Publications by authors named "Mojtaba Salouti"

25 Publications

  • Page 1 of 1

Immunogenicity evaluating of the SLNs-alginate conjugate against Pseudomonas aeruginosa.

J Immunol Methods 2021 Jan 28;488:112938. Epub 2020 Nov 28.

Nanobiotechnology Research Center, Zanjan Branch, Islamic Azad University, Zanjan, Iran. Electronic address:

P. aeruginosa is of particular importance due to its numerous pathogens and the spread of its multidrug-resistant strains around the world. Hence there is a need to develop an effective vaccine to prevent the diseases with P. aeruginosa. The aim of present study was to evaluate the immunogenicity of alginate (Alg) antigen in conjugation with SLN as a candidate for nanovaccine against P. aeruginosa in mouse model. Alginate is a weak immunogen, but the immune responses produced by alginate are effective in killing Pseudomonas bacteria. To increase the immunogenicity of alginate, SLN was used that is useful in drug delivery and can boost prolonged effectiveness. The results of ELISA and opsonophagocytosis tests showed that Alg-SLN conjugate has a better ability to stimulate the immune system to produce more immunoglobulins with better performance compared to alginate antigen alone. The challenge test also demonstrated that the Alg-SLN treated mice showed a higher level of immunity than the mice treated with pure alginate against infections caused by P. aeruginosa. Overally the findings showed the efficacy of new prepared vaccine to induce immunogenicity, and therefore it can be considered as a candidate for a strong vaccine against P. aeruginosa.
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http://dx.doi.org/10.1016/j.jim.2020.112938DOI Listing
January 2021

Enhanced sensitivity and efficiency of detection of based on modified magnetic nanoparticles by photometric systems.

Artif Cells Nanomed Biotechnol 2020 Dec;48(1):810-817

Department of Pathology, Zanjan University of Medical Sciences, Zanjan, Iran.

is an important infectious factor in the food industry and hospital infections. Many methods are used for detecting bacteria but they are mostly time-consuming, poorly sensitive. In this study, a nano-biosensor based on iron nanoparticles (MNPs) was designed to detect . MNPs were synthesized and conjugated to Biosensors. Then was lysed and nano-biosensor (MNP-TiO-AP-SMCC-Biosensors) was added to the lysed bacteria. After bonding the bacterial genome to the nano-biosensor, MNPs were separated by a magnet. Bacterial DNA was released from the surface of nano-biosensor and researched by Nano-drop spectrophotometry. The results of SEM and DLS revealed that the size of MNPs was 20-25 nm which increased to 38-43 nm after modification and addition of biosensors. The designed nano-biosensor was highly sensitive and specific for the detection of . The limit of detection (LOD) was determined as 230 CFU mL. There was an acceptable linear correlation between bacterial concentration and absorption at 3.7 × 10-3.7× 10 whose linear diagram and regression was  = 0.242 + 2.08 and = .996. Further, in the presence of other bacteria as a negative control, it was absolutely specific. The sensitivity of the designed nano-biosensor was investigated and compared through PCR.
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http://dx.doi.org/10.1080/21691401.2020.1748638DOI Listing
December 2020

Preparation of a nanovaccine against M16 based on PLGA nanoparticles and oligopolysaccharide antigen.

Artif Cells Nanomed Biotechnol 2019 Dec;47(1):4248-4256

Faculty of Sciences, Department of Microbiology, Islamic Azad University, Zanjan, Iran.

Brucellosis is one of the most common and important diseases between humans and animals. Herein, we developed a nanovaccine against based on oligopolysaccharide (OPS) antigen and PLGA nanoparticles. The conjugation of extracted OPS with poly lactic-co-glycolic acid was performed. The antigenicity evaluation was conducted in 4 groups of 5 female BALB/c mice including OPS-PLGA conjugate, OPS alone, PLGA alone and PBS as a control. The mice were vaccinated intra-peritoneal three times with two-week intervals. To determine the immune response and functional capacity of the antibodies, the enzyme linked immunosorbent, opsonophagocytosis and challenge tests were performed. For checking the immunization ability of the nanovaccine, the challenge test was performed. The results showed a significant increase in the total IgG and IgM antibody titres in the mice vaccinated with OPS-PLGA conjugate in comparison with other groups. The sera of animals immunized with OPS-PLGA conjugate promoted efficient opsonophagocytosis of bacteria. The results of challenge assay showed that the immunization with OPS-PLGA conjugate gave a high level of protection in comparison with other groups. These findings showed that the new nanovaccine can be considered as a candidate for immunization of animals and humans against the diseases caused by that needs further investigations.
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http://dx.doi.org/10.1080/21691401.2019.1687490DOI Listing
December 2019

Highly selective and sensitive detection of Staphylococcus aureus with gold nanoparticle-based core-shell nano biosensor.

Mol Cell Probes 2018 10 25;41:8-13. Epub 2018 Jul 25.

Biology Research Center, Zanjan Branch, Islamic Azad University, Zanjan, Iran.

Staphylococcus aureus is a gram-positive and opportunistic pathogen that is one of the most common causes of nosocomial infections; therefore, its rapid diagnosis is important and valuable. Today, the use of nanoparticles is expanding due to their unique properties. The purpose of the present study is the determination of S. aureus by a colorimetric method based on gold nanoparticles (AuNPs). Firstly, S. aureus was cultured on both LB media (broth and agar) and their chromosomal DNA was extracted. Afterwards, primers and biosensor were designed based on Protein A sequence data in the gene bank. PCR assay was performed under optimal conditions and the PCR product was electrophoresed on 2-percent agarose gel. The synthesized biosensors were conjugated with AuNPs and, eventually, a single-stranded genome was added to the conjugated AuNPs and hybridization was performed. The results were evaluated based on color change detected by the naked eye, optical spectrophotometry, and transient electron microscopy. Finally, the sensitivity and specificity of the AuNP-biosensor were determined. The results of the present study showed a 390 bp band on the agarose electrophoresis gel, which confirmed the presence of Protein A genes on the chromosome of the bacteria. The PCR and colorimetric methods were compared with each other. The sensitivity of the PCR and colorimetric methods were 30 ng μL and 10 ng μL, respectively. The limit of detection (LOD) equaling 8.73 ng μL was determined and the specificity of the method was confirmed by the DNA of other bacteria. According to the results, the present method is rapid and sensitive in detecting S. aureus.
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http://dx.doi.org/10.1016/j.mcp.2018.07.004DOI Listing
October 2018

Spectrophotometric, colorimetric and visually detection of Pseudomonas aeruginosa ETA gene based gold nanoparticles DNA probe and endonuclease enzyme.

Spectrochim Acta A Mol Biomol Spectrosc 2018 Jun 24;199:421-429. Epub 2018 Mar 24.

Department of Biology, Science and Research Branch (IAU), Islamic Azad University, Tehran, Iran.

Colorimetric DNA detection is preferred over other methods for clinical molecular diagnosis because it does not require expensive equipment. In the present study, the colorimetric method based on gold nanoparticles (GNPs) and endonuclease enzyme was used for the detection of P. aeruginosa ETA gene. Firstly, the primers and probe for P. aeruginosa exotoxin A (ETA) gene were designed and checked for specificity by the PCR method. Then, GNPs were synthesized using the citrate reduction method and conjugated with the prepared probe to develop the new nano-biosensor. Next, the extracted target DNA of the bacteria was added to GNP-probe complex to check its efficacy for P. aeruginosa ETA gene diagnosis. A decrease in absorbance was seen when GNP-probe-target DNA cleaved into the small fragments of BamHI endonuclease due to the weakened electrostatic interaction between GNPs and the shortened DNA. The right shift of the absorbance peak from 530 to 562nm occurred after adding the endonuclease. It was measured using a UV-VIS absorption spectroscopy that indicates the existence of the P. aeruginosa ETA gene. Sensitivity was determined in the presence of different concentrations of target DNA of P. aeruginosa. The results obtained from the optimized conditions showed that the absorbance value has linear correlation with concentration of target DNA (R: 0.9850) in the range of 10-50ngmL with the limit detection of 9.899ngmL. Thus, the specificity of the new method for detection of P. aeruginosa was established in comparison with other bacteria. Additionally, the designed assay was quantitatively applied to detect the P. aeruginosa ETA gene from 10 to 10CFUmL in real samples with a detection limit of 320CFUmL.
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http://dx.doi.org/10.1016/j.saa.2018.03.056DOI Listing
June 2018

Enhanced antibacterial effect of azlocillin in conjugation with silver nanoparticles against .

IET Nanobiotechnol 2017 Dec;11(8):942-947

Department of Genetic, Faculty of Sciences, Zanjan Branch, Islamic Azad University, Zanjan, Iran.

In recent years, the problems associated with bacterial resistance to antibiotics caused nanodrugs to be considered as a new way for infectious diseases treatment. The main purpose of this study was to develop a new agent against a very difficult bacterium to treat, based on azlocillin antibiotic and silver nanoparticles (AgNPs). Azlocillin was conjugated with AgNPs by chemical methods and its antimicrobial activity was studied against . using well diffusion agar method. Then, minimum inhibitory concentration and minimum bactericidal concentration of the new conjugate was specified with macro-dilution method. The animal study showed the considerable enhanced antibacterial effect of azlocillin in conjugation with AgNPs against . in comparison with azlocillin alone, AgNPs alone and azlocillin in combination with AgNPs.
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http://dx.doi.org/10.1049/iet-nbt.2017.0009DOI Listing
December 2017

BBN conjugated GNPs: a new targeting contrast agent for imaging of breast cancer in radiology.

IET Nanobiotechnol 2017 Aug;11(5):604-611

Department of Radiology, Faculty of Paramedical Sciences, Zanjan University of Medical Sciences, Zanjan, Iran.

Using of targeted contrast agents in X-ray imaging of breast cancer can improve the accuracy of diagnosis, staging, and treatment planning by providing early detection and superior definition of tumour volume. This study demonstrates a new class of X-ray contrast agents based on gold nanoparticles (GNPs) and bombesin (BBN) for imaging of breast cancer in radiology. GNPs were synthesised in spherical shape in the size range of 15 ± 2 nm and conjugated with BBN followed by coating with polyethyleneglycol (PEG). The and behaviour of PEG-coated GNPs-BBN conjugate was investigated performing cytotoxicity, binding, and internalisation assays as well as biodistribution and X-ray imaging studies in mouse bearing breast tumour. Cytotoxicity study showed biocompatibility of the prepared bioconjugate. The binding and internalisation studies using T47D cell line approved the targeting ability of new agent. The biodistribution study showed the considerable accumulation of prepared conjugate in breast tumour in mouse model. The breast tumour was clearly visualised in X-ray images taken from the mouse model. The results showed the potential of PEG-coated GNPs-BBN conjugate as a contrast agent in X-ray imaging of breast tumour in humans that need further investigations.
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http://dx.doi.org/10.1049/iet-nbt.2016.0191DOI Listing
August 2017

Fluorescence bio-barcode DNA assay based on gold and magnetic nanoparticles for detection of Exotoxin A gene sequence.

Biosens Bioelectron 2017 Jun 19;92:679-686. Epub 2016 Oct 19.

Department of Biology, Science and Research Branch, Islamic Azad University (IAU), Tehran, Iran.

Bio-barcode DNA based on gold nanoparticle (bDNA-GNPs) as a new generation of biosensor based detection tools, holds promise for biological science studies. They are of enormous importance in the emergence of rapid and sensitive procedures for detecting toxins of microorganisms. Exotoxin A (ETA) is the most toxic virulence factor of Pseudomonas aeruginosa. ETA has ADP-ribosylation activity and decisively affects the protein synthesis of the host cells. In the present study, we developed a fluorescence bio-barcode technology to trace P. aeruginosa ETA. The GNPs were coated with the first target-specific DNA probe 1 (1pDNA) and bio-barcode DNA, which acted as a signal reporter. The magnetic nanoparticles (MNPs) were coated with the second target-specific DNA probe 2 (2pDNA) that was able to recognize the other end of the target DNA. After binding the nanoparticles with the target DNA, the following sandwich structure was formed: MNP 2pDNA/tDNA/1pDNA-GNP-bDNA. After isolating the sandwiches by a magnetic field, the DNAs of the probes which have been hybridized to their complementary DNA, GNPs and MNPs, via the hydrogen, electrostatic and covalently bonds, were released from the sandwiches after dissolving in dithiothreitol solution (DTT 0.8M). This bio-barcode DNA with known DNA sequence was then detected by fluorescence spectrophotometry. The findings showed that the new method has the advantages of fast, high sensitivity (the detection limit was 1.2ng/ml), good selectivity, and wide linear range of 5-200ng/ml. The regression analysis also showed that there was a good linear relationship (∆F=0.57 [target DNA]+21.31, R=0.9984) between the fluorescent intensity and the target DNA concentration in the samples.
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http://dx.doi.org/10.1016/j.bios.2016.10.030DOI Listing
June 2017

In vitro evaluation of actively targetable superparamagnetic nanoparticles to the folate receptor positive cancer cells.

Mater Sci Eng C Mater Biol Appl 2016 Dec 1;69:1147-58. Epub 2016 Aug 1.

Faculty of Biomedical Engineering, Universiti Teknologi Malaysia, Skudai, 81310 Johor Bahru, Johor, Malaysia.

Engineering of a physiologically compatible, stable and targetable SPIONs-CA-FA formulation was reported. Initially fabricated superparamagnetic iron oxide nanoparticles (SPIONs) were coated with citric acid (CA) to hamper agglomeration as well as to ameliorate biocompatibility. Folic acid (FA) as a targeting agent was then conjugated to the citric acid coated SPIONs (SPIONs-CA) for targeting the specific receptors expressed on the FAR+ cancer cells. Physiochemical characterizations were then performed to assure required properties like stability, size, phase purity, surface morphology, chemical integrity and magnetic properties. In vitro evaluations (MTT assay) were performed on HeLa, HSF 1184, MDA-MB-468 and MDA-MB-231cell lines to ensure the biocompatibility of SPIONs-CA-FA. There were no morphological changes and lysis in contact with erythrocytes recorded for SPIONs-CA-FA and SPIONs-CA. High level of SPIONs-CA-FA binding to FAR+ cell lines was assured via qualitative and quantitative in vitro binding studies. Hence, SPIONs-CA-FA was introduced as a promising tool for biomedical applications like magnetic hyperthermia and drug delivery. The in vitro findings presented in this study need to be compared with those of in vivo studies.
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http://dx.doi.org/10.1016/j.msec.2016.07.076DOI Listing
December 2016

Gentamicin-gold nanoparticles conjugate: a contrast agent for X-ray imaging of infectious foci due to Staphylococcus aureus.

IET Nanobiotechnol 2016 Aug;10(4):190-4

Department of Radiology, Faculty of Paramedical and Health Sciences, Zanjan University of Medical Science, Zanjan, Iran.

There is no optimal imaging method for the detection of unknown infectious foci in some diseases. This study introduces a novel method in X-ray imaging of infection foci due to Staphylococcus aureus by developing a contrast agent based on gold nanoparticles (GNPs). GNPs in spherical shape were synthesised by the reduction of tetrachloroauric acid with sodium citrate. Then gentamicin was bound directly to citrate functionalised GNPs and the complex was stabilised by polyethylene glycol. The interaction of gentamicin with GNPs was confirmed by ultraviolet-visible and Fourier transform infrared spectroscopies. The stability of complex was studied in human blood up to 6 h. The stability of conjugate was found to be high in human blood with no aggregation. The biodistribution study showed localisation of gentamicin-GNPs conjugate at the site of Staphylococcal infection. The infection site was properly visualised in X-ray images in mouse model using the gentamicin-GNPs conjugate as a contrast agent. The results demonstrated that one may consider the potential of new nanodrug as a contrast agent for X-ray imaging of infection foci in human beings which needs more investigations.
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http://dx.doi.org/10.1049/iet-nbt.2015.0034DOI Listing
August 2016

Synergistic Antibacterial Activity of Plant Peptide MBP-1 and Silver Nanoparticles Combination on Healing of Infected Wound Due to Staphylococcus aureus.

Jundishapur J Microbiol 2016 Jan 2;9(1):e27997. Epub 2016 Jan 2.

Department of Microbiology, Faculty of Sciences, Zanjan Branch, Islamic Azad University, Zanjan, IR Iran.

Background: Wound infection is a common problem in hospitals and is typically caused by the antibiotic-resistant Staphylococcus aureus, which is a major pathogen for skin and soft tissue infections worldwide.

Objectives: The aim of this study was to investigate the synergistic antibacterial effect of plant peptide MBP-1 and silver nanoparticles on infected wounds caused by S. aureus.

Materials And Methods: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of MBP-1 and silver nanoparticles both on their own and in combination form were determined against S. aureus via macrodilution and microdilution methods. The synergistic antibacterial effect of silver nanoparticles and MBP-1 was investigated on infected wounds caused by S. aureus in a mouse model.

Results: The MIC and MBC of MBP-1 were found to be 0.6 and 0.7 mg/mL, respectively. MIC and MBC of silver nanoparticles were determined to be 6.25 and 12.5 mg/L, respectively. MIC and MBC of the silver nanoparticles and MBP-1 combination were found to be 3.125 mg/mL, 0.5 mg/L; and 6.25 mg/mL, 0.6 mg/L, respectively. The infected wound healed properly after the combined use of MBP-1 and silver nanoparticles.

Conclusions: The synergistic effect was found on the healing of infected wounds caused by S. aureus by using an MBP-1 and silver nanoparticles combination in a mouse model.
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http://dx.doi.org/10.5812/jjm.27997DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4834128PMC
January 2016

Synthesis, characterization and in vitro evaluation of exquisite targeting SPIONs-PEG-HER in HER2+ human breast cancer cells.

Nanotechnology 2016 Mar 10;27(10):105601. Epub 2016 Feb 10.

Department of Bioprocess Engineering, Faculty of Chemical Engineering, Universiti Teknologi Malaysia, Skudai 81310, Johor Bahru, Johor, Malaysia.

A stable, biocompatible and exquisite SPIONs-PEG-HER targeting complex was developed. Initially synthesized superparamagnetic iron oxide nanoparticles (SPIONs) were silanized using 3-aminopropyltrimethoxysilane (APS) as the coupling agent in order to allow the covalent bonding of polyethylene glycol (PEG) to the SPIONs to improve the biocompatibility of the SPIONs. SPIONs-PEG were then conjugated with herceptin (HER) to permit the SPIONs-PEG-HER to target the specific receptors expressed over the surface of the HER2+ metastatic breast cancer cells. Each preparation step was physico-chemically analyzed and characterized by a number of analytical methods including AAS, FTIR spectroscopy, XRD, FESEM, TEM, DLS and VSM. The biocompatibility of SPIONs-PEG-HER was evaluated in vitro on HSF-1184 (human skin fibroblast cells), SK-BR-3 (human breast cancer cells, HER+), MDA-MB-231 (human breast cancer cells, HER-) and MDA-MB-468 (human breast cancer cells, HER-) cell lines by performing MTT and trypan blue assays. The hemolysis analysis results of the SPIONs-PEG-HER and SPIONs-PEG did not indicate any sign of lysis while in contact with erythrocytes. Additionally, there were no morphological changes seen in RBCs after incubation with SPIONs-PEG-HER and SPIONs-PEG under a light microscope. The qualitative and quantitative in vitro targeting studies confirmed the high level of SPION-PEG-HER binding to SK-BR-3 (HER2+ metastatic breast cancer cells). Thus, the results reflected that the SPIONs-PEG-HER can be chosen as a favorable biomaterial for biomedical applications, chiefly magnetic hyperthermia, in the future.
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http://dx.doi.org/10.1088/0957-4484/27/10/105601DOI Listing
March 2016

Breast cancer photothermal therapy based on gold nanorods targeted by covalently-coupled bombesin peptide.

Nanotechnology 2015 May 22;26(19):195101. Epub 2015 Apr 22.

Department of Biology, University of Guilan, Rasht, Iran. Department of Chemical and Biomolecular Engineering, Tulane University, New Orleans, Louisiana, USA.

Photothermal therapy, a minimally invasive treatment method for killing cancers cells, has generated a great deal of interest. In an effort to improve treatment efficacy and reduce side effects, better targeting of photoabsorbers to tumors has become a new concept in the battle against cancer. In this study, a bombesin (BBN) analog that can bind to all gastrin-releasing peptide (GRP) receptor subtypes was bound covalently with gold nanorods (GNRs) using Nanothinks acid as a link. The BBN analog was also coated with poly(ethylene glycol) to increase its stability and biocompatibility. The interactions were confirmed by ultraviolet-visible and Fourier transform infrared spectroscopy. A methylthiazol tetrazolium assay showed no cytotoxicity of the PEGylated GNR-BBN conjugate. The cell binding and internalization studies showed high specificity and uptake of the GNR-BBN-PEG conjugate toward breast cancer cells of the T47D cell line. The in vitro study revealed destruction of the T47D cells exposed to the new photothermal agent combined with continuous-wave near-infrared laser irradiation. The biodistribution study showed significant accumulation of the conjugate in the tumor tissue of mice with breast cancer. The in vivo photothermal therapy showed the complete disappearance of xenographted breast tumors in the mouse model.
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http://dx.doi.org/10.1088/0957-4484/26/19/195101DOI Listing
May 2015

Synthesis and characterization of Bombesin-superparamagnetic iron oxide nanoparticles as a targeted contrast agent for imaging of breast cancer using MRI.

Nanotechnology 2015 Feb 2;26(7):075101. Epub 2015 Feb 2.

Nanostructure Lab, Physics Department, University of Guilan, Rasht, Iran.

The targeted delivery of superparamagnetic iron oxide nanoparticles (SPIONs) as a contrast agent may facilitate their accumulation in cancer cells and enhance the sensitivity of MR imaging. In this study, SPIONs coated with dextran (DSPIONs) were conjugated with bombesin (BBN) to produce a targeting contrast agent for detection of breast cancer using MRI. X-ray diffraction, transmission electron microscopy, and vibrating sample magnetometer analyses indicated the formation of dextran-coated superparamagnetic iron oxide nanoparticles with an average size of 6.0 ± 0.5 nm. Fourier transform infrared spectroscopy confirmed the conjugation of the BBN with the DSPIONs. A stability study proved the high optical stability of DSPION-BBN in human blood serum. DSPION-BBN biocompatibility was confirmed by cytotoxicity evaluation. A binding study showed the targeting ability of DSPION-BBN to bind to T47D breast cancer cells overexpressing gastrin-releasing peptide (GRP) receptors. T2-weighted and T2*-weighted color map MR images were acquired. The MRI study indicated that the DSPION-BBN possessed good diagnostic ability as a GRP-specific contrast agent, with appropriate signal reduction in T2*-weighted color map MR images in mice with breast tumors.
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http://dx.doi.org/10.1088/0957-4484/26/7/075101DOI Listing
February 2015

Anti protein A antibody-gold nanorods conjugate: a targeting agent for selective killing of methicillin resistant Staphylococcus aureus using photothermal therapy method.

J Microbiol 2015 Feb 28;53(2):116-21. Epub 2015 Jan 28.

Department of Microbiology, Sciences and Research Branch, Islamic Azad University, Fars, Iran.

The high prevalence of methicillin resistant Staphylococcus aureus (MRSA) and developing resistance to antibiotics requires new approaches for treatment of infectious diseases due to this bacterium. In this study, we developed a targeting agent for selective killing of MRSA using photothermal therapy method based on anti protein A antibody and gold nanorods (GNRs). Polystyrene sulfonate (PSS) coated GNRs were conjugated with anti protein A antibody. The FT-IR and UV-vis analyses approved the formation of anti protein A antibody-gold nanorods conjugate. In vitro study of photothermal therapy showed 82% reduction in the MRSA cells viability which was significantly greater than the ablation effect of free GNRs and laser alone. Significant accumulation of anti protein A antibody-GNRs in the infected muscle in comparison with normal muscle approved the targeting ability of new agent. In vivo study of photothermal therapy resulted in a significant reduction (73%) in the bacterial cells viability in the infected mouse model. These results demonstrated the ability of anti protein A antibody-GNRs conjugate in combination with NIR laser energy for selective killing of MRSA in mouse model.
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http://dx.doi.org/10.1007/s12275-015-4519-4DOI Listing
February 2015

Bactericidal Effect of Silver Nanoparticles on Intramacrophage Brucella abortus 544.

Jundishapur J Microbiol 2014 Mar 1;7(3):e9039. Epub 2014 Mar 1.

Biology Research Center, Zanjan Branch, Islamic Azad University, Zanjan, IR Iran.

Background: Brucellosis is an infectious disease that is caused by Brucella spp. As Brucella spp. are intramacrophage pathogens, the treatment of this infection is very difficult. On the other hand, due to the side effects of the brucellosis treatment regime, it is necessary to find new antimicrobial agents against it.

Objectives: The aim of this study was to investigate the antimicrobial effect of silver nanoparticles against Brucella abortus 544 in the intramacrophage condition.

Materials And Methods: The antimicrobial effect of silver nanoparticles was determined by an agar well diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of silver nanoparticles against B. abortus 544 were determined by a broth macrodilution method. The effect of time on the antimicrobial activity of silver nanoparticles was analyzed. The effect of silver nanoparticles on the intramacrophage survival of B. abortus 544 was studied on mice peritoneal macrophages.

Results: The well diffusion agar study showed that silver nanoparticles have an antimicrobial effect on B. abortus 544. The MIC and MBC of silver nanoparticles against B. abortus 544 were; 6 ppm and 8 ppm, respectively. The silver nanoparticles showed antibacterial effects within 40 minutes. The results of the macrophage culture indicated that silver nanoparticles have antibacterial activity against intramacrophage B. abortus 544, and the highest efficiency was observed at a concentration of 8-10 ppm of silver nanoparticles.

Conclusions: The results showed that silver nanoparticles have an antimicrobial effect against intramacrophage B. abortus 544.
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http://dx.doi.org/10.5812/jjm.9039DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4138654PMC
March 2014

Enhanced delivery of gentamicin to infection foci due to Staphylococcus aureus using gold nanorods.

Drug Deliv 2016 24;23(1):49-54. Epub 2014 Apr 24.

b Department of Microbiology, Faculty of Sciences , Zanjan Branch, Islamic Azad University , Zanjan , Iran.

Bacterial infections continue to be one of the major causes of morbidity and mortality. Although many methods for diagnosing and treating of infectious diseases currently exist, there is an urgent need for new and improved approaches for bacterial destruction. The present study focuses on the conjugation of gold nanorods (GNRs) with gentamicin via the Nanothink acid linker and its application in delivery of gentamicin to infection foci due to Staphylococcus aureus. The interaction between gentamicin and gold nanorods was confirmed by FT-IR spectroscopy. The high performance liquid chromatography (HPLC) and atomic absorption spectroscopy analyses showed that 2050 gentamicin molecules were attached to each gold nanorod. The minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of gentamicin-GNRs conjugate showed the enhancement of antibacterial effect of gentamicin. The biodistribution study demonstrated localization of the complex at the site of Staphylococcal infection with high sensitivity in mouse model.
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http://dx.doi.org/10.3109/10717544.2014.903533DOI Listing
October 2016

Gold nanorods-bombesin conjugate as a potential targeted imaging agent for detection of breast cancer.

J Photochem Photobiol B 2014 Jan 15;130:40-6. Epub 2013 Nov 15.

Biology Research Center, Zanjan Branch, Islamic Azad University, Zanjan, Iran. Electronic address:

Photoacoustic imaging (PAI) is a hybrid biomedical imaging modality that offers both strong optical absorption contrast and high ultrasonic resolution. PAI is capable of in vivo molecular imaging, thus facilitating further molecular and cellular characterization of cancer. In this study, Gold nanorods (GNRs) were synthesized and coated with polyethyleneglycol (PEG). Then, the PEG-GNRs were conjugated with bombesin (BBN), a cancer seeking peptide, for production of a potential photoacoustic targeting imaging agent for detection of breast cancer. The optical property, biocompatibility, stability and in vitro/in vivo binding affinities of GNR-PEG-BBN for breast cancer cells were investigated. UV-vis spectroscopy confirmed the conjugation of bombesin with PEG-coated GNRs. The stability assessment proved high optical stability of GNR-PEG-BBN in human blood serum up to 12h. Cytotoxicity study showed biocompatibility of GNR-PEG-BBN conjugate. Molecular targeting ability was approved in cells over expressing gastrin-releasing peptide (GRP) receptor (breast cancer cell line) in comparison with cells that do not express GRP receptor (skin fibroblast cell line). The selective accumulation of GNR-PEG-BBN was demonstrated in breast tumor in comparison with unconjugated gold nanorods, following the intravenous administration of GNR-PEG-BBN to breast tumor-bearing mice. This study demonstrated the potential of GNR-PEG-BBN as a photoacoustic imaging agent that can provide improved specificity and sensitivity for breast cancer detection.
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http://dx.doi.org/10.1016/j.jphotobiol.2013.10.019DOI Listing
January 2014

Nanoemulsions and nonwoven fabrics carrying AgNPs: antibacterial but may be cytotoxic.

Artif Cells Nanomed Biotechnol 2014 Dec 15;42(6):392-9. Epub 2013 Oct 15.

Division of Nanotechnology and Nanomedicine, Hacettepe University , Ankara , Turkey.

Abstract The aim of this study is to prepare nonwoven fabrics carrying silver nanoparticles (AgNPs), and to investigate their antibacterial activities and cytotoxicities in parallel. AgNPs were impregnated from their nanoemulsions onto two commercially available nonwoven fabrics: pure-cotton fabrics (PCF) and polyester/viscous fabrics (PVF), by a simple adsorption (dipping) and were then heat stabilized. PCF exhibited stronger antibacterial effects on both Staphylococcus aureus and Escherichia coli. In-vitro cell culture studies demonstrated that AgNPs nanoemulsions and also fabrics carrying them were cytotoxic on L929-fibroblasts in all concentrations used here (6.25-400 ppm) in different extends. Only the fabrics loaded with AgNPs using nanoemulsion with the lowest concentration of 6.25 ppm exhibited low cytotoxicity but were still antibacterial.
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http://dx.doi.org/10.3109/21691401.2013.834908DOI Listing
December 2014

Development of gentamicin-gold nanospheres for antimicrobial drug delivery to Staphylococcal infected foci.

Drug Deliv 2013 ;20(1):34-9

Department of Microbiology, Faculty of Sciences, Zanjan Branch, Islamic Azad University, Zanjan, Iran.

Even though the therapeutic efficacy of numerous antimicrobial drugs has been well established, inefficient delivery can result in an inadequate therapeutic index. Gold nanoparticles have unique physicochemical properties such as large surface area to mass ratio and functionalizable structure. These properties can be applied to facilitate the administration of antimicrobial drugs, thereby overcoming some of the limitations in traditional antimicrobial therapeutics. In this study, gold nanospheres were used as a drug carrier system for gentamicin delivery to Staphylococcal infected foci. Conjugation of gentamicin with gold nanospheres was performed in HEPES buffer. The attachment of gentamicin to gold nanospheres was confirmed by UV/Vis spectroscopy. The HPLC and atomic absorption spectrometer analyses showed that 347 gentamicin molecules were attached to each gold nanosphere. Minimum inhibitory concentration and minimum bactericidal concentration studies showed the enhanced antibacterial effect of gentamicin-gold nanospheres complex in comparison with free gentamicin. The biodistribution study showed the localization of the complex at the site of Staphylococcal infection foci with high sensitivity in mouse model.
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http://dx.doi.org/10.3109/10717544.2012.746402DOI Listing
October 2013

Preparation and biological evaluation of (177)Lu conjugated PR81 for radioimmunotherapy of breast cancer.

Nucl Med Biol 2011 Aug 6;38(6):849-55. Epub 2011 May 6.

Department of Biology, Faculty of Sciences, Zanjan Branch, Islamic Azad University, Zanjan 45156-58145, Iran.

Aim: PR81 is a monoclonal antibody that binds with high affinity to MUC1 antigen that is over expressed in 80% of breast cancers. In this study, we developed a method for indirect labeling of PR81 with lutetium-177 and performed all preclinical qualifications in production of a biologic agent for radioimmunotherapy of breast cancer.

Materials And Methods: The radiochemical purity and in vitro stability of (177)Lu labeled PR81 was determined by instant thin layer chromatography. The immunoreactivity and cell toxicity of the complex were tested on MCF7 cell line. The biodistribution and scintigraphy studies were performed in BALB/c mice with breast tumor.

Results: The radiochemical purity was 91.2±3.8% after 2 h. The in vitro stabilities in phosphate buffer and human blood serum were 83.1±3.4% and 76.2±3.6% at 96 h, respectively. The immunoreactivity of the complex was 83.4±2.4%. The cell toxicity study showed that the complex inhibited 85.2±3.4% growth of MCF7 cells at a concentration of 2500 ng/ml after 96 h. The biodistribution and scintigraphy studies showed the accumulation of the complex at the site of tumors with high sensitivity and specificity.

Conclusion: The results showed that one may consider (177)Lu-DOTA-PR81 as a potential radiopharmaceutical for therapy of human breast cancer, which needs further investigations.
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http://dx.doi.org/10.1016/j.nucmedbio.2011.02.009DOI Listing
August 2011

Comparison of (99m)Tc-labeled PR81 and its F(ab')₂ fragments as radioimmunoscintigraphy agents for breast cancer imaging.

Ann Nucl Med 2011 Feb;25(2):87-92

Department of Biology, Faculty of Sciences, Islamic Azad University, Zanjan Branch, 45156-58145 Zanjan, Iran.

Objective: We digested anti-MUC1 monoclonal antibody PR81 to produce F(ab')₂ fragments. A comparison was performed between the two radiolabeled PR81 and F(ab')₂ fragments for breast tumor imaging in a mouse model.

Methods: The optimum conditions for pepsin digestion of PR81 were investigated in terms of enzymes: antibody ratio, digestion time duration and preserved immunoreactivity of the produced fragments. The F(ab')₂ fragments were labeled with Technetium-99m using HYNIC as a chelator and tricine as a co-ligand. The immunoreactivity of the complexes was assessed by radioimmunoassay using MCF7 cells. Biodistribution and imaging studies were performed in female BALB/c mice with breast tumor xenograft at 4, 8 and 24 h post-administration. The PR81 was labeled with technetium-99m in the same way for comparison.

Results: The optimum time duration for PR81 digestion was found to be 28 h at an enzyme:antibody weight ratio of 1:20 that resulted in 95.2 ± 4.7% purity. The labeling of intact PR81 and its F(ab')₂ fragments were 87.6 ± 4.2 and 76.1 ± 3.3% after 1 h, respectively (p value <0.05). The percentage of immunoreactivity of F(ab')₂ fragments and intact PR81 were 75.4 ± 2.1% and 85.7 ± 2.9%, respectively (p value <0.05). The biodistribution and imaging studies demonstrated localization of the fragments at 4 h post-administration with high sensitivity and specificity.

Conclusion: The results showed that F(ab')₂ fragment of PR81 is more suitable than intact PR81 for safer and more rapid detection of human breast cancer.
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http://dx.doi.org/10.1007/s12149-010-0434-2DOI Listing
February 2011

Radioimmunotherapy of MCF7 breast cancer cell line with 131I-PR81 monoclonal antibody against MUC1: comparison of direct and indirect radioiodination methods.

Hum Antibodies 2010 ;19(1):15-25

Department of Clinical Biochemistry, School of Medical Sciences, Tarbiat Modarres University, Tehran, Iran.

PR81 is a monoclonal antibody that binds with high affinity to MUC1, which is over expressed on breast and other tumors. The objective of this study was to compare the two labeling methods (direct and indirect radioiodination) for application of this antibody against MUC1 as a radioimmunotherapeutical agent.Monoclonal antibody (PR81) against the tandem repeat of the core protein (MUC1) was prepared, characterized, purified, and labeled with 131I using the direct (chloramin-T) and indirect (Fmoc-D-Tyr (tBu)-D-Tyr (tBu)-D-Lys (Boc)-OH (YYK) attached to N-hydroxysuccinimide as a linker between PR81 and 131I) methods. The immunoreactivity of 131I-PR81 and 131I-TP-PR81 complexes with MUC1 (the native protein), BSA-P20 (a 20 amino acid corresponding the tandem repeat of MUC1) and MCF7 cell line were performed by RIA. In vitro stability of 131I-PR81 and 131I-YYK-peptide-PR81 complexes in human serum was determined by thin layer chromatography (TLC). Cell toxicity and in vitro internalization studies were performed with the MCF7 cell line, and the tissue biodistribution of the 131I-PR81 and 131I- YYK-peptide -PR81 complexes was evaluated in normal BALB/c mice at 4, 24 and 48 hrs. The labeling efficiency was determined by measuring the percentage recovery of radioactivity in the final product relative to the initial activity in the shipment vial, was found to be 59.9% +/- 7.9% for direct and 50% +/- 3.2% for indirect methods. 131I-PR81 and 131I- YYK- peptide -PR81 complexes showed high immunoreactivity towards MUC1 protein, BSA-P20 and MCF7 cell line. In vitro stability of the labeled products in human serum which was measured by thin layer chromatography (TLC) was found to be more than 50% over 24 hr for 131I-PR81 and 70% for 131I- YYK-peptide -PR81 complexes. Cell toxicity and in vitro internalization studies showed that the 131I-PR81 and 131I- YYK-peptide -PR81 complexes inhibited 80% growth of the MCF7 cultured cell lines in vitro in a high concentration and up to 40% of the 131I-PR81 and 60% of the 131I- YYK-peptide -PR81 complexes internalized after 24 h. Biodistribution studies were performed in normal BALB/c mice at 4, 24 and 48 hrs post-injection. Thyroid and stomach levels from PR81 labeled with 131I- YYK-peptide were two- to three- fold less than those with directly labeled 131I-PR81, suggesting low recognition of its D-iodotyrosine residue by endogenous deiodinase. These results show that the indirect labeling was better than the indirect labeling and 131I- YYK-peptide -PR81 may be considered as a promising candidate for therapy of breast cancer.
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http://dx.doi.org/10.3233/HAB-2010-0216DOI Listing
October 2010

Radiolabeling of trastuzumab with 177Lu via DOTA, a new radiopharmaceutical for radioimmunotherapy of breast cancer.

Nucl Med Biol 2009 May;36(4):363-9

Department of Medical Physics, Tarbiat Modares University, Tehran, Iran.

Aim: Trastuzumab is a monoclonal antibody that is used in treating breast cancer. We labeled this monoclonal antibody with lutetium-177 and performed in vitro quality control tests as a first step in the production of a new radiopharmaceutical.

Material And Methods: Trastuzumab was labeled with lutetium-177 using DOTA as chelator. Radiochemical purity and stability in buffer and human blood serum were determined using thin layer chromatography. Immunoreactivity and toxicity of the complex were tested on MCF7 breast cancer cell line.

Results: The radiochemical purity of the complex was 96+/-0.9%. The stabilities in phosphate buffer and in human blood serum at 96 h postpreparation were 93+/-1.2% and 85+/-3.5%, respectively. The immunoreactivity of the complex was 89+/-1.4%. At a concentration of 1 nM, the complex killed 70+/-3% of MCF7 cells. At 1.9 nM, 90+/-5% of the cells were killed.

Conclusions: The results showed that the new complex could be considered for further evaluation in animals and possibly in humans as a new radiopharmaceutical for use in radioimmunotherapy against breast cancer.
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http://dx.doi.org/10.1016/j.nucmedbio.2009.01.015DOI Listing
May 2009

Breast tumor targeting with (99m)Tc-HYNIC-PR81 complex as a new biologic radiopharmaceutical.

Nucl Med Biol 2008 Oct;35(7):763-8

Department of Medical Physics, Tarbiat Modares University, Tehran, Iran.

Human epithelial mucin, MUC1, is commonly overexpressed in adenocarcinoma that includes more than 80% of breast cancers. The PR81 is a murine anti-MUC1 monoclonal antibody (MAb) that was prepared against the human breast cancer. We developed an indirect method for labeling of this antibody with (99m)Tc in order to use the new preparation in immunoscintigraphy studies of BALB/c mice bearing breast tumors. The (99m)Tc-PR81 complex was prepared using the HYNIC as a chelator and tricine as a coligand. The labeling efficiency determined by instant thin-layer chromatography (ITLC) was 89.2%+/-4.7%, and radiocolloides measured by cellulose nitrate electrophoresis were 3.4%+/-0.9%. The in vitro stability of labeled product was determined at room temperature by ITLC and in human serum by gel filtration chromatography - 88.3%+/-4.6% and 79.8%+/-5.7% over 24 h, respectively. The integrity of labeled MAb was checked by means of sodium dodecyl sulfate polyacrylamide gel electrophoresis, and no significant fragmentation was seen. The results of cell binding studies showed that both labeled and unlabeled PR81 were able to compete for binding to MCF 7 cells. Biodistribution studies performed in female BALB/c mice with breast tumor xenografts at 4, 16 and 24 h after the (99m)Tc-HYNIC-PR81 injection demonstrated a specific localization of the compound at the site of tumors and minimum accumulation in non target organs. The tumor imaging was performed in BALB/c mice with breast xenograft tumors at 4, 8, 12, 16, 20, 24, 28, 32 and 36 h after the complex injection. The tumors were visualized with high sensitivity after 8 h. The findings showed that the new radiopharmaceutical is a promising candidate for radioimmunoscintigraphy of the human breast cancer.
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http://dx.doi.org/10.1016/j.nucmedbio.2008.07.002DOI Listing
October 2008