MiR-320a upregulation contributes to the effectiveness of pemetrexed by inhibiting the growth and invasion of human lung cancer cell line (Calu-6).

BACKGROUND: Lung cancer is a common and deadly disease. Chemotherapy is the most common treatment, which inhibits cancer cell growth. Pemetrexed (PMX) is often used with other drugs. Environmental stress can affect regulatory non-coding RNAs such as MicroRNAs that modify gene expression. This study investigates the effect of PMX on the hsa-miR-320a-3p expression in the Calu-6 lung cancer cell line. METHODS AND RESULT: Calu-6 cells were cultured in an incubator with 37 °C, 5% CO2, and 98% humidity. The MTT test was performed to determine the concentration of PMX required to inhibit 50% of cell growth. To examine growth inhibition and apoptosis, release of lactate dehydrogenase (LDH), cell assays and caspase 3 and 7 enzyme activity were used. Finally, molecular studies were conducted to compare the expression of hsa-miR-320a-3p and genes including VDAC1, DHFR, STAT3, BAX and BCL2 before and after therapy. RESULTS: According to a study, it has been observed that PMX therapy significantly increases LDH release after 24 h. The study found that PMX's IC50 on Calu-6 is 8.870 µM. In addition, the treated sample showed higher expression of hsa-miR-320a-3p and BAX, while the expression of VDAC1, STAT3, DHFR and BCL2 decreased compared to the control sample. CONCLUSION: According to the findings of the current research, hsa-miR-320a-3p seems to have the potential to play an important role in the development of novel approaches to the treatment of lung cancer.

Tailored design and preparation of magnetic nanocomposite particles for the isolation of exosomes.

Here, we prepared a magnetic nanocomposite system composed of a cluster of magnetite nanoparticles coated with silica shell (MSNPs) with an average diameter of 140 ± 20 nm and conjugated with CD9 antibody (AntiCD9) using different strategies including adsorption or chemical conjugation of antibody molecules to either aminated MSNPs (AMSNPs) or carboxylated MSNPs (CMSNPs). Then, MSNPs were employed to isolate exosomes from ultracentrifuge-enriched solution, PC3 cell-culture medium, or exosome-spiked simulated plasma samples. Quantitative tests using nanoparticle-tracking analysis confirmed antibody-covalently conjugated MSNPs, i.e. the AntiCD9-AMSNPs and AntiCD9-CMSNPs enabled >90% recovery of exosomes. Additionally, the exosomes isolated with AntiCD9-CMSNPs showed higher recovery efficiency compared to the AntiCD9-AMSNPs. For both nanoadsorbents, lower protein impurities amounts were obtained as compared to that of exosomes isolated by ultracentrifugation and Exocib kit. The mean diameter assessment of the isolated exosomes indicates that particles isolated by using AntiCD9-AMSNPs and AntiCD9-CMSNPs have smaller sizes (136 ± 2.64 nm and 113 ± 11.53 nm, respectively) than those obtained by UC-enriched exosomes (140.9 ± 1.6 nm) and Exocib kit (167 ± 10.53 nm). Such promising results obtained in the isolation of exosomes recommend magnetic nanocomposite as an efficient tool for the simple and fast isolation of exosomes for diagnosis applications.

Biotransformation of benzaldehyde into L-phenylacetylcarbinol using magnetic nanoparticles-coated yeast cells.

OBJECTIVES: The yeast cells were coated with Fe3O4 magnetic nanoparticles and employed as biocatalyst for the microbial biotransformation of benzaldehyde into L-phenylacetylcarbinol (L-PAC). RESULTS: Saccharomyces cerevisiae CEN.PK113-7D yeast cells were coated with magnetic nanoparticles to facilitate the cells separation process. Transmission electron microscopy, powder XRD diffraction, and vibrating sample magnetometer were used to characterize magnetic nanoparticles and magnetic nanoparticle-coated yeast cells. Then the reusability of magnetically recoverable cells in production of L-PAC was investigated. Results show that coating yeast cells with magnetic nanoparticles does not affect their size and structure. Coated cells were also used in seven consecutive batch cycles and no significant reduction for L-PAC titer was observed in any of the cycles. CONCLUSION: Coating yeast cells with magnetic nanoparticles enabled rapid separation and reuse of cells in several successive batch cycle without affecting their ability to produce L-PAC.

Study of Molecular Conformation and Activity-Related Properties of Lipase Immobilized onto Core-Shell Structured Polyacrylic Acid-Coated Magnetic Silica Nanocomposite Particles.

A facile approach for the preparation of core-shell structured poly(acrylic acid) (PAA)-coated Fe3O4 cluster@SiO2 nanocomposite particles as the support materials for the lipase immobilization is reported. Low- or high-molecular-weight (1800 and 100,000, respectively) PAA molecules were covalently attached onto the surface of amine-functionalized magnetic silica nanoacomposite particles. The successful preparation of particles were verified by scanning transmission electron microscopy (STEM), X-ray diffraction (XRD), vibrating sample magnetometer (VSM), thermogravimetric analysis (TGA), zeta potential measurement, and Fourier-transform infrared (FTIR) techniques. Once lipase is covalently immobilized onto the particles with an average diameter of 210 ± 50 nm, resulting from high binding sites concentrations on the low- and high-molecular-weight PAA-coated particles, high lipase immobilization efficiencies (86.2% and 89.9%, respectively), and loading capacities (786 and 816 mg g(-1), respectively) are obtained. Results from circular dichroism (CD) analysis and catalytic activity tests reveal an increase in the ß-sheet content of lipase molecules upon immobilization, along with an enhancement in their activities and stabilities. The lipases immobilized onto the low- and high-molecular-weight PAA-coated particles show maximum activities at 55 and 50 °C, respectively, which are ∼28% and ∼15% higher than that of the free lipase at its own optimum temperature (40 °C), respectively. The immobilized lipases exhibit excellent performance at broader temperature and pH ranges and high thermal and storage stabilities, as well as superior reusability. These prepared magnetic nanocomposite particles can be offered as suitable support materials for efficient immobilization of enzymes and improvement of the immobilized enzymes properties.

Curcumin-Loaded Amine-Functionalized Mesoporous Silica Nanoparticles Inhibit α-Synuclein Fibrillation and Reduce Its Cytotoxicity-Associated Effects.

This study aimed to develop a drug carrier based on amine-functionalized mesoporous silica nanoparticles (AAS-MSNPs) for a poorly water-soluble drug, curcumin (CUR), and to study its effects on α-synuclein (α-Syn) fibrillation and cytotoxicity. Here, we show that AAS-MSNPs possess high values of loading efficiency and capacity (33.5% and 0.45 mg drug/mg MSNPs, respectively) for CUR. It is also revealed that α-Syn species interact strongly with the CUR-loaded AAS-MSNPs, leading to a significant inhibition of the fibrillation process. Furthermore, these samples reduce the toxic effects of CUR. However, drug-loaded AAS-MSNPs do not affect the cytotoxic properties of the formed fibrils considerably. In addition, CUR loaded onto AAS-MSNPs shows enhanced stability in comparison with that of the free drug. These remarkable properties introduce AAS-MSNPs as a promising tool for the formulation of poorly water-soluble drugs such as CUR.

Desulfurization activity and reusability of magnetite nanoparticle-coated Rhodococcus erythropolis FMF and R. erythropolis IGTS8 bacterial cells.

The application of Fe3 O4 nanoparticles to the separation of desulfurizing bacterial cells and their influence on the desulfurization activity and reusability of the two bacterial strains Rhodococcus erythropolis FMF and R. erythropolis IGTS8 were investigated. Magnetite nanoparticles were synthesized via the reverse coprecipitation method. Transmission electron microscopy (TEM) images showed that the magnetite nanoparticles had sizes of 5.35 ± 1.13 (F1 nanoparticles) and 8.74 ± 1.18 nm (F2 nanoparticles) when glycine was added during the synthesis of nanoparticles and when it was absent from the reaction mixture, respectively. Glycine was added after the synthesis of both F1 and F2 nanoparticles to stabilize the nanoparticle dispersion. TEM images of cells treated with magnetite nanoparticles indicated that F1 nanoparticles were immobilized on the surface of bacterial cells more evenly than the F2 nanoparticles. Desulfurization activities of the F1 magnetite nanoparticle-coated R. erythropolis FMF and R. erythropolis IGTS8 cells (with sulfur-removal percentage values of 70 ± 4 and 73 ± 3, respectively), as examined with the spectrophotometric Gibbs assay (based on dibenzothiophene degradation and sulfur-removal percentage), were not significantly different from those for the free bacterial cells (67 ± 3 and 69 ± 4, respectively). These results indicate that magnetite nanoparticles cannot affect the desulfurization activity of cells examined in this work. Isolation of bacterial cells from the suspension using a magnet and evaluation of desulfurization activity of separated cells showed that Fe3 O4 nanoparticles can provide a high-efficiency recovery of bacterial cells from a suspension, with the reused magnetite nanoparticle-coated bacterial cells being able to maintain their desulfurization activity efficiently.

Effect of the Structure of Magnetic Nanocomposite Adsorbents on the Lysozyme Separation Efficiency.

In this study, we prepared various anionic magnetic adsorbents through the carboxyl functionalization of core/shell-structured Fe3O4/SiO2 (FS) particles by either succinic anhydride (FSC), low-molecular-weight (MW 1800) polyacrylic acid (PAA) (FSP1), or high-molecular-weight (MW 100,000) PAA (FSP2), and then, investigated the effect of the structure of adsorbents and operational parameters on their performance for the lysozyme separation. The type and size of functional molecules have significant effects on the surface concentration of functional carboxyl groups onto the adsorbent particles (increase in the order of FSP2 > FSP1 > FSC), and consequently on the adsorption efficiency (AE) (∼100, 98, and 62%, respectively) and adsorption capacity (AC) (∼1000, 980, and 621 mg·g-1, respectively) of the adsorbents. However, the loss of the antibacterial activity of separated lysozyme molecules due to the molecular conformational change increased in the order of FSP2 > FSP1 = FSC, as compared to the free lysozyme. The application of basic buffer solutions for the elution of adsorbed enzyme molecules resulted in more adverse effects on the enzyme activity. The obtained results recommend that FSP1 can be used as a suitable anionic adsorbent for the isolation of positively charged proteins, owing to its high adsorption capacity, excellent reusability, and structural stability, as well as the high purity, structural stability, and activity recovery of the isolated proteins.

pH Shock-promoted lysozyme corona for efficient pathogenic infections treatment: Effects of surface chemistry of mesoporous silica nanoparticles and loading method.

The emergence of antibiotic resistant bacteria because of the antibiotics abusement was the motivation to develop the effective alternatives to traditional antibiotics. Hence, various lysozyme corona were prepared through the physical and covalent attachment of lysozyme molecules onto either the bare or carboxyl-functionalized mesoporous silica particles. The prepared samples were characterized by STEM, TGA/DTA, zeta potential, FTIR, UV-vis and CD spectroscopic methods. All the prepared lysozyme-coated particles exhibited an efficient antibacterial activity against Listeria monocytogenes, as a case study, in vitro with no cytotoxicity. The minimal inhibition concentration (MIC) of the lysozyme-physically adsorbed bare and carboxyl-functionalized mesoporous silica nanoparticles (L-MS and L-ads-CMS, respectively) and the lysozyme-covalently attached carboxyl-functionalized MS particles (L-cov-CMS) was 2, 5.3 and 1.7 folds lower than that of the free lysozyme, respectively. Additionally, for the first time, it was reported that the pretreatment of lysozyme corona of L-ads-CMS through inducing a pH-shock can lead to the enhancement of antibacterial properties thereof. This behavior was associated to the controlled release of the immobilized lysozyme molecules and their conformational stability. These natural antibacterial lysozyme-coated silica nanoparticles showing the "pH-shock enhanced activity" could be of utmost interest for design of the highly active enzyme-modified nanoparticles.

Ultrasound-mediated gene delivery into suspended plant cells using polyethyleneimine-coated mesoporous silica nanoparticles.

Sonoporation, ultrasound-mediated membrane perforation can potentially puncture plasma membrane and rigid cell wall on presumably reversible basis which benefit gene transfection and plant biotechnology. Herein, positively charged poly-ethyleneimine (PEI)-coated mesoporous silica nanoparticles (MSNs) with an average diameter of 100 ± 8.7 nm was synthesized for GUS-encoding plasmid delivery into the suspended tobacco cells using the ultrasound treatment. The overall potential of PEI-MSN for DNA adsorption was measured at 43.43 µg DNA mg-1 PEI-MSNs. It was shown that high level of sonoporation may adversely upset the cell viability. Optimal conditions of ultrasonic treatment are obtained as 8 min at 3 various intensities of 160, 320 and 640 W. Histochemical staining assay was used to follow the protein expression. It was shown that PEI-coated MSNs efficiently transfer the GUS-encoding plasmid DNA into the tobacco cells. The results of this study showed that ultrasonic treatment provides an economical and straightforward approach for gene transferring into the plant cells without any need to complicated devices and concerns about safety issues.

Mesoporous silica nanoparticles-based formulations of a chimeric proteinous vaccine candidate against necrotic enteritis disease.

To develop a nanoparticle-based vaccine against necrotic enteritis, a chimeric antigen (rNA) consisting of the main antigens of Clostridium perfringens, NetB, and Alpha toxin, was prepared. Then, the rNA molecules were loaded onto the functionalized mesoporous silica nanoparticles (MSNPs) using physical adsorption or covalent conjugation methods. The characterization of synthesized nanoparticles was performed by scanning electron microscopy, dynamic light scattering, zeta potential measurement, Fourier transform infrared spectroscopy, and thermogravimetry techniques. The results revealed that the spherical nanoparticles with an average diameter of 90 ±â€¯12 nm and suitable surface chemistries are prepared. MSNPs-based formulations did not show any significant toxicity on the chicken embryo fibroblast cells. The results of the challenge experiments using subcutaneous or oral administration of the as-prepared formulations in the animal model showed that the as-prepared nanosystems, similar to those formulated with a commercial adjuvant (Montanide), present stronger humoral immune responses as compared to that of the free proteins. It was also indicated that the best protection is obtained in groups vaccinated with MSNPs-based nanovaccine, especially those who orally received covalently conjugated nanovaccine candidates. These results recommend that the MSNPs-based formulated chimeric proteinous vaccine candidates can be considered as an effective immunizing system for the oral vaccination of poultry against gastrointestinal infectious diseases.

Comparison of oncolytic virotherapy and nanotherapy as two new miRNA delivery approaches in lung cancer.

Lung cancer is known as the second leading cause of cancer death. Finding ways to detect early-stage lung cancer can remarkably increase the survival rate. Biomarkers such as microRNAs can be helpful in cancer diagnosis, predicting its prognosis, and patients' chances of survival. Numerous studies have confirmed the correlation between microRNA expression and the likelihood of patients surviving after treatment. Consequently, it is necessary to study the expression profile of microRNAs during and after treatment. Oncolytic virotherapy and nanotherapy are two neoteric methods that use various vectors to deliver microRNAs into cancer cells. Although these treatments have not yet entered into the clinical trials, much progress has been made in this area. Analyzing the expression profile of microRNAs after applying nanotherapy and oncolytic virotherapy can evaluate the effectiveness of these methods. This review refers to the studies conducted about these two approaches. The advantages and disadvantages of these methods in delivery and affecting microRNA expression patterns are discussed below.

Highly efficient detection of cancer-derived exosomes using modified core-shell electrospun nanofibers as a capture substrate and antibody immobilized-graphene quantum dots as a signaling agent.

In the past few years graphene quantum dots (GQDs) have been used as a signaling agent for medical diagnosis. They can be modified and labeled with different macromolecules to give them potential to be attached to a specific target. Herein GQDs were labeled with an antibody which is specific for cancer-derived exosomes, isolated from blood serum by using a specialized PCL-gelatin core-shell NFM. This membrane showed excellent sensitivity for isolating exosomes from a complex mixture such as serum, and the GQD-antibody complex detected the isolated exosomes with great sensitivity. The final results allow this method to be considered as one that can be used to quantify the concentration of a desired analyte in a mixture.

The fate of mesenchymal stem cells is greatly influenced by the surface chemistry of silica nanoparticles in 3D hydrogel-based culture systems.

Polymeric hydrogel-based 3D scaffolds are well-known structures, being used for cultivation and differentiation of stem cells. However, scalable systems that provide a native-like microenvironment with suitable biological and physical properties are still needed. Incorporation of nanomaterials into the polymeric systems is expected to influence the physical properties of the structure but also the stem cells fate. Here, alginate/gelatin hydrogel beads incorporated with mesoporous silica nanoparticles (MSNs) (average diameter 80.9 ±â€¯10 nm) and various surface chemistries were prepared. Human adipose-derived mesenchymal stem cells (hASCs) were subsequently encapsulated into the alginate/gelatin/silica hydrogels. Incorporation of amine- and carboxyl-functionalized MSNs (A-MSNs and C-MSNs) significantly enhances the stability of the hydrogel beads. In addition, the expression levels of Nanog and OCT4 imply that the incorporation of A-MSNs into the alginate/gelatin beads significantly improves the proliferation and the stemness of encapsulated hASCs. Importantly, our findings show that the presence of A-MSNs slightly suppresses in vivo inflammation. In contrast, the results of marker gene expression analyses indicate that cultivation of hASCs in alginate beads incorporated with C-MSNs (10% w/w) leads to a heterogeneously differentiated population of the cells, i.e., osteocytes, chondrocytes, and adipocytes, which is not appropriate for both cell culture and differentiation applications.

Preventing protein adsorption from a range of surfaces using an aqueous fish protein extract.

We utilize an aqueous extract of fish proteins (FPs) as a coating for minimizing the adsorption of fibrinogen (Fg) and human serum albumin (HSA). The surfaces include stainless steel (SS), gold (Au), silicon dioxide (SiO(2)), and poly(styrene) (PS). The adsorption processes (kinetics and adsorbed mass) are followed by quartz crystal microbalance with dissipation (QCM-D). Complementary surface information is provided by X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). QCM-D shows no mass increases to any of the FP-coated surfaces upon treating with Fg or HSA. Also, when Fg- or HSA-coated surfaces are exposed to the FPs, a significant increase in adsorbed mass occurs because the FPs are highly surface-active displacing Fg. Additionally, fluorescence microscopy confirms that very little Fg adsorbs to the FP-coated surfaces. We propose that FP coatings prevent protein adsorption by steric stabilization and could be an alternative method for preventing unwanted bioadhesion on medical materials.

Investigation of Desulfurization Activity, Reusability, and Viability of Magnetite Coated Bacterial Cells.

BACKGROUND: Magnetic separation using magnetic nanoparticles can be used as a simple method to isolate desulfurizing bacteria from a biphasic oil/water system. OBJECTIVES: Magnetite nanoparticles were applied to coat the surface of Rhodococcus erythropolis IGTS8 and Rhodococcus erythropolis FMF desulfurizing bacterial cells, and the viability and reusability of magnetite-coated bacteria evaluated by using various methods. MATERIAL AND METHODS: Magnetite nanoparticles were synthesized through a reverse co-precipitation method. Glycine was added during and after the synthesis of magnetite nanoparticles to modify their surface and to stabilize the dispersion of the nanoparticles. The glycine-modified magnetite nanoparticles were immobilized on the surface of both oil-desulfurizing bacterial strains. Reusability of magnetite-coated bacterial cells was evaluated via assessing the desulfurization activity of bacteria via spectrophotometry using Gibb's assay, after the separation of bacterial cells from 96h-cultures with the application of external magnetic field. In addition, CFU and fluorescence imaging were used to investigate the viability of magnetite-coated and free bacterial cells. RESULTS: TEM micrographs showed that magnetite nanoparticles have the size approximately 5.35±1.13 nm. Reusability results showed that both magnetite-coated bacterial strains maintain their activity even after 5 × 96h-cycles. The viability results revealed glycine-modified magnetite nanoparticles did not negatively affect the viability of two bacterial strains R. erythropolis IGTS8 and R. erythropolis FMF. CONCLUSIONS: In conclusion, the glycine-modified magnetite nanoparticles have great capacity for immobilization and separation of desulfurizing bacteria from suspension.

Silica nanoparticle surface chemistry: An important trait affecting cellular biocompatibility in two and three dimensional culture systems.

Great advantages bestowed by mesoporous silica nanoparticles (MSNs) including high surface area, tailorable pore diameter and surface chemistry, and large pore volume render them as efficient tools in biomedical applications. Herein, MSNs with different surface chemistries were synthesized and investigated in terms of biocompatibility and their impact on the morphology of bone marrow-derived mesenchymal stem cells both in 2D and 3D culture systems. Bare MSNs (BMSNs) were synthesized by template removing method using tetraethylorthosilicate (TEOS) as a precursor. The as-prepared BMSNs were then used to prepare amine-functionalized (AMSNs), carboxyl-functionalized (CMSNs) and polymeric amine-functionalized (PMSNs) samples, consecutively. These nanoparticles were characterized by scanning electron microscopy, zeta potential measurement, dynamic light scattering, BET (Brunauer, Emmett, Teller) analysis, and FTIR technique. In a 3D culture system, stem cells were encapsulated in alginate hydrogel in which MSNs of different functionalities were incorporated. The results showed good biocompatibility for both BMSNs and AMSNs in 2D and 3D culture systems. For these samples, the viability of about 80% was acquired after 2 weeks of 3D culture. When compared to the control, CMSNs caused higher cell proliferation in the 2D culture; while they showed cytotoxic effects in the 3D culture system. Interestingly, polymeric amine-functionalized silica nanoparticles (PMSNs) resulted in disrupted morphology and very low viability in the 2D cell culture and even less viability in 3D environment in comparison to BMSNs and AMSNs. This significant decrease in cell viability was attributed to the higher uptake values of highly positively charged PMSNs by cells as compared to other MSNs. This up-regulated uptake was evaluated by using an inductively coupled plasma optical emission spectroscopy instrument (ICP-OES). These results uncover different interactions between cell and nanoparticles with various surface chemistries. Building on these results, new windows are opened for employing biocompatible nanoparticles such as BMSNs and AMSNs, even at high concentrations, as potential cargos for carrying required growth and/or differentiation factors for tissue engineering applications.

Mesoporous silica nanoparticles carrying multiple antibiotics provide enhanced synergistic effect and improved biocompatibility.

Treatment of polymicrobial infections requires combination therapy with drugs that have different antimicrobial spectra and possibly work in synergy. However, the different pharmacokinetics and adverse side effects challenge the simultaneous delivery of multiple drugs at the appropriate concentrations to the site of infection. Formulation of multiple drugs in nano-carrier systems may improve therapeutic efficacy by increasing the local concentration and lowering the systemic concentration, leading to fewer side effects. In this study, we loaded polymyxin B and vancomycin on bare and carboxyl-modified mesoporous silica nanoparticles (B-MSNs and C-MSNs, respectively) to achieve simulataneous local delivery of antibiotics against Gram-positive and -negative bacteria. Polymyxin B adsorbed preferentially to nanoparticles compared to vancomycin. The total antibiotic loading was 563 µg and 453 µg per mg B-MSNs or C-MSNs, respectively. Both B-MSNs and C-MSNs loaded with antibiotics were effective against Gram-negative and Gram-positive bacteria. The antibiotics had synergistic interactions against Gram-negative bacteria, and the antimicrobial efficacy was higher for antibiotic-loaded C-MSNs compared to free antibiotics at the same concentration even though the cytotoxicity was lower. Our study shows that formulations of existing antibiotics in nanocarrier systems can improve their therapeutic efficiency, indicating that combination therapy with drug-loaded silica nanoparticles may provide a better treatment outcome for infections that require high concentrations of multiple drugs.

Loading of polymyxin B onto anionic mesoporous silica nanoparticles retains antibacterial activity and enhances biocompatibility.

Polymyxin B is a polycationic antibiotic used as the last line treatment against antibiotic-resistant Gram negative bacteria. However, application of polymyxin B is limited because of its toxicity effects. Herein, we used bare and surface modified mesoporous silica nanoparticles (MSNs) with an average diameter of 72.29 ±â€¯8.17 nm as adsorbent for polymyxin B to improve its therapeutic properties. The polymyxin B adsorption onto MSN surfaces was explained as a function of pH, type of buffer and surface charge of nanoparticles, according to the ζ-potential of silica nanoparticles and adsorption kinetics results. The highest value of the adsorption capacity (about 401 ±â€¯15.38 mg polymyxin B/ g silica nanoparticles) was obtained for the bare nanoparticles in Tris buffer, pH 9. Release profiles of polymyxin B showed a sustained release pattern, fitting Power law and Hill models. The antibiotic molecules-loaded nanoparticles showed enhanced antibacterial activity compared to free antibiotic against different Gram negative bacteria. Biocompatibility evaluation results revealed that loading of polymyxin B onto MSNs can decrease the cytotoxicity effects of the drug by reducing ROS generation. Our results suggest that formulation of drugs by adsorption onto MSNs may offer a way forward to overcome the adverse effects of some antibiotics such as polymyxin B without compromising their antimicrobial properties.

EspA-loaded mesoporous silica nanoparticles can efficiently protect animal model against enterohaemorrhagic E. coli O157: H7.

In the present study, the application of mesoporous silica nanoparticles (MSNPs) loaded with recombinant EspA protein, an immunogen of enterohaemorrhagic E. coli, was investigated in the case of BALB/c mice immunization against the bacterium. MSNPs of 96.9 ± 15.9 nm in diameter were synthesized using template removing method. The immunization of mice was carried out orally and subcutaneously. Significant immune responses to the antigen were observed for the immunized mice when rEspA-loaded MSNPs were administered in both routes in comparison to that of the antigen formulated using a well-known adjuvant, i.e. Freund's. According to the titretitre of serum IL-4, the most potent humoral responses were observed when the mice were immunized subcutaneously with antigen-loaded MSNPs (244, 36 and 14 ng/dL of IL-4 in the serum of mice immunized subcutaneously or orally by antigen-loaded MSNPs, and subcutaneously by Freund's adjuvant formulated-antigen, respectively). However, the difference in serum IgG and serum IgA was not significant in mice subcutaneously immunized with antigen-loaded MSNPs and mice immunized with Freund's adjuvant formulated-antigen. Finally, the immunized mice were challenged orally by enterohaemorrhagic E. coli cells. The amount of bacterial shedding was significantly reduced in faecesfaeces of the animals immunized by antigen-loaded MSNPs in both subcutaneous and oral routes.