strength was unaffected by the etching order and the period of water storage. Scanning electron microscopic examination of randomly selected fractured peroxide-treated specimens indicated that the failure occurred primarily at the bonding resin-enamel interface and that it was associated with areas of resin nonattachment and an alteration in resin quality. It is suspected that these changes were caused by the presence of residual peroxide or peroxide-related substances at or near the enamel surface.

Adhesion of composite resin to bleached and unbleached bovine enamel.

Cylinders of microfil and small-particle light-cured composite resin were bonded to the flattened labial enamel surface of young bovine incisor teeth which had previously been subjected to four different treatments: (1) teeth immersed in 35% hydrogen peroxide (HP) for 60 min and etched (E) with 37% phosphoric acid gel for 60 sec; (2) teeth immersed in saline (S) for 60 min and E for 60 sec; (3) teeth E for 60 sec and immersed in HP for 60 min; and (4) teeth E for 60 sec, immersed in S for 60 min. Specimens were stored in water at 37 degrees for one and seven days prior to tension- and shear-testing. A total of 256 teeth was tested--eight teeth in each group for each day, for each resin, and for each test. Statistical analysis of the results indicated that there was a highly significant reduction in adhesive bond strength of the resins when the enamel was exposed to HP as compared with S. SEM examination of randomly selected fractured test specimens indicated that this reduction in adhesive bond strength occurred primarily at the bonding resin-enamel interface. Less significant differences in bond strength were noted in the control specimens, with regard to resin type, time of storage, and the etching order.