Development of poly (I:C) modified doxorubicin loaded magnetic dendrimer nanoparticles for targeted combination therapy

The objective of this study was to develop and evaluate the anticancer activity and the safety of a combinational drug delivery system using polyamidoamine (PAMAM) dendrimer-coated iron oxide nanoparticles for doxorubicin and poly I:C delivery in vitro. Dendrimer-coated magnetic nanoparticles (DcMNPs) are suitable for drug delivery system as nanocarriers with their following properties, such as surface functional groups, symmetry perfection, internal cavities, nano-size and magnetization. These nanoparticles could be targeted to the tumor site under a magnetic field since they have a magnetic core. DcMNPs were found as a convenient vehicle for targeted doxorubicin delivery in cancer therapy. Poly (I:C) binding on doxorubicin loaded DcMNPs (DcMNPs-Dox) was reported for the first time in the literature. It was also demonstrated that loading of doxorubicin into the cavities of DcMNPs increases the binding efficiency of poly (I:C) to the surface functional groups of dendrimer up to 10 times. When we compare the in vitro cytotoxic properties of doxorubicin, poly (I:C) and poly (I:C) bound doxorubicin loaded DcMNPs (PIC-DcMNPs-Dox), it was observed that PIC-DcMNPs-Dox show the highest cytotoxic effect by passing the cell resistance mechanisms on doxorubicin resistant MCF7 (MCF7/Dox) cells. Results demonstrated that applying PIC-DcMNPs-Dox would improve the efficacy by increasing the biocompatibility of system in blood stream and the toxicity inside tumor cells. These results provide invaluable information and new insight for the design and optimization of a novel combinational drug delivery system for targeted cancer therapy. (C) 2014 Elsevier Masson SAS. All rights reserved

Doksorubisin Bağlı Kitosan Kaplı Manyetik Nanoparçacıkların Meme Kanseri Hücre Hatları Üzerindeki Etkinliğinin İmmünositokimya Yöntemi İle Belirlenmesi

Nanoparçacıkların kanser tedavisi, ilaç hedefleme ve kontrollü ilaç salım sistemlerinde kullanımı çalışmaları son zamanlarda artış göstermektedir. Nanoparçacıklar küçük boyutları sayesinde hücre içine kolay alınabilmekte ve çeşitli malzemelerle kaplanarak yüzey özellikleri değiştirilebilmektedir. Tümör hücrelerine hedeflendirilebilen manyetik nanoparçacıklar (MNP) kanser tanı ve tedavisinde kullanılmaktadır. Manyetik özellik taşıyan nanoparçacıklar, manyetik alan varlığında istenilen bölgeye hedeflenebilme özellikleri sayesinde ilaç hedefleme ve kontrollü salıma yeni bir boyut kazandırmıştır. Dışarıdan uygulanan manyetik alan tarafından kolayca kontrol edilebilen bu tür nanoparçacıklar, yeterli miktarda anti-kanser ilacın hücrelere taşınmasında etkili olmaktadır. Bu çalışmada manyetik özellikli kitosan kaplı nanoparçacıkların sentezlenmesi, anti kanser bir ilaç olan Doksorubisin’in nanoparçacıklara yüklenmesi ve in vitro koşullarda nanoparçacıkların MCF-7 meme kanseri hücre hatlarına uygulanması amaçlanmaktadır. Sentezlenen kitosan kaplı nanoparçacıkların merkezinde bulunan manyetit (Fe3O4) demiroksit nanoparçacıklara manyetik özellik sağlayacaktır. Kitosan polimeri ile kaplı olarak sentezlenen MNP’lerin karakterizasyonu yapıldıktan sonra, MNP’lere doksorubisin ilacı yüklenecektir. Oluşturulan yapının karakteristik özellikleri belirlenecek ve in vitro koşullarda istenilen hücrelere yönlendirilebilme özellikleri incelenecektir. MCF-7 meme kanseri hücre hatları üzerindeki toksik etkileri sitotoksisite analizleri ile araştırılacaktır. Projenin son kısmında, Doksorubisin yüklü nanoparçacıkların MCF-7 hücre hatları üzerindeki etkinliği immünositokimya yöntemi ile belirlenecektir

Doxorubicin loading, release, and stability of polyamidoamine dendrimer-coated magnetic nanoparticles

WOS: 000319071200016PubMed ID: 23558592Nanotechnology is a promising alternative to overcome the limitations of classical chemotherapy. As a novel approach, dendrimer-coated magnetic nanoparticles (DcMNPs) maintain suitable drug delivery system because of their buildup of functional groups, symmetry perfection, nanosize, and internal cavities. They can also be targeted to the tumor site in a magnetic field. The aim of this study is to obtain an effective targeted delivery system for doxorubicin, using polyamidoamine (PAMAM) DcMNPs. Different generations (G2, G3, G4, and G7) of PAMAM DcMNPs were synthesized. Doxorubicin loading, release, and stability efficiencies in these nanoparticles (NPs) were studied. The results showed that low-generation NPs obtained in this study have pH-sensitive drug release characteristics. G4DcMNP, which releases most of the drug in lower pH, seems to be the most suitable generation for efficient Doxorubicin delivery. Furthermore, application of doxorubicin-loaded G4DcMNPs may help to overcome doxorubicin resistance in MCF-7 cells. On the contrary, G2 and G3DcMNPs would be suitable for the delivery of drugs such as vinca alkaloids (Johnson IS, Armstrong JG, Gorman M, Burnett JP. 1963. Cancer Res 23:13901427.) and taxenes (Clarke SJ, Rivory LP. 1999. Clin Pharmacokinet 36(2):99114.), which show their effects in cytoplasm. The results of this study can provide new insights in the development of pH-sensitive targeted drug delivery systems to overcome drug resistance during cancer therapy. (c) 2013 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:18251835, 2013TUBITAKTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [TBAG-109T949, TBAG-2215]; Middle East Technical UniversityMiddle East Technical University [BAP-07-02-2010-06]This study was supported by TUBITAK (TBAG-109T949 and TBAG-2215) and Middle East Technical University (BAP-07-02-2010-06)

PAMAM dendrimer-coated iron oxide nanoparticles: synthesis and characterization of different generations

This study focuses on the synthesis and characterization of different generations (G(0)-G(7)) of polyamidoamine (PAMAM) dendrimer-coated magnetic nanoparticles (DcMNPs). In this study, superparamagnetic iron oxide nanoparticles were synthesized by co-precipitation method. The synthesized nanoparticles were modified with aminopropyltrimethoxysilane for dendrimer coating. Aminosilane-modified MNPs were coated with PAMAM dendrimer. The characterization of synthesized nanoparticles was performed by X-ray diffraction, X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), dynamic light scattering, and vibrating sample magnetometry (VSM) analyses. TEM images demonstrated that the DcMNPs have monodisperse size distribution with an average particle diameter of 16 +/- 5 nm. DcMNPs were found to be superparamagnetic through VSM analysis. The synthesis, aminosilane modification, and dendrimer coating of iron oxide nanoparticles were validated by FTIR and XPS analyses. Cellular internalization of nanoparticles was studied by inverted light scattering microscopy, and cytotoxicity was determined by XTT analysis. Results demonstrated that the synthesized DcMNPs, with their functional groups, symmetry perfection, size distribution, improved magnetic properties, and nontoxic characteristics could be suitable nanocarriers for targeted cancer therapy upon loading with various anticancer agents

Synthesis optimization and characterization of chitosan-coated iron oxide nanoparticles produced for biomedical applications

WOS: 000310608100037The chitosan-coated magnetic nanoparticles (CS MNPs) were in situ synthesized by cross-linking method. In this method; during the adsorption of cationic chitosan molecules onto the surface of anionic magnetic nanoparticles (MNPs) with electrostatic interactions, tripolyphosphate (TPP) is added for ionic cross-linking of the chitosan molecules with each other. The characterization of synthesized nanoparticles was performed by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS/ESCA), Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), dynamic light scattering (DLS), thermal gravimetric analysis (TGA), and vibrating sample magnetometry (VSM) analyses. The XRD and XPS analyses proved that the synthesized iron oxide was magnetite (Fe3O4). The layer of chitosan on the magnetite surface was confirmed by FTIR. TEM results demonstrated a spherical morphology. In the synthesis, at higher NH4OH concentrations, smaller sized nanoparticles were obtained. The average diameters were generally between 2 and 8 nm for CS MNPs in TEM and between 58 and 103 nm in DLS. The average diameters of bare MNPs were found as around 18 nm both in TEM and DLS. TGA results indicated that the chitosan content of CS MNPs were between 15 and 23 % by weight. Bare and CS MNPs were superparamagnetic. These nanoparticles were found non-cytotoxic on cancer cell lines (SiHa, HeLa). The synthesized MNPs have many potential applications in biomedicine including targeted drug delivery, magnetic resonance imaging (MRI), and magnetic hyperthermia.[TUBITAK-TBAG(1001)/109T949]The support of Assist. Prof. Dr Bora Mavis for FTIR is gratefully acknowledged, as well as financial support by TUBITAK-TBAG(1001)/109T949

Polyinosinic: polycytidylic acid loading onto different generations of PAMAM dendrimer-coated magnetic nanoparticles

WOS: 000322593200053Poly (I:C), which is a synthetic double-stranded RNA, have significant toxicity on tumor cells. The immobilization of Poly (I:C) onto nanoparticles is important for the fabrication of targeted delivery systems. In this study, different generations of newly synthesized PAMAM dendron-coated magnetic nanoparticles (DcMNP) which can be targeted to the tumor site under magnetic field were efficiently loaded for the first time with Poly (I:C). Different generations of DcMNPs (G(2), G(3), G(4), G(5), G(6), and G(7)) were synthesized. Poly (I:C) activation was achieved in the presence of EDC and 1-methylimidazole. Loading of Poly (I:C) onto DcMNPs was followed by agarose gel electrophoresis. Acidic reaction conditions were found as superior to basic and neutral for binding of Poly (I:C). In addition, having more functional groups at the surface, higher generations (G(7), G(6), and G(5)) of PAMAM DcMNPs were found more suitable as a delivery system for Poly (I:C). Further in vitro and in vivo analyses of Poly (I:C)/PAMAM magnetic nanoparticles may provide new opportunities for the selective targeting and killing of tumor cells.TUBITAKTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [TBAG-109T949, TBAG-2215]; Middle East Technical UniversityMiddle East Technical University [BAP-07-02-2010-06]This study was supported by TUBITAK (TBAG-109T949 and TBAG-2215), and Middle East Technical University (BAP-07-02-2010-06)

Polyhydroxybutyrate-Coated Magnetic Nanoparticles for Doxorubicin Delivery: Cytotoxic Effect Against Doxorubicin-Resistant Breast Cancer Cell Line

WOS: 000345012600018PubMed ID: 25137407In this study, polyhydroxybutyrate (PHB)-coated magnetic nanoparticles (MNPs) were prepared by coprecipitation of iron salts (Fe2+ and Fe3+) by ammonium hydroxide. Characterizations of PHB-coated MNPs were performed by Fourier transform infrared spectroscopy, x-ray diffraction, dynamic light scattering, thermal gravimetric analysis, vibrating sample magnetometry, and transmission electron microscopy analyses. Doxorubicin was loaded onto PHB-MNPs, and the release efficiencies at different pHs were studied under in vitro conditions. The most efficient drug loading concentration was found about 87% at room temperature in phosphate-buffered saline (pH 7.2). The drug-loaded MNPs were stable up to 2 months in neutral pH for mimicking physiological conditions. The drug release studies were performed with acetate buffer (pH 4.5) that mimics endosomal pH. Doxorubicin (60%) released from PHB-MNPs within 65 hours. Doxorubicin-loaded PHB-MNPs were about 2.5-fold more cytotoxic as compared with free drug on resistant Michigan Cancer Foundation-7 (human breast adenocarcinoma, MCF-7) cell line (1 mu M doxorubicin) in vitro. Therefore, doxorubicin-loaded PHB-MNPs lead to overcome the drug resistance.Ahi Evran University, Kirsehir, TurkeyAhi Evran University [BAP-PYO-KMY.4001.12.007]Supported by Ahi Evran University (BAP-PYO-KMY.4001.12.007), Kirsehir, Turkey

Synthesis of Doxorubicin loaded magnetic chitosan nanoparticles for pH responsive targeted drug delivery

WOS: 000340301500028PubMed ID: 24931189Targeted drug delivery is a promising alternative to overcome the limitations of classical chemotherapy. In an ideal targeted drug delivery system carrier nanoparticles would be directed to the tumor tissue and selectively release therapeutic molecules. As a novel approach, chitosan coated magnetic nanoparticles (CS MNPs) maintain a pH dependent drug delivery which provides targeting of drugs to the tumor site under a magnetic field. Among various materials, chitosan has a great importance as a pH sensitive, natural, biodegradable, biocompatible and bioadhesive polymer. The aim of this study was to obtain an effective targeted delivery system for Doxorubicin, using chitosan coated MNPs. Different sized CS MNPs were produced by in situ synthesis method. The anti-cancer agent Doxorubicin was loaded onto CS MNPs which were characterized previously. Doxorubicin loading was confirmed by FTIR. Drug loading and release characteristics, and stability of the nanoparticles were investigated. Our results showed that the CS MNPs have pH responsive release characteristics. The cellular internalization of Doxorubicin loaded CS MNPs were visualized by fluorescent microscopy. Doxorubicin loaded CS MNPs are efficiently taken up by MCF-7 (MCF-7/S) and Doxorubicin resistant MCF-7 (MCF-7/1 mu M) breast cancer cells, which increases the efficacy of drug and also maintains overcoming the resistance of Doxorubicin in MCF-7/Dox cells. Consequently, CS MNPs synthesized at various sizes can be effectively used for the pH dependent release of Doxorubicin in cancer cells. Results of this study can provide new insights in the development of pH responsive targeted drug delivery systems to overcome the side effects of conventional chemotherapy. (C) 2014 Elsevier B.V. All rights reserved

Chitosan magnetic nanoparticles for pH responsive Bortezomib release in cancer therapy

WOS: 000342667800019PubMed ID: 24880680The use of nanotechnology in cancer treatment offers exciting opportunities, including the possibility of destroying tumors with minimal damage to healthy tissue by novel targeted drug delivery systems. pH differences between healthy and tumor microenvironment provide pH responsive release of drugs at tumor site via smart nanoparticles. In this study, chitosan coated superparamagnetic iron oxide nanoparticles (CS MNPs) were in situ synthesized by ionic crosslinking method as nanocarrier systems and loaded with the drug Bortezomib (Velcade (R)). The drug loading capacity, drug release and stability of CS MNPs were analyzed. CS MNPs were visualized inside the cells by fluorescence microscopy. The cytotoxicity of Bortezomib, CS MNPs and Bortezomib loaded CS MNPs were tested by XTT analyses in vitro. Gene expression analyses revealed that pro-apoptotic PUMA and NOXA genes were upregulated while anti-apoptotic BCL-2, SURVIVIN and cIAP-2 genes were downregulated at Bortezomib loaded CS MNP treated cells. Immunocytochemical analyses demonstrated an increase in p53 tumor suppressor protein levels at treated cells, which supports the upregulation of PUMA and NOXA genes, while Survivin protein level did not significantly change. This study points out that the pH responsive magnetic targeting of Bortezomib is more efficacious than free drug treatment. Moreover, targeted delivery of Bortezomib would reduce the frequency of drug administration by lowering the required amount of drug dose. (C) 2014 Elsevier Masson SAS. All rights reserved

Effect of gemcitabine and retinoic acid loaded PAMAM dendrimer-coated magnetic nanoparticles on pancreatic cancer and stellate cell lines

WOS: 000342668300009PubMed ID: 25108345Gemcitabine is an anticancer drug used in the treatment of different cancer types, including pancreatic ductal adenocarcinoma. The maximum tolerated dose in humans is restricted by its side effects on healty cells. Furthermore, the fibrotic stroma produced by the pancreatic stellate cells prevents effective delivery of chemotherapeutic agents providing a safe-haven for the cancer cells. This becomes more of a problem considering the short half-life of this drug. Magnetic nanoparticle-based targeted drug delivery systems are a promising alternative to overcome the limitations of classical chemotherapies. The aim of this study is to obtain an effective targeted delivery system for gemcitabine using magnetic nanoparticles (MNPs) and all-trans retinoic acid (ATRA). This dual approach targets the tumor cells and its infrastructure - stellate cells - simultaneously. Gemcitabine and ATRA were loaded onto the PAMAM dendrimer-coated magnetic nanoparticles (DcMNPs), which were synthesized and characterized previously. Drug loading and release characteristics, and stability of the nanoparticles were investigated. Gemcitabine and ATRA loaded MNPs are efficiently taken up by pancreatic cancer and stellate cells successfully targeting and eliminating both cells. Results of this study can provide new insights on pancreatic cancer therapy where tumor is seen as a system with its stroma insead of epithelial cells alone. (C) 2014 Elsevier Masson SAS. All rights reserved.TUBITAK (The Scientific And Technological Research Council of Turkey)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [2219]This study is supported by TUBITAK (The Scientific And Technological Research Council of Turkey)-2219 International Postdoctoral Research Fellowship Programme