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Ellagic acid as a polyphenol or micronutrient which can be naturally found in different vegetables and fruits, have gained considerable attention for cancer therapy due to considerable biological activities and different molecular targets. Ellagic acid with low hydrolysis and lipophilic and hydrophobic nature is not able to be absorbed in circulation. So, accumulation inside the intestinal epithelial cells or metabolization to other urolithins lead to limitation of direct evaluation of EA effects in clinical studies. This review focuses on the studies which supported anticancer activity of pure or fruit-extracted ellagic acid through in vitro, in vivo, in silico and drug delivery methods. The results demonstrate ellagic acid modulates the expression of various genes incorporated in cancer-related process of apoptosis and proliferation, inflammation related-gens and oxidative-related gens. Moreover, ellagic acid formulation in carriers composed of lipid, silica, chitosan, iron- bovine serum albumin nanoparticles obviously enhanced the stable release and confident delivery with minimum loss. Also, in silico analysis proved that ellagic acid was able to be placed at position of co-crystal ADP, in the deep cavity of protein target, and tightly interact with binding pocket residues leading to suppression of substrate availability of protein and its activation inhibition. This article is protected by copyright. All rights reserved.

The principal impediment to gene therapy is the development of efficient, nontoxic gene carriers that can handle and deliver foreign genetic materials into various cell types, including healthy and cancerous cells. Poly-l-lysine (PLL) polymers are one of the most favorable gene carriers among nonviral vectors, and PLL had low transfection and safety issues. The purpose of this study was to measure cellular toxicity, DNA damage, and apoptotic effects of PLL nanoparticles. Neuro2A mammalian cells were cultured and exposed to PLL/DNA complexes at different polymer/DNA ratios ( C/ P ratio 2 and 6) for 24 h. To evaluate metabolic activity, genotoxicity, and apoptotic influences of PLL nanoparticle, the following experimental methods were employed, in order: 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), DNA damage (COMET analysis) assay, and sub-G1 peak apoptosis assay. Our data indicate that toxicity is concentration dependent and a high concentration of polymer declined the metabolic activity. In addition, largest complexes ( C/ P 6 in HEPES buffered saline buffer) have slighter negative impact on metabolic activity. In agreement with our cytotoxicity data, apoptotic assay result represented that increase in size of PLL/DNA complexes decrease the number of apoptotic cells. Also, there was a remarkable increase in percent tail DNA of Neuro2A cells treated with higher concentration of PLL and its polyplexes. The present study demonstrated that PLL/DNA complexes caused cytotoxic, apoptotic, and genotoxic effects in a dose-dependent and weight ratio-dependent manner, which also affected the size of polyplexes.

Cerium oxide nanoparticles (CNPs) with desired particle size and spherical morphology were prepared from cerium nitrate in bio media of Salvia macrosiphon Boiss seeds extract, as a green synthesis route. Then they were characterized by XRD, UV–Vis and FTIR spectroscopies, FESEM and TGA. Band gap energy of the prepared powders was also determined which was found in the range of 2.5–3.5 eV. Determination of DLS and zeta potential were showed that CNPs had the small size and unique colloidal stability, respectively. Then the photo-catalytic activity of them was investigated by degradation of Rhodamine B (RhB) dye as a model of waste water pollutants, under UV-irradiation and optimum conditions were evaluated. Results showed that decreasing the particle size increased the rate of photo-catalytic reaction remarkably but ascending the band gap energy, in contrast. The photo-catalytic mechanism was also studied by using different scavengers.

Aims The in vivo targeted diagnostic applications of biosynthetic Cerium oxide nanoparticles (CeO2-NPs), prepared by applying chitosan as a stabilizer, was explored by evaluating the cytotoxicity through MTT assay on WEHI 164 cell line, the Hemolytic activity of CeO2-NPs and biodistribution in rats. Main methods The CeO2-NPs were characterized through the use of TGA/DTG, PXRD, FESEM, FTIR, and UV–Vis spectroscopy. The biodistribution of CeO2-NPs were determined by directly labeled nanoparticles with Technetium-99 m (99mTc) radioisotope (99mTc-CeO2-NPs). The labeling efficiency and stability of 99mTc-CeO2-NPs were also measured with Instant Thin Layer Chromatography (ITLC) method. The saturation study was investigated by 1 mCi of 99mTc-CeO2-NPs using different concentrations of WEHI 164 cells after 4 h of incubation. In vivo biodistribution study was performed by intravenous injection of 600 μCi/200 μL 99mTc-CeO2-NPs through rat's tail. Key findings CeO2-NPs seemed to have a low cytotoxic effect on WEHI 164 cell line and did not result in hemolysis. The biodistribution of CeO2-NPs has shown that a huge amount of 99mTc-CeO2-NPs was amassed in the living human organs, including liver, lung, spleen, stomach, and thyroid which shows the in vivo stability of the labeled conjugate. Herein, we have developed a facile, economical, and greener synthetic procedure applying Chitosan template. This green approach is comparable to conventional methods that utilize hazardous materials which are would be a suitable alternative to circumvent synthetic issues related to these materials. Significance The bio-applications of nano-sized CeO2-NPs were explored to find new horizon to use nanotechnology as the diagnostic tool.

Carrageenan hydrogel as a “greener” and a vegetable-based stabilizing agent has the potential for many biosyntheses of different nanoparticles by sol-gel method. Herein, we describe for the first time an economic and eco-friendly preparation of cerium oxide nanoparticles (CeO2-NPs) using carrageenan. When carrageenan hydrogel comes in contact with a cerium nitrate solution, cerium ions anchor themselves to the –SO3- groups into the carrageenan and after the gelation process, have fewer opportunities escaping from the polymeric network. The CeO2-NPs were well-prepared and successfully characterized by PXRD, FTIR, FESEM, UV–Vis, and TGA-DTA. The calcined CeO2-NPs showed strong UV absorption (λmax = 328 nm) with the calculated band gap of 2.69 eV. The results obtained from FESEM images indicate that CeO2-NPs obtained at 600 °C ranges from 18 to 60 nm and have a mean diameter of ~34 nm. The in vitro cytotoxicity study on WEHI 164 cell line has mentioned low toxic and non-toxic CeO2-NPs in a range of concentrations (0.97–250 μg/ml), thus, we reckon that the greener synthesized CeO2-NPs will have persistent utilization in various fields of medical applications.

Context: Polypropylenimine (PPI), a cationic dendrimer with defined structure and positive surface charge, is a potent non-viral vector. Dexamethasone (Dexa) conveys to the nucleus through interaction with its intracellular receptor. Objective: This study develops efficient and non-toxic gene carriers through conjugation of Dexa at various percentages (5, 10 and 20%) to the fourth and the fifth generation PPIs (PPIG4s and PPIG5s). Materials and methods: The 21-OH group of Dexa (0.536 mmol) was modified with methanesulfonyl chloride (0.644 mmol) to activate it (Dexa-mesylate), and then it was conjugated to PPIs using Traut's reagent. After dialysis (48 h) and lyophilization, the physicochemical characteristics of products (PPI-Dexa) including zeta potential, size, buffering capacity and DNA condensing capability were investigated and compared with unmodified PPIs. Moreover, the cytotoxicity and transfection activity of the Dexa-modified PPIs were assessed using Neuro2A cells. Results: Transfection of PPIG4 was close to PEI 25 kDa. Although the addition of Dexa to PPIG4s did not improve their transfection, their cytotoxicity was improved; especially in the carrier to DNA weight ratios (C/P) of one and two. The Dexa conjugation to PPIG5s enhanced their transfection at C/P ratio of one in both 5% (1.3-fold) and 10% (1.6-fold) Dexa grafting, of which the best result was observed in PPIG5-Dexa 10% at C/P ratio of one. Discussion and conclusions: The modification of PPIs with Dexa is a promising approach to improve their cytotoxicity and transfection. The higher optimization of physicochemical characteristics, the better cell transfection and toxicity will be achieved.

Aims Gene therapy is a promising strategy for the treatment of various diseases. Polyethylenimine (PEI) has received considerable attention for gene delivery applications due to their appropriate properties. However, their toxicity has raised concerns which cause to be used with cautious. This study aimed to prepare different complexes of PEI/DNA and evaluate their parameters affecting in vitro cytotoxicity. Also, apoptosis rate was measured to determine the mechanism of cell toxicity. Materials and methods The complexes were prepared through conjugation and characterized using dynamic light scattering, MTT and flow cytometry techniques. Key findings The particles' size was from 81 nm to 2785 nm and was increased in the HBS buffer compared to HBG buffer. In the case of branched PEIs, the size of particles was inversely associated with molecular weight. The cytotoxicity results showed that linear 250 KDa PEI was non-toxic whereas branched PEIs with lower molecular weights showed toxicity effects in a concentration dependent manner. Also, the cytotoxicity effects of branched PEIs were proportional with carrier/plasmid (C/P) ratio and were more for the polyplexes prepared in HBG buffer compared to HBS buffer after 24 h incubation. Flow cytometry results confirmed that apoptosis is the main mechanism of cell toxicity produced by polyplexes. Significance The results showed the effect of PEI size on its cytotoxicity. Also, the toxicity effects of PEI-derived polyplexes in vivo environment was evaluated.

With high potency in treatment of various diseases, gene therapy is mainly hindered by lack of safe and efficient gene delivery vectors. The current study was aimed to develop an efficient non-viral vector with adequate cytotoxicity. To this end, alkylcarboxylate chains (6C, 10C, 16C) were exploited to ameliorate the characteristics of PEI 750 kDa. Briefly, alkylcarboxylate chains with three different lengths (6C, 10C, 16C) were chemically grafted to the primary amine groups of PEI 750 kDa in three percentages (10, 50, and 100%). After evaluating the physicochemical properties of prepared vectors including surface charge, size, buffering capacity, and DNA condensing, their transfection efficiency and cytotoxicity were investigated in Neuro2A cells. The polyplexes size were 158.9–264.5 nm and their zeta potentials were 14–30 mV, while their buffering capacity and DNA condensing were not significantly decreased. The highest transfection efficiency in term of C/P ratio was observed in PEI750-10C-68%, PEI750-10C-7%, and PEI750-6C-7% at C/P ratios of 2, 4, and 6, respectively. Altogether, the decanoylcarboxylate-modified PEI with medium grafting percentages showed promising results as gene delivery vector. To sum up, the modification of high molecular weight PEIs by alkylcarboxylate chains is an efficient approach for development of more efficient non-viral vectors.

Nuclear transportation of genetic materials is a critical step in gene therapy. Human gene therapy is not routine enough in clinic because an effective and safe gene carrier does not exist yet. It is reported that glucocorticoids such as dexamethasone (Dexa) facilitate nuclear transportation of plasmid DNA or other polynucleotide molecules to the nucleus. This study is aimed to investigate the influences of Dexa on physicochemical properties, cell transfection and toxicity of polyallylamine (PAA). After activation, Dexa mesylate was conjugated to both unmodified PAA and hexyl acrylate-conjugated PAAs (PAA-6c Acr) through a one-step reaction. PAA-Dexa was purified with dialysis against distilled water and then freeze-dried. The physicochemical properties of the synthesized vectors such as buffering capacity, DNA condensing ability, size, and zeta potential were determined. The PAA 15 kDa-based polyplexes showed better cell transfection than those based on PAA65 kDa in Neuro2A cell line. To sum up, transfection activity was improved in two types of the modified vectors including PAA15-6c Acr30%-Dexa5% at lower carrier to pDNA (C/P) ratios and PAA15-6c Acr50%-Dexa10% at higher C/P ratios. Altogether, the modification of PAA and PAA-6c Acr with Dexa significantly reduced their cytotoxicity, but had not guided their cell transfection to a desired point.

Targeted nano-carriers are highly needed to promote nucleic acid delivery into the specific cell for therapeutic approaches. Pullulan as a linear carbohydrate has an intrinsic liver targeting property interacting with asialoglycoprotein receptor (ASGPR) found on liver cells. In the present study, we developed polyamidoamine (PAMAM)-pullulan conjugates and investigated their targeting activity in delivering gene into liver cells. The particle size, zeta potential, buffering capacity and ethidium bromide exclusion assays of the conjugates were evaluated. The cytotoxicity and transfection efficiency of new derivatives were assessed following in vitro transfection of HepG2 (receptor positive) and N2A (receptor negative) cell lines. Size of conjugated polymers ranged between 118 and 184 nanometers and their cytotoxicity were similar to PAMAM. Among six produced nanocarriers, G4PU4 and G5PU4 enhanced transfection efficiency in HepG2 cells compared to unmodified PAMAM. Therefore, the PAMAM-pullulan derivatives seem to improve delivery of nucleic acids into the liver cells expressing asialoglycoprotein receptor with minimal transfection in non-targeted cells.

In this research, silver nanoparticles (AgNPs) were prepared via a ‘green’ procedure using an aqueous extract of Handelia trichophylla. The formation of AgNPs was confirmed by its light brown colour. The AgNPs were formed in silver nitrate (1 mM) via a bioreduction process in spherically shaped NPs with a mean diameter in the range of 20–50 nm. Moreover, the green synthesized AgNPs seemed to demonstrate a higher antibacterial activity against human pathogenic bacteria. In addition, the in vitro cytotoxicity effect of biosynthesized AgNPs was also investigated, which was detected to be up to $$15.62 \,\upmu \hbox {g}\, \hbox {ml}^{-1}$$ in the treated Neuro2A cells. Low toxicity and high antibacterial activity of biosynthesized silver nanoparticles can be utilized in different biological, biomedical and industrial applications.

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.

Polyethylenimine (PEI) and poly (l-lysine) (PLL) are among the most investigated non-viral gene carriers. However, both polymers contain deficiencies that restrict their applications. In the present study, we synthesized PLL-alkyl-PEI conjugates via 6-carbon alkyl linker and investigated their possible advantages in gene delivery. Four PLL copolymers were synthesized with different molecular weights and ratios of PEI. The physiochemical properties of synthesized conjugates such as size, zeta potential, DNA condensation ability, buffering capacity and cytotoxicity were investigated. Renilla luciferase assay was employed to evaluate the gene transfection efficiency of pDNA-polymer to Neuro2A cell line. DNA condensation and particle size measurements showed that new PLL-PEI conjugates could form polyplexes in nano-scale size in the range of 99-122 nm and were able to condense DNA at low concentration. While cytotoxicity reduced in some groups, the transfection efficiency increased about 2.8 and 4 fold as compared to the unmodified PEI 1.8 kDa and 10 kDa, respectively. The results of the present study showed that the chemical modifications of PEI with PLL could significantly improve transfection efficiency and PLP10-10% shows the most promise as a new gene carrier.

Peptide Brevinin-2R (B2R) has derived from frog skin secretions and possesses cytotoxic effects on cancer cells. Beside, cerium oxide nanoparticle (CNP) has antioxidant properties and could be used in anticancer studies. The purpose of this study is to investigate antioxidant and cytotoxicity of cerium oxide nanoparticle conjugated with B2R. First, cerium oxide nanoparticles were amine functionalized and peptide attached to it through establishment of peptide bond. Then, (1,1-diphenyl-2-picrylhydrazyl) DPPH and 2, 2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) radical scavenging activities of CNP-B2R were determined. MTT assay were used in order to compare cytotoxicity of CNP- B2R on two different cell lines. HFLF-pI5 cell and A549 cell lines were selected as representative of normal and cancer cells, respectively. Also, the cytotoxic effects of CNP, B2R and CNP-B2R were investigated on A549 cell line. Results of antioxidant evaluations showed that the antioxidant activity of CNP-Peptide increased at higher concentration of CNP-B2R with IC50 of 0.2 and 0.54 (mg/mL) for DPPH and ABTS assays, respectively. The results of cytotoxicity effects showed that CNP-B2R was more potent in killing tumor cell line in comparison with normal cell line. Cytotoxicity of CNP, B2R and CNP-B2R demonstrated that CNP-B2R and B2R had the lower cell viability effects compared to CNP. Our findings showed cytotoxicity of CNP-B2R against cancer cell lines in comparison with normal cells indicating the potential anticancer properties of CNP-B2R.

Theranostic nanoparticles (NPs) are promising for opening new windows toward personalized disease management. Using a single particle capable of both diagnosis and drug delivery, is the major benefit of such particles. In the present study, chitosan NPs were used as a dual action carrier for doxorubicin (DOX; chemotherapeutic agent) and superparamagnetic iron oxide nanoparticles (SPIONs; imaging agent). SPIONs and DOX were loaded at different concentrations within poly-l-arginine-chitosan-triphosphate matrix (ACSD) using the ionic gelation method. NPs' size were in the range of 184.33 ± 4.4 nm. Drug release analysis of DOX loaded NPs (NP-DOX) showed burst release at pH 5.5 (as in tumor environment) and slow release at pH 7.4 (physiological condition), demonstrating pH-sensitive drug release profile. NP-DOX internalization was confirmed by flowcytometry and fluorescent microscopy. Uptake process results were corroborated by accumulation of drug in the intracellular space. Iron content was evaluated by inductively coupled plasma and prussian blue staining. In vitro magnetic resonance imaging (MRI) showed a decline in T 2 relaxation times by increasing iron concentration. MRI analysis also confirmed uptake of NPs at the optimum concentration in C6 glioma cells. In conclusion, ACSD NPs could be utilized as a promising theranostic formulation for both diagnosis and treatment of glioblastoma.

Menthol is an organic compound with diverse medicinal and commercial applications, and is made either synthetically or through extraction from mint oils. The aim of the present study was to investigate menthol levels in selected menthol-producing species belonging to the Lamiaceae family, and to determine phylogenetic relationships of menthol dehydrogenase gene sequence among these species. Three genus of Lamiaceae, namely Mentha, Salvia, and Micromeria, were selected for phytochemical and phylogenetic analyses. After identification of each species based on menthol dehydrogenase gene in NCBI, BLAST software was used for the sequence alignment. MEGA4 software was used to draw phylogenetic tree for various species. Phytochemical analysis revealed that the highest and lowest amounts of both essential oil and menthol belonged to Mentha spicata and Micromeria hyssopifolia, respectively. The species Mentha spicata and Mentha piperita, which were assigned to one cluster in the dendrogram, contained the highest amounts of essential oil and menthol while Micromeria species, which was in the distinct cluster and placed in the farther evolutionary distance, contained the lowest amount of essential oil and menthol. Phylogenetic and phytochemistry analyses showed that essential oil and menthol contents of menthol-producing species are associated with menthol dehydrogenase gene sequence.

Cationic polymers such as polyallylamine (PAA) having primary amino groups are poor transfection agents and possess a high cytotoxicity index when used without any chemical modification. In this study, PAA was modified with cholesterol in order to improve transfection efficiency and to reduce cytotoxicity. PAA polymers with molecular weights of 15 and 65 kDa were selected and grafted with cholesterol at percentages of 5, 10, 15, 30, and 50. After purification, the efficacy of the synthetic vectors was evaluated in terms of DNA condensation using the ethidium bromide test, buffering capacity, particle size, zeta potential, transfection efficiency, and cytotoxicity assay in Neuro2A cell lines. According to the ethidium bromide test, these vectors can condense DNA at moderate and high carrier to plasmid (C/P) ratios. The buffering capacity of the prepared vector in both molecular weights was less than unmodified PAA. Particle size measurements demonstrated that modified PAAs were able to form nanoparticles ranging in size from 125 to 530 nm. The vectors based on PAA 15 kDa demonstrated a better transfection efficiency than the vectors made of PAA 65 kDa. Cytotoxicity studies showed that toxicity of all vectors was less than PAA. Some cholesterol modified polymers composed of PAA (15 kDa) were suitable vectors for gene delivery with low cytotoxicity.

Non-viral vectors such as polymers and liposomes have been used as 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 tried to design non-viral gene delivery vectors that mimic the viral vectors using the benefits of both cationic liposomes and cationic polymer vectors along with targeting glucocorticoid receptors to enhance cellular trafficking of vectors. Cationic liposomes containing DOTAP and cholesterol were prepared by thin-film hydration following extrusion method. Dexamethasone mesylate was synthesized and then conjugated to polyethylenimine through a one-step reaction. A novel gene delivery system, Lipopolyplex was developed by premixing liposome and different molecular weight of bPEI-Dexa as carriers followed by addition of plasmid at three different carrier/pDNA (C/P) weight ratios. The resulted complexes were characterized for their size, zeta potential and ability of DNA condensation. Transfection efficiency of vectors in neuro2A was determined by Luciferase reporter gene assay. 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 their size by increasing the molecular weight of PEIs. All complexes had positive surface charge. Complete condensation of DNA was occurred at C/P ratio of one for all complexes. Lipopolyplexes were more efficient than polyplexes and lipoplexes alone and transfection efficiency was improved by adding dexamethasone. The complexes containing liposome, PEI 10 kDa and dexamethasone (PEI10:Lipo:Dexa(0.05)) had the highest transfection activity about 40-fold and 3.6-fold in comparison with PEI10 and PEI10:Lipo, respectively. Furthermore, the non-viral vectors described in this study showed low cytotoxicity. The results of this study confirmed that PEI in combination with liposome forms lipopolyplex with low toxicity and enhanced transfection efficiency. Moreover, using dexamethasone, in combination with lipopolyplex might be useful to increase the gene delivery potential of these lipopolyplexes.

The applications of dendrimer-based vectors seem to be promising in non-viral gene delivery because of their potential for addressing the problems with viral vectors. In this study, generation 3 poly(propyleneimine) (G3-PPI) dendrimers with 1, 4-diaminobutane as a core initiator was synthesized using a divergent growth approach. To increase the hydrophobicity and reduce toxicity, 10% of primary amines of G3-PPI dendrimers were replaced with bromoalkylcarboxylates with different chain lengths (6-bromohexanoic and 10-bromodecanoic). Then, to retain the overall buffering capacity and enhance transfection, the alkylcarboxylate–PPIs were conjugated to 10 kDa branched polyethylenimine (PEI). The results showed that the modified PPI was able to form complexes with the diameter of less than 60 nm with net-positive surface charge around 20 mV. No significant toxicity was observed in modified PPIs; however, the hexanoate conjugated PPI–PEI (PPI-HEX-10% PEI) and the decanoate conjugated PPI–PEI (PPI-DEC-10%-PEI) showed the best transfection efficiency in murine neuroblastoma (Neuro-2a) cell line, even PPI-HEX-10%-PEI showed transfection efficiency equal to standard PEI 25 kDa with reduced toxicity. This study suggested a new series of hyperbranched (PEI)–dendrimer (PPI) architectural copolymers as non-viral gene delivery vectors with high transfection efficiency and low toxicity.

Superparamagnetic iron oxide nanoparticles (SPIONs; Fe3O4) were prepared by the covalent binding of starch onto the surface of magnetic Fe3O4 nanoparticles, by a “green” co–precipitation method. The scanning electron microscopy (SEM) illustrated the regular spheres of Fe3O4 particles that were coated by starch molecules to be regular spheres with a mean diameter of about 40 nm. Powder X–ray diffraction patterns (PXRD) indicated that the prepared samples were pure Fe3O4 with a spinel structure while the coating of starch did not result in a phase change. The starch coating onto the Fe3O4 nanoparticles was also demonstrated by Fourier transform infrared (FTIR) spectra. Magnetic measurements revealed that the saturated magnetization of the starch–SPIONs reached 42.1 emu/g, having nanoparticles with the characteristics of superparamagnetism, while a dose dependent toxicity with non–toxic effect of concentration below 62.5 mg/mL was observed in the studies of in vitro cytotoxicity on neuro2A cells. & 2014 Elsevier Ltd and Techna Group S.r.l. All rights reserved.

The use of food-directed and natural products for the synthesis of different nanoparticles ( e.g ., metal and metal oxide) is of enormous interest to modern nanoscience and nanotechnology. We have developed a facile and green chemistry method with bio-directed, and low cost materials for the synthesis of cerium oxide nanoparticles (CeO 2 -NPs) using honey. In this method, the conversion of cerium cations into CeO 2 -NPs was achieved via a sol–gel process in aqueous honey solutions. The synthesized CeO 2 -NPs were characterized by the following title: UV–vis spectroscopy, field emission scanning electron microscopy (FESEM), Fourier transform infrared spectroscopy (FT-IR), thermogravimetric (TGA–DTA) analysis, Energy dispersive spectrum (EDS), and powder X-ray diffraction (PXRD). Spherical CeO 2 -NPs were synthesized at different calcination temperatures and FESEM imaging along with its corresponding particles size distribution indicated the formation of nanoparticles in size of about 23 nm. The PXRD analysis revealed fluorite cubic structure for CeO 2 -NPs with preferential orientation at (111) reflection plane. In vitro cytotoxicity studies on neuro2A cells, a dose dependent toxicity with non-toxic effect of a concentration below about 25 µg/mL was illustrated. The synthesis of CeO 2 -NPs in aqueous honey solutions was found to be comparable to those obtained from conventional reduction methods that uses hazardous materials proving to be an excellent alternative for the preparation of CeO 2 -NPs, using food and bio-derived materials.

Green and Biological synthetic methods for preparing of different nanoparticles (e.g., metal and metal oxide) using microorganisms, enzymes, and plant extracts have been suggested as a possible low cost and eco-friendly alternative to chemical and physical methods. In this paper, we would like to report the green and biosynthesis of nanoceria particles (NCs), using gum tragacanth by both chemical and biological methods. The NCs were synthesized at different calcination temperatures and structural, morphological, and optical properties of the synthesized NCs have been characterized by using UV–vis spectrophotometer, FESEM, PXRD analysis. FESEM images display that the NCs prepared were mono-dispersed and the average size was ranged from about 20 to 40 nm. In vitro viability studies on neuro2A cells, a dose dependent toxicity with non-toxic effect of concentrations below 30 µg/mL is presented. The synthesis of NCs using gum tragacanth was found to be comparable to those obtained from conventional reduction methods using hazardous polymers or surfactants.

Crystalline nanoceria powders have been synthesized by green-mediated sol–gel method in aqueous solution. The cerium nitrate hexahydrate as precursor and starch were taken in water and precipitated with ammonium hydroxide at pH 10. To terminate the growth of nanoceria particles and also to stabilize them, long-chain starch molecules were utilized. The sample was then calcined for 2 h in a temperature range of 120–600 °C and characterized by the number of techniques, including powder X-ray diffraction (PXRD), UV–vis spectrophotometry, and high-magnification transmission electron microscopy (TEM). The particle size obtained from TEM was in the range of 6 nm with uniform shape and narrow particle size distribution. The diffraction pattern completely indexed with the cubic fluorite structure of CeO 2 . The calcined nanoceria powders showed strong UV absorption. In vitro cytotoxicity studies on Neuro2A cells showed a dose dependent toxicity with non-toxic effect of concentration up to 175 µg/mL. The results show that starch is an interesting material that can be used as a stabilizer in the sol–gel processes for preparing small nanoceria particles.

Aims The development of highly efficient and low toxic non-viral gene delivery vectors is the most challenging issues for successful application of gene therapy. A particular focus has been on understanding structure-activity relationships for transfection activity and toxicity of polyethylenimine (PEI). During the last decade, the use of cerium oxide nanoparticles (CeO2-NPs) in biomedicine has attracted much attention due to their pH-dependent antioxidant activity. CeO2-NPs provide protection normal cells from various forms of reactive oxygen species, but possess innate cytotoxicity and apoptosis to cancer cells. The purpose of this study was to design a new class of gene carriers by low molecular weight PEI (B-PEI 10 kDa) coordination onto CeO2-NPs. Main methods B-PEI 10 kDa was conjugated to CeO2-NPs by Epichlorohydrin linker. Transfection efficiency, cytotoxic and apoptotic effects of pDNA-PEI-CeO2 NPs were evaluated on WEHI 164 cancer cells and normal L929 cells lines. Key findings PEI-CeO2 NPs was able to condense the pDNA at carrier/plasmid (C/P) weight ratios of 0.5. The size and zeta potential of pDNA-PEI-CeO2 NPs were 124 ± 7 nm and 22 ± 2 mV, respectively. The transfection efficacy of synthesized pDNA-PEI-CeO2 NPs improved and the cytotoxicity was decreased compared to pDNA-PEI. Moreover, pDNA-PEI-CeO2 NPs induced more apoptosis than unmodified PEI and CeO2-NPs control groups. pDNA-PEI-CeO2 NPs displayed more transfection, cytotoxicity, and apoptosis in WEHI 164 cancer cells than normal L929 cells. Significance In conclusion, PEI-CeO2 nanocarriers could act as a potential candidate for gene and drug delivery to cancerous and tumor cells.

The use of plant extract in the synthesis of nanomaterials can be a cost effective and eco-friendly approach. In this work we report the “ green” and biosynthesis of zinc oxide nanoparticles (ZnO-NPs) using gum tragacanth . Spherical ZnO-NPs were synthesized at different calcination temperatures. Transmission electron microscopy (TEM) imaging showed the formation most of nanoparticles in the size range of below 50 nm. The powder X-ray diffraction (PXRD) analysis revealed wurtzite hexagonal ZnO with preferential orientation in (101) reflection plane. In vitro cytotoxicity studies on neuro2A cells showed a dose dependent toxicity with non-toxic effect of concentration below 2 µg/mL. The synthesized ZnO-NPs using gum tragacanth were found to be comparable to those obtained from conventional reduction methods using hazardous polymers or surfactants and this method can be an excellent alternative for the synthesis of ZnO-NPs using biomaterials.

Cerium oxide nanoparticles (CeO 2 -NPs) were synthesized via the sol–gel method in gelatin media. Long-chain gelatin compounds were utilized to terminate the growth of CeO 2 -NPs and stabilize them. The CeO 2 -NPs were characterized by a number of techniques, including X-ray diffraction analysis (XRD), UV–vis spectrophotometry, and high-magnification transmission electron microscopy (TEM). The CeO 2 -NPs calcined at different temperatures exhibited a cubic fluoride structure with sizes less than approximately 10 nm. The influence of the calcination temperature on the morphology of CeO 2 -NPs was also investigated. In vitro cytotoxicity studies on neuro2A cells showed a dose-dependent toxicity with non-toxic effect of concentration below 10 μg/mL. The results indicated that gelatin is an interesting material that can be used as a stabilizer in the sol–gel processes for preparing small CeO 2 -NPs.

Background Carboxyalkylation of branched 25 kDa polyethylenimine (PEI) was considered to reduce the positive surface charge of the polymer without reducing its ‘proton sponge’ buffering capacity, and to provide alkylene domains for hydrophobic interactions, thus generating optimized novel PEI carriers for efficient delivery of small interfering RNA (siRNA). Methods Substitution of PEI was evaluated in the range of 6 to > 50 mole percentage of primary amines. Additionally, variation of the carboxyalkyl chain (one to 15 methylene groups) was explored to modulate the carrier hydrophobicity. Carriers were characterized in their buffering capacity, capability of siRNA polyplex formation, and cytotoxicity. Marker gene-silencing efficacy was evaluated using Neuro2A-eGFPLuc neuroblastoma cells. Results Carboxyalkylation strongly reduced cytotoxicity of PEI and improved siRNA mediated luciferase gene knockdown. An optimum silencing activity was observed at an alkylcarboxylation degree of 6–9 mole percentage of primary amines and with a broad range of carboxyalkylene chains (containing one to 15 methylene groups). Strongly enhanced gene-silencing efficacy also was observed when the biocompatible polymers were separately added at 1 h after transfection with tolerated doses of standard PEI25/siRNA polyplexes. Conclusions Carboxyalkylation of branched 25 kDa PEI resulted in polymers with strongly reduced cytotoxicity and improved silencing efficacy. Mechanistic studies demonstrated that the presence of a surplus of free carboxyalkylated polymer is responsible for the improved siRNA delivery. Copyright © 2010 John Wiley & Sons, Ltd.

In this study, a series of alkyl-oligoamine derivatives of low-toxicity 10 kDa polyethylenimine (PEI) were synthesized to enhance the hydrophobicity of PEI while preserving most of its primary amine content. PEI was reacted with a series of ω-bromoalkylcarboxylic acids with different chain lengths (2-bromoacetic, 6-bromohexanoic, 10-bromodecanoic and 16-bromohexadecanoic acids) to modify hydrophobicity followed by coupling to various oligoamines (spermine, spermidine, ethylendiamine or diethylentriamine) to partially restore primary amine density. These modifications were designed to influence hydrophobic–hydrophilic balance as well as maintain the proton sponge effect in order to create an efficient vector with low toxicity. Ethidium bromide exclusion assays and dynamic light scattering studies showed that the modified PEIs could bind to plasmid DNA and form nanoparticles in the range of 100 nm. The transfection efficiency of modified PEIs complexed with a luciferase reporter gene (pCMV-luc) in N2A murine neuroblastoma cells was increased to a level comparable to that of 25,000 Da PEI. These results indicate that hydrophobic modification of low-toxicity PEI without reduction in primary amine content is an effective strategy for improving transfection efficiency of polycation-based non-viral vectors while maintaining low toxicity.

The applications of polyamidoamine (PAMAM) dendrimers have attracted much attention in biomedicine specially non-viral gene delivery because of thier unique characteristics including hyperbranching, multivalency, and well-defined uniform globular three-dimensional structures. In the current study, in order to enhance the transfection efficiency and reduce the cytotoxicity of PAMAMs, bromoalkylcarboxylates with different chain length (2-bromoacetic, 6-bromohexanoic, 10-bromodecanoic and 16-bromohexadecanoic acids) were covalently conjugated with 10% and 30% of primary amines of generation 4 and 5 (G4 and G5) of PAMAM dendrimers to increase the hydrophobicity of the carrier. At the next stage, the alkylcarboxylate-PAMAMs were pegylaed to further modify the PAMAM structures for biological applications. Obtained results demonstrated that the prepared PAMAM derivatives had particle size around 140 nm with net-positive surface charge. None of the prepared PAMAM-based non-viral vactors exhibited significant hemolytic activity and also cytotoxicity. Meanwhile decahexanoate–PAMAM G4 [G4(16C-10%)] and decanoate–PAMAM G4 conjugated to polyethylene glycol (PEG) (G4[(10C-30%)(10C-PEG)]) showed highest transfection efficiency in murine neuroblastoma (Neuro-2a) cell line, interestingly only the latter had improved transfection efficiency in mesenchymal stem cells (MSCs). This study proved the potential utility of alkylcarboxylate-grafted PAMAM dendrimers (G4 and G5) with or without PEG modification for efficient gene transfer into cancerous cells as well as MSCs.

Introduction of an efficient gene delivery vector is still the main challenge of gene therapy. Both polyethylenimine (PEI) and poly(l-lysine) (PLL) comprise disadvantages which limited their application. To explore whether their deficiencies could be compensated by preparing copolymers consisting of both PLL and PEI, we generated several combinations of PLL-alkyl-PEI copolymers conjugated to aptamer and evaluated their both gene delivery efficiency and down-regulation of Bcl-XL, an anti-apoptotic gene, in lung cancer cell line. PLL was conjugated to either 10% or 50% of PEI by grafting different percentages of PEI to alkylated-PLL as core. The properties of modified polymers including size, surface charge density, DNA condensation ability, buffering capacity and cytotoxicity were evaluated. According to transfection results, aptamer conjugated PLL-alkyl-10%-PEI (PLPE8%) was selected for further gene silencing study by plasmid shRNA. Decrease in Bcl-XL gene expression was estimated by both RT-PCR and western-blot experiments. The obtained results revealed that the new copolymers had appropriate nano-scale size (117-128 nm) even after aptamer conjugation (168-183 nm). Moreover, they exhibited increased transfection efficiencies by up to 1.8-5 folds and acceptable cytotoxicity. The apoptosis was induced in transfected cells by shRNA-aptamer-copolymer due to the down-regulation of mRNA and protein levels. This study suggested a new vector for targeted non-viral gene delivery with high transfection efficiency in lung cancer or pulmonary systems.

Nowadays gene delivery is a topic in many research studies. Non-viral vectors have many advantages over viral vectors in terms of safety, immunogenicity and gene carrying capacity but they suffer from low transfection efficiency and high toxicity. In this study, polyallylamine (PAA), the cationic polymer, has been modified with hydrophobic branches to increase the transfection efficiency of the polymer. Polyallylamine with molecular weights of 15 and 65 kDa was selected and grafted with butyl, hexyl and decyl acrylate at percentages of 10, 30 and 50. The ability of the modified polymer to condense DNA was examined by ethidium bromide test. The complex of modified polymer and DNA (polyplex) was characterized for size, zeta potential, transfection efficiency and cytotoxicity in Neuro2A cell lines. The results of ethidium bromide test showed that grafting of PAA decreased its ability for DNA condensation but vectors could still condense DNA at moderate and high carrier to DNA ratios. Most of polyplexes had particle size between 150 and 250 nm. The prepared vectors mainly showed positive zeta potential but carriers composed of PAA with high percentage of grafting had negative zeta potential. The best transfection activity was observed in vectors with hexyl acrylate chain. Grafting of polymer reduced its cytotoxicity especially at percentages of 30 and 50. The vectors based of PAA 15 kDa had better transfection efficiency than the vectors made of PAA 65 kDa. In conclusion, results of the present study indicated that grafting PAA 15 kDa with high percentages of hexyl acrylate can help to prepare vectors with better transfection efficiency and less cytotoxicity.

Curcumin might be a good natural therapeutic agent; it could inhibit intercellular inflammation pathways and reduce a lot of unfavorable molecular events in chronic inflammatory diseases.Copyright © 2014, Razavi Hospital; Published by Kowsar Corp. This is an open-access article distributed under the terms of the Creative Commons Attribution Li-cense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Nonviral gene delivery systems are considered to be a safe alternative to the viral system used in gene therapy. Although polyethylenimine (PEI) is among the most promising gene-carrier candidates for efficient nonviral gene delivery, safety concerns regarding its toxicity remain challenging. The aim of this study was to evaluate the less considered aspects of toxicity including genotoxicity effects of branched PEI with different molecular weights (1800 Da, 25 kDa, 750 kDa). Neuro2A mammalian cells were cultured in DMEM with 10% FBS and exposed to PEI/DNA complexes at different polymer/DNA ratios (known as the C/P ratio). To evaluate metabolic activities and genotoxicity, the treated cells were subjected to MTT and comet assays, respectively. Intracellular reactive oxygen species were also determined by using the fluorescent dye dichlorofluorescin diacetate. Concentration with relation to toxicity, genotoxicity, and ROS production was observed in Neuro2A cells treated with PEI/DNA nanoparticles. The difference among toxicities induced by different molecular weights of PEI at higher C/P ratios was significant, whereas no significant toxicity effect was induced by PEI at C/P ratios of 1 and lower. We found concentration-dependent genotoxicity and a direct correlation between nanoparticle concentrations and DNA damage, which cannot be ignored and should be considered for the design of new generations of PEI derivatives with enhanced delivery properties and reduced toxicity.

In this study, l-thyroxine was covalently grafted on 25 kDa branched polyethylenimine (PEI), and the ability of the nano-sized polyplexes for transferring plasmid encoding interleukin-12 (IL-12) gene was evaluated. As there are several problems in systemic administration of recombinant IL-12 protein, local expression of the plasmid encoding IL-12 gene inside the tumor tissue has been considered as an effective alternative approach. The l-thyroxine-conjugated PEI polyplexes were prepared using pUMVC3-hIL12 plasmid, and their transfection activity was determined in HepG2 human liver carcinoma and Neuro2A neuroblastoma cell lines. The polyplexes characterized in terms of DNA condensation ability, particle size, zeta potential, and buffering capacity as well as cytotoxicity and resistance to enzyme digestion. The results revealed that l-thyroxine conjugation of PEI increased gene transfer ability by up to two fold relative to unmodified 25 kDa PEI, the gold standard for non-viral gene delivery, with the highest increase occurring at degrees of conjugation around 10 %. pDNA condensation tests and dynamic light scattering measurements exhibited the ability of PEI conjugates to optimally condense the plasmid DNA into polyplexes in the size range around 200 nm. The modified polymers showed remarkable buffering capacity and protection against enzymatic degradation comparable to that of unmodified PEI. These results suggest that l-thyroxine conjugation of PEI is a simple modification strategy for future investigations aimed at developing a targeting gene vehicle.

Background Various strategies have been examined to improve both transfection efficiency and cytotoxicity of polyethylenimine (PEI), a widely used polycationic nonviral gene vector. In the present study, we sought to improve PEI transfection efficiency by combining the osmotic burst mechanism for lysing endocytotic vesicles with the lipid depletion mechanism, which was accomplished by maintaining buffering capacity at the same time as adding a lipid-absorbing hydrophobic shell. Methods PEI was altered via the substitution of various percentages of its primary amines with carboxylate-terminated short, moderate and long alkyl chains, by reaction with bromoacetic, 6-bromohexanoic, 10-bromodecanoic and 16-bromohexadecanoic acids. Modified polymers were complexed with plasmid and the particle size and zeta potential of the polyplexes were determined. Ethidium bromide dye exclusion was used to show the DNA-binding ability of the polymers and their transfection activity and cytotoxicity was evaluated in Neuro2A mammalian cells. Results Decreased DNA-binding ability resulted from increases in either the degree of substitution or hydrocarbon chain length. Particle size and zeta potential measurements demonstrated that modified PEI polymers were able to form nanoparticles in the size range 60–195 nm, and surface charge decreased with an increasing degree of substitution. Higher degrees of substitution resulted in decreased cytotoxicity of polymers. Alkylcarboxylate substitution of primary amines on PEI enhanced transfection efficiencies by up to approximately five-fold relative to underivatized PEI, with the greatest increases occurring with 6-bromohexanoic acid derivatives at degrees of substitution below 10%. Conclusions The results obtained suggest that an appropriate balance between cationic and hydrophobic regions of alkylated PEI yields the optimal nonviral vector with high transfection efficiency and low toxicity. Copyright © 2009 John Wiley & Sons, Ltd.

The lack of efficient delivery systems is still limiting the full therapeutic potential of siRNA. For the purpose of nucleic acid transfer, among other synthetic carrier systems, polycations have been applied. Favorable characteristics of suitable polymers include nucleic acid binding, compaction, protection, and biocompatibility. However the lack of nucleic acid transfer activity in transfection-based screening often abandons promising candidates. Here we present that functionalization may turn polycations with poor delivery activity into efficient carriers:  for example, polylysine, on its own lacking nucleic acid transfer activity, displayed high efficiency in siRNA delivery after modification with polyethylene glycol and a pH-responsive endosomolytic peptide. Hence these findings have implication for the selection process of polymeric carriers for siRNA.

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 EC 50 for Soxhlet extracts in dimethylsulfoxide was 0.30 mg/mL. The EC 50 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.