The evaluation of anticancer activity by synthesizing 5FU loaded albumin nanoparticles by exposure to UV light.

In this study, as a new synthesis method, UV light was employed as a type of cross-linking agent to control drug storage and to produce nanoparticles of different sizes and to stabilize the nanoparticles for the first time. We showed that the exposure time of the 5FU albumin solution to UV light produces differences in the size and characterization of the nanoparticles and also produces different cytotoxic effects on MCF-7 breast cancer cells. While the 5FU-A1 nanoparticles we synthesized with 1 h UV storage were approximately 43 nm, the 5FU-A2 nanoparticles we synthesized with UV storage for 3 h increased to an average of 300 nm. 5FU-A1 (IC50 value: 2.5 µg/mL) was approximately 16 times more cytotoxic than free 5FU (IC50 value 39.39 µg/mL) on MCF-7 cancer cells. Moreover, when normal HUVEC cells are treated with 5FU-A1 at a concentration of 2.5 µg/mL, more than 80% of these normal cells remain viable. In addition, we examined the rate of early-to-late apoptosis and necrosis in MCF-7 cancer cells using the Annexin V/PI flow cytometry assay. According to our results, 5FU-A1 promoted the apoptosis pathway. Finally, we examined P-gp activity with MDR1/ABCB1 antibody by flow cytometry and Rhodamine123 with fluorescent dye.

New oxovanadium(IV) complexes overcame drug resistance and increased in vitro cytotoxicity by an apoptotic pathway in breast cancer cells.

In order to examine the anticancer potential of oxovanadium(IV) complexes with thiosemicarbazone, two new complexes were prepared starting from 2-thenoyltrifluoroacetone-S-methylthiosemicarbazone. The complexes with tetradentate thiosemicarbazone ligand were characterized by elemental analysis, IR, ESI MS, and single-crystal X-ray diffraction analysis. Cytotoxicity on breast cancer cells, MDA-MB-231 and MCF-7, was determined by MTT assay. Cisplatin was positive control and the results were compared with those of the normal cells, HUVEC and 3T3. The complexes exhibited greater activity on cancer cells than cisplatin, but they were cytotoxic at several times higher concentrations in the healthy cells. In our study, the presence of thiophene and fluoro groups in the oxovanadium(IV) complexes with thiosemicarbazone increased greatly the cytotoxic activity of the complexes on breast cancer cells. Moreover, the complexes induced apoptosis-mediated cell death and also reduced the expression of MDR-1 or P-glycoprotein and ABCG2. As a result, the findings indicated that the complexes have selective cytotoxicity on breast cancer cells and can overcome multidrug resistance. These properties of the complexes make it possible to be a potential anticancer drug candidate for breast cancer treatment.

New thiosemicarbazone-based Zinc(II) complexes. In vitro cytotoxicity competing with cisplatin on malignant melanoma A375 cells and its relation to neuraminidase inhibition.

New thiosemicarbazone-based zinc(II) complexes were synthesized to study their cytotoxicity on A375 malignant melanoma cells. The complexes containing salicylidene (Zn1a), 3-methoxy-salicylidene (Zn1b) or 4-methoxy-salicylidene (Zn1c) moiety were characterized by analytical and spectroscopic methods. Anticancer potential of the complexes was determined by MTT test and HUVEC endothelial cells line was used to comprehend the effect on normal cells. Zn1b with an IC50 of 13 µM was found to be highly cytotoxic against A375 cancer cells, more effective than cisplatin (IC50: 37 µM). Zn1a and Zn1c did not have a negative effect on cell viability in the normal cells and gave the impression that they are more advantageous than cisplatin in this respect. Further, the ability of Zn1a-c to inhibit neuraminidase enzyme and its role in cytotoxicity was discussed. The test revealed that the Zn1b with 3-methoxy substituent exhibited higher inhibition activity against the neuraminidase than the Zn1a and Zn1c as analogical to the cytotoxicity results. In neuraminidase inhibition, IC50 values of Zn1b and Zn1c were 14 and 66 µM, respectively. These concentrations were very close to the cytotoxicity concentrations for Zn1b and Zn1c. The findings may indicate the role of neuraminidase enzyme inhibition in cell death for Zn1b and Zn1c.

Biochemical assay and spectroscopic analysis of oxidative/antioxidative parameters in the blood and serum of substance use disorders patients. A methodological comparison study.

Substance abuse such as opioids, cannabis, and alcohol causes activation on the immune system and the release of reactive oxygen species (ROS) into the blood and serum. These substances cause an effect on oxidant and antioxidant status in patients with substance abuse. Mainly, wide-open to the ROS are lipids and proteins included blood, which suffers peroxidation. In this study, for the first-time differentiation of the effects of cannabis, alcohol and other synthetic substances on blood and serum samples, were performed. For this purpose, the level of the malondialdehyde (MDA) and glutathione (GSH) in serum and red blood cells, was measured using biochemical assay methods and Fourier Transform InfraRed spectroscopy (FTIR). The results showed, that peroxidation which is dignified as the production of MDA was increased for substance use disorder (SUD) patients (18.01 ±â€¯2.97) compared to the control group (10.75 ±â€¯2.28) (p < 0.001) and for antioxidant capacity, GSH level were significantly increased for SUD patients (p < 0.001). For the discrimination of protein and lipid region obtained from FTIR spectroscopy, we extracted features by principal component analyze (PCA) of protein (1800 cm-1 to 900 cm-1) and lipid (3200 cm-1 to 2800 cm-1) regions for blood and serum samples collected from patients with different types of SUD and healthy control (HC) participants. For the consideration of lipid oxidation, lipid saturation, lipid desaturation and protein aggregation the peak heights at 1740 cm-1 to 2960 cm-1, 2920 cm-1 to 2960 cm-1, 3012 cm-1 to 2960 cm-1, and 1630 cm-1 to 1650 cm-1 regions were studied for SUD and HC. Moreover, more visible changes were noticed for proteins region, than for lipids. The most notice structural changes were observed in amide II in serum spectra. Then we classified protein and lipid region's PCA results of blood and serum by Linear discriminant analysis (LDA) and Support vector machine (SVM). Confidence of the specificity, sensitivity and accuracy of blood and serum were obtained as 100%, 100% and 100% individually. This is the first comparative study on the spectrochemical tool and biochemical assay on SUD. Our results presented 100% discrimination of disorder region compared to healthy subjects.

The interaction of light-activatable 2-thioxanthone thioacetic acid with ct-DNA and its cytotoxic activity: Novel theranostic agent.

In this study, a thioxanthone derivative, 2-Thioxanthone Thioacetic Acid (TXSCH2COOH) was used to analyze the type of binding to calf thymus DNA in a physiological buffer (Tris-HCl buffer solution, pH:7.0). Several spectroscopic techniques were employed including UV-Vis absorption and fluorescence emission spectroscopy and viscosity measurements were also used to clarify the binding mode of TXSCH2COOH to ct-DNA. The intrinsic binding constant Kb of TXSCH2COOH-ct-DNA was found as 2.5 × 103 M-1 from the absorption studies. Increasing of fluorescence emission intensity was found approximately 74.4% by adding ct-DNA to the TXSCH2COOH solution. Fluorescence microscopy was employed to display imaging of the TXSCH2COOH-ct-DNA solution. Increasing of the iodide quenching effect was observed when TXSCH2COOH was added to the double stranded DNA and the calculated quenching constants of TXSCH2COOH and TXSCH2COOH-ct-DNA were found to be 1.89 × 103 M-1 and 1.19 × 104 M-1, respectively. Additionally, the iodide quenching experiment was conducted with single stranded DNA which led to a high Ksv value. All the experimental results including the viscosity values of ct-DNA with TXSCH2COOH demonstrated that the binding of TXSCH2COOH to ct-DNA was most likely groove binding. Furthermore, TXSCH2COOH was found to be an A-T rich minor groove binder. This was confirmed by the displacement assays with Hoechst 33258 compared to Ethidium Bromide. The in vitro cytotoxic activity measurements were performed by MTT assay on HT29 cell line for 72 h. TXSCH2COOH exhibited notable cytotoxic activities compared to the standard chemotherapy drugs, fluorouracil (5-FU), cisplatin in tumorigenic HT29 cell line. The 50% growth-inhibitory concentration (IC50) for TXSCH2COOH was 19,8 µg/mL while 5-FU and cisplatin were 28.9 µg/mL, 20 µg/mL, respectively. The increase in cytotoxic effect when TXSCH2COOH is activated by light indicates the potential of being theranostic cancer drug candidate.

Silver nanoparticle/capecitabine for breast cancer cell treatment.

This study aimed to evaluate antiproliferative and proapoptotic effects of Capecitabine bonded silver particles on human breast cancer cells (MCF-7). Different sizes of Ag NPs (in sizes 5, 10, 15, 30 nm) were synthesized. The characterization of silver and drug-bonded silver nanoparticles was performed through UV-VIS, FTIR, and SEM analysis. Silver and drug-bonded silver nanoparticles were measured by zetasizer. Antiproliferative and proapoptotic effects of capecitabine, silver and drug-bonded silver nanoparticles were evaluated using XTT, Anneksin V, respectively. According to the results, silver nanoparticles of 10 nm size have shown the lowest toxic effect. Drug-bonded nanoparticles significantly increased the number of early and late apoptotic cells on MCF-7 cells.

Characterization and Isolation of Very Small Embryonic-like (VSEL) Stem Cells Obtained from Various Human Hematopoietic Cell Sources.

Stem cell transplantation is one of the available treatments for leukemia, lymphoma, hereditary blood diseases and bone marrow failure. Bone marrow (BM), peripheral blood progenitor cells (PBPC), and cord blood (CB) are the predominant sources of stem cells. Recently a new type of stem cell with a pluripotent potential has been identified. These cells were named "very small embryonic like stem cells (VSELs)". It is claimed that VSEL stem cells can be found in adult BM, peripheral blood (PB), CB and other body tissues. This study is designed to characterize and isolate VSEL stem cells from different human hematopoietic sources; CB, PB and apheresis material (PBPC). VSEL stem cells were isolated from MNC and erythrocyte layers for all materials by using centrifugation and ficoll gradient method. We determined embryonic markers by flow cytometry, immunofluorescence and western blotting methods. Results from western blotting and immunofluorescence show high level of NANOG and OCT4 protein expression in PB, apheresis material and CB. Immunofluorescence images showed cytoplasmic and nuclear presence of these proteins. Flow cytometry results exhibited a higher expression of VSELs markers on debris area than CD45- population and higher expression on CB than PB. As a result, these findings have shown that it is necessary to investigate the function of pluripotent stem cell markers in differentiated adult cells. We further conclude that erythrocyte lysis method had the highest cell recovery amount among erythrocyte lysis and ficoll gradient methods. Consequently, this study gives us new information and viewpoints about expression of pluripotent stem cell (PSC) markers in adult tissues.

Novel palladium (II) complexes with tetradentate thiosemicarbazones. Synthesis, characterization, in vitro cytotoxicity and xanthine oxidase inhibition.

In vitro cytotoxicity and xanthine oxidase inhibition capabilities were investigated for five palladium (II) chelate complexes. The palladium complexes were synthesized by starting from S-alkyl-thiosemicarbazones where the alkyl component is methyl, ethyl, propyl or butyl. The solid complexes are characterized by elemental analysis and spectroscopic techniques (UV-visible, IR and 1H NMR). In order to be able to verify the N2O2-type thiosemicarbazidato ligand (L2-) structure in the square planar geometry, complex 1 has been studied as a representative by using single crystal X-ray crystallography. The in vitro cytotoxic activity measurements were carried out in HepG2 and Hep3B hepatocellular carcinomas, HCT116 colorectal carcinoma, and 3 T3 mouse fibroblast cell lines. The palladium complexes exhibited notable cytotoxic activities in all cell lines at lower µM concentrations compared to the standard chemicals, cisplatin and allopurinol. IC50 values were determined between 0.42 ± 0.01 and 12.01 ± 0.37 µg/ml in examining the antixanthine oxidase abilities of the complexes. Two complexes with S-methyl group exhibited a high inhibition activity on the xanthine oxidase. The results indicated that these complexes could be used as active pharmaceutical ingredients.

Cellular interactions with bacterial cellulose: Polycaprolactone nanofibrous scaffolds produced by a portable electrohydrodynamic gun for point-of-need wound dressing.

Electrospun nanofibrous scaffolds are promising regenerative wound dressing options but have yet to be widely used in practice. The challenge is that nanofibre productions rely on bench-top apparatuses, and the delicate product integrity is hard to preserve before reaching the point of need. Timing is critically important to wound healing. The purpose of this investigation is to produce novel nanofibrous scaffolds using a portable, hand-held "gun", which enables production at the wound site in a time-dependent fashion, thereby preserving product integrity. We select bacterial cellulose, a natural hydrophilic biopolymer, and polycaprolactone, a synthetic hydrophobic polymer, to generate composite nanofibres that can tune the scaffold hydrophilicity, which strongly affects cell proliferation. Composite scaffolds made of 8 different ratios of bacterial cellulose and polycaprolactone were successfully electrospun. The morphological features and cell-scaffold interactions were analysed using scanning electron microscopy. The biocompatibility was studied using Saos-2 cell viability test. The scaffolds were found to show good biocompatibility and allow different proliferation rates that varied with the composition of the scaffolds. A nanofibrous dressing that can be accurately moulded and standardised via the portable technique is advantageous for wound healing in practicality and in its consistency through mass production.

The effects of 17ß-estradiol on blood brain barrier integrity in the absence of the estrogen receptor alpha; an in-vitro model.

The blood-brain barrier (BBB), which saves the brain from toxic substances, is formed by endothelial cells. It is mainly composed of tight junction (TJ) proteins existing between endothelial cells. Estrogen is an important regulatory hormone of BBB permeability. It protects the BBB before menopause, but may increase BBB permeability with aging. In addition, nitric oxide modulates BBB permeability. Alcohol impairs the integrity of the BBB with oxidants and inflammatory mediators such as iNOS. We investigated the effects of estrogen on BBB integrity in an in vitro BBB model created with ERα-free HUVEC (human umbilical vein endothelial-like cells) to mimics the menopausal period. In vitro BBB model is created with HUVEC/C6 (rat glioma cells) co-culture. The effect of 17ß-estradiol on ethanol-induced BBB disruption and change/or increase of iNOS activity, which modulate BBB integrity, were evaluated. Inducibility and functionality of BBB were investigated using transendothelial electrical resistance (TEER) and the expression of proteins TJ proteins (occludin and claudin-1) and iNOS activity by immunostaining. Our results revealed that 17ß-estradiol treatment before and after ethanol decrease expression of occludin and claudin-1 and value of TEER which are BBB disrupt indicators. In addition, ethanol and 17ß-estradiol separately and pre- and post-ethanol 17ß-estradiol treatment increased iNOS expression. Thus our study suggests caution in the use of 17ß-estradiol after menopause because 17ß-estradiol at this time may both increase the inflammatory process as well as damage the BBB. We think that beneficial effects of 17ß-estradiol may be through ERα but it needs further studies.

Chitosan-co-Hyaluronic acid porous cryogels and their application in tissue engineering.

In this work, the usability of chitosan-co-hyaluronic acid cryogels as a tissue-engineering scaffold was investigated. Chitosan-co-hyaluronic acid cryogels were synthesized at subzero temperature. Cryogels which were composed of various compositions of chitosan and hyaluronic acid (0, 10, 20, 30 and 50wt% hyaluronic acid) was prepared. Morphological studies showed that the macroporous cryogels have been developed with 90-95% porosity. Particularly, the mechanical and biomaterial property of pure chitosan was improved by making copolymer with hyaluronic acid in different concentration. The MTT cell viability results demonstrated that the cryogels have no significant cytotoxicity effect on 3T3 fibroblast and SAOS-2 cells.

Fabrication of naturel pumice/hydroxyapatite composite for biomedical engineering.

BACKGROUND: We evaluated the Bovine hydroxyapatite (BHA) structure. BHA powder was admixed with 5 and 10 wt% natural pumice (NP). Compression strength, Vickers micro hardness, Fourier transform infrared spectroscopy, scanning electron microscopy (SEM) and X-ray diffraction studies were performed on the final NP-BHA composite products. The cells proliferation was investigated by MTT assay and SEM. Furthermore, the antimicrobial activity of NP-BHA samples was interrogated. RESULTS: Variances in the sintering temperature (for 5 wt% NP composites) between 1000 and 1300 °C, reveal about 700 % increase in the microhardness (~100 and 775 HV, respectively). Composites prepared at 1300 °C demonstrate the greatest compression strength with comparable result for 5 wt% NP content (87 MPa), which are significantly better than those for 10 wt% and those that do not include any NP (below 60 MPa, respectively). CONCLUSION: The results suggested the optimal parameters for the preparation of NP-BHA composites with increased mechanical properties and biocompatibility. Changes in micro-hardness and compression strength can be tailored by the tuning the NP concentration and sintering temperature. NP-BHA composites have demonstrated a remarkable potential for biomedical engineering applications such as bone graft and implant.

Decreased IFN-γ and IL-12 levels in panic disorder.

OBJECTIVE: The aim of this study is to assess the measures of proinflammatory cytokines in patients with panic disorder in comparison with the healthy subjects. METHODS: Twenty three patients with panic disorder with or without agoraphobia and twenty three controls were recruited for the study. Plasma samples of all subjects were analyzed for TNF-α, IFN-γ, IL-1ß, IL-2, IL-6, and IL-12 concentrations and NK-cell activity is measured in the peripheral blood samples of the subjects. RESULTS: We found significant differences on the mean values of IL-12 (p=0.01) and IFN-γ (p=0.02) between the panic disorder and control groups. In a logistic regression analysis, IFN-γ values were significant statistical predictors of the presence of panic disorder (B=-0.07, SE=0.03, p=0.04). CONCLUSION: The most important implication of our results is to suggest a relation between panic disorder and low levels of IFN-γ, compatible with the results of the animal studies showing that IFN-γ plays a role by acting to regulate the development of anxiety-like behaviors.

Cell growth on in situ photo-cross-linked electrospun acrylated cellulose acetate butyrate.

In this study, electrospinning was combined with UV curing technology for producing in situ photo cross-linked fibers from methacrylated cellulose acetate butyrate (CABIEM). ECV304 and 3T3 cells were seeded on electrospun fibrous scaffolds. Collagen modified CABIEM fibers were also prepared for improving cell adhesion and proliferation. Cross-linking and the morphology of the fibers were characterized by ATR-FTIR spectrometry and environmental scanning electron microscopy (ESEM). The cytotoxicity of the fibers was examined using the MTT cytotoxicity assay. According to the results, electrospun fibrous scaffolds are non-toxic and cell viability depends on the amount of collagen. It was found that cell adhesion and cell growth were enhanced as the collagen percentage was increased.

Preparation of biocompatible, UV-cured fumarated poly(ether-ester)-based tissue-engineering hydrogels.

The aim of this study was to develop biodegradable, photo-polymerizable in situ gel-forming systems prepared from a fumaric acid monoethyl ester (FAME) modified poly(lactide-co-glycolide) (PLGA) co-polymer. By reacting lactide and glycolide in the presence of stannous octoate as a catalyst and 2-ethyl,2-hydroxymethyl 1,3-propanediol as an initiator, hydroxyl terminated branched PLGA was synthesized. Afterwards, at room temperature hydroxyl terminated branched PLGA was reacted with fumaric acid monoethyl ester (FAME). N,N'-dicyclohexylcarbodiimide and triethylamine were used as a coupling agent and catalyst, respectively. The gel percentage, equilibrium mass swelling, degradation profile and polymerization kinetics of the hydrogels were investigated. All of the results were influenced by the amount of FAME modified PLGA co-polymer. Biocompatibility of the hydrogels was examined by using MTT cytotoxicity assay. According to the results, hydrogels are biocompatible and cell viability percentage depends on the amount of PLGA co-polymer. While the amount was 15% in hydrogel composition, cell viability was 100%, but after increasing the PLGA co-polymer amount to 30% the viability reduced to 78%.

Cytotoxic effects of leaf, stem and root extracts of Nerium oleander on leukemia cell lines and role of the p-glycoprotein in this effect.

Nerium oleander (No), is a toxic plant. In recent studies, it was determined that the extracts of this plant are effective to treat some types of cancer, but these studies are limited and do not include human leukemia. In the present study, firstly we aimed to investigate in vitro the cytotoxic effects of No on the HL60 and K562 leukemia cell lines. The cells were incubated with six different concentrations of each three extracts. MTT assay was employed as a cytotoxicity test. It was observed that concentrations of 1000, 500 and 50 microg/ml from each extract possess marked antileukemic effects. No leaf and root extracts were seen to be more cytotoxic than the stem extract according to LC50. Secondly, in order to understand the role of P-gp in cytotoxicity, P-gp levels of K562 resistant and sensitive cells were measured by flow cytometry before treatment extracts, and then, the cells were incubated with No leaf, stem and root extracts in 500 microg/ml concentrations overnight. After incubation, measurements showed decreased levels of P-gp in the cells. Hence, it is possible to think contributes to their cytotoxic effects that inhibiting of the P-gp pump by No extracts on leukemia cell lines.