Shlomo Margel

Publications of Shlomo Margel

• Magnetic Scaffolds Enriched with Bioactive Nanoparticles for Tissue Engineering

  Authors: Hadas Skaat, Ofra Ziv-Polat, Abraham Shaha, David Last, Yael Mardor, Shlomo Margel

  Advanced Healthcare Materials. 03/2012; 1(2-DOI: 10.1002/adhm.201100056):133-238.

• Novel magnetic fibrin hydrogel scaffolds containing thrombin and growth factors conjugated iron oxide nanoparticles for tissue engineering.

  Authors: Ofra Ziv-Polat, Hadas Skaat, Abraham Shahar, Shlomo Margel

  International journal of nanomedicine. 01/2012; 7:1259-74.

  Novel tissue-engineered magnetic fibrin hydrogel scaffolds were prepared by the interaction of thrombin-conjugated iron oxide magnetic nanoparticles with fibrinogen. In addition, stabilization ofNovel tissue-engineered magnetic fibrin hydrogel scaffolds were prepared by the interaction of thrombin-conjugated iron oxide magnetic nanoparticles with fibrinogen. In addition, stabilization of basal fibroblast growth factor (bFGF) was achieved by the covalent and physical conjugation of the growth factor to the magnetic nanoparticles. Adult nasal olfactory mucosa (NOM) cells were seeded in the transparent fibrin scaffolds in the absence or presence of the free or conjugated bFGF-iron oxide nanoparticles. The conjugated bFGF enhanced significantly the growth and differentiation of the NOM cells in the fibrin scaffolds, compared to the same or even five times higher concentration of the free bFGF. In the presence of the bFGF-conjugated magnetic nanoparticles, the cultured NOM cells proliferated and formed a three-dimensional interconnected network composed mainly of tapered bipolar cells. The magnetic properties of these matrices are due to the integration of the thrombin- and bFGF-conjugated magnetic nanoparticles within the scaffolds. The magnetic properties of these scaffolds may be used in future work for various applications, such as magnetic resonance visualization of the scaffolds after implantation and reloading the scaffolds via magnetic forces with bioactive agents, eg, growth factors bound to the iron oxide magnetic nanoparticles.

• Enhancement of the growth and differentiation of nasal olfactory mucosa cells by the conjugation of growth factors to functional nanoparticles.

  Authors: Hadas Skaat, Ofra Ziv-Polat, Abraham Shahar, Shlomo Margel

  Bioconjugate chemistry. 12/2011; 22(12):2600-10.

  Growth factors are critical components in the tissue engineering approach. Basic fibroblast growth factor (bFGF), a representative growth factor, stimulates the cellular functions of various cellsGrowth factors are critical components in the tissue engineering approach. Basic fibroblast growth factor (bFGF), a representative growth factor, stimulates the cellular functions of various cells and has been used extensively for the repair and regeneration of tissues. The in vivo half-life time of free bFGF is short, about 3-10 min, due to rapid enzymatic degradation. Stabilization of the bFGF was accomplished by the covalent or physical conjugation of this factor to fluorescent maghemite (γ-Fe(2)O(3)) nanoparticles. In the present study, nasal olfactory mucosa (NOM) cells from adult rats were cultured in suspension on chitosan microcarriers (MCs) in the presence of the nonconjugated or bFGF-conjugated nanoparticles, or the free factor. The floating cells/nonconjugated, conjugated, or free bFGF/MCs aggregates were then seeded in a viscous gel. In this manuscript, we are the first to report that the stabilization of the factor by its conjugation to these nanoparticles significantly improved NOM cell-proliferation properties (migration, growth, and differentiation), compared to the same concentration, or even five times higher, of the free factor. This novel approach may significantly contribute to the advancement of the tissue engineering field.

• Synthesis, fluorescence and biodistribution of a bone-targeted near-infrared conjugate.

  Authors: Dana M Mizrahi, Ofra Ziv-Polat, Benny Perlstein, Eran Gluz, Shlomo Margel

  European journal of medicinal chemistry. 08/2011; 46(10):5175-83.

  Enhanced imaging of early-stage bone abnormalities, such as primary tumors or metastases is highly required as the widely-used bone scan frequently lacks the desired sensitivity. Near IR (NIR)Enhanced imaging of early-stage bone abnormalities, such as primary tumors or metastases is highly required as the widely-used bone scan frequently lacks the desired sensitivity. Near IR (NIR) fluorescence imaging affords high contrast and enhanced sensitivity, as body tissue expresses minimal autofluorescence at NIR range (600-1200 nm). Indocyanine green (ICG), a biocompatible NIR dye, is widely used in the imaging of various organs, such as liver, heart and blood circulation. We report the preparation and in-vivo testing of a bone-targeting ICG derivative, in comparison to the parent molecule(s). Since ICG itself is chemically unreactive, and could not form conjugates, we prepared two novel ICG conjugatable derivatives. The overall ICG structure was maintained while only a replacement of one or two sulfonate groups with carboxylic acids resulted in new linkers for covalent binding to biomolecules. These derivatives were evaluated for their fluorescence and biodistribution in comparison to ICG and were found to be comparable. One of the novel ICG-derivatives was conjugated to a bone-targeting moiety and this new compound was found to bind to growing regions of the skeleton, and emit fluorescence for as long as two weeks in young mice.

• Synthesis and characterization of magnetic poly(divinyl benzene)/Fe3O4, C/Fe3O4/Fe, and C/Fe onionlike fullerene micrometer-sized particles with a narrow size distribution.

  Authors: Ron Snovski, Judith Grinblat, Shlomo Margel

  Langmuir : the ACS journal of surfaces and colloids. 08/2011; 27(17):11071-80.

  Magnetic poly(divinyl benzene)/Fe(3)O(4) microspheres with a narrow size distribution were produced by entrapping the iron pentacarbonyl precursor within the pores of uniform porous poly(divinylMagnetic poly(divinyl benzene)/Fe(3)O(4) microspheres with a narrow size distribution were produced by entrapping the iron pentacarbonyl precursor within the pores of uniform porous poly(divinyl benzene) microspheres prepared in our laboratory, followed by the decomposition in a sealed cell of the entrapped Fe(CO)(5) particles at 300 °C under an inert atmosphere. Magnetic onionlike fullerene microspheres with a narrow size distribution were produced by annealing the obtained PDVB/Fe(3)O(4) particles at 500, 600, 800, and 1100 °C, respectively, under an inert atmosphere. The formation of carbon graphitic layers at low temperatures such as 500 °C is unique and probably obtained because of the presence of the magnetic iron nanoparticles. The annealing temperature allowed control of the composition, size, size distribution, crystallinity, porosity, and magnetic properties of the produced magnetic microspheres.

• Hydrophobic microspheres for in situ removal of 2-phenylethanol from yeast fermentation.

  Authors: Yigal Achmon, Jenny Goldshtein, Shlomo Margel, Ayelet Fishman

  Journal of microencapsulation. 07/2011; 28(7):628-38.

  The commercial production of the fragrance compound 2-phenylethanol (2-PE) from phenylalanine by yeast is limited by the accumulation of the toxic product, and therefore, in situ product removalThe commercial production of the fragrance compound 2-phenylethanol (2-PE) from phenylalanine by yeast is limited by the accumulation of the toxic product, and therefore, in situ product removal techniques are required. We describe the use of hydrophobic polymethylmethacrylate (PMMA) microspheres of narrow size distribution of 1.53 ± 0.10 µm diameter for continuous removal of 2-PE from the fermentation medium by a mechanism of swelling. In shake flask experiments with conditions simulating 2-PE stress, a 10-fold increase in productivity was measured for systems containing >10% (w/v) microspheres. A 1 L fed-batch fermentation with 8% (w/v) of PMMA microspheres resulted in a total 2-PE concentration of 7.05 g/L, from which 5.40 g/L was incorporated inside the resin, implying 76% encapsulation. This ratio of 0.07 g/g of product per resin is among the highest reported to date. Scanning electron microscopy revealed a concomitant increase in sphere diameter confirming that swelling occurred.

• First multimodal embolization particles visible on x-ray/computed tomography and magnetic resonance imaging.

  Authors: Soenke H Bartling, Johannes Budjan, Hagit Aviv, Stefan Haneder, Bettina Kraenzlin, Henrik Michaely, Shlomo Margel, Steffen Diehl, Wolfhard Semmler, Norbert Gretz, Stefan O Schönberg, Maliha Sadick

  Investigative radiology. 03/2011; 46(3):178-86.

  Embolization therapy is gaining importance in the treatment of malignant lesions, and even more in benign lesions. Current embolization materials are not visible in imaging modalities. However, it isEmbolization therapy is gaining importance in the treatment of malignant lesions, and even more in benign lesions. Current embolization materials are not visible in imaging modalities. However, it is assumed that directly visible embolization material may provide several advantages over current embolization agents, ranging from particle shunt and reflux prevention to improved therapy control and follow-up assessment. X-ray- as well as magnetic resonance imaging (MRI)-visible embolization materials have been demonstrated in experiments. In this study, we present an embolization material with the property of being visible in more than one imaging modality, namely MRI and x-ray/computed tomography (CT). Characterization and testing of the substance in animal models was performed. To reduce the chance of adverse reactions and to facilitate clinical approval, materials have been applied that are similar to those that are approved and being used on a routine basis in diagnostic imaging. Therefore, x-ray-visible Iodine was combined with MRI-visible Iron (Fe3O4) in a macroparticle (diameter, 40-200 μm). Its core, consisting of a copolymerized monomer MAOETIB (2-methacryloyloxyethyl [2,3,5-triiodobenzoate]), was coated with ultra-small paramagnetic iron oxide nanoparticles (150 nm). After in vitro testing, including signal to noise measurements in CT and MRI (n = 5), its ability to embolize tissue was tested in an established tumor embolization model in rabbits (n = 6). Digital subtraction angiography (DSA) (Integris, Philips), CT (Definition, Siemens Healthcare Section, Forchheim, Germany), and MRI (3 Tesla Magnetom Tim Trio MRI, Siemens Healthcare Section, Forchheim, Germany) were performed before, during, and after embolization. Imaging signal changes that could be attributed to embolization particles were assessed by visual inspection and rated on an ordinal scale by 3 radiologists, from 1 to 3. Histologic analysis of organs was performed. Particles provided a sufficient image contrast on DSA, CT (signal to noise [SNR], 13 ± 2.5), and MRI (SNR, 35 ± 1) in in vitro scans. Successful embolization of renal tissue was confirmed by catheter angiography, revealing at least partial perfusion stop in all kidneys. Signal changes that were attributed to particles residing within the kidney were found in all cases in all the 3 imaging modalities. Localization distribution of particles corresponded well in all imaging modalities. Dynamic imaging during embolization provided real-time monitoring of the inflow of embolization particles within DSA, CT, and MRI. Histologic visualization of the residing particles as well as associated thrombosis in renal arteries could be performed. Visual assessment of the likelihood of embolization particle presence received full rating scores (153/153) after embolization. Multimodal-visible embolization particles have been developed, characterized, and tested in vivo in an animal model. Their implementation in clinical radiology may provide optimization of embolization procedures with regard to prevention of particle misplacement and direct intraprocedural visualization, at the same time improving follow-up examinations by utilizing the complementary characteristics of CT and MRI. Radiation dose savings can also be considered. All these advantages could contribute to future refinements and improvements in embolization therapy. Additionally, new approaches in embolization research may open up.

• Enhancement of the Growth and Differentiation of Nasal Olfactory Mucosa Cells by the Conjugation of Growth Factors to Functional Nanoparticles

  Authors: Hadas Skaat, Ofra Ziv-Polat, Abraham Shahar, Shlomo Margel

  Bioconjugate Chemistry. 01/2011; 22(12-DOI: 10.1021/bc200454k):2600-2610.

  Growth factors are critical components in the tissue engineering approach. Basic fibroblast growth factor (bFGF), a representative growth factor, stimulates the cellular functions of various cellsGrowth factors are critical components in the tissue engineering approach. Basic fibroblast growth factor (bFGF), a representative growth factor, stimulates the cellular functions of various cells and has been used extensively for the repair and regeneration of tissues. The in vivo half-life time of free bFGF is short, about 3–10 min, due to rapid enzymatic degradation. Stabilization of the bFGF was accomplished by the covalent or physical conjugation of this factor to fluorescent maghemite (γ-Fe2O3) nanoparticles. In the present study, nasal olfactory mucosa (NOM) cells from adult rats were cultured in suspension on chitosan microcarriers (MCs) in the presence of the nonconjugated or bFGF-conjugated nanoparticles, or the free factor. The floating cells/nonconjugated, conjugated, or free bFGF/MCs aggregates were then seeded in a viscous gel. In this manuscript, we are the first to report that the stabilization of the factor by its conjugation to these nanoparticles significantly improved NOM cell-proliferation properties (migration, growth, and differentiation), compared to the same concentration, or even five times higher, of the free factor. This novel approach may significantly contribute to the advancement of the tissue engineering field.

• Convection-enhanced delivery of methotrexate-loaded maghemite nanoparticles.

  Authors: Enav Corem-Salkmon, Zvi Ram, Dianne Daniels, Benny Perlstein, David Last, Sharona Salomon, Gregory Tamar, Ran Shneor, David Guez, Shlomo Margel, Yael Mardor

  International journal of nanomedicine. 01/2011; 6:1595-602.

  Convection-enhanced delivery (CED) is a novel approach for delivering drugs directly into brain tumors by intracranial infusion, enabling the distribution of high drug concentrations over largeConvection-enhanced delivery (CED) is a novel approach for delivering drugs directly into brain tumors by intracranial infusion, enabling the distribution of high drug concentrations over large tissue volumes. This study was designed to present a method for binding methotrexate (MTX) to unique crystalline, highly ordered and superparamagnetic maghemite nanoparticles via human serum albumin (HSA) coating, optimized for CED treatments of gliomas. Naked nanoparticles and HSA- or polyethylene glycol (PEG)-coated nanoparticles with/without MTX were studied. In vitro results showed no toxicity and a similar cell-kill efficacy of the MTX-loaded particles via HSA coating to that of free MTX, while MTX-loaded particles via PEG coating showed low efficacy. In vivo, the PEG-coated nanoparticles provided the largest distributions in normal rat brain and long clearance times, but due to their low efficacy in vitro, were not considered optimal. The naked nanoparticles provided the smallest distributions and shortest clearance times. The HSA-coated nanoparticles (with/without MTX) provided good distributions and long clearance times (nearly 50% of the distribution volume remained in the brain 3 weeks post treatment). No MTX-related toxicity was noted. These results suggest that the formulation in which HSA was bound to our nanoparticles via a unique precipitation method, and MTX was bound covalently to the HSA, could enable efficient and stable drug loading with no apparent toxicity. The cell-kill efficacy of the bound MTX remained similar to that of free MTX, and the nanoparticles presented efficient distribution volumes and slow clearance times in vivo, suggesting that these particles are optimal for CED.

• Fully synthetic phage-like system for screening mixtures of small molecules in live cells.

  Authors: Gerardo Byk, Shirly Partouche, Aryeh Weiss, Shlomo Margel, Raz Khandadash

  Journal of combinatorial chemistry. 03/2010; 12(3):332-45.

  A synthetic "phage-like" system was designed for screening mixtures of small molecules in live cells. The core of the system consists of 2 mum diameter cross-linked monodispersed microspheres bearingA synthetic "phage-like" system was designed for screening mixtures of small molecules in live cells. The core of the system consists of 2 mum diameter cross-linked monodispersed microspheres bearing a panel of fluorescent tags and peptides or small molecules either directly synthesized or covalently conjugated to the microspheres. The microsphere mixtures were screened for affinity to cell line PC-3 (prostate cancer model) by incubation with live cells, and as was with phage-display peptide methods, unbound microspheres were removed by repeated washings followed by total lysis of cells and analysis of the bound microspheres by flow-cytometry. Similar to phage-display peptide screening, this method can be applied even in the absence of prior information about the cellular targets of the candidate ligands, which makes the system especially interesting for selection of molecules with high affinity for desired cells, tissues, or tumors. The advantage of the proposed system is the possibility of screening synthetic non-natural peptides or small molecules that cannot be expressed and screened using phage display libraries. A library composed of small molecules synthesized by the Ugi reaction was screened, and a small molecule, Rak-2, which strongly binds to PC-3 cells was found. Rak-2 was then individually synthesized and validated in a complementary whole cell-based binding assay, as well as by live cell microscopy. This new system demonstrates that a mixture of molecules bound to subcellular sized microspheres can be screened on plated cells. Together with other methods using subcellular sized particles for cellular multiplexing, this method represents an important milestone toward high throughput screening of mixtures of small molecules in live cells and in vivo with potential applications in the fields of drug delivery and diagnostic imaging.

• Synthesis and characterization of functionalized magnetic maghemite nanoparticles with fluorescent probe capabilities for biological applications.

  Authors: Benny Perlstein, Tammy Lublin-Tennenbaum, Inbal Marom, Shlomo Margel

  Journal of biomedical materials research. Part B, Applied biomaterials. 11/2009;

  Nanoparticles with innovative optical, chemical, and magnetic properties combined in a single nanoparticle may be useful as biosensors, targeting agents, and therapeutic agents in the biomedicalNanoparticles with innovative optical, chemical, and magnetic properties combined in a single nanoparticle may be useful as biosensors, targeting agents, and therapeutic agents in the biomedical field. This study describes new magnetic nanoparticles (MNPs) containing the fluorescent dye rhodamine isothiocyanate (RITC) covalently encapsulated within the nanoparticles. These nanoparticles have been prepared by nucleation followed by controlled growth of iron oxide layers onto iron oxide/gelatin-RITC nuclei. The formed RITC labeled MNPs (R-MNPs) are of narrow size distribution, exhibit the fluorescent spectrum of RITC, yet are more photostable. Because of the covalent encapsulation of RITC within the MNPs no detectable leakage of the fluorescent dye into the aqueous continuous phase was observed. This manuscript also demonstrates that the surface of the R-MNPs retains similar ligand binding efficiency as the equivalent nonfluorescent MNPs. Specific cell labeling was obtained by incubating glia cells with R-MNPs conjugated to glial cell line-derived neurotrophic factor (GDNF) protein. We further showed that the R-MNPs may be used for pH sensing between the pH range of 5 and 9. This feature may enable the use of the R-MNPs as a pH sensor of animal tissues and cell compartments. Thus, these functional narrow size distribution R-MNPs with both magnetic and fluorescent properties may provide an important research tool for biological sensing. (c) 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2010.

• Enhancement of incisional wound healing by thrombin conjugated iron oxide nanoparticles.

  Authors: Ofra Ziv-Polat, Moris Topaz, Tamar Brosh, Shlomo Margel

  Biomaterials. 10/2009;

  Thrombin has been clinically used for topical hemostasis and wound management for more than six decades. The half-life of thrombin in human plasma is shorter than 15s due to close control byThrombin has been clinically used for topical hemostasis and wound management for more than six decades. The half-life of thrombin in human plasma is shorter than 15s due to close control by inhibitors. In order to stabilize the thrombin, it was bound to maghemite (gamma-Fe(2)O(3)) nanoparticles, as demonstrated in previous work. The aim of the present study was to examine the efficiency of the bound thrombin for wound healing applications compared to the free thrombin. For this purpose incisional wounds on rat skin were treated with a mixture of fibrinogen, CaCl(2) solution and free or bound thrombin. The wounds' edges were then approximated by skin staples. The control incisional wounds were closed with staples only. In the course of 28 days of healing the highest values of skin tensile strength were observed following treatment with the bound thrombin. Significantly lower values of tensile strength were observed following treatment with the free thrombin, and the lowest values were obtained following treatment with staples only. The histological findings correlate with the mechanical strength measurements, which demonstrate the most advanced stages of healing following treatment with the bound thrombin.

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• Radiopaque iodinated copolymeric nanoparticles for X-ray imaging applications.

  Authors: Hagit Aviv, Sonke Bartling, Fabian Kieslling, Shlomo Margel

  Biomaterials. 08/2009;

  Recently we described iodinated homopolymeric radiopaque nanoparticles of 28.9+/-6.3nm dry diameter synthesized by emulsion polymerization of 2-methacryloyloxyethyl(2,3,5-triiodobenzoate) (MAOETIB).Recently we described iodinated homopolymeric radiopaque nanoparticles of 28.9+/-6.3nm dry diameter synthesized by emulsion polymerization of 2-methacryloyloxyethyl(2,3,5-triiodobenzoate) (MAOETIB). The nanoparticle aqueous dispersion, however, was not stable and tended to agglomerate, particularly at weight concentration of dispersed nanoparticles above approximately 0.3%. The agglomeration rate increases as the concentration of nanoparticles in aqueous phase rises and prevents the potential in vivo use as contrast agent for medical X-ray imaging. Here we describe efforts to overcome this limitation by synthesis of iodinated copolymeric nanoparticles of 25.5+/-4.2nm dry diameter, by emulsion copolymerization of the monomer, MAOETIB, with a low concentration of glycidyl methacrylate (GMA). The surface of resulting copolymeric nanoparticles is far more hydrophilic than that of polyMAOETIB (PMAOETIB) nanoparticles. Therefore, P(MAOETIB-GMA) nanoparticles are significantly more stable against agglomeration in aqueous continuous phase. After intravenous injection of P(MAOETIB-GMA) nanoparticles in rats and mice (including those with a liver cancer model) CT-imaging revealed a significant enhanced visibility of the blood pool for 30min after injection. Later, lymph nodes, liver and spleen strongly enhanced due to nanoparticle uptake by the reticuloendothelial system. This favorably enabled the differentiation of cancerous from healthy liver tissue and suggests our particles for tumor imaging in liver and lymph nodes.

• Synthesis and characterization of fluorinated magnetic core-shell nanoparticles for inhibition of insulin amyloid fibril formation.

  Authors: Hadas Skaat, Georges Belfort, Shlomo Margel

  Nanotechnology. 07/2009; 20(22):225106.

  Maghemite (gamma-Fe2O3) magnetic nanoparticles of 15.0 +/- 2.1 nm are formed by nucleation followed by controlled growth of maghemite thin films on gelatin-iron oxide nuclei. Uniform magneticMaghemite (gamma-Fe2O3) magnetic nanoparticles of 15.0 +/- 2.1 nm are formed by nucleation followed by controlled growth of maghemite thin films on gelatin-iron oxide nuclei. Uniform magnetic gamma-Fe2O3/poly (2,2,3,3,4,4,4-heptafluorobutyl acrylate) (gamma-Fe2O3/PHFBA) core-shell nanoparticles are prepared by emulsion polymerization of the fluorinated monomer 2,2,3,3,4,4,4-heptafluorobutyl acrylate (HFBA) in the presence of the maghemite nanoparticles. The kinetics of the insulin fibrillation process in the absence and in the presence of the gamma-Fe2O3/PHFBA core-shell nanoparticles are elucidated. A significant direct slow transition from alpha-helix to beta-sheets during insulin fibril formation is observed in the presence of the gamma-Fe2O3/PHFBA nanoparticles. This is in contradiction to our previous manuscript, which illustrated that the gamma-Fe2O3 core nanoparticles do not affect the kinetics of the formation of the insulin fibrils, and to other previous publications that describe acceleration of the fibrillation process by using various types of nanoparticles. These core-shell nanoparticles may therefore be also useful for the inhibition of conformational changes of other amyloidogenic proteins that lead to neurodegenerative diseases such as Alzheimer's, Parkinson's, Huntington's, mad cow and prion diseases.

• Synthesis of fluorescent-maghemite nanoparticles as multimodal imaging agents for amyloid-beta fibrils detection and removal by a magnetic field.

  Authors: Hadas Skaat, Shlomo Margel

  Biochemical and biophysical research communications. 07/2009;

  Early diagnosis in Alzheimer's disease (AD), before the onset of marked clinical symptoms, is critical in preventing the irreversible neuronal damage that eventually leads to dementia and ultimatelyEarly diagnosis in Alzheimer's disease (AD), before the onset of marked clinical symptoms, is critical in preventing the irreversible neuronal damage that eventually leads to dementia and ultimately death. Therefore, there is an urgent need for in vivo imaging agents, which are valuable as specific biomarkers to demonstrate the location and density of amyloid plaques in the living human brain. The present manuscript describes a novel method for selective marking of Abeta(40) fibrils by non-fluorescent gamma-Fe(2)O(3) and fluorescent-magnetic gamma-Fe(2)O(3)-rhodamine or gamma-Fe(2)O(3)-Congo red nanoparticles, and the complete removal of the magnetized fibrils from the aqueous continuous phase by a magnetic field. These fluorescent-maghemite nanoparticles as multimodal imaging agents have a great advantage due to the combination of the magnetic and fluorescence imaging into one nanostructured system. This hybrid system, which selectively marks Abeta(40) fibrils, might enable the early detection of plaques using both MRI and fluorescence microscopy, and therefore may be applied in in vivo AD diagnosis studies. These fluorescent-magnetic nanoparticles may also be useful as selective biomarkers to detect the location and the removal of other amyloid plaques derived from different amyloidogenic proteins that lead to neurodegenerative diseases, e.g., Parkinson's, Huntington's, mad cow, and prion diseases.

• Synthesis and characterization of recombinant factor VIIa-conjugated magnetic iron oxide nanoparticles for hemophilia treatment.

  Authors: Gilead Shafir, Anna Galperin, Shlomo Margel

  Journal of biomedical materials research. Part A. 01/2009;

  Maghemite (gamma-Fe(2)O(3)) nanoparticles of 15.0 +/- 2.1 nm in diameter were prepared by nucleation, followed by controlled growth of magnetic iron oxide thin films onto gelatin nuclei.Maghemite (gamma-Fe(2)O(3)) nanoparticles of 15.0 +/- 2.1 nm in diameter were prepared by nucleation, followed by controlled growth of magnetic iron oxide thin films onto gelatin nuclei. Functionalization of these magnetic nanoparticles with activated double bonds was accomplished by interacting divinyl sulfone with the gelatin coating of the gamma-Fe(2)O(3) nanoparticles. The activated double bonds were then used for covalent binding, via Michael addition reaction, of recombinant factor VIIa and human serum albumin to the surface of these nanoparticles. Recombinant factor VIIa was also physically bound to the magnetic nanoparticles by interacting this factor with the human serum albumin conjugated gamma-Fe(2)O(3) nanoparticles. The influence of factor VIIa concentration on the immobilization yield has been elucidated. Leakage of the bound factor VIIa into PBS containing 4% albumin was insignificant. The coagulant activity of the physically adsorbed recombinant factor VIIa was similar to that of the free one and was significantly better than that of the covalently bound. The blood half-life of free factor VIIa is short, about 2-3 h, because of digestion by proteolytic enzymes and inhibitory effects. Stabilization of factor VIIa against trypsin (a model proteolytic enzyme) and chloromethyl ketone-type inhibitor was accomplished by conjugation of the factor to the gamma-Fe(2)O(3) nanoparticles. This stabilization may extend the blood half-life of factor VIIa. Therefore, IV injection of factor VIIa conjugated gamma-Fe(2)O(3) nanoparticles instead of free factor may avoid the frequent dosing and reduce the cost of hemophilia treatment. (c) 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2009.

• Synthesis and Characterization of Thrombin Conjugated gama-Fe2O3 Magnetic Nanoparticles for Hemostasis

  Authors: Ofra Ziv, Tammy Lublin-Tennenbaum, Shlomo Margel

  Advanced Biomaterials. 01/2009; 11:251-260.

  Thrombin is the final protease produced in the clotting pathways. Thrombin has been used in the clinic more than six decades for topical hemostasis and wound management. In human plasma the half-lifeThrombin is the final protease produced in the clotting pathways. Thrombin has been used in the clinic more than six decades for topical hemostasis and wound management. In human plasma the half-life of thrombin is shorter than 15 seconds due to close control by inhibitors. In order to stabilize thrombin, this enzyme was conjugated covalently and physically to gama-Fe2O3 magnetic nanoparticles. The physical conjugation was accomplished through adsorption of thrombin to BSA coating on the nanoparticles. The coagulant activity of the covalently bound thrombin was significantly lower than that of the physically adsorbed thrombin. Leakage of the physically bound thrombin into PBS containing 4% HSA was negligible. The physical conjugation of thrombin onto the nanoparticles stabilized the thrombin against its major inhibitor antithrombin III and improved its storage stability. At optimal CaCl2 concentration, the clotting time by the bound thrombin is shorter than that of the free enzyme. This novel conjugated thrombin may be an efficient candidate for topical hemostasis and wound healing.

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• Effect of maghemite nanoparticles on insulin amyloid fibril formation: Selective labeling, kinetics, and fibril removal by a magnetic field.

  Authors: Hadas Skaat, Mirco Sorci, Georges Belfort, Shlomo Margel

  Journal of biomedical materials research. Part A. 12/2008;

  Maghemite (gamma-Fe(2)O(3)) magnetic nanoparticles of 15.0 +/- 2.1 nm were formed by nucleation followed by controlled growth of maghemite thin films on gelatin-iron oxide nuclei. Human insulinMaghemite (gamma-Fe(2)O(3)) magnetic nanoparticles of 15.0 +/- 2.1 nm were formed by nucleation followed by controlled growth of maghemite thin films on gelatin-iron oxide nuclei. Human insulin amyloid fibrils were formed by incubating the monomeric insulin dissolved in aqueous continuous phase at pH 1.6 and 65 degrees C. Magnetic human insulin amyloid fibrils/gamma-Fe(2)O(3) nanoparticle assemblies were prepared by interacting the gamma-Fe(2)O(3) nanoparticles with the insulin amyloid fibrils during or after their formation. The nanoparticles attached selectively to the insulin fibrils in both cases. The kinetics of the insulin fibrillation process in the absence and the presence of the gamma-Fe(2)O(3) nanoparticles was elucidated. The insulin amyloid fibrils/gamma-Fe(2)O(3) nanoparticle assemblies were easily extracted from the aqueous phase via a magnetic field. We hypothesize that this selective extraction approach may also be applicable for the removal of other amyloidogenic proteins that lead to neurodegenerative diseases (e.g., Alzheimer's, Parkinson's, Huntington's, mad cow, and prion diseases) from their continuous phase, e.g. milk, blood, neurological fluid, etc. (c) 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2008.

• Synthesis and characterization of magnetic and non-magnetic core-shell polyepoxide micrometer-sized particles of narrow size distribution.

  Authors: Melany Omer-Mizrahi, Shlomo Margel

  Journal of colloid and interface science. 10/2008;

  Core polystyrene microspheres of narrow size distribution were prepared by dispersion polymerization of styrene in a mixture of ethanol and 2-methoxy ethanol. Uniform polyglycidylCore polystyrene microspheres of narrow size distribution were prepared by dispersion polymerization of styrene in a mixture of ethanol and 2-methoxy ethanol. Uniform polyglycidyl methacrylate/polystyrene core-shell micrometer-sized particles were prepared by emulsion polymerization at 73 degrees C of glycidyl methacrylate in the presence of the core polystyrene microspheres. Core-shell particles with different properties (size, surface morphology and composition) have been prepared by changing various parameters belonging to the above seeded emulsion polymerization process, e.g., volumes of the monomer glycidyl methacrylate and the crosslinker monomer ethylene glycol dimethacrylate. Magnetic Fe(3)O(4)/polyglycidyl methacrylate/polystyrene micrometer-sized particles were prepared by coating the former core-shell particles with magnetite nanoparticles via a nucleation and growth mechanism. Characterization of the various particles has been accomplished by routine methods such as light microscopy, SEM, FTIR, BET and magnetic measurements.

• Immunogenicity of bioactive magnetic nanoparticles: natural and acquired antibodies.

  Authors: Ofra Ziv, Ramy R Avtalion, Shlomo Margel

  Journal of biomedical materials research. Part A. 07/2008; 85(4):1011-21.

  Uniform magnetic nanoparticles (MNP) were prepared by nucleation followed by controlled growth of maghemite thin films onto porcine gelatin nuclei. The formed gelatin containing MNP (Gel-MNP) wereUniform magnetic nanoparticles (MNP) were prepared by nucleation followed by controlled growth of maghemite thin films onto porcine gelatin nuclei. The formed gelatin containing MNP (Gel-MNP) were then coated with dextran (Gel-MNP-Dex) followed by human serum albumin (Gel-MNP-Dex-HSA). Since these MNP are designated for clinical applications, studies concerning the immunogenicity of their antigenic components (porcine gelatin, dextran, and HSA) have been performed in BALB-C mice. These studies demonstrated that plasma of nonimmunized mice already contains basal levels of natural antibodies against all of these antigenic components. This work also demonstrated that the conjugated gelatin is a weak immunogen: Intraperitoneal injection of the various MNP (Gel-MNP, Gel-MNP-Dex dispersed in PBS emulsified with Incomplete Freund's Adjuvant (IFA) mineral oil and Gel-MNP-Dex-HSA dispersed in PBS) did not increase significantly the acquired anti-gelatin antibody titers. Exceptional behavior was observed following immunization with Gel-MNP-Dex-HSA dispersed in PBS emulsified with IFA, which exhibited an adjuvant effect and turned gelatin into a stronger immunogen. In contrast to gelatin, the conjugated HSA and dextran were found to be strong immunogens. The possibility that the various MNP will not induce an autoimmune response as a result of their clinical use is discussed in the contest of the protective role of the natural antibodies.

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