Combination therapy application of Abemaciclib with Doxorubicin in triple negative breast cancer cell line MDA-MB-231.

Due to lack of clinical biomarkers, Triple Negative Breast Cancer (TNBC) is more likely to have spread to other tissues at time of diagnosis and therapy planning generally involves use of cytotoxic chemotherapy agents, such as Doxorubicin. We aimed to investigate possible advantages of using combination strategy using Doxorubicin alongside Abemaciclib. After determining the IC50 values for Doxorubicin (DOX) and Abemaciclib (ABE); CompuSyn and ComBenefit software were used to reveal the effect resulting from the combination of two drugs. Following the determined effect, cell death was revealed by fluorescence microscopy and a colony forming assay was performed to see the potential of even a single cancer cell with adhesive character to survive over time and form a clone of itself. Detection of changing antioxidant activity following DOX, ABE and DOX+ABE combination therapy in MDAMB231 cells was determined by measuring MDA, SOD and GSH activities. The expression of Cleaved Caspase 3, PARP, Cleaved PARP, Cdk2 and Bax, which changed as a result of DOX, ABE and DOX+ABE application, was shown by Western Blotting.Cyclin-dependent kinase inhibitors appear as promising agents in therapy planning for breast cancer due to their prominent role in cell cycle regulation, where the number of studies interrogating its efficiency in the treatment of cancer such as TNBC is limited. For this reason, in this study, we aimed to determine the impact of the combined use of the CDK4/6 inhibitors ABE and DOX on the cytotoxicity, apoptotic homeostasis, alterations in antioxidative mechanisms, and the molecular pathways that they utilize. Our results showed that when used in combination, Doxorubicin and Abemaciclib showed a synergistic effect on TNBC cell line MDA-MB-231.

Investigation of cytotoxic antiproliferative and antiapoptotic effects of nanosized boron phosphate filled sodium alginate composite on glioblastoma cancer cells.

BACKGROUND: The effects of nanosized boron phosphate-filled sodium alginate composite gel (SA/BP) on the biological characteristics of three types of glioblastoma multiforme (GBM) cells (C6, U87MG and T98G) were examined in this study. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay was used to determine the cytotoxicity of the composite gel on GBM, which was then compared to L929 healthy cells. Furthermore, wound healing, apoptosis, and colony formation capacities were evaluated. The investigation revealed that the SA/BP composite gel was successful in all GBM cells and could be used as a treatment agent for GBM and/or other invasive cancer types. METHODS AND RESULTS: According to the results, the SA/BP composite gel had no effect on healthy fibroblast cells but had a lethal effect on all glioblastoma cells. Additionally, the wound healing method was used to examine the effect of the SA/BP composite gel on cell migration. It was discovered that the wound closed in 24 h in untreated control group cells, while the SA/BP composite gel closed up to 29.62%, 26.77% and 11.31% of the wound for C6, U87MG and T98G cell lines respectively. SA/BP significantly reduced cell migration in cancer cells. The effect of the generated SA/BP composite gel on cell colony development was assessed using a colony formation assay, and the cells reduced colony formation for all GBMs. It was roughly 45% for 24 h and 30% for 48 h when compared to the control group for C6 cells, 33%(24 h) and 40%(48 h) for U87MG cells, 40%(24 h) and 43%(48 h) for T98G cells. DAPI(4',6-Diamidino-2-phenylindole) and JC-1(5,5',6,6'-Tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine, iodide) staining to evaluate apoptosis revealed that the SA/BP composite gel dramatically enhanced the frequency of all GBMs undergoing apoptosis. CONCLUSIONS: In line with experimental findings, it was observed that the SA/BP composite gel system did not affect healthy fibroblast cells but had a cytotoxic effect on glioblastoma cells, significantly reduced cell migration and colony-forming capacity of cells, and significantly increased apoptosis and depolarization of cell membranes. Based on all these findings, it can be said that SA/BP composite gel has cytotoxic, antiproliferative and antiapoptotic effects on different glioblastoma cells.

Antioxidant Activity and Hemocompatibility Study of Quercetin Loaded Plga Nanoparticles.

Quercetin (QU) is an important flavonoid compound presenting lots of biological activities, but its application has been limited due to its low aqueous solubility and instability. In this study, conducted to improve these properties of the quercetin, quercetin-encapsulated PLGA nanoparticles were prepared, characterized, and evaluated for antioxidant and hemolytic activity. Nanoparticles were produced by single emulsion solvent evaporation method. Four different process parameters initial QU amount, PVA concentration, PVA volume, and initial PLGA amount were investigated to obtain the nanoparticles which have minimum particle size and maximum entrapment efficiency. Synthesized nanoparticles were evaluated for particle size, entrapment efficiency, and reaction yield. Additionally, antioxidant properties and in-vitro hemolytic activity of quercetin loaded nanoparticles with different particle size were also evaluated for the first time in the literature. The antioxidant activity results showed that nanoparticles have different antioxidant activity, depending on the amount of quercetin release from nanoparticles at different particle sizes. The hemolytic activity results show that all nanoparticles exhibited favorable compatibility to red blood cells and no significant hemolytic effect was observed.

Cytotoxic and mutagenic potential of juglone: a comparison of free and nano-encapsulated form.

Despite its evidenced beneficial herbicidal, antibacterial, antiviral, antifungal, and antioxidant effects, the application of juglone (5-hydroxy-1,4,-naphthoquinone) is limited due to its low water solubility and allelopathic and toxic effects. In recent years, research has aimed to overcome these limitations by increasing its solubility and controlling its release through nanoparticular systems. This is the first study to have synthesised and characterised juglone-loaded polymeric nanoparticles and compared them with free juglone for cytotoxicity in mouse (L929 fibroblasts) and alfalfa cells and for mutagenic potential in Salmonella typhimurium TA98/100. Mouse and plant cells treated with free and nano-encapsulated juglone showed a decrease in cell viability in a dose and time-dependent manner, but this effect was significantly lower with the nano-encapsulated form at lower doses. In the TA98 strain with S9, nano-encapsulated juglone did not exhibit mutagenic effects, unlike the free form. Since all results show that juglone encapsulation with polymeric nanoparticles reduced the toxic and mutagenic effects, it has a promising potential to be applied in medicine, food safety, and agriculture.

Catechin Loaded Poly(lactic-co-glycolic acid) Nanoparticles: Characterization, Antioxidant and Cytotoxic Activity Against MCF-7 Breast Cancer Cells.

Here we evaluate the cytotoxic and antioxidant properties of Catechin (Cat) loaded nanoparticles (CatNps) on breast cancer cell lines. CatNps were prepared by the modified single emulsion solvent evaporation method. The structural, physical and chemical properties of the CatNps including PLGA-Cat interactions, and surface characteristics, were analyzed by dynamic light scattering (DLS), Fourier transform infrared (FT-IR), and Scanning Electron Microscopy (SEM). The average particle size of CatNps was 190.5±1.762 nm with an encapsulation efficiency of 10% and zeta potential value of 13.7±1.258 mV. The CatNps had reaction yield of 43.47±1.1% and loading capacity of 3.710±2.6% Treating MCF-7 cells with CatNps for 48 h led to a decrease in cell viability with an of IC50 22.59 µg/mL. The antioxidant behavior of CatNps was evaluated by the DPPH method in various pH environments (pH 3-, 6-, 7.4-, 9- and 12). Despite the low encapsulation efficiency, CatNps showed significant antioxidant efficacy against DPPH radicals relative to Cat. Our results revealed that CatNps could be used as promising anti-cancer agents due to their enhanced cytotoxic activity, increased bioavailability, and antioxidant properties.

Inhibitory effects of aptamer targeted teicoplanin encapsulated PLGA nanoparticles for Staphylococcus aureus strains.

Emergence of resistance to traditional antibiotic treatments necessitates alternative delivery systems. Teicoplanin is a glycopeptide antibiotic used in the treatments of serious infections caused by Gram-positive bacteria, including Methicillin Resistant Staphylococcus aureus (MRSA). One strategy to keep up with antibiotic resistance development is to limit dose and amount during treatments. Targeted delivery systems of antibiotics have been suggested as a mechanism to slow-down the evolution of resistance and to increase efficiency of the antimicrobials on already resistant pathogens. In this study, we report teicoplanin delivery nanoparticles of Poly Lactic-co-Glycolic Acid (PLGA), which are functionalized with S. aureus specific aptamers. A 32-fold decrease in minimum inhibitory concentration (MIC) values of teicoplanin for S. aureus was demonstrated for susceptible strains and about 64-fold decline in MIC value was achieved for moderately resistant clinical isolates of MRSA upon teicoplanin treatment with aptamer-PLGA nanoparticles. Although teicoplanin delivery in PLGA nanoparticles without targeting demonstrated eightfold decrease in MIC of susceptible strains of S. aureus and S. epidermidis and twofold in MIC of resistant strains, the aptamer targeting specifically decreased MIC for S. aureus, but not for S. epidermidis. Therefore, aptamer-targeted PLGA delivery of antibiotic can be an attractive alternative to combat with some of the multi-drug resistant bacterial pathogens.

Quercetin-loaded nanoparticles enhance cytotoxicity and antioxidant activity on C6 glioma cells.

Quercetin (Qu) is a natural flavonoid present in many commonly consumed food items. The dietary phytochemical quercetin prevents tumor proliferation and is a potent therapeutic cancer agent. The purpose of this study was to synthesize and characterize quercetin-loaded poly(lactic-co-glycolic acid) nanoparticles (Qu1NP, Qu2NP, and Qu3NP) with different size and encapsulation properties and to evaluate their in vitro activity on C6 glioma cells. Nanoparticles were synthesized by single emulsion solvent evaporation method. Then, particle size, zeta potential, polydispersity index and encapsulation efficiency of nanoparticles were determined. Particle size of Qu1NP, Qu2NP, and Qu3NPs were determined as 215.2 ± 6.2, 282.3 ± 7.9, and 584.5 ± 15.2 nm respectively. Treating C6 glioma cells with all nanoparticle formulations effectively inhibited the cell proliferation. Qu1NPs were showed the lowest IC50 value in 48 h with 29.9 µg/ml and achieved higher cellular uptake among other nanoparticles and Qu. Additionally, 48-h treatment with Qu1NPs significantly decreased MDA level (14.90 nmol/µg protein) on C6 glioma cells which is related to reduced oxidative stress in cells. Findings of this study revealed that quercetin's cellular uptake and anti-oxidant activity is improved by small-sized Qu1NPs in C6 glioma cells.

Comparative evaluation of hesperetin loaded nanoparticles for anticancer activity against C6 glioma cancer cells.

The aim of this study was to evaluate anti-cancer properties of hesperetin (Hsp) and hesperetin-loaded poly(lactic-co-glycolic acid) nanoparticles (HspNPs) for glioblastoma treatment. Nanoparticles prepared by single emulsion method had a size of less than 300 nm with 70.7 ± 3.9% reaction yield and 26.4 ± 1.1% Hsp loading capacity. Treatment of C6 glioma cells with HspNPs for 24 and 48 h resulted in dose- and time-dependent decrease in cell viability, with approximate IC50 of 28 and 21 µg/mL, respectively (p = .036 for 24 h, p = .025 for 48 h). The percentage of PCNA positive cells decreased to 20% and 10%, respectively, for Hsp- and HspNP-treated cells at concentration of 100 µg/mL. Treatment with increasing concentrations of HspNPs (25, 50, 75 and 100 µg/mL) resulted in 9.1-, 7-, 12.5- and 12.7-fold in increase in apoptotic cell number. Optimum doses of Hsp and HspNPs were found to increase oxidative damage in C6 glioma cells. MDA levels, an indicator of lipid peroxidation, were found to be significantly elevated at 75 and 100 µg/mL exposure concentration of HspNPs with (p = .002) and (p = .018), respectively for 48-h treatment. The results obtained with this study showed biocompatible polymeric nanoparticle systems has great advantages to enhance anti-cancer activity and poor solubility of therapeutic agents. Overall our findings suggest that Hsp-loaded PLGA nanoparticle systems showed significant anti-cancer activity and HspNPs could be used as promising novel anti-cancer agent.

Synthesis of hesperetin-loaded PLGA nanoparticles by two different experimental design methods and biological evaluation of optimized nanoparticles.

Hesperetin was effectively encapsulated into poly (d,l-lactic-co-glycolic acid) nanoparticles by using experimental design methods. A seven-factor Plackett-Burman design was used in order to determine the major process parameters. A significant linear equation, which shows the effect of each process parameter on encapsulation efficiency was developed, and then the most effective factors were determined. Further investigation and optimization was carried out by applying the three-factor three-level Box-Behnken design. Significant second-order mathematical models were developed by regression analysis of the experimental data for both responses: encapsulation efficiency and nanoparticle size. The two step experimental design allowed the synthesis of the desired nanoparticle formulations with maximum encapsulation efficiency (80.5 ± 4.9%) and minimum particle size (260.2 ± 16.5 nm) at optimum process conditions: 0.5% polyvinyl alcohol (PVA) concentration, 5.13 water:organic phase ratio, and 3.59 ml min-1 flow rate of the emulsified solution into 0.1% PVA. Furthermore, the biological activity of these optimized nanoparticles were determined with antimicrobial activity and cytotoxicity studies; results were then compared to the free hesperetin. The cytotoxicity result revealed that hesperetin and hesperetin-loaded nanoparticles were biocompatible with normal cell line L929 fibroblast cells up to 184.83 and 190.88 µg ml-1 for 24 h, and up to 133.24 and 134.80 µg ml-1 for 48 h, respectively. In the antimicrobial study, the optimized nanoparticle showed inhibition activity (minimal inhibitory concentration (MIC) values were 125 µg ml-1 for Escherichia coli, and 200 µg ml-1 for Staphylococcus aureus), while the free hesperetin did not demonstrate activity in both strains (MIC value >200 µg ml-1). These in vitro results may provide useful information for the investigation of hesperetin-loaded nanoparticles in diagnostic and therapeutic applications.

An In vivo study: Adjuvant activity of poly-n-vinyl-2-pyrrolidone-co-acrylic acid on immune responses against Melanoma synthetic peptide.

Peptides have been studied as an important class of components in medicine to control many major diseases with vaccination. Polymers as adjuvants are capable of enhancing the vaccine potential against various diseases by improving the delivery of antigens, and they reduce the booster doses of vaccines. In brief, polymers are promising candidates for peptide-based vaccine delivery platforms. The purpose of the present study was to create a possible alternative approach in the treatment of malignant melanoma and/or to prevent metastasis of melanoma. The study was designed as both an experimental and an in vivo study. We prepared a complex and covalent conjugate of MAGE-3 121-134 (L-L-K-Y-R-A-R-E-P-V-T-K-A-E) T-cell epitope as a vaccine candidate for melanoma. These conjugates were able to generate an immune response in mice after a single immunization, without the help of any external adjuvant. The peptide-polymer complexes activated the immune system in the best way and formed the highest antigen specific immune response. These results indicate the adjuvant activity of Poly(N-vinyl-2- pyrrolidone-co-acrylic acid) [P(VP-co-AA)] and the potential use of P(VP-coAA)-peptide based vaccine prototypes for future melanoma cancer vaccine formulations.

Enhancement of Antifungal Activity of Juglone (5-Hydroxy-1,4-naphthoquinone) Using a Poly(d,l-lactic-co-glycolic acid) (PLGA) Nanoparticle System.

This study aimed to synthesize and characterize juglone-entrapped poly(d,l-lactic-co-glycolic acid) (PLGA) nanoparticles and compare the antifungal properties of free juglone with its PLGA nanoparticle formulation for the first time. The juglone-loaded nanoparticles prepared using the oil-in-water (o/w) single-emulsion solvent evaporation method were characterized by the reaction yield (RY), encapsulation efficiency (EE), polydispersity index (PDI), particle size, zeta potential (ZP), FT-IR, and in vitro release properties and evaluated for their morphological features using SEM. The nanoparticle formulation had size, RY, ZP, EE, and PDI values of 212 nm, 66.91 ± 2.4%, -16.3 ± 0.7 mV, 70.66 ± 3.1%, and 0.083 ± 0.024, respectively. In vitro release showed a triphasic pattern with initial burst followed by sustained release and dormant phase over the study period, releasing about 72.8% in total after 42 days. The antifungal studies against Aspergillus flavus, Candida albicans, and Fusarium spp. using agar dilution and top agar dilution methods indicated that the juglone-encapsulated nanoparticle was more effective than free juglone. This study showed that the top agar method, which was applied for the first time on antifungal activity, is more suitable for the nanoparticular system based on sustained release. Therefore, PLGA nanoparticle formulations may be an important tool for application in many areas for the effective and beneficial use of hydrophobic compounds such as juglone.

Comparative evaluation of antibacterial activity of caffeic acid phenethyl ester and PLGA nanoparticle formulation by different methods.

The aim of the present study was to evaluate the antimicrobial activity of nanoparticle and free formulations of the CAPE compound using different methods and comparing the results in the literature for the first time. In parallel with this purpose, encapsulation of CAPE with the PLGA nanoparticle system (CAPE-PLGA-NPs) and characterization of nanoparticles were carried out. Afterwards, antimicrobial activity of free CAPE and CAPE-PLGA-NPs was determined using agar well diffusion, disk diffusion, broth microdilution and reduction percentage methods. P. aeroginosa, E. coli, S. aureus and methicillin-resistant S. aureus (MRSA) were chosen as model bacteria since they have different cell wall structures. CAPE-PLGA-NPs within the range of 214.0 ± 8.80 nm particle size and with an encapsulation efficiency of 91.59 ± 4.97% were prepared using the oil-in-water (o-w) single-emulsion solvent evaporation method. The microbiological results indicated that free CAPE did not have any antimicrobial activity in any of the applied methods whereas CAPE-PLGA-NPs had significant antimicrobial activity in both broth dilution and reduction percentage methods. CAPE-PLGA-NPs showed moderate antimicrobial activity against S. aureus and MRSA strains particularly in hourly measurements at 30.63 and 61.25 µg ml(-1) concentrations (both p < 0.05), whereas they failed to show antimicrobial activity against Gram-negative bacteria (P. aeroginosa and E. coli, p > 0.05). In the reduction percentage method, in which the highest results of antimicrobial activity were obtained, it was observed that the antimicrobial effect on S. aureus was more long-standing (3 days) and higher in reduction percentage (over 90%). The appearance of antibacterial activity of CAPE-PLGA-NPs may be related to higher penetration into cells due to low solubility of free CAPE in the aqueous medium. Additionally, the biocompatible and biodegradable PLGA nanoparticles could be an alternative to solvents such as ethanol, methanol or DMSO. Consequently, obtained results show that the method of selection is extremely important and will influence the results. Thus, broth microdilution and reduction percentage methods can be recommended as reliable and useful screening methods for determination of antimicrobial activity of PLGA nanoparticle formulations used particularly in drug delivery systems compared to both agar well and disk diffusion methods.

Selenium status in blood, urine, and hair samples of newly diagnosed pediatric cancer patients.

BACKGROUND/AIM: Selenium (Se) is a trace element that has multiple functions. Low Se amounts in serum and hair have been reported in pediatric and adult cancer patients. The aim of our study was to evaluate Se levels in the serum, urine, and hair of pediatric cancer patients with leukemia, lymphoma, and solid tumors when compared with healthy children. MATERIALS AND METHODS: The concentrations of Se in the serum, hair, and urine of 32 Turkish children as healthy controls and 88 Turkish children diagnosed with acute leukemia (58), lymphoma (16), and solid tumors (14) were measured using inductively coupled plasma mass spectroscopy. RESULTS: Se levels in the serum and hair of the children with cancer were significantly lower than those of the controls. There were no differences between the leukemia, lymphoma, and solid tumors group. On the other hand, the Se levels of the urine samples were slightly elevated in cancer patients compared with the control group. There was no marked difference in the Se levels of patients with different types of cancer. CONCLUSION: Se deficiency might be associated with the development of pediatric cancer. Especially in children, additional studies are needed to define whether low levels of Se may play a role in cancer pathogenesis.

Protective effect of chemically modified SOD on lipid peroxidation and antioxidant status in diabetic rats.

Reactive oxygen species mediated oxidative stress play an important role on the injury of tissue damage and increased attention has been focused on the role of free radicals in diabetes mellitus (DM). In the present study firstly superoxide dismutase (SOD) enzyme was chemically modified with two different polymer and physicochemical properties of these conjugates clearly analyzed. Then, the stability of carboxymethylcellulose-SOD (CMC-SOD) and poly methyl vinyl ether-co-maleic anhydride-SOD (PMVE/MA-SOD) conjugates was investigated against temperature and externally added H2O2. Moreover, we investigated the effect of chemically modified SOD enzyme on lipid peroxidation and antioxidant status in streptozotocin (STZ)-induced diabetic rats. PMVE/MA-SOD conjugate treatment significantly reduced MDA level compared with the control groups, native and CMC-SOD conjugate treated groups in brain, kidney and liver tissue. GSH and SOD enzyme activity in diabetic groups was significantly increased by treatment of CMC-SOD and PMVE/MA-SOD conjugates. The protective effects on degenerative changes in diabetic rats were also further confirmed by histopathological examination. This study provides the preventative activity of SOD-polymer conjugates against complication of oxidative stress in experimentally induced diabetic rats. These results suggest that chemically modified SOD is effective on the oxidative stress-associated disease and offer a therapeutic advantage in clinical use.