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The interaction between osteogenic and angiogenic cells through a coculturing system in biocompatible materials has been considered for successfully engineering vascularized bone tissue equivalents. In this study, we developed a hydrogel-blended scaffold consisted of gelatin methacryloyl (GelMA) and alginate enriched with hydroxyapatite nanoparticles (HAP) to model an in vitro prevascularized bone construct. The hydrogel-based scaffold revealed a higher mechanical stiffness than those of pure (GelMA), alginate, and (GelMA+ HAP) hydrogels. In the present study, we generated a green fluorescent protein (GFP) knock-in umbilical vein endothelial cells (HUVECs) cell line using the CRISPR/Cas9 technology. The GFP was inserted into the human-like ROSA locus of HUVECs genome. HUVECs expressing GFP were cocultured with OB-like cells (MG-63) within three-dimensionally (3D) fabricated hydrogel to investigate the response of cocultured osteoblasts and endothelial cells in a 3D structure. Cell viability under the 3D cocultured gel was higher than the 3D monocultured. Compared to the 3D monocultured condition, the cells were aligned and developed into the vessel-like structures. During 14 days of culture periods, the cells displayed actin protrusions by the formation of spike-like filopodia in the 3D cocultured model. Angiogenic and osteogenic-related genes such as CD31, vWF, and osteocalcin showed higher expression in the cocultured versus the monocultured. These results have collectively indicated that the 3D cocultured hydrogel facilitates interaction among cells, thereby having a greater effect on angiogenic and osteogenic properties in the absence of induction media.

Liposomal peptide-based vaccines can potentially suppress cancer cells proliferation in the host. To enhance the effectiveness of vaccination against cancer, additional strategies should also be employed. One strategy to promote peptide-based vaccine efficacy and induce powerful immune responses, is simultaneous activation of CD4+ and CD8+ T cells. To address this problem, we tested the efficacy of a nano-liposomal vaccine containing P5 peptide, a cytotoxic T lymphocytes (CTL) specific peptide derivative of rat HER2/neu protein, Pan HLA-DR (PADRE) peptide, a universal CD4+ T helper cell epitope and monophosphoryl lipid A (MPL) a toll-like receptor 4 ligand. We observed potent CD8+ T cell immune responses in TUBO mice vaccinated with liposomal P5 peptide in combination with PADRE and MPL. Also, this formulation remarkably improved anti-tumor effects against cells overexpressing HER2 in BALB/c mice compared to liposomal vaccine containing P5 only. Furthermore, we found that vaccination with Lip-P5- Integrated PADRE-MPL formulation significantly induced IFN-γ production, increased CD8+ T cells numbers and enhanced survival compared to other groups of treated mice. In conclusion, our study indicated that Lip-P5-Integrated PADRE-MPL, after further confirmatory investigations, could be employed as a promising vaccine to generate potent CTL anti-tumor immune responses that could be beneficial to treatment of HER2+ breast cancer.

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.

The preparation of silver nanoparticles (Ag-NPs) has been performed through a ‘green’ procedure by utilizing the aqueous flowering shoots of Allium giganteum extract at room temperature. The UV-vis...

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.

Nanotechnology as a multidisciplinary and scientific innovation plays an important role in numerous biomedical applications, such as molecular imaging, biomarkers and biosensors and also drug delivery. A wide range of studies have been conducted on using of nanoparticles for early diagnosis and targeted drug therapy of various diseases. In fact, the small size, customized surface, upgraded solubility, or multi-functionality of nanoparticles enabled them to interact with complex cellular functions in new ways which opened many doors and created new biomedical applications. These studies demonstrated that nanotechnology vehicles can formulate biological products effectively, and this nano-formulated products with a potent ability against different diseases, were represented to have better biocompatibility, bioaccessibility and efficacy, under in vitro and in vivo conditions.

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.

Introduction: Cationic polymers have the potential to be modified to achieve an ideal gene vector lacking viral vector defects. The aim of the present study was to improve polyallylamine (PAA) transfection efficiency and to reduce cytotoxicity by incorporating of cell-penetrating peptides (CPPs). Methods: To prepare the peptide-based polyplexes, PAA (15 kDa) was modified with 2 peptides (TAT and CyLoP-1) by covering the 0.5% and 1% of amines. Buffer capacity and DNA condensation ability of modified polymer, particle size and zeta potential of nanoparticles, cell viability, and transfection activity of vectors were evaluated. Results: In low carrier to plasmid (C/P) weight ratios such as 0.5 and 1, the unmodified polymer was more capable to condense the DNA compared to the synthesized vectors. In C/P ratio of 2, the plasmid was fully condensed in all vectors. The size of polyplexes ranged from 195 to 240 nm. The zeta potential was almost as the same as PAA and varied from 25 to 27 mV. All polyplexes increased the buffer capacity compared to PAA. The transfection efficiency was improved compared to unmodified polymer especially in the vectors modified with 1% of TAT or CyLoP-1 peptides in C/P ratio of 2. The cytotoxicity of prepared vectors was less than PAA. In most ratios, the cytotoxicity of the CyLoP-1 modified samples was less than the TAT modified ones. Conclusion: Modification of PAA with CPPs improved the transfection activity of vector.

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.

Bio-based synthesis of Nano-Ceria (NC) was performed in Linum usitatissimum L. (Lu) seeds extract as capping agent. Obtained gel was calcined at 400, 500, and 600 °C to investigate the effect of temperature on the size and morphology of the particles. All samples had spherical morphology which their crystallite size was decreased in higher temperature. Products were characterized by TGA/DTA, UV–vis, FESEM, FTIR and XRD. The band gap of the prepared samples was calculated through Tauc plot in the range of 3.2–3.4 eV. The results of MTT assay confirmed that NC has been shown no significant toxicity on A549 cell line. 2′,7′–dichlorofluorescin diacetate (DCFDA) was used to determine antioxidant properties of NC on A549 cell and the results showed that all concentrations of NC could reduce reactive oxygen species (ROS). NC was labeled with technetium (99mTc) for in vivo bio-distribution study in Wistar rat. Radiolabeled NC was stable in different environments of PBS buffer and human serum with radiochemical purity of more than 95% according to the instant thin layer chromatography (ITLC) method. DLS analysis showed radiolabeled particles had small size and pleasant colloidal stability. Bio-distribution of radiolabeled NC illustrated the highest accumulation in kidneys (1 h: 5.94 ± 0.77%ID/g, 4 h: 10.95 ± 5.99%ID/g, 24 h 7.94 ± 0.36%ID/g). Moreover, low uptake of 99mTc-NC in stomach confirmed the in vivo stability of 99mTc-NC. Accordingly, NC could be a worthy candidate for biological purposes and in vivo studies.

In this study, we used cerium oxide nanoparticles and evaluated their anti-cancer effects in a mouse model of fibrosarcoma. For evaluation of anti-cancer effects of nanoceria, tumor volume measurement, TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assay, quantitative real-time PCR (qPCR) for Bax and Bcl2 genes, a panel of liver and kidney function tests and hematoxylin-eosin staining were done. Nanoceria dominantly accumulated in the tumor and it could significantly decrease tumor growth and volume in tumor-bearing mice that received nanoceria for four weeks. Cerium oxide nanoparticle showed potential anti-cancer properties against fibrosarcoma.

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.

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.

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.

Ceria nanoparticles (CN) were produced via green route in Cydonia oblonga miller (Com) seeds extract as capping and stabilizing agent. They were characterized by a variety of physico-chemical methods such as XRD, FT-IR, UV-Vis, FESEM, TGA/DTA, and photoluminescence (PL). Crystalline size of CN was increased with ascending the calcination temperature while band gap energy of them was descending from 2.4 to 3.1 eV. Cell viability was determined by MTT assay and results showed that CN had no significance toxicity on A549 cell line. Also, antioxidant effect of CN on the same cell line was performed via 2′,7′–dichlorofluorescin diacetate (DCFDA). All concentration of CN specially 15.6 μg/ml could neutralize the oxygen reactive species (ROS). Photo-catalytic study was performed to evaluate dyes degradation ability of CN under UV-A irradiation. Results illustrated that samples calcined in lower temperature showed better photo-catalytic activity. Also, PL study showed that the photoluminescence emission intensity of samples was diminished with increasing the calcination temperature. In conclusion, enhance the photo-catalytic activity and photoluminescence emission was related to presence of oxygen defects in CN structure which were increased in lower calcination temperature and responsible for improving the optical properties of CN.

The present study aim to design a liposomal electrochemical sensor using 1, 2-dioleoyl-3-trimethylammoniumpropane (DOTAP) and dioleoylphosphatidylethanolamine(DOPE), chimeric probes and p19, it has been considered as a caliper molecule as well. Also the competitor structural hybrid (RNA) was used to detect three types of miRs in one screen printed electrode modified by gold nanoparticle (SCPE/GNP). In this purpose, the sensor signal stabilized when the cationic DOTAP-DOPE with hybrids of the chimeric probes (Stem, M-linear) sandwiched in order to detect 221–124a miRs. Given the lack of accessibility to RNA-miRs segments of chimeric probes, p19 inhibited the electrochemical reaction and shifted signal to off. After that p19 connected with the free hybrid of T-linear/21miR (just RNA) as competing for structure and the signal was shifted to ON, again. In this study, the electrochemical measurements were performed between the potentials at −0.4 V and +0.4 V with 1 mM [Fe(CN)6]-3-/4 which DOTAP-DOPE acted as an enhancer layer in the electrostatically reaction. This sensor determines as low as 0.4 fM of miRNA with high selectivity and specificity for sequential analysis of 124a-221-21 miRs in just 2 h.

Due to large surface area, tunable pore size, easy surface manipulation, and low-toxicity mesoporous silica nanoparticles (MSNs) may act as a suitable vector for gene delivery. In order to make MSNs as a suitable gene delivery system, we modified the surface of phosphonated MSNs (PMSN) with polyethyleneimine (PEI) 10 and 25 KDa. Then nanoparticles were loaded with chloroquine (CQ) (a lysosomotropic agent) and complexed with plasmid DNA. The transfection efficiency and cytotoxicity of these nanoparticles was examined using green fluorescent protein plasmid (pGFP) and cytotoxicity assay. All PEI coated nanoparticles showed positive zeta potential and mean size was ranged between 170 and 215 nm with polydispersity index bellow 0.35. PEI-coated MSNs significiantly enhanced GFP gene expression in Neuro-2 A cells compared to PEI 10 and 25 KDa. The results of the cytotoxicity assays showed that these nanoparticles have an acceptable level of viability but CQ loaded nanoparticles showed higher cytotoxicity and lower transfection activity than CQ free nanoparticles.

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.

In this work, we designed a liposomal electrochemical sensor with DOTAP-DOPE liposome, chimeric probes, p19 as a caliper molecule, and the competitor structural hybrid (just RNA) for detection of three micro-RNAs in one SPCE/GNP electrode. The sensor is stabled when the cationic spherical DOTAP-DOPE liposomes sandwich with hybrids of the different sandwiched of probes (T-M-linear, Stem) and 21-124a-221miRs. With the addition of P19, in the presence of a sandwiched competitor (T-linear/21miR), the system is stable (ON) and is shut off in the presence of structural sandwiched hydrides of M-linear+124a/Stem+221 miR due to the lack of adequate access to segments of RNA-miRs of chimeric probes. For the first time in this study, three probes were sandwiched on the separate liposome for sequential identification of 21-124a-221 or multiplex detection of miRs (221 or 124a with 21) with high specificity and sensitivity (as low as 0.1 fM). Electrochemical impedance (EIS) were performed for sensing three miRs in PBS containing 1 mM [Fe(CN)6]−3/−4 which DOTAP-DOPE liposome acted as an enhancing intermediate layer in the electrochemical reactions. Transmission Electron Microscopy (TEM)), Atomic Force Microscopy (AFM), Dynamic Light Scattering (DLS) and Ultraviolet–Visible (UV) spectroscopic techniques are used to understand the interactions between the DOPE-DOTAP, AuNP, different probes, miRs and p19.

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.

RNAi-based gene therapy has been recently considered as a promising approach against cancer. Targeted delivery of drug, gene or therapeutic RNAi-based systems to tumor cells is one of the important issues in order to reduce side effects on normal cells. Several strategies have been developed to improve the safety and selectivity of cancer treatments including antibodies, peptides and recently aptamers with various attractive characteristics including higher target specificity, affinity and reduced toxicity. Here we described a novel targeted delivery platform comprising modified PAMAM with 10-bromodecanoic acid (10C) and 10C-PEG for improvement of transfection efficiency, AS1411 aptamer for targeting nucleolin ligand on target cancer cells and shRNA plasmid for specific knockdown of Bcl-xL protein. Modified vector could significantly improve the transfection efficiency even after covalent or non-covalent aptamer binding compared to the non-targeted vector in A549 cells. The results of gene silencing and apoptosis assay indicated that our targeted shRNA delivery system could efficiently down-regulate the Bcl-xL expression up to 25% and induce 14% late apoptosis in target cancer cells with strong cell selectivity. This study proposed a novel targeted non-viral system for shRNA-mediated gene-silencing in cancer cells.

Gene therapy has emerged as an influential tool for treating the genetic and specific acquired disorders. Among all kinds of gene delivery systems, the cationic polymer polyethyleneimine (PEI) is considered as a promising non-viral gene delivery vector, although there are still concerns about its magnitude of cytotoxicity. While any cell insult leads to unfolded/misfolded protein accumulation and its consequent unfold protein response, evaluating the expression profile of ER-stress genes would be a sensitive indicator of cell stress. Beside cytotoxicity assays, real-time RT-PCR was used to investigate the effects of PEI nanoparticles on the endoplasmic reticulum. Treating Neuro2A cells revealed that PEI can induce cell toxicity in a concentration-dependent manner. Also, It increased the transcript levels of Grp78 (Bip), Atf4 and Chop, and splicing of Xbp1. To further optimize the transfection properties in Neuro2A cells, PEI was used to deliver a plasmid DNA containing GFP reporter. While different PEI/plasmid ratios revealed similar transfection efficiency, increasing the PEI/plasmid ratio led to induction of ER-stress markers. These results underscored that beside the effectiveness of PEI, using the lowest possible ratio of PEI/plasmid would minimize the detrimental effects of PEI on cells and confer it a beneficial therapeutic importance in nucleic acid delivery.

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.

Polyethyleneimine (PEI) is one of the most widely used polymers in gene delivery. Cell penetrating peptides (CPPs) are a group of peptides that can transfer materials from the membrane to the cytoplasm and nucleus. The aim of this study was to modify PEI with three different CPPs as a cationic polymer in order to investigate and compare their efficiency in gene delivery. The CPP-modified polymers were evaluated in terms of plasmid condensation, buffering capacity and number of primary amines. The prepared nanoparticles were examined for size, zeta potential, transfection efficiency and cytotoxicity in LNCAP cells. The resulting nanoparticles had a particle size between 183 and 313 nm. Measuring the zeta potential of the vectors indicated that the charge of the nanoparticles is between 21 and 25 mV. Vectors can condense plasmid in carrier-plasmid ratio of 4. Transfection results on the LNCAP cell line showed that the vector containing the BR2 peptide had the best transfection activity. In the cytotoxicity test, it was observed that vector containing CyLoP1 in carrier-plasmid ratio of 4 had the highest cell cytotoxicity. In conclusion, conjugation of CPPs to PEI due to physicochemical changes of the polymer resulted in improved transfection activity.

The present study aim to design a liposomal electrochemical sensor using 1, 2-dioleoyl-3-trimethylammoniumpropane (DOTAP) and dioleoylphosphatidylethanolamine(DOPE), chimeric probes and p19, it has been considered as a caliper molecule as well. Also the competitor structural hybrid (RNA) was used to detect three types of miRs in one screen printed electrode modified by gold nanoparticle (SCPE/GNP). In this purpose, the sensor signal stabilized when the cationic DOTAP-DOPE with hybrids of the chimeric probes (Stem, M-linear) sandwiched in order to detect 221–124a miRs. Given the lack of accessibility to RNA-miRs segments of chimeric probes, p19 inhibited the electrochemical reaction and shifted signal to off. After that p19 connected with the free hybrid of T-linear/21miR (just RNA) as competing for structure and the signal was shifted to ON, again. In this study, the electrochemical measurements were performed between the potentials at −0.4 V and +0.4 V with 1 mM [Fe(CN)6]-3-/4 which DOTAP-DOPE acted as an enhancer layer in the electrostatically reaction. This sensor determines as low as 0.4 fM of miRNA with high selectivity and specificity for sequential analysis of 124a-221-21 miRs in just 2 h.