Beruflich Dokumente
Kultur Dokumente
1. Magnetoliposomes
om
es
Figure 1. Illustration of aqueous magnetoliposomes (on the left) and dry magnetoliposomes (on the right).
1
Magnetoliposomes: synthesis and applications MAP-fis
The flow of the MLs in blood results from the competition between magnetic
forces of the magnetic nanoparticles in blood circulation and magnetic forces applied by
the external magnetic field. The magnetic fluid is localized in the site of interest when
the magnetic forces are exerted on linear blood flow of the arteries or capillaries [8]. To
improve cell uptake, cationic amphiphiles (composed of biodegradable moieties and
specifically designed to exert minimal cytotoxic effects) can be inserted into
magnetoliposomes coat [9]. In drug delivery, after the localization of the
magnetoliposomes in the therapeutic site, drug liberation can be induced by different
physico-chemical environment variations, such as pH or temperature changes.
The magnetic force, important for magnetophoresis applications and the spin
spin relaxation time depend on the nanoparticles concentration, and to how close they
are from each other [10]. This way, controlling the number and the time of encapsulated
magnetic nanoparticles in the magnetoliposomes is a key issue.
1.1 Liposomes
Liposomes act as encapsulation and transport system that can incorporate different
substances as nutrients, genes and drugs. Due to their amphipathic composition,
2
Magnetoliposomes: synthesis and applications MAP-fis
incorporated substances can be either hydrophilic and/or hydrophobic, the first ones will
be incorporated in the aqueous cavity and the second inserted or adsorbed on the
membrane by solubilization in the lipid bilayer or by covalent linkage to active surface
groups in the liposome membrane. Anti-tumor drugs can also be transported by
magnetoliposomes using bifunctional reagents [12], taking advantage of magnetic
targeting.
Liposomes are structurally flexible in size, composition and fluidity [13]. The
choice of phospholipids determines the rigidity and the charge of the bilayer. Saturated
phospholipids with long acyl chains form a rigid, rather impermeable bilayer structure,
while unsaturated species from natural sources (egg or soy bean phosphatidylcholine)
give much more permeable but less stable vesicles. Phase transition of the liposomes is
also an important characteristics. It is affected by the hydrocarbon length, unsaturation,
charge and headgroup species of the phospholipids. It defines the temperature required
to induce a change in the physical state of the liposomes, from the ordered gel phase,
where the hydrocarbon chains are fully extended and closely packed, to the disordered
liquid crystalline phase, where the hydrocarbon chains are randomly oriented and fluid.
According to size and number of lamellae liposomes are divided into different
subcategories (table 1 and figure 3).
3
Magnetoliposomes: synthesis and applications MAP-fis
Table 1. Liposome classification based on structural parameters [14]. Figure 3. Schematic representation
of different liposomes.
Liposomes are very important for biomedical applications as they can overcome
many of the problems associated with other systems used for therapy, such as those
involving solubility, pharmacokinetics, in vivo stability and toxicity [15, 16].
For in vivo applications liposomes must be very small (100 nm) in order to
reduce the recognition and phagocytosis, thereby increasing the probability of
penetration into the tissues of interest [20]. Recent studies have revealed that the
functionalization of the liposomes surface improves their efficacy. Functionalize the
lipid bilayer with FAB portions (a part of the antibody molecule that binds antigen),
make liposomes to be uptake only by cells that have specific antigens. In order to make
the system less recognizable by the immune system, a PEG (polyethylene glycol) crown
can be added. This will also provide stealth ability (long-circulating) and substance
accumulation in the site of interest [6]. It has also been reported that cationic liposomes
preferably target vasculature [21], while anionic ones are captured by monocytes/blood
4
Magnetoliposomes: synthesis and applications MAP-fis
neutrophils [3]. This last behavior has been used to target the brain through the blood
brain barrier (BBB) [22].
Since it is now accepted that magnetic fields are not especially contraindicated
for humans, the potential of magnetism has been recognized in many biological
applications [4]. Magnetic materials have attracted great interest in recent years because
of novel effects that arise due to size reduction, as is the case of superparamagnetism.
This type of magnetism is present in magnetic nanoparticles due to their size and it only
occurs below critical sizes in which nanoparticles are single domain. Critical sizes (dc)
of superparamagnetic nanoparticles depend on the material; iron nanoparticles have
superparamagnetic behavior below 20nm, while in nickel nanoparticles it occurs below
30nm [24, 25].
5
Magnetoliposomes: synthesis and applications MAP-fis
Figure 4. Cobalt hysteresis for different size samples; the larger particles shows hysteresis while the smaller are
superparamagnetic and do not have hysteresis [24].
6
Magnetoliposomes: synthesis and applications MAP-fis
to reorient under the influence of a specified magnetic field in 100 s (a typical timescale
for a measurement). This temperature is important because it represents the maximum
in susceptibility and the lower limit of superparamagnetic behavior [24]. Blocking
temperature is an important characteristic for many magnetic applications that depend
on particle size. A wide particle size distribution will result in a wide range of blocking
temperatures, and so, in a non-ideal magnetic behavior for the magnetic fluid.
Iron and nickel are considered metals of biological interest as they present
magnetic properties at room temperature [29]. However, particles of these metals have
some issues as its toxicity, high reactivity and also the fact that they are easily degraded
due to high surface/volume ratio. In order to overcome these problems and make them
compatible for biological applications, core-shell structures are used. Core-shell
structure consists of a metal or metallic oxide core, encapsulated in an inorganic or a
polymeric coating; silica is commonly used (figure 5). Silica coating will separate the
particles, thereby preventing a cooperative switching. It will also adjust the magnetic
properties of nanoparticles, as the extent of dipolar coupling is related to the distance
between particles and this, in turn, depends on the thickness of the inert silica shell.
With this structure, the undesirable effects are avoided and the properties of the
magnetic nanoparticles, such as thermal and chemical stability and solubility, are
improved. Core-shell structures also allow the conjugation of other molecules [30]. For
in vivo applications, nanoparticles must have high magnetization so that their movement
in blood circulation can be easily controlled. Nanoparticles with superparamagnetic
behavior are preferred because the risk of forming agglomerates is negligible at room
temperature [31, 32]. They can be guided to a therapeutic site and are not subject to
7
Magnetoliposomes: synthesis and applications MAP-fis
strong magnetic interactions in the dispersion. Also, they are readily stabilized in
physiological conditions [28].
The high biocompatibility and versatile nature of liposomes have made these
systems keystone components in many biomedical research areas. Liposomes can be
combined with a large variety of nanomaterials, such as magnetic nanoparticles which
have greatest applications in biomedicine (figure 6).
Because the unique features of both the magnetic colloid and the versatile lipid
bilayer can be joined, the resulting so-called magnetoliposomes can be exploited in a
great array of biomedical applications. In therapy, the most promising applications of
magnetoliposomes are magnetic controlled drug delivery and hyperthermia [6, 7].
Otherwise, in diagnosis, magnetic nanoparticles have been used as contrast agents in
MRI (magnetic resonance imaging) [5].
8
Magnetoliposomes: synthesis and applications MAP-fis
Cancer is a disease that causes more than six million deaths per year worldwide
[19]. In pharmaceutical industry new cytotoxic agents for cancer treatment have been
produced. However, the administration of these drugs continues to be a problem
because of toxicity to healthy cells. To overcome this problem, magnetoliposomes have
been proposed as carrier systems that protect and transport the drug to the target site by
means of an external magnetic field.
9
Magnetoliposomes: synthesis and applications MAP-fis
[42, 43] and hyperthermia makes use of magnetic nanoparticles to deliver toxic amount
of thermal energy to targeted tumors. Therefore, magnetic nanoparticles are used as
hyperthermia agents. For a better temperature control, nanoparticles should be uniform
in size and shape. Also, they should be subdomain in order to absorb much more power
at tolerable AC (alternating current) magnetic fields [44, 45].
Body tissues contains lots of water, so, when a magnetic field is applied, the
average magnetic moment of the many water protons become aligned with the direction
of the field. When a radio frequency current is briefly turned on, an electromagnetic
field with resonance frequency is absorbed, flipping the spin of the protons in the
magnetic field. When the field is turned off, the spins of the protons return to the
10
Magnetoliposomes: synthesis and applications MAP-fis
thermodynamic equilibrium and the bulk magnetization is reorganized with the static
magnetic field. Protons in different tissues return to their equilibrium state at different
relaxation rates. The image is obtained from different variables, as spin density and T1
and T2 relaxation times (T1 is the spin-lattice or longitudinal relaxation time, and T2 is
the spin-spin or transverse relaxation time) that are used to create a MRI contrast which
relies on the differential uptake of different tissues.
2. Preparation techniques
11
Magnetoliposomes: synthesis and applications MAP-fis
Simple soft chemical methods have been widely used as they do not demand
extreme pressure or temperature control, are easy to handle, and do not require special
or expensive equipment. Nickel nanoparticles can be obtained with different soft
chemical methods, namely by the reduction of nickel chloride with hydrazine (N2H4):
12
Magnetoliposomes: synthesis and applications MAP-fis
In this process, reagents are separated in two different microemulsions that are
then mixed under agitation. After mixing, the reverse micelles collide among
themselves to exchange the reactants solubilized in the nanoreactors (individual reverse
micelles) and then again break apart (figure 8). This way, the reagents undergo
homogeneous mixing and the polydispersity of the particles decrease. Decoalescence
ensures the presence of the protective coating for the controlled nucleation and growth
and it also prevents aggregation [63].
Figure 8. Mechanism showing the intermicellar exchange for the formation of nanoparticles.
The main problem associated with the synthesis of small magnetic nanoparticles
are the magnetic attractions of the metallic nanoparticles. This contributes to
agglomeration/coalescence into larger particles and their subsequent settling out of the
reaction environment. A novel route for the synthesis of smaller magnetic nanoparticles
13
Magnetoliposomes: synthesis and applications MAP-fis
has been proposed. This method is based on the reduction of metal chloride ionic
clusters in the confined space of the anionic surfactant AOT (bis(2-
ethylhexyl)sulfosuccinate) reverse micelles [64].
From this method, smaller nanoparticles, with diameters lower than 100 nm, were
obtained (figure 9). The resulting particles are covered with an AOT layer that prevents
particles agglomeration, due to electrostatic repulsions of the reverse micelles.
Figure 9. SEM images of Ni nanoparticles synthesized in the confined space of AOT reversed micelles.
14
Magnetoliposomes: synthesis and applications MAP-fis
method to synthesize iron oxides from aqueous Fe2+ and Fe3+ salt solutions by the
addition of a base under inert atmosphere at room temperature or at elevated
temperature:
Fe+ +
2 + Fe3 + 8HO Fe3 O4 + 4H2 O
Figure 10. Fe3O4 nanoparticles synthesized by co-precipitation method, the scaler bar is 30 nm [65].
15
Magnetoliposomes: synthesis and applications MAP-fis
The disadvantages of this method are the low yield and poor crystallinity of the
nanoparticles, which limit its practical use. Also, the large amount of organic solvents
used would lead to not only higher costs, but also a non-friendly impact in the
environment.
Figure 11. Fe3O4 nanoparticles prepared by thermal decomposition of iron oleate Fe(OA)3 [65].
16
Magnetoliposomes: synthesis and applications MAP-fis
17
Magnetoliposomes: synthesis and applications MAP-fis
In the ethanolic injection method (figure 13), a lipid solution in ethanol is rapidly
injected in a buffer solution under vortex. The buffer solution must be at a temperature
18
Magnetoliposomes: synthesis and applications MAP-fis
above the transition temperature of the lipid [76-78]. Small liposomes in a range of 30-
110 nm are obtained. The advantages of ethanolic injection include its potential for
scale up, the simplicity of the procedure, low cost and low expenditure of time [79]. On
the other hand, the drawbacks of this method are that the population is poorly
homogeneous, liposomes are dilute and it is also difficult to remove all the ethanol [54].
From the alkaline co-precipitation of iron salts in the presence of the lipid DOPG
(1,2-Dioleoyl-sn-glycero-3-[phospho-rac-(1-glycerol)]) phospholipid molecules,
magnetite nanoparticles covered by a monolayer of DOPG molecules were synthesized
[8]. Dry magnetoliposomes are based on the growth of a second lipid layer around these
nanoparticles. Adding slowly another volume of lipid solution, equal to that used in the
synthesis of those nanoparticles, a second phospholipid layer is formed around the iron
oxide core [8]. This way, a lipid bilayer is created around the nanoparticles and dry
magnetoliposomes are obtained. Dry magnetoliposomes can also be obtained by mixing
SUV liposomes (obtained by strong sonication with tip) with MNPs, followed by
dialysis [6].
19
Magnetoliposomes: synthesis and applications MAP-fis
References
[1] M. Mezei, V. Gulasekharam, Liposomes - a selective drug delivery system for the topical route of
administration. Life Sciences, 26 (1980) 1473-1477.
[2] R. L. Juliano, Liposomes as a drug delivery system. Trends in Pharmacological Sciences, 2 (1981)
3941.
[3] G. Poste, C. Cucana, A. Raz, P. Bugelski, R. Kirsj, I. J. Fidler, Analysis of the fate of systemically
administered liposomes and implication for their use in drug delivery. Cancer Research, 42 (1982)
1412-1422.
[4] S. Mornet, S. Vasseur, F. Grasset, E. Duguet, Magnetic nanoparticle design for medical diagnosis and
therapy. Journal of Materials Chemistry, 14 (2004) 2161-2175.
[11] A. D. Bangham, M. M. Standish, J. C. Watkins, Diffusion of univalent ions across the lamellae of
swollen phospholipids. Journal of Molecular Biology, 13 (1965) 238-252.
[12] J-B. Sun, J-H. Duan, S-L. Dai, J. Ren, L. Guo, W. Jiang, Preparation and Anti-Tumor Efciency
Evaluation of Doxorubicin-Loaded Bacterial Magnetosomes: Magnetic Nanoparticles as Drug Carriers
Isolated From Magnetospirillum gryphiswaldense. Biotechnology and Bioengineering, 101 (2008)
13131320.
[13] Y. Malam, M. Loizidou, A. M. Seifalian, Liposomes and nanoparticles: nanosized vehicles for drug
delivery in cancer. Trends in Pharmacological Sciences, 30 (2009) 592-599.
20
Magnetoliposomes: synthesis and applications MAP-fis
[20] G. Gregoriadis, Engineering liposomes for drug delivery: progress and problems. Trends in
Biotechnology, 13 (1995) 527-537.
[24] D. L. Huber, Synthesis, Properties, and Applications of Iron Nanoparticles. Small, 1 (2005) 482
501.
[28] C. Xu, S. Sun, Monodisperse magnetic nanoparticles for biomedical applications. Polymer
International, 56 (2007) 821826.
[29] L. B. Bangs, New developments in particle-based immunoassays: Introduction. Pure and Applied
Chemistry, 68 (1996) 1873-1879.
[30] N. Sounderya, Y. Zhang, Use of core/shell structured nanoparticles for biomedical applications.
Recent Patents on Biomedical Engineering, 1 (2008) 34-42.
[31] E. H. Frei, S. Shtrikman, D. Treves, Critical size and nucleation field of ideal ferromagnetic
particles. Physical Review, 106 (1957) 446455.
[34] R. Langer, New methods of drug delivery. Science, 249 (1990) 1527-1533.
[36] M. Arruebo, R. F. Pacheco, M. R. Ibarra, J. Santamara, Magnetic nanoparticles for Drug delivery.
Nanotoday, 2 (2007) 22-32.
21
Magnetoliposomes: synthesis and applications MAP-fis
[41] P. Hlig, M. Bach, T. Vlkel, T. Nahde, S. Hoffmann, R. Mller, R. E. Kontermann, Novel RGD
lipopeptides for the targeting of liposomes to integrin-expressing endothelial and melanoma cells.
Protein Engineering, Design and Selection, 17 (2004) 433-441.
[43] G. Fleissner, B. Stahl, P. Thalau, G. Falkenberg, G. Fleissner. A novel concept of Fe-mineral based
magnetoreception: histological and physicochemical data from the upper beak of homing pigeons.
Naturwissenschaften, 94 (2007) 631642.
[44] U. Hafeli, G. Pauer, S. Failing, G. Tapolsky, Radiolabeling of magnetic particles with rhenium-188
for cancer therapy. Journal of Magnetism and Magnetic Materials, 225 (2001) 7378.
[47] M-S. Martina, J-P. Fortin, C. Menager, O. Clement, G. Barratt, C. G-Madelmont, F. Gazeau, V.
Cabuil, S. Lesieur, Generation of Superparamagnetic Liposomes Revealed as Highly Efficient MRI
Contrast Agents for in Vivo Imaging. Journal of the American Chemical Society, 127 (2005) 10676-
10685.
[49] R. C. Semelka, T. K. G. Helmberger, Contrast agents for MR imaging of the liver. Radiology, 218
(2001) 2738.
[50] W. S. Enochs, G. Harsh, F. Hochberg, R. Weissleder, Improved delineation of human brain tumors
on MR images using a long-circulating, superparamagnetic iron oxide agent. Journal of Magnetic
Resonance Imaging, 9 (1999) 228 232.
[53] C. M. Niemeyer, Nanoparticles, Proteins, and Nucleic Acids: Biotechnology Meets Materials
Science. Angewandte Chemie, 40 (2001) 41284158.
22
Magnetoliposomes: synthesis and applications MAP-fis
[56] Y. Lee, J. Lee, C. J. Bae, J. G. Park, H. J. Noh, Large-scale synthesis of uniform and crystalline
magnetite nanoparticles using reverse micelles as nanoreactors under reflux conditions.
Advanced Functional Materials, 15 (2005) 503-509.
[59] S. H. Chung, A. Hoffmann, S. D. Bader, C. Liu, B. Kay, Biological sensors based on Brownian
relaxation of magnetic nanoparticles. Applied Physics Letters, 85 (2004) 2971-2973.
[64] P. Calandra, Synthesis of Ni nanoparticles by reduction of NiCl2 ionic clusters in the confined space
of AOT reversed micelles. Materials Letters, 63 (2009) 2416-2418.
[65] X. Peng, H. Chen, J. Huang, H. Mao, D. M. Shin, Targeted magnetic iron oxide nanoparticles for
tumor imaging and therapy. International Journal of Nanomedicine, 3 (2008) 311-321.
[66] A-H. Lu, E. L. Salabas, F. Schuth, Magnetic Nanoparticles: Synthesis, Protection, Functionalization,
and Application. Angewandte Chemie International Edition, 46 (2007) 12221244.
[67] L. Vayssires, C. Chanac, E. Tronc, J. P. Jolivet, Size Tailoring of Magnetite Particles Formed by
Aqueous Precipitation: An Example of Thermodynamic Stability of Nanometric Oxide Particles. Journal
of Colloid and Interface Science, 205 (1998) 205212.
[68] R. Y. Hong, B. Feng, X. Cai, G. Liu, H. Z. Li, J. Ding, Y. Zheng, D. G. Wei, Double-miniemulsion
preparation of Fe3O4/poly(methyl methacrylate) magnetic latex. Journal of Applied Polymer Science,
112 (8998) 2009.
[69] A. B. Chin, I. I. Yaacob, Synthesis and characterization of magnetic iron oxide nanoparticles via w/o
microemulsion and Massart's procedure. Journal of Materials Processing Technology, 191 (235-237)
2007.
[70] C. Liu, B. Zou, A. J. Rondinone, Z. J. Zhang, Reverse Micelle Synthesis and Characterization of
Superparamagnetic MnFe2O4 Spinel Ferrite Nanocrystals. Journal of Physical Chemistry B, 104
(2000) 1141-1145.
[71] S-W. Lee, D-H. Chang, M-S. Shim, B-O. Kim, S-O. Kim, M-H. Seo, Ionically Fixed Polymeric
Nanoparticles as a Novel Drug Carrier. Pharmaceutical Research, 24 (2007) 1508-1516.
23
Magnetoliposomes: synthesis and applications MAP-fis
[72] K. Butter, A. P. Philipse, G. J. Vroege, Synthesis and properties of iron ferrofluids. Journal of
Magnetism and Magnetic Materials, 252 (2002) 1-3.
[73] J. Wan, W. Cai, X. Meng, E. Liu, Monodisperse water-soluble magnetite nanoparticles prepared
by polyol process for high-performance magnetic resonance imaging. Chemical Communications, 47
(2007) 5004-5006.
[74] A. G.-Hens, J. M. F.-Romero, Analytical methods for the control of liposomal delivery systems.
Trends in Analytical Chemistry, 25 (2006) 167178.
[77] M .L. Steigerwald, L. E. Brus, la3 bination as opposed to improving the minority carrier. Accounts
of Chemical Research, 23 (1990) 183-188.
24