Greater Cytotoxic Effect Research Articles

Amino acid derivative of probenecid potentiates apoptosis-inducing effects of vinblastine by increasing oxidative stress in a cancer cell-specific manner

Many chemotherapeutic drugs suffer from multidrug resistance (MDR). Efflux transporters, namely ATP-binding cassettes (ABCs), that pump the drugs out of the cancer cells comprise one major reason behind MDR. Therefore, ABC inhibitors have been under development for ages, but unfortunately, without clinical success. In the present study, an l-type amino acid transporter 1 (LAT1)-utilizing derivative of probenecid (PRB) was developed as a cancer cell-targeted efflux inhibitor for P-glycoprotein (P-gp), breast cancer resistant protein (BCRP) and/or several multidrug resistant proteins (MRPs), and its ability to increase vinblastine (VBL) cellular accumulation and apoptosis-inducing effects were explored. The novel amino acid derivative of PRB (2) increased the VBL exposure in triple-negative human breast cancer cells (MDA-MB-231) and human glioma cells (U-87MG) by 10–68 -times and 2-5-times, respectively, but not in estrogen receptor-positive human breast cancer cells (MCF-7). However, the combination therapy had greater cytotoxic effects in MCF-7 compared to MDA-MB-231 cells due to the increased oxidative stress recorded in MCF-7 cells. The metabolomic study also revealed that compound 2, together with VBL, decreased the transport of those amino acids essential for the biosynthesis of endogenous anti-oxidant glutathione (GSH). Moreover, the metabolic differences between the outcomes of the studied breast cancer cell lines were explained by the distinct expression profiles of solute carriers (SLCs) that can be concomitantly inhibited. Therefore, attacking several SLCs simultaneously to change the nutrient environment of cancer cells can serve as an adjuvant therapy to other chemotherapeutics, offering an alternative to ABC inhibitors.

Total Antioxidant Potential, Total Phenolic Profile and Cytotoxic Activity Against Brain Cancer: Melocan and Galdirik§.

Brain cancer is known to be one of the most difficult types of cancer to cure. It has a serious impact on the lives of diagnosed people due to the insufficient treatment options and their side effects. The search for new alternative treatments is therefore ongoing. Melocan (Smilax excelsa L.) and galdirik (Trachystemon orientalis) are of great importance in both traditional culinary culture and traditional medicine around the Black Sea; however, the knowledge about their antioxidant and cytotoxic effects remains fairly limited. The aim of this study is to determine the antioxidant and cytotoxic activity of Smilax excelsa and Trachystemon orientalis on the C6 glioblastoma cell line. The plants of Smilax excelsa and Trachystemon orientalis were dried and extracted and then their total phenolic content (TPC) and phenolic profiles were studied. In addition, their total antioxidant status (TAS) and total oxidant status (TOS) were determined using an assay kit. We also analysed the total antioxidant activity (TAA) using the DPPH radical scavenging assay and the cytotoxic effect on the glioma cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay. According to the results, the water extracts of Smilax excelsa and Trachystemon orientalis had higher TPC (expressed in gallic acid equivalents on dry mass basis: 1158.17 and 262 mg/100 g, respectively) than the ethanol extracts. TAA expressed in Trolox equivalents on dry mass basis was 192.86 and 131.92 mg/100 g for Smilax excelsa and Trachystemon orientalis, respectively. The MTT assay showed that Trachystemon orientalis had a greater cytotoxic effect. In conclusion, the findings of the current study are promising for the development of new drugs. This is the first study that aims to evaluate the potential cytotoxic activity of two local Turkish plants, Smilax excelsa and Trachystemon orientalis, against C6 glioblastoma cells. The results confirm that both plants could be used as good therapeutic agents for the treatment of cancer in the future.

Synthesis, structure characterization and antitumor activities of copper and cobalt thiocyanate complexes with 3-acetlpyridine ligand

Complexes of [Cu(SCN)2(3-Acpy)2], (1) and [Co(SCN)2(H2O)2(3-Acpy)2], (2) were synthesized at ambient temperature by adding an aqueous solution of copper(II) or cobalt(II) chloride to H2O/CH3CN solution of 3-acetlpyridine (3-Acpy) and potassium thiocyanate. Elemental analysis, FT-IR and UV–vis spectra were used to comprehensively characterize complexes 1 and 2. Single crystal X-ray diffraction was used to analyze the crystal structure of complexes 1 and 2. In complex 1, the copper is bonded to two 3-Acpy ligands and two thiocyanates groups in trans mode giving square planer geometry. While, in complex 2, the cobalt ion is coordinated to two water molecules, two 3-Acpy ligands and two thiocyanates ions in trans mode giving an octahedral structure. Through hydrogen bonding, the discrete units of complexes 1 and 2 extend to form a 3D supramolecular network. Additionally, the luminescence spectra of complexes 1 and 2 were investigated. Complexes 1 and 2 were evaluated to determine whether they were able to inhibit the growth of liver cancer (HepG2 cells) and breast cancer (MCF-7 cells). Complex 1 exhibits lower IC50 values than complex 2 against both of the two human cell lines. In HepG2 cells treated with complexes 1 and 2 at dosages roughly half of the IC50, apoptosis of complexes 1 and 2 is triggered by 18% and 22%, respectively. It was found that copper complex 1 has a greater cytotoxic effect on MCF-7 and influences on late apoptosis of HepG2.

Changes in Length and Positive Charge of Palindromic Sequence RWQWRWQWR Enhance Cytotoxic Activity against Breast Cancer Cell Lines.

Breast cancer is one of the main causes of premature death in women; current treatments have low selectivity, generating strong physical and psychological sequelae. The palindromic peptide R-1-R (RWQWRWQWR) has cytotoxic activity against different cell lines derived from cancer and selectivity against noncancerous cells. To determine if changes in the charge/length of this peptide increase its activity, six peptides were obtained by SPPS, three of them with addition of Arg at the N, C-terminal or both and three with deletion of Arg at the N, C-terminal or both. The cytotoxic and selective activities were evaluated against MCF-7, MDA-MB-231, and MCF-12 cell lines and fibroblast primary cell culture, evidencing that the RR-1-R peptide with the inclusion of Arg in the N-terminal end maintained selectivity and increased cytotoxicity against lines derived from breast cancer. The effect of this addition regarding the type of induced cell death was evaluated by flow cytometry, showing very low rates of necrosis and a significant majority of apoptotic events with activation of both Caspase 8 and Caspase 9. This work allowed us to find a modification that generates a peptide with greater cytotoxic effects and can be considered a promising molecule for other approaches to improve anticancer peptides.

ODP276 THYROID CANCER CELLS ARE MORE RESISTANT THAN NORMAL CELLS TO BISPHENOL-A CYTOTOXIC EFFECT

Abstract Bisphenol-A (BPA) is a well-known endocrine disruptor that produces important effects on thyroid metabolism, can interfere with TSH transcriptional activity, alter the transcription of T3-associated genes and, thus, modulate its activity in different cell lines. However, its direct effects on thyroid cells, especially on mutated thyroid cells, are still poorly understood. We tested 14 different concentrations of BPA, ranging from 0.4µg/mL to 400µg/mL, including LME (Specific Migration Limit =1μg/mL, recommended as acceptable by the Brazilian National Health Surveillance Agency-ANVISA) on two human thyroid cell lines: Nthy-ori 3-1 (from normal thyroid follicular cells) and TPC-1 (from papillary carcinoma). Cell viability analysis was made using the Trypan blue assay on cells exposed to BPA for 24h and 48h in technical and biological triplicate. We observed a non-monotone dose-response curve, characteristic of disruptors’ effect, on both cell lines, with greater cytotoxic effect with higher doses. In fact,100μg/mL already killed almost all Nthy-ori 3-1 cells (99% at 24h and 97% at 48h), whereasthe doses of 0.78µg/mL; 4µg/mL and 12.5µg/mL caused 50% kill. Optical microscopy revealed some irregular cells lost on the plate, and few debris with50μg/mL. Concentrations of 100 to 400µg/mL produced many dark small cells detached from the plate and clumps of dead cells. The SCI dose (1µg/mL - allowed in daily life), resulted in 79% of Nthy-ori 3-1 cell death after 24h of exposure. However, only 7% of the cell were dead after 48h of exposure to the SCI dose. This reduction in death rate after longer exposure to BPA was observed at all concentrations, suggesting a potent cellular recovery mechanism. The TPC-1 strain showed a different behavior, with a much more linear dose-dependent cytotoxic effect and much more resistance to the harmful effects of BPA at low doses. In fact, 0.4μg/mL to 3.125μg/mL produced less than 10% cell death, and microscopy confirmed the maintenance of the integrity of virtually all cells. On the contrary, BPA concentrations from 100 to 400µg/mL killed almost all TPC-1 cells at both 24h and 48h times. We conclude that this thyroid papillary carcinoma cell line is more resistant than the normal thyroid lineage to low BPA concentrations (from 0.4 to 50µg/mL) but this endocrine disruptor is highly cytotoxic at higher concentrations, causing more than 90% cell deaths on both normal and mutated thyroid cell lines. Presentation: No date and time listed

Folate decorated lipid chitosan hybrid nanoparticles of 5-fluorouracil for enhanced anticancer efficacy against colon cancer

The study aimed to develop folate decorated lipid chitosan hybrid nanoparticles for targeted delivery of 5-fluorouracil in colon cancer by utilizing the overexpressed folate receptors on the surface of HT-29 and HCT 116 cancer cell lines. The developed formulations were prepared by the ionic gelation method with slight modifications. The developed formulations exhibited spherical morphology, smaller particle size (158 to 225 nm), zeta potential (32.24 to 35.95 mV), PDI (0.19 to 0.35), and high encapsulation efficiency (85.3 % to 94.2 %) with optimal physicochemical characteristics. The in vitro release showed a biphasic release pattern with an initial burst release followed by a sustained release for 48 h. Moreover, the in vitro cell line study revealed that FA-CLPN-2 exhibited an enhanced cellular uptake and greater cytotoxic effect in HT-29 and HCT 116 cell lines compared to non-targeted CLPN-2 and free drug solution due to the folate receptor facilitated endocytosis process. The in vivo toxicity study revealed the safety and biocompatibility of the developed formulations in biological systems. The stability study demonstrates the stability of the developed formulations. Overall, these results suggest that the folate decorated lipid chitosan hybrid nanoparticles could be used as a potential delivery system for tumor-targeted therapy with reduced side effects.

In vitro antifungal activity of Morinda citrifolia (noni) extract against Candida albicans

Candida albicans is the main agent of the most common fungal infection, Candidiasis. It is an opportunistic and dangerous pathogen, especially in immunosuppressed patients. The biological properties of Morinda citrifolia (noni) make it a potent antifungal. In this study, antifungal effect of M. citrifolia was evaluated to verify its effect on human cells. Extract of M. citrifolia was used against strains of C. albicans (cEC 1291). Glucose consumption in C. albicans biofilm was determined at different concentrations of M. citrifolia, and germ tube formation was evaluated in the presence and absence of M. citrifolia. Fungicidal activity was determined by the kinetics of fungal cell death. THP-1 and HeLa cells were used for cell viability and apoptosis, and cell proliferation assays, respectively. Cells treated with M. citrifolia maintained higher concentration of glucose than the control group (p < 0.05). Germ tube formation was inhibited in cells treated with M. citrifolia (p < 0.05). M. citrifolia exerted a cytotoxic effect on C. albicans cells with 99.99% lethality after 6.82 h (1:1 and 1:2), and reduced the viability of THP-1 cells by 25% and 67% after 12 and 36 h, respectively. Annexin V expression in THP-1 increased in groups that received higher concentrations of M. citrifolia (p < 0.05), reducing the proliferation of THP-1 and HeLa cells (2.8-fold). A greater cytotoxic effect was observed in fungal cells. These results indicate that M. citrifolia exerts biological activity against C. albicans and reduces the viability and proliferation of human cells.

Efficacy of Gold and Nickel on the Anticancer Activity and Physical Properties of Pemetrexed Loaded on Fullerene C60 Buckysomes

Objective: This study aims to create a new delivery system for pemetrexed, employing a metal-C60 fullerene combination for the first time. Additionally, a lung cancer cell line study will be used to examine the impact of this combination on the release, behavior, and drug’s cytotoxicity. Methods: To utilize a laser to irradiate hydrocarbons, fullerene C60 (Buckysomes) was produced and then nanosized by adding different volumes of isopropyl alcohol and ultrasonication. Gold Nanoparticles (AuNPs) and nickel nanoparticles (NiNPs) were also prepared each one separately and added to the previous mixture together with Pemetrexed (PMX) with further ultrasonication for 20 minutes. The mixture is kept in the refrigerator for 20 hours and then filtered. The precipitate was dried and characterized for particle size, invitro release, and anticancer activity in comparison to pemetrexed loaded on C60 fullerene previously prepared in our laboratory. Results: The evaluation techniques revealed the successful loading of pemetrexed on metals- fullerene C60 nanocarrier, SG10 was the optimum sample with % of yield reach to 95.36. The release profile shows that the percentage release of pemetrexed after 240 minutes is equal to 40% from pure PMX, while the release after 240 minute was found to be 87.8 % from pemetrexed loaded gold nanoparticles (AuNPs) fullerene C60 nanocarrier (SG10), while NiNPs not significantly improved the release profile of Pemetrexed from Pemetrexed loaded NiNPs – fullerene (SN7). Pemetrexed loaded on gold nanoparticles (AuNPs), when compared to pure PMX (3.1 M) and SN7 (11.5 M), fullerene nanocarrier (SG10) exhibits a lower IC50 (1.55 M) and a greater cytotoxic effect (high IR percent) on A549 cancer cells. Pemetrexed’s cytotoxicity in A549 was enhanced after being loaded onto gold nanoparticles (AuNPs)-fullerene nanocarriers (90.4% cell death for SG10) as opposed to pure drug (60% cell death). This finding suggests that the loading process enhanced the drug’s cytotoxic activity for the tumor cells, which may have sped up the onset of action (30% cell death after 24 hours). Conclusion: This study successfully prepared Metals- fullerene C60 Buckysomes loaded with pemetrexed utilizing gold and nickel, which may serve (especially Au- fullerene) as a suitable nanocarrier for pemetrexed, resulting in an improvement to solubility, the release pattern, and cytotoxicity.

Оцінка потенційної токсичності мийно-дезінфекційних засобів на культурі клітин in vitro

Detergents with disinfectants properties today are widely used in various fields of human activity for cleaning surfaces, equipment, tools while destroying infectious agents. An active substance, as disinfectants, it contains various chemical compounds with antimicrobial, antiviral and antifungal properties. To establish the safety of these substances, along with toxicological studies on animals, experiments are conducted on alternative in vitro test systems, in particular on cell culture, which allows to quickly assess and predict the potential toxicity of detergents to humans and other living organisms. The purpouse of the study was assessment of the potential danger of detergents and disinfectants for humans by determining their cytotoxic effects in experiments on cell culture in vitro. Material and methods. Studies were performed to evaluate the cytotoxic effect of detergents with disinfectants properties : "Bioblysk", "Blysk Plus" and "Blysk" (manufactured by PE "HYGIENICS", Poland) on the lung cancer cells (line A-549) using cytochemical methods: MTT- with methyltetrazolium blue and CRB - test with sulforodamine B. Results. It was found that all detergents have the greatest cytotoxic effect on A-549 cells at a concentration of 1.0% of the native solution. The lowest concentrations 0.03% and 0.015% have no toxic effect on cells. With the help of probit analysis based on the obtained results, LC50 values were calculated: for Bioblysk - 0.15% (MTT test) and 0.15% (CRP test); for Blysk Plus LC50 - 0.25% (MTT test) and 0.2% (CRP test); for Blysk - 0.2% (MTT test and CRP test). LC50 values for detergents were 10 times lower than the recommended minimum concentrations of working solutions Conclusions. The obtained data indicate that the citotoxicity effect of detergents on A-549 cells was manifested by the effect on mitochondrial function and protein synthesis. The severity of studied drugs toxic effect depended on its chemical composition and pH, concentration in the incubation medium. The calculated LC50 values indicate their possible danger to the respiratory system, so they require appropriate protection.

Antibacterial and Cytotoxic Effects of Multi-Walled Carbon Nanotubes Functionalized with Iodine

The effect of multi-walled carbon nanotubes (MWCNTs) functionalized with iodine (15, 10, and 5 wt %) is studied for bacteria Escherichia coli and MCF-7 breast adenocarcinoma tumor cells. A significant increase in the antimicrobial activity of MWCNTs is obtained when they are modified with 15 and 10 wt % iodine: at concentrations of 0.1 and 0.01 g/L, the survival of bacteria was less than 2%, while for MWCNTs without iodine, the indicator was 7 and 30%, respectively. The modification of MWCNTs with 5-wt % iodine also enhances the antibacterial properties of nanotubes, but less significantly. MCF-7 cells also have the greatest cytotoxic effect on MWCNTs functionalized with 15- and 10-wt % iodine; however, unlike bacteria, the difference in the survival rates between composites and compared with pure MWCNTs was no more than 3%. Comparison of the effects of iodine in MWCNTs and pure iodine showed that functionalized MWCNTs at concentrations above 0.001 g/L are more toxic to bacteria, while pure iodine solutions are more cytotoxic to cells.

Bee Venom Effect on Glioblastoma Cells Viability and Gelatinase Secretion.

BackgroundThe involvement of MMP-2 and MMP-9 in the pathogenesis of various kinds of cancers including glioblastoma is well documented. The evaluation of the anticancer potential of honey bee (Apis mellifera) venom (BV) consisting of the inhibition of MMP-2 and MMP-9 secretion in a glioblastoma cell culture model was the aim of the study.Methods8-MG-BA and GAMG human primary glioblastoma cell lines vs. HT-22 mouse hippocampal neuronal cells were applied for the study. The BV dose (0.5, 1.0, 1.25, 1.5, 1.75, 2.0, 2.5, and 5.0 μg/ml) and time-dependent (24, 48, 72 h) cytotoxicity was evaluated with the tetrazolium-based colorimetric assay (MTT test). MMP-2 and MMP-9 activities in the cell culture medium under different BV concentrations were determined by gelatin zymography.ResultsA dose and time-dependent BV effect on cytotoxicity of both glioblastoma cell lines and hippocampus line was observed. The weakest, but statistically important effect was exerted by BV on HT-22 cells. The greatest cytotoxic effect of BV was observed on the 8-MG-BA line, where a statistically significant reduction in viability was observed at the lowest BV dose and the shortest incubation time. The reduction of both gelatinases secretion was observed at 8-MG-BA and GAMG lines without significant effect of HT-22 cell line.ConclusionIn vitro studies indicate that BV has both cytotoxic and inhibitory effects on the secretion of MMP-2 and MMP-9 in selected lines of glioma, suggesting anticancer properties of BV.

Coumarin Derivatives Exert Anti-Lung Cancer Activity by Inhibition of Epithelial-Mesenchymal Transition and Migration in A549 Cells.

A series of coumarin derivatives and isosteres were synthesized from the reaction of triflic intermediates with phenylboronic acids, terminal alkynes, and organozinc compounds through palladium-catalyzed cross-coupling reactions. The in vitro cytotoxic effect of the compounds was evaluated against two non-small cell lung carcinoma (NSCLC) cell lines (A-549 and H2170) and a normal cell line (NIH-3T3) using cisplatin as a reference drug. Additionally, the effects of the most promising coumarin derivative (9f) in reversing the epithelial-to-mesenchymal transition (EMT) in IL-1β-stimulated A549 cells and in inhibiting the EMT-associated migratory ability in A549 cells were also evaluated. 9f had the greatest cytotoxic effect (CC50 = 7.1 ± 0.8 and 3.3 ± 0.5 μM, respectively against A549 and H2170 cells) and CC50 value of 25.8 µM for NIH-3T3 cells. 9f inhibited the IL-1β-induced EMT in epithelial cells by inhibiting the F-actin reorganization, attenuating changes in the actin cytoskeleton reorganization, and downregulating vimentin in A549 cells stimulated by IL-1β. Treatment of A549 cells with 9f at 7 µM for 24 h significantly reduced the migration of IL-1β-stimulated cells, which is a phenomenon confirmed by qualitative assessment of the wound closure. Taken together, our findings suggest that coumarin derivatives, especially compound 9f, may become a promising candidate for lung cancer therapy, especially in lung cancer promoted by NSCLC cell lines.

Phytochemical analysis and anticancer activity of Falcaria vulgaris Bernh growing in Moghan plain, northwest of Iran

BackgroundFalcaria vulgaris Bernh among the most important member of Apiaceae family has been used for medical investigation in Iran and some regions in the world. This plant possesses a range of coumarin and flavonoids compounds that have many therapeutic properties such as gastrointestinal and liver diseases, skin ulcers, gastric ulcers, and intestinal inflammation. It has also been found that these compounds lead to cytotoxic effects.ObjectiveThis study contains concentrates on the cytotoxic effect and induction of apoptosis on cancerous cells (SW-872) through various extracts and essential oil of Falcaria vulgaris Bernh. It considers the volatile compounds of effective samples.MethodsThe shoot of the plant was extracted by the Soxhlet apparatus and its essential oil was taken by the Clevenger apparatus. The cytotoxicity of the samples was evaluated by the MTT method and the mechanism of cancer cell death by flow cytometry and finally, the volatile compounds of essential oils and effective extracts were identified by GC-MS.ResultsThe results demonstrated that n-Hexane extract and 40% VLC fraction had the greatest cytotoxic effect on SW-872 cells. While, the most abundant volatile compounds in essential oil and 40% VLC fraction of n-Hexane extract were terpenoid compounds like (+) spathulenol and caryophyllene oxide, in n-Hexane extract tetradecan, and spathulenol were the most, respectively.ConclusionThe fraction of 40% n-Hexane was in a concentration-dependent manner and significantly with controlling cells inhibited the growth of cancer cells. A plausible explanation could be made to account for this effect. This inhibition was made through induction of apoptosis and due to the presence of effective volatile compounds such as terpenoids and non-terpenoids which could be considered as valuable natural sources for the isolation of anti-cancer compounds.

Phenolic content, antioxidant, cytotoxic and antiproliferative effects of fractions of Vigna subterraenea (L.) verdc from Mpumalanga, South Africa

Consistent intake of legumes has been correlated with decreased possibility of developing colorectal cancer (CRC) due to the content of some phytochemicals like polyphenols. Bambara groundnut (BGN) is an underutilized crop with a rich nutritional profile, but have not been exploited for its nutraceutical and medicinal benefits. In this study, total polyphenol, flavonoid (flavonol and flavanol) content, antioxidant activity and cytotoxicity/antiproliferative properties of 70% ethanolic extracts of whole BGN, cotyledon and seed coat on Caco-2 and HT-29 colon cancer cells were evaluated. Seed coat had a significantly (p < 0.05) higher composition of total polyphenol, flavonol and flavan-3-ol (flavanol) compared to whole seed and cotyledon. Antioxidant activity determined with ferric reducing antioxidant power (FRAP), 2,2- azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and oxygen radical absorbance capacity (ORAC) assays, showed that seed coat with higher polyphenolic content had significantly (p < 0.05) greater antioxidant activity. BGN fractions demonstrated cytotoxic and antiproliferative effects against HT-29 and Caco-2 colon cancer cells in a dose-dependent manner, with seed coat and whole seed exhibiting greater cytotoxicity and higher antiproliferative activity and colon cancer cell inhibition. Extracts of the cotyledon also showed cytotoxic activity and hindered cancer cell growth/division but to a significantly (p < 0.05) lower magnitude. BGN parts indicated a greater cytotoxic effect and potential to slow down Caco-2 colon cancer cell growth and division over HT-29. This result provides new knowledge on the possible health benefits of BGN, as well as the potential for product development and may influence its consumption and utilisation.

Design and Characterization of Liposomal Methotrexate and Its Effect on BT-474 Breast Cancer Cell Line.

Background: Breast cancer is the most common type of cancer among women worldwide. Traditional treatments, including chemotherapy, surgery, mastectomy, and radiotherapy, are commonly used. Because of the limitation of the aforementioned methods, novel treatment strategies are needed. Methotrexate is a chemotherapeutic drug, which is commonly used to treat breast cancer. Because of the side effects of the free drug, the liposomal form of the drug is suggested. Methods: Liposomal methotrexate was prepared and the encapsulation efficiency was measured. Moreover, the particle size and the zeta potential were measured. The liposome morphology was confirmed using transmission electron microscopy. The MTT assay was done to examine the cytotoxicity of free and encapsulated methotrexate on BT-474 cell line. The Annexin-V/PI dual staining assay was performed to assess the apoptosis in BT-474 breast cancer cells via the flow cytometry method. Results: The transmission electron microscopy results confirmed the integrated and spherical structure of the nanoparticles. The results of drug release showed that in acidic pH (5.4), more than 90% of the drug was released after 24 hours, which was higher than 2 other pHs. Furthermore, the IC50 value of liposomal methotrexate was determined as 2.15 and 0.82 mg/mL for 24 and 48 hours. The flow cytometry results confirmed that liposomal methotrexate had a greater cytotoxic effect on cancer cells compared with free methotrexate. Conclusion: Because of the advantages of liposomal based nanocarriers, in this study, liposomal methotrexate could be suggested as an appropriate candidate to treat breast cancer.

Preclinical Assessment of Paclitaxel- and Trastuzumab-Delivering Magnetic Nanoparticles Fe3O4 for Treatment and Imaging of HER2-Positive Breast Cancer.

Objective: The purpose of this study was to investigate the anticancer activity and the potential imaging use of the innovative combination of magnetic nanoparticles (MNPs)-Fe3O4, paclitaxel (PTX), and trastuzumab (Herceptin) in HER2-positive breast cancer.Methods: MNPs-Fe3O4 was synthesized and underwent water phase transfer and hydrophobic molecular loading, and its surface was then coupled with Herceptin mono-antibody. The morphological characteristics of MNPs-Fe3O4 were observed under transmission electron microscopy (TEM). Effects of PTX-Herceptin-MNPs-Fe3O4 on breast cancer cells were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,4-diphenyltetrazolium bromide assay and the flow cytometric apoptosis assay. To establish a xenograft model, we injected breast cancer SK-BR-3 cells into the left thighs of nude mice. We measured the effect of PTX-Herceptin-MNPs-Fe3O4 on tumor growth by measuring tumor size and calculating inhibition rate with immunohistochemistry analysis further performed, and analyzed MNPs-Fe3O4 accumulation in tumor lesions using in vivo magnetic resonance imaging and in vivo fluorescence imaging.Results: Most MNPs were in spherical shape of about 10 nm in diameter observed under TEM. PTX-Herceptin-MNPs-Fe3O4 showed greater cytotoxic effects, and induced a higher apoptosis rate of SK-BR-3 cells than all the other groups, with corresponding changes of apoptosis-related proteins. Meanwhile, the in vivo tumor xenograft model showed that tumor inhibition rate in the PTX-Herceptin-MNPs-Fe3O4 group was higher than in the PTX-Herceptin group. Furthermore, PTX-Herceptin-MNPs-Fe3O4 enhanced the T2 imaging contrast enhancement effect on tumors in tumor-bearing mice.Conclusion: The novel PTX-Herceptin-MNPs-Fe3O4 combination may represent a promising alternative breast cancer treatment strategy and may facilitate tumor imaging.

Research of cytotoxic activity of drinking water using methods in vitro

Aim. The aim of this work is to assess the quality of drinking water of various origins by its cytotoxic effect on human and animal cell cultures in in vitro experiments. Methods. The cytotoxic effect of control water obtained in accordance with the requirements of DSTU 4174: 2003, tap water, pump room and packaged Evian water was studied. The studies were carried out on NEC-293 cells (human kidney cells) and L929 (mouse fibroblasts) cells from the Institute of Microbiology and Virology named after V.I. D.K. Zabolotny National Academy of Sciences of Ukraine. Results. Studies in the MTT test of cytotoxicity of these water samples showed that tap water and bottled water exhibited the greatest toxic activity against human kidney cells of the NEC-293 line (the number of viable cells under their influence was 61.84% and 79.06%, respectively). Under the influence of water from the pump room - 84.55%, the control water showed the least influence - 93.13%. In the test with sulforhodamine B, the number of living cells under the influence of control water was 100.00%, tap water 80.15%, water from pump rooms 97.11%, Evian 84.70%. Conclusions. The greatest cytotoxic effect according to the MTT test and the test with sulforhodamine B on human kidney cells of the HEK-293 line and fibroblasts of the L929 mouse was exhibited by tap and bottled water. The least influence on the viability of the cells was exerted by the water from the pump room and the control. The negative effect of water on cell viability was manifested in the dysfunction of mitochondria and protein synthesis.&#x0D; Keywords: cytotoxicity, cell culture, drinking water.

Pluronic F127/Doxorubicin microemulsions: Preparation, characterization, and toxicity evaluations

The development of drug delivery systems minimizing the side effects of conventional chemotherapy is one of the major challenges in the field of biomaterials for cancer treatment. This work reports the formulation and characterization of oil-in-water Pluronic F127 microemulsions to enhance the bioavailability of doxorubicin (DOX). The density functional theory (DFT) calculations at the M06-2X level of theory were done to study the interaction details of DOX with ethyl butyrate, sodium caprylate, and one unit of the polymeric chain of surfactant Pluronic F127 in water solution, which are used in the synthesis process. Specifically, the quantum theory of atoms in molecules (QTAIM) analysis was performed to determine the nature of interactions. The highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) energies were calculated to show the direction of charge transfer within each complex. Furthermore, the natural bond orbital (NBO) analysis was performed on the studied systems. The size of F127/DOX microemulsion was about 7.0 nm by dynamic light scattering analysis. In vitro toxicity of standard DOX and DOX-loaded microemulsions were assessed against MCF-7 and C152 (malignant) and HUVEC (non-malignant) cell lines. Intracellular lactate dehydrogenase (LDH) leakage was evaluated as an indicator of membrane integrity. In vitro assessments revealed that Pluronic F127/DOX microemulsions caused substantial morphological changes and greater cytotoxic effects than standard DOX. Pluronic F127/DOX microemulsions were injected intraperitoneally at 12 and 24 mg/kg into rats. The free (bulk) DOX group induced severe histopathological changes and significant increases in serum kidney markers and serum liver enzymes. The 24 mg/kg dose of Pluronic F127/DOX microemulsions also induced fatty changes and elevation of serum liver enzymes and creatinine. Overall, this new drug delivery system formulation shows promise for cancer treatment and deserves to be further studied in the future.

Anticancer effects against colorectal cancer models of chloro(triethylphosphine)gold(I) encapsulated in PLGA\u2013PEG nanoparticles

Chloro(triethylphosphine)gold(I), (Et3PAuCl hereafter), is an Auranofin (AF)-related compound showing very similar biological and pharmacological properties. Like AF, Et3PAuCl exhibits potent antiproliferative properties in vitro toward a variety of cancer cell lines and is a promising anticancer drug candidate. We wondered whether Et3PAuCl encapsulation might lead to an improved pharmacological profile also considering the likely reduction of unwanted side-reactions that are responsible for adverse effects and for drug inactivation. Et3PAuCl was encapsulated in biocompatible PLGA–PEG nanoparticles (NPs) and the new formulation evaluated in colorectal HCT-116 cancer cells in comparison to the free gold complex. Notably, encapsulated Et3PAuCl (nano-Et3PAuCl hereafter) mostly retains the cellular properties of the free gold complex and elicits even greater cytotoxic effects in colorectal cancer (CRC) cells, mediated by apoptosis and autophagy. Moreover, a remarkable inhibition of two crucial signaling pathways, i.e. ERK and AKT, by nano-Et3PAuCl, was clearly documented. The implications of these findings are discussed.

Inclusion complex of clausenidin with hydroxypropyl-β-cyclodextrin: Improved physicochemical properties and anti-colon cancer activity

The long-term objective of the present study was to prepare, physicochemically characterize and determine the anticancer of clausenidin/hydroxypropyl-β-cyclodextrin (Clu/HPβCD) inclusion complex. We used differential scanning calorimetry, X-ray diffractometer, fourier transform infrared spectroscopy, ultraviolet–visible spectrophotometer and 13C and 1H nuclear magnetic resonance followed by in vitro anticancer assays. The orientation and intermolecular interactions of Clausenidin within cyclodextrin cavity were also ascertained by molecular docking simulation accomplished by AutoDock Vina. The guest molecule was welcomed by the hydrophobic cavity of the host molecule and sustained by hydrogen bond between host/guest molecules. The constant drug release with time, and increased solubility were found after successful complexation with HPβCD as confirmed by physicochemical characterizations. Clausenidin had greater cytotoxic effect on colon cancer HT29 cells when incorporated into HPβCD cavity than dissolved in DMSO. Also, from a comparison of cell viability between normal and cancer cells, a reduced side effect was observed. The Clu/HPβCD inclusion complex triggered reactive oxygen species-mediated cytotoxicity in HT29 cells. The inclusion complex-treated HT29 cells showed cell cycle arrest and death by apoptosis associated with caspases activation. The presence of HPβCD seems to aid the anticancer activity of clausenidin.