In vivo evaluation of Sono-chemo therapy via hollow gold nanoshells conjugated to mitoxantrone on breast cancer.

Objectives: Conventional methods of cancer treatment include surgery, chemotherapy, radiation therapy, and immunotherapy. Chemotherapy, as one of the main methods of cancer treatment, due to the lack of targeted distribution of the drug in tumor tissues, is not able to destroy cancer cells and also affects healthy tissues and causes serious side effects in patients. Sonodynamic therapy (SDT) is a promising strategy for non-invasive treatment of deep solid cancer tumors. In this study, for the first time, the sono-sensitive activity of mitoxantrone was investigated and then mitoxantrone (MTX) was conjugated to hollow gold nanostructure (HGN) to improve the efficiency of in vivo SDT. Materials and Methods: Firstly, after the synthesis of hollow gold nanoshells and the PEGylation process, conjugation of MTX was performed. Then, after evaluating the toxicity of the treatment groups in vitro, in order to perform an in vivo study, 56 male Balb/c mice that had been tumorized by subcutaneous injection of 4T1 cells were divided into eight groups of breast tumor model. Ultrasonic irradiation (US) conditions including intensity of 1.5 W/cm2 (with a frequency of 800 kHz, 5 min), MTX concentration of 2 µM, and HGN dose of 2.5 mg/kg (unit of animal weight) were used. Results: The results show that administration of PEG-HGN-MTX caused a slight reduction in tumor size and growth compared with free MTX. Ultrasound also improved the therapeutic effect of the gold nanoshell in treated groups, and the HGN-PEG-MTX-US treated groups were able to significantly reduce and control tumor size and growth. Conclusion: The findings also show that MTX and HGN can be used as sonosensitizers in SDT. Also, HGN-PEG-MTX can act as a sono-chemotherapy agent for the combination of sonodynamic therapy and chemotherapy for in vivo breast tumors.

Removal of paraquat from aqueous solutions by plant extracts as an ecofriendly approach.

Paraquat is a very water-soluble herbicide widely used in agricultural and nonagricultural lands. The removal of paraquat from polluted water is very essential. Hence, three experiments were arranged separately to investigate the ability of plant extracts include Aloe vera, Portulaca grandiflora, kalanchoe daigremontiana, Sedum rubotinctum, Brassica rapa var. rapa, Helxine soleirolii, Brassica oleracea var. capitata, Crassula ovate, Aptenia cordifolia, Tradescantia albiflora to the removal of paraquat (PQ) from water. In the 1st experiment, High-performance liquid chromatography was used to determine the removal rate of PQ by using plant extracts. In the 2nd and 3rd experiments, PQ solutions were subjected to plant extracts treatments and then spray on the leaves of littleseed canarygrass (Phalaris minor) under greenhouse and field conditions, respectively. In general, the results of these experiments revealed that all plant extracts resulted in higher PQ removal from water compared to control. The maximum removal of PQ was observed at pH 11, 25 °C, 30 Min, and 0.2 mL in the presence of plant extracts. In the presence of P. grandiflora, C. ovate, k. daigremontiana, A. cordifolia, H. soleirolii, Aloe vera, S. rubotinctum, B. oleracea var. capitata, B. rapa var. rapa, and T. albiflora plant extracts, PQ removal from water increased to 68.34%, 65.45%, 63.97%, 59.81%, 59.29%, 55.44%, 52.06%, 50.34%, 48.86%, and 46.44% under normal conditions and 79.71%, 78.61%, 78.00%, 75.83%, 74.66%, 72.12%, 71.57%, 71.02%, 69.35%, and 68.73% under optimum conditions, respectively. Results of the 2nd and 3rd experiments were demonstrated a decrease in PQ performance on littleseed canarygrass control. It can be concluded that plant extracts can reduce the residual of PQ in water bodies.

Docetaxel encapsulation in nanoscale assembly micelles of folate-PEG-docetaxel conjugates for targeted fighting against metastatic breast cancer in vitro and in vivo.

Due to the high frequency and mortality of breast cancer, developing efficient targeted drug delivery systems for frightening against this malignancy is among cancer research priorities. The aim of this study was to synthesize a targeted micellar formulation of docetaxel (DTX) using DTX, folic acid (FA) and polyethylene glycol (PEG) conjugates as building blocks. In the current study, two therapeutic polymers consisting of FA-PEG-DTX and PEG-DTX conjugates were synthesized and implemented to form folate-targeted and non-targeted micelles. Dissipative particle dynamics (DPD) method was used to simulate the behavior of the nanoparticle. The anti-cancer drug, DTX was loaded in to the micelles via solvent switching method in order to increase its solubility and stability. The cytotoxicity of the targeted and non-targeted formulations was evaluated against 4T1 and CHO cell lines. In vivo therapeutic efficiency was studied using ectopic tumor model of metastatic breast cancer, 4T1, in Female BALB/c mice. The successful synthesis of therapeutic polymers, FA-PEG-DTX and PEG-DTX were confirmed implementing 1HNMR spectral analysis. The size of DTX-loaded non-targeted and targeted micelles were 176.3 ± 8.3 and 181 ± 10.1 nm with PDI of 0.23 and 0.17, respectively. Loading efficiencies of DTX in non-targeted and targeted micelles were obtained to be 85% and 82%, respectively. In vitro release study at pH = 7.4 and pH = 5.4 showed a controlled and continuous drug release for both formulations, that was faster at pH = 5.4 (100% drug release within 120 h) than at pH = 7.4 (80% drug release within 150 h). The targeted formulation showed a significant higher cytotoxicity against 4T1 breast cancer cells (high expression of folate receptor) within the range of 12.5 to 200 µg/mL in comparison with no-targeted one. However, there was no significant difference between the cytotoxicity of the targeted and non-targeted formulations against CHO cell line as low-expressed cell line. In accordance with in vitro investigation, in vivo studies verified the ideal anti-tumor efficacy of the targeted formulation compared to Taxotere and non-targeted formulation. Based on the obtained data, FA-targeted DTX-loaded nano-micelles significantly increased the therapeutic efficacy of DTX and therefore can be considered as a new potent platform for breast cancer chemotherapy.

A comparative study on generating hydroxyl radicals by single and two-frequency ultrasound with gold nanoparticles and protoporphyrin IX.

Sonodynamic therapy (SDT) is a new manner of killing cancer cells based on the cytotoxic interactions of ultrasound with sonosensitizing agents. It is shown that gold nanoparticles (GNPs) increase the efficiency of cavitation activity of ultrasound. In this study the influence of a single and/or two frequencies of ultrasound waves to generate hydroxyl radicals (·OH) was assessed in the presence of protoporphyrin IX (PpIX) and/or GNPs. Ultrasound cavitation activity was determined by recording fluorescence signals from chemical terephthalic acid (TA) dosimeters with or without PpIX and/or GNPs at the frequencies of 0.8 and 2.4 MHz individually and aggregately. To study hydroxyl radicals, experiments were performed with and without hydroxyl radical scavengers mannitol, histidine, and sodium azide. Cavitation activity was amplified by increasing ultrasound intensity and exposure time. The cavitation activity induced by dual ultrasound frequency was remarkably higher than the summation of effects produced by individual frequencies. All three scavengers reduced the fluorescence signal level. The effect of GNPs on intensifying cavitation activity at higher frequency was greater than that of lower frequency. PpIX showed a more effective sonosensitizing property at the lower frequency. Also, estimated synergism at dual frequency irradiation was improved in the presence of GNPs. We found that GNPs increased hydroxyl radical production at 2.4 MHz and that PpIX increased hydroxyl radical production at 0.8 MHz. Dual frequency exposure was more effective than single frequency exposure. PpIX at low frequency and gold nanoparticles at high frequency both enhance sonodynamic treatment efficacy.

Sonophotodynamic therapy mediated by liposomal zinc phthalocyanine in a colon carcinoma tumor model: Role of irradiating arrangement.

OBJECTIVES: Low penetration depth of light is the main defect of photodynamic therapy (PDT), which could be improved by sonodynamic therapy (SDT). In this study, a combination of PDT and SDT known as sonophotodynamic therapy (SPDT) was investigated using two reverse arrangements in CT26 tumor model. MATERIALS AND METHODS: The liposomal zinc phthalocyanine was synthesized and characterized. It was then administered to CT26 tumor models as a sensitizer. The animal models were subjected to PDT, SDT, and the combined treatment in different groups. The doubling time for the survival of tumors and animals was considered as a measure to evaluate treatments efficacy. RESULTS: In all treatment groups there was a significant decline in tumor volume 15 days after treatment compared to the main control group, but the optimum response was observed in the group receiving a combined treatment with the priority of PDT. 120 days after treatment, in the groups treated by PDT and SDT, the tumor shrank by 20%, while in the group receiving SPDT with PDT priority, 80% of tumors was recovered. No case of complete tumor progression was observed in SPDT group with SDT priority. This could be due to the pores created in cell membranes during ultrasound irradiation of the tumor, which removed the sensitizer molecules from the cells and reduced PDT efficacy in SPDT group with SDT priority. CONCLUSION: It seems that SPDT with PDT priority offers a more efficient alternative than each of PDT, SDT individually or SPDT with the reverse arrangement.

A simple, sensitive and rapid isocratic reversed-phase high-performance liquid chromatography method for determination and stability study of curcumin in pharmaceutical samples.

OBJECTIVE: This study was designed to develop and validate a new reversed-phase high-performance liquid chromatography (RP-HPLC) method based on Q2 (R1) International Conference on Harmonization (ICH) guideline for determination of curcumin in pharmaceutical samples. MATERIALS AND METHODS: The HPLC instrument method was optimized with isocratic elution with acetonitrile: ammonium acetate (45:55, v/v, pH 3.5), C18 column (150 mm×4.6 mm×5 µm particle size) and a flow rate of 1 ml/min in ambient condition and total retention time of 17 min. The volume of injection was set at 20 µl and detection was recorded at 425 nm. The robustness of the method was examined by changing the mobile phase composition, mobile phase pH, and flow rate. RESULTS: The method was validated with respect to precision, accuracy and linearity in a concentration range of 2-100 µg/ml. The limit of detection (LOD) and limit of quantification (LOQ) were 0.25 and 0.5 µg/ml, respectively. The percentage of recovery was 98.9 to 100.5 with relative standard deviation (RSD) < 0.638%. CONCLUSION: The method was found to be simple, sensitive and rapid for determination of curcumin in pharmaceutical samples and had enough sensitivity to detect degradation product of curcumin produced under photolysis and hydrolysis stress condition.

Combined multispectroscopic and molecular dynamics simulation investigation on the interaction between cyclosporine A and ß-lactoglobulin.

ß-Lactoglobulin (ß-LG), the major whey protein in milks of many mammals, has a high affinity for a wide range of compounds. Cyclosporine A (CsA), is an immunosuppressant drug mainly prescribe in organ transplantation to prevent rejection. In this study, the interaction of CsA with ß-LG was investigated using various spectroscopic techniques (UV-visible and fluorescence) in an aqueous medium at two temperatures of 298 and 310K in combination with a molecular dynamics simulation study. The titration results indicated that CsA quenched the fluorescence intensity of ß-LG through a static mechanism. The binding constants for the binding of CsA to ß-LG at two different temperatures 298 and 310K were obtained 1.12×105 and 0.87×105M-1, respectively. Thermodynamic data indicated that the hydrophobic interactions and hydrogen bonds dominate in the binding site. Results of fluorescence resonance energy transfer (FRET) measurements suggest that resonance energy transfer occurs between ß-LG and CsA. Moreover, MD simulation results implied that CsA can interact with ß-LG, without affecting the secondary structure of ß-LG. Experimental and MD simulations data reciprocally supported each other.

Dextran-poly lactide-co-glycolide polymersomes decorated with folate-antennae for targeted delivery of docetaxel to breast adenocarcinima in vitro and in vivo.

In this study, a tumor-targeted, docetaxel encapsulated dextran-poly lactide-co-glycolide (DEX-PLGA) polymersomes was fabricated for breast cancer chemotherapy. To attain active cancer targeting, docetaxel encapsulated polymersomes was conjugated with folate for folate receptor guided delivery which is overexpressed in various cancer cells including breast adenocarcinoma. Docetaxel was encapsulated in the bilayer of DEX-PLGA or folate-conjugated DEX-PLGA (FA-DEX-PLGA) polymersomes via nanoprecipitation method in order to provide the controlled drug release. The size of polymersomes obtained were 178.53±2.5nm and offered drug encapsulation efficiency and loading content of 78.85±3.81% and 9.32±0.27, respectively. The data demonstrate that the prepared polymersome formulations sustained docetaxel release for a period of 6days. Cellular uptake study and MTT assay showed that the developed folate-targeted docetaxel-loaded polymersomes had higher cytotoxicity than non-targeted ones as well as free form of drug and was accumulated in 4T1 and MCF-7 cells in vitro. The in vivo tumor inhibitory effect of folate-conjugated docetaxel-loaded DEX-PLGA polymersomes (FA-DEX-PLGA-DTX-NPs) demonstrated an increased therapeutic potency of targeted formulation over both non-targeted (DEX-PLGA-DTX-NPs) and free drug. The represented data reveal that the prepared targeted drug delivery system was able to control the docetaxel release, and also enhance the antitumor potency of docetaxel. This study may open new direction for preparation of folate receptor targeted polymersomes based on DEX-PLGA copolymers for translational purposes.

Investigation into the interaction of losartan with human serum albumin and glycated human serum albumin by spectroscopic and molecular dynamics simulation techniques: A comparison study.

The interaction between losartan and human serum albumin (HSA), as well as its glycated form (gHSA) was studied by multiple spectroscopic techniques and molecular dynamics simulation under physiological conditions. The binding information, including the binding constants, effective quenching constant and number of binding sites showed that the binding partiality of losartan to HSA was higher than to gHSA. The findings of three-dimensional fluorescence spectra demonstrated that the binding of losartan to HSA and gHSA would alter the protein conformation. The distances between Trp residue and the binding sites of the drug were evaluated on the basis of the Förster theory, and it was indicated that non-radiative energy transfer from HSA and gHSA to the losartan happened with a high possibility. According to molecular dynamics simulation, the protein secondary and tertiary structure changes were compared in HSA and gHSA for clarifying the obtained results.

Progress in the development of lipopolyplexes as efficient non-viral gene delivery systems.

Efficient gene therapy is mainly dependent on the gene transfer capability of gene delivery vectors. Non-viral vectors have become the research interest of many researchers because these vectors are safer than viral vectors. Acquiring the advantages of both polyplexes and lipoplexes, the lipopolyplex (LPP) is a ternary nanocomplex composed of cationic liposome, polycation, and nucleic acid. Considering the polycationic component, ternary complexes (LPPs) are divided into cationic polymer-based LPPs and cationic peptide-based LPPs. Considering the capability of rational design, LPP is an interesting field of research to design a more potent nucleic acid carrier. With the promising transfection activity and safety observed in the LPPs, many researchers have formulated various types of lipids and polycations to achieve an efficient and safe carrier for gene therapy. Here we provide a review on the designed LPPs for efficient delivery of different nucleic acids such as plasmid DNA, siRNA, shRNA, and DNA vaccines.

Nicotine content of domestic cigarettes, imported cigarettes and pipe tobacco in iran.

BACKGROUND: There are many different kinds of cigarettes and tobacco available in the market. Since nicotine content of various brands of cigarettes are very variable, therefore evaluation and comparison of nicotine content of different brands of cigarettes is important. The goal of the present study was to determine and compare nicotine content of various domestic and imported cigarettes available in the area. METHODS: Fourteen popular imported brands and nine popular domestic brands of cigarettes and three available brands of tobaccos were investigated for the amounts of nicotine content. Nicotine was extracted from each cigarette and tobacco samples and was analyzed by high performance liquid chromatography (HPLC) method. FINDINGS: The amount of nicotine in each cigarette was from 6.17 to 12.65 mg (1.23 ± 0.15 percent of tobacco weight in each cigarette) in domestic cigarettes. It was between 7.17-28.86 mg (1.80 ± 0.25 percent of tobacco weight in each cigarette) for imported cigarette, and between 30.08- 50.89 mg (3.82 ± 1.11 percent) for the pipe nicotine. There was significant difference in nicotine amount between imported and domestic brands of cigarettes. There was also no significant difference in nicotine content between light and normal cigarettes in imported brands. CONCLUSION: Nicotine content of all tested cigarettes, imported and domestic brands, were higher than the international standard.

A novel nanosonosensitizer for sonodynamic therapy: in vivo study on a colon tumor model.

OBJECTIVES: The particles in a liquid decrease the ultrasonic intensity threshold needed for cavitation onset. In this study, a new nanoconjugate composed of protoporphyrin IX and gold nanoparticles was used as a nucleation site for cavitation. The nonradiative relaxation time of protoporphyrin IX in the presence of gold nanoparticles is longer than the similar time without gold nanoparticles. METHODS: This study was conducted on colon carcinoma tumors in BALB/c mice. The tumor-bearing mice were randomly divided into 6 groups (each containing 15 mice): (1) control, (2) protoporphyrin IX, (3) gold nanoparticle-protoporphyrin IX conjugate, (4) ultrasound alone, (5) ultrasound + protoporphyrin IX, and (6) ultrasound + gold nanoparticle-protoporphyrin IX conjugate. In the respective groups as indicated above, protoporphyrin IX or the gold nanoparticle-protoporphyrin IX conjugate was injected into the tumors. Ultrasound irradiation was performed on the tumors 24 hours after injection. Antitumor effects were estimated by evaluation of the relative tumor volume, doubling time, and 5-folding time for the tumors after treatment. The cumulative survival fraction of the mice and percentage of the lost tissue volume (treated) were also assessed in the different groups. RESULTS: A significant difference in the average relative volumes of the tumors 13 days after treatment was found between the ultrasound + gold nanoparticle-protoporphyrin IX group and the other groups (P < .05). The longest doubling and 5-folding times were observed in the ultrasound + gold nanoparticle-protoporphyrin IX and ultrasound + protoporphyrin IX groups. CONCLUSIONS: Protoporphyrin IX conjugated to gold nanoparticles has been introduced as a promising compound and a new sonosensitizer for improving the tumor response to sonodynamic therapy by reducing the relative tumor volume and increasing the cumulative survival fraction.