Viral vector mimicking and nucleus targeted nanoparticles based on Dexamethasone conjugated Polyethylenimine nano-liposomes: Preparation and evaluation of transfection efficiency

Non-viral vectors such as polymers and liposomes have been used for gene delivery systems to overcome intrinsic problems of viral vectors, but transfection efficiency of these vectors is lower than viral vectors. In the present study, we conjugated dexamethasone (Dexa) to branched polyethylenimine (b-PEI), in combinations with cationic liposomes as a nonviral system to improve transfection efficiency. Cationic liposomes containing DOTAP and cholesterol were prepared by thin-film hydration following extrusion method. Dexamethasone mesylate was synthesized and conjugated to polyethylenimine by a one-step reaction. A novel gene delivery system, Lipopolyplex was developed by premixing liposome and different Mw of bPEI-Dexa as carriers following by addition of plasmid at three different C/P ratios. The resultant complexes were characterized for their size, zeta potential and ability of DNA condensation. Luciferase reporter gene was used for determination of transfection efficiency in neuro2A cells. Also, the toxicity of gene carriers was investigated in this cell line. Mean particle size of prepared complexes was less than 200 nm and there was no significant difference in size by increasing molecular weight of PEIs. All complexes had positive surface charge. Complete condensation of DNA was occurred at C/P ratio of 1 for all complexes. lipopolyplexes were more efficient than polyplexes and lipoplexes alone and transfection efficiency was improved by adding dexamethasone. Complexes containing liposome, PEI 10 kDa and dexamethasone had the highest transfection activity. Furthermore, non-viral vectors described in this study showed low cytotoxicity. The results of this study confirm that PEI in combination with liposome as lipopolyplex have low toxicity and may enhance transfection efficiency. Moreover, conjugation of dexamethasone to PEI, in combination with cationic liposome might be useful for the gene deliver

Recent Advances in Lung Cancer Therapy Based on Nanomaterials: A Review

Lung cancer is one of the commonest cancers with a significant mortality rate for both genders, particularly in men. Lung cancer is recognized as one of the leading causes of death worldwide, which threatens the lives of over 1.6 million people every day. Although cancer is the leading cause of death in industrialized countries, conventional an-ticancer medications are unlikely to increase patients' life expectancy and quality of life significantly. In recent years, there are significant advances in the development and applications of nanotechnology in cancer treatment. The superiority of nanostructured approaches is that they act more selectively than traditional agents. This progress led to the development of a novel field of cancer treatment known as nanomedicine. Various formulations based on nanocarriers, including lipids, polymers, liposomes, nanoparticles and dendrimers have opened new horizons in lung cancer therapy. The application and expan-sion of nano-agents lead to an exciting and challenging research era in pharmaceutical science, especially for the delivery of emerging anti-cancer agents. The objective of this review is to discuss the recent advances in three types of nanoparticle formulations for lung cancer treatments modalities, including liposomes, polymeric micelles, and den-drimers for efficient drug delivery. Afterward, we have summarized the promising clinical data on nanomaterials based therapeutic approaches in ongoing clinical studies

Cationic Liposomes Modified with Polyallylamine as a Gene Carrier: Preparation, Characterization and Transfection Efficiency Evaluation

Purpose: Cationic polymers and cationic liposomes have shown to be effective non-viral gene delivery vectors. In this study, we tried to improve the transfection efficiency by employing the advantages of both. Methods: For this purpose, modified polyallylamines (PAAs) were synthesized. These modifications were done through the reaction of PAA (15 KDa) with acrylate and 6-bromoalkanoic acid derivatives. Liposomes comprising of these cationic polymers and cationic lipid were prepared and extruded through polycarbonate filters to obtain desired size. Liposome-DNA nanocomplexes were prepared in three carrier to plasmid (C/P) ratios. Size, zeta potential and DNA condensation ability of each complex were characterized separately and finally transfection efficiency and cytotoxicity of prepared vectors were evaluated in Neuro2A cell line. Results: The results showed that mean particle size of all these nanocomplexes was lower than 266 nm with surface charge of 22.0 to 33.9 mV. Almost the same condensation pattern was observed in all vectors and complete condensation was occurred at C/P ratio of 1.5. The lipoplexes containing modified PAA 15 kDa with 10% hexyl acrylate showed the highest transfection efficacy and lowest cytotoxicity in C/P ratio of 0.5. Conclusion: In some cases nanocomplexes consisting of cationic liposome and modified PAA showed better transfection activity and lower cytotoxicity compared to PAA

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

Curcumin improved liposomal mitomycin-induced cell toxicity in bladder cancer cell

Objective (s): This study aimed at investigation of preparation and stability study of Mitomycin and Mitomycin loaded nanoliposems and efficacy of the combination of Mitomycin -loaded nanoliposomes and Curcumin versus Mitomycin in suppressing HTB-9 and L929 cell lines, in vitro.Materials and Methods: An HPLC method was validated based on Q2 (R1) International Conference on Harmonization (ICH) guideline for determination of Mitomycin in pharmaceutical samples. Soybean phosphatidylcholine (SPC) or hydrogenated soybean phosphatidylcholine (HSPC) mixed with cholesterol at 2:1 molar ratio respectively in two different groups to prepare Mitomycin nanoliposomes. Cell toxicity of free Mitomycin, Mitomycin loaded liposomes and curcumin was measured using MTT assay in bladder cancer cell line for treatment group and Fibroblast-like cell as control group. Also, propidium iodide staining was done to determine the level of DNA fragmentation.Results: The validated HPLC method showed stability based on the ICH guideline (n=3 and RS

Evaluation of leishmanicidal effect of Euphorbia petiolata extract by in vivo anti-leishmanial assay using promastigotes of Leishmania major

Objective: The extract of different species of Euphorbia has been successfully used as a remedy for treatment of cutaneous leishmaniasis. The aim of this study was to assess the in vitro leishmanicidal effect of Euphorbia petiolata (E. petiolata) extract. Materials and Methods: Ethanolic percolated and methanolic Soxhlet extract of E. petiolata on promastigotes of L. major at different concentrations of extracts, one positive control group and one negative control group as well as 1 solvent control were prepared and placed in 24-well plates that contained 40,000 parasites/well. Afterwards, plates were incubated at 25 ˚C for six days and number of parasites in each well were determined on days 2, 4 and 6 of the experiment. Results: Both percolated and Soxhlet extracts in methanol and DMSO solvents had significant effects (equal to that of amphotericin B) on promastigote form of parasite at the concentration of 1 mg/ml. At lower concentrations, the extracts of E. petiolata had favorable leishmanicidal activity and killed L. major promastigotes dose-dependently. Conclusion: Our results support the possibility of E. petiolata extracts application as an anti-leishmanial agent with similar effects to amphotericin B

Chemical composition, antimicrobial activity and antiviral activity of essential oil of Carum copticum from Iran

Objectives: Evaluation of therapeutic effects of Carum copticum (C. copticum) has been the subject of several studies in recent years. Thymol the major component of C. copticum is a widely known anti-microbial agent. In this study, the antibacterial and anti viral activities of essential oil of C. copticum fruit were determined. Materials and Methods: Essential oil of C. copticum was analyzed by means of gas chromatography-mass spectrometry (GC-MS). The antimicrobial activity of the oil was evaluated against six Gram (+/-) bacteria and fungi, using the micro broth dilution technique. Antiviral activity of the essential oil was evaluated using a Bacillus phage CP51. Results: From the ten identified constituents, representing 98.7% of the oil, thymol (72.3%), terpinolene (13.12%) and o-cymene (11.97%) were the major components. It was found that the oil exhibited strong antimicrobial activity againstStaphylococcus aureus (S. aureus) andBacillus subtilis (B. subtilis) (MIC, 0.00025% v/v). Furthermore, the antiviral activity of the oil was evaluated by plaque reduction assay. Conclusion: The essential oil showed an antiviral activity against phage when phage was pre-incubated with the essential oil prior to its exposure to B. cereus and without any pre-incubation with the phage, suggesting that the oil directly inactivated virus particles

Evaluation of leishmanicidal effect of Euphorbia erythadenia extract by in vitro leshmanicidal assay using promastigotes of Leishmania major

Objective: To evaluate leishmanicidal effects of Euphorbia erythadenia plant extract. Methods: Extraction was done using methanolic Soxhlet of dried and ground aerial parts of the plant. Then, five different extract concentrations, in addition of positive, negative and solvent controls were prepared and added to a 24-well plate containing 40 000 parasites/well. The extract concentrations were 1, 0.5, 0.25, 0.125 and 0.062 5 mg/mL. Amphotricin B (0.5 mg/mL) was used as positive control while negative control contained only culture medium. After 3 d incubation at 25 °C the amount of parasites in each well was determined on each day of experiment microscopially using Neubar chamber. Results: Soxhlet extract as well as amphotricin B killed all parasites at concentration of 1 mg/mL. The leshmanicidal activity of lower doses of extract was dose-dependent. The EC50 for Soxhlet extracts in dimethylsulfoxide was 0.30 mg/mL. The EC50 for Soxhlet extracts in methanol was 0.23 mg/mL. No obvious effects from the control solvent on the Leishmania major promastigotes were observed. Conclusions: The Soxhlet extract of Euphorbia erythadenia showed suitable leishmanicidal activity, especially in higher concentration fractions