RU2619854C2 - Stand for research of process of magnetocontrollable nanoparticle delivery into vascular system (versions) - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: stand for research of the process of magnetocontrollable nanoparticle delivery to the vascular system comprises a Y-shaped tube with single end located between the magnet and the recorder, and connected to a storage tank, a pump, a flow meter, as well as a pressure sensor, and a nanoparticle delivery element. The storage tank output is connected through the pump and the flow meter to one of the forked ends of the Y-shaped tube, forming a closed loop that simulates the circulatory system. At that, the said forked end of the Y-shaped tube is also connected to the pressure sensor and the nanoparticle delivery element. The nanoparticle delivery element is a syringe, according to the first version, and an electromagnetic probe connected to a power source, according to the second version.

EFFECT: opportunity to research the process of magnetocontrollable nanoparticle delivery to the vascular system.

3 cl, 7 dwg, 1 tbl, 1 ex

Description

The invention relates to medicine, in particular to experimental medicine, and can be used to study the process of accumulation of magnetic nanoparticles in a given section of the vascular system under the influence of an external magnetic field.

In recent years, nanoparticles with magnetic properties have been actively studied and are used in medical imaging, diagnostics, and therapy. The use of contrast agents based on magnetic nanoparticles (MNPs) in magnetic resonance imaging allows visualization of inflammation of various localization in the nervous system, kidneys, intestines, lungs, liver, bone and adipose tissues [Wu Y., Briley-Saebo K., Xie J. et al. Inflammatory bowel disease: MR- and SPECT / CT-based macrophage imaging for monitoring and evaluating disease activity in experimental mouse model-pilot study // Radiology. - 2014 .-- Vol. 271, No. 2. - P. 400-406; Luciani A., Dechoux S., Deveaux V. et al. Adipose tissue macrophages: MR tracking to monitor obesity-associated inflammation // Radiology. - 2012. - Vol. 263, No. 3. - P. 786-878]. In addition, MNPs in experimental studies are used in the following areas: for targeted delivery of anticancer drugs, targeted thermosensitive chemotherapy [Pradhan P., Giri J., Rieken F. et al. Targeted temperature sensitive magnetic liposomes for thermo-chemotherapy // J Control Release. - 2010 .-- Vol. 142, No. 1. - P. 108-129], magnetic photodynamic therapy [Cinteza L.O., Ohulchanskyy T.Y., Sahoo Y. et al. Diacyllipid micelle-based nanocarrier for magnetically guided delivery of drugs in photodynamic therapy // Mol Pharm. - 2006. - Vol. 3, No. 4. - P. 415-437] and fluorescence imaging [Yang L., Mao H., Cao Z. et al. Molecular imaging of pancreatic cancer in an animal model using targeted multifunctional nanoparticles // Gastroenterology. - 2009. - Vol. 136, No. 5 - P. 1514-1538].

The effectiveness of the use of anti-ischemic drugs immobilized on the surface of magnetic nanoparticles is presented in the literature in isolated articles. At the same time, magnetically controlled drug delivery to the ischemic zone can potentially provide an increase in therapeutic effect while reducing the severity of systemic side effects [Galagudza M., Korolev D., Postnov V. et al. Passive targeting of ischemic-reperfused myocardium with adenosine-loaded silica nanoparticles // Int J Nanomedicine. - 2012. - No. 7. - P. 1671-1678]. In the work of Zhang et al. [Zhang Y., Li W., Ou L. et al. Targeted delivery of human VEGF gene via complexes of magnetic nanoparticle-adenoviral vectors enhanced cardiac regeneration // PLoS One. - 2012. - Vol. 7, No. 7. - P. e39490], the beneficial effect of using magnetic nanoparticles as a transporter of vascular endothelial growth factor in a rat coronary artery ligation model was demonstrated.

To establish the prospects of using magnetic nanoparticles as a transporter of drugs, it is necessary to solve a number of problems, in particular, to find out the optimal magnetic field intensity to ensure the accumulation of nanoparticles in the focus of ischemia, to study the biocompatibility of nanoparticles, and also to study the cardioprotective effect of drugs immobilized on the surface of nanoparticles.

However, the study of the process of magnetically controlled delivery is more appropriate to begin under in vitro model conditions. This requires the development of a specialized test bench that simulates a section of the circulatory system and allows you to apply a magnetic effect, as well as register the accumulation of nano-objects. Similar problems were partially solved in the design of auxiliary circulatory systems. The peak of research in this area came at the end of the 70s, the beginning of the 80s of the last century [Description of the invention to copyright certificate No. 685294. Stand-simulator of the circulatory system of the body. / M.A. Lokshin, Yu.N. Gavrilov, V.I. Kovin, publ. 09/15/79; Description of the invention to copyright certificate No. 936922. Stand for modeling the circulatory system. / A.P. Osipov, V.M. Mordashev, V.A. Kremnev, Yu.M. Kiselev, publ. 06/23/82; Description of the invention to copyright certificate No. 939013. Device for modeling hemodynamic phenomena in the circulatory system. / B.C. Bednenko, A.S. Nekhaev, A.N. Kozlov, publ. 06/30/82; Description of the invention to the copyright certificate SU 1029961 A. Stand for testing an artificial heart. / M.A. Lokshin, V.I. Kopin, V.G. Severin, A.V. Vrubel, V.A. Stasenkov, publ. 07.23.83].

However, the disadvantages of these stands should include their limited functionality.

Conducted by the authors of patent research did not reveal a prototype of the invention.

The problem to which the invention is directed, is to study the process of accumulation of magnetic nanoparticles in a given section of the vascular system under the influence of an external magnetic field.

This problem is solved due to the fact that in the first embodiment, the design of the stand for studying the process of magnetically controlled delivery of nanoparticles to the vascular system includes a U-shaped tube, the unit end of which, located between the magnet and the recorder, is connected to the storage tank, the output of the latter is connected through the pump and flow meter with one of the bifurcated ends of the U-shaped tube, forming a closed loop simulating the circulatory system, and said bifurcated end of the U-shaped tube is also connected to the sensor pressure and the nanoparticle delivery element, which is a syringe.

This problem is also solved due to the fact that in the second embodiment, the design of the stand for studying the process of magnetically controlled delivery of nanoparticles into the vascular system includes a U-shaped tube, the unit end of which, located between the magnet and the recorder, is connected to the storage tank, the output of the latter is connected through a pump and a flowmeter with one of the bifurcated ends of the U-shaped tube, forming a closed loop that simulates the circulatory system, and said bifurcated end of the tube is also connected to the sensor luminescence, and at the other forked end of the U-shaped tube is placed the nanoparticle delivery element, which is an electromagnetic probe connected to a power source.

In the second embodiment, this problem is also solved by the construction of an electromagnetic probe made in the form of a solenoid with a length significantly exceeding its diameter.

The technical result provided by the given set of features of the claimed group of inventions is the creation of a universal stand for studying the process of magnetically controlled delivery of nanoparticles into the vascular system, combining a model of a portion of the circulatory system, a magnetic exposure application system, and a nanoparticle accumulation monitoring system.

The invention is illustrated by drawings, where:

in FIG. 1 shows a block diagram of a stand for studying the process of magnetically controlled delivery of nanoparticles into the vascular system (option 1);

in FIG. 2 is a block diagram of a bench for studying the process of magnetically controlled delivery of nanoparticles to the vascular system (option 2);

in FIG. 3 shows the design of the magnetic probe (option 2);

in FIG. 4 is a graph of the volumetric flow rate of the model fluid versus pump power;

in FIG. 5 is a micrograph of a magnetite nanopowder;

in FIG. 6 is a graph of the magnetic susceptibility of magnetic nanoparticles;

in FIG. 7 - accumulation of magnetic nanoparticles in the zone of motion of the model fluid: A - photograph of the accumulation of magnetic nanoparticles; B is a schematic representation of the topography of the magnetic field (distribution of magnetic induction in space).

The design of the stand for studying the process of magnetically controlled delivery of nanoparticles to the vascular system in the first embodiment (Fig. 1) contains: a U-shaped tube 1, the unit end of which, located between the magnet 2 and the recorder 3, is connected to the storage tank 4, the output of the latter is connected through a pump 5 and a flow meter 6 with one of the bifurcated ends of the U-shaped tube 1, said bifurcated end of the U-shaped tube 1 is also connected to a pressure sensor 7 and a nanoparticle delivery element 8, which is a syringe.

The design of the stand for studying the process of magnetically controlled delivery of nanoparticles to the vascular system in the second embodiment (Fig. 2) contains: a U-shaped tube 1, the unit end of which, located between the magnet 2 and the recorder 3, is connected to the storage tank 4, the output of the latter is connected through a pump 5 and a flow meter 6 with one of the bifurcated ends of the U-shaped tube 1, forming a closed loop simulating the circulatory system, and said bifurcated end of the U-shaped tube 1 is also connected to the pressure sensor 7, and in the other zdvoenny end of the U-shaped tube 1 is inserted nanoparticle delivery element 8, which is an electromagnetic probe connected to a power source 9.

The U-shaped tube 1 (Figs. 1 and 2) can be made, for example, of pyrex glass.

As the recorder 3 (Fig. 1 and 2) can be applied industrial video camera or digital camera. It is also possible to use fluorescent dyes in conjunction with the corresponding filters and radiation sources.

The hydraulic part of the simulating circulatory system (Fig. 1 and 2) includes: a storage tank 4, which is a receiver, a pump 5, which can be used, for example, a pump from an artificial kidney apparatus, and a flow meter 6. Flow and pressure in the system are measured just before entering the U-shaped tube.

As an element for the delivery of nanoparticles 8 according to option 2 (Fig. 2), an electromagnetic probe in the form of a solenoid (Fig. 3) was designed with a length significantly exceeding the diameter and providing a calculated magnetic field induction of 6.55 mT. The parameters of the electromagnetic probe are shown in the table.

The developed design of the electromagnetic probe makes it possible to study the balance of magnetic confinement - turbulent separation forces for various operating modes and to identify the parameters of guaranteed confinement for a certain type of MNP.

There are two fundamentally different delivery methods for MNPs, respectively, and two different delivery elements. The first option (Fig. 1) involves the introduction of a suspension of nanoparticles using a syringe 8 and subsequent accumulation in the zone of action of the magnetic field created by the magnet 2 located along the axis of the glass tube. The second option (Fig. 2) involves the introduction of nanoparticles held on an electromagnetic probe 8 directly into a U-shaped tube in the area of its direct passage along the axis. The power of the electromagnetic probe 8 is provided from a DC power source 9.

The volumetric flow rate of the model fluid in the experimental zone can vary in the range from 40 to 80 l / h, which corresponds to a preset power of 10 to 100% (Fig. 4).

Distilled water was used as a model fluid.

The work of the stand for studying the process of magnetically controlled delivery of nanoparticles to the vascular system in the first embodiment (Fig. 1) is as follows.

The model fluid from the storage tank 4 is fed into the Y-shaped tube 1 by means of a pump 5, where a suspension of magnetic nanoparticles is introduced using a syringe 8. The retention of magnetic nanoparticles in the Y-shaped tube is carried out using a magnet 2. The accumulation of magnetic nanoparticles is controlled by the recorder 3. The flow rate of the model fluid is measured using a flowmeter 6, pressure - pressure sensor 7. Thus, magnetically controlled drug delivery associated with magnetic nanoparticles to local areas of the vascular bed. An example of such sites are the limbs. In this case, upon delivery of a medicinal substance, for example, neotone (creotine phosphate, phosphocreatine) to an extremity, muscle tissue can be regenerated. As a magnet, you can use mixed magnetic thermoplastic elastomers for medical devices [Kisel L.O., Krasovsky V.N. Korolev D.V., Suvorov K.A. Compound magnetic thermoplastic elastomers (TEP) for medical devices // Rubber and rubber. - 1999, No. 1. - S. 11-13].

The work of the stand for studying the process of magnetically controlled delivery of nanoparticles to the vascular system in the second embodiment (Fig. 2) is as follows.

A model fluid from the storage tank 4 is fed into the Y-shaped tube 1 by means of a pump 5. Magnetic nanoparticles are introduced into the Y-shaped tube by means of an electromagnet 8. The electromagnet is powered from a source 9, which allows setting various values of current and voltage. The retention of magnetic nanoparticles in the Y-shaped tube, detached from the electromagnet 8, is carried out using a magnet 2. The accumulation of magnetic nanoparticles is monitored by the recorder 3. Measurement of the flow rate of the model fluid is carried out using a flow meter 6, pressure - pressure sensor 7. This circuit allows you to simulate the process magnetically controlled delivery directly to the vasculature using delivery vehicles similar to stent delivery vehicles used for stenting coronary arteries th.

EXAMPLE

An experiment on the study of magnetically controlled delivery was carried out for a suspension of magnetite nanoparticles with a size of 7-10 nm (Fig. 5) and specific surface area, measured by the simplified BET method at the Klyachko-Gurvich facility, 95 m ² / g. The calculated specific surface area of the nanomaterial is 130 m ² / g [Afonin M.V., Evreinova N.V., Korolev D.V. et al. Investigation of the physical properties and biodegradation of magnetite nanoparticles in vitro // Biotechnosphere. - 2015. - No. 2 (38). - S. 32-34]. As can be seen from the micrograph (Fig. 5), the nucleation of future MNPs begins with the formation of gamma-iron oxide. This is evidenced by the pronounced needle shape of the nanoparticles.

A plot of magnetic susceptibility obtained with a Lake Shore 7410 vibrating magnetometer (Lake Shore Cryotronics Inc., USA) in air at standard temperature is shown in FIG. 6. The absence of a hysteresis loop indicates that the samples studied belong to superparamagnets or to soft ferromagnetic materials with high magnetic permeability. This coincides with the classical ideas about the magnetic properties of nanomaterials [Pozdnyakov VA. Physical material science of nanostructured materials: Textbook. allowance. - M .: MGIU, 2007. - 423 p.]. Variants of using such MNPs are proposed by the authors in the Patent for invention RU 2525430 C2. Carrier for drugs and biologically active substances for the treatment and diagnosis and method for its preparation. / Korolev D.V., Afonin M.V., Galagudza M.M. et al., publ. 08/10/2014, Bull. Number 22.

The synthesis of nanomaterial was carried out as follows [Korolev D.V., Galagudza M.M., Afonin M.V. et al. Rationale for the use of magnetic nanoparticles for targeted delivery of drugs to ischemic skeletal muscle // Biotechnosphere. - 2012 .-- 1 (19). - S. 2-6]. To a solution containing a mixture of iron (II) sulfates, iron (III) in a molar ratio of 2: 1 and a volume of 700 ml, with constant stirring at a speed of 4 ml / min, a mixture of 25% solution of ammonium hydroxide and 1% solution of ammonium acetate was added. Thus, the ratio of iron to ammonium acetate was 2: 1: 0.1. The synthesis was carried out until a saturated black color was fixed and a pH value of 8–9 was established. The next day, the resulting colloidal product was separated by centrifugation and washed 4 times with distilled water. To prepare a dry sample, the obtained MNPs were filtered off and subjected to freeze drying at a temperature of -48 ° C for 48 hours.

A colloidal solution of nanoparticles in physiological saline had a concentration of MNP of 2 mg / ml and was prepared on an ultrasonic disperser UZD-2 for 5 minutes.

A study of magnetically controlled delivery was carried out on the claimed stand according to option 1. A suspension of nanoparticles in an amount of 5 ml with a concentration of 2 mg / ml in physiological solution was injected using a syringe for 10 s, as shown in FIG. 1. The pumping speed of the model fluid was 80 l / h (100% power). Distilled water was used as a model fluid. The retention of nanoparticles from the fluid flow was carried out using a constant magnetic field of a class N35 neodymium magnet measuring 30 × 20 × 10 mm.

The experiment showed that within 5 minutes from the flow of the circulating model fluid, 100% MNPs settle in the area of application of the magnetic field (Fig. 7A). Thus, during the experiment, at a given flow rate of the model fluid, 10 mg of nanoparticles of magnetic material accumulated in the tube. The predominant accumulation of MNPs was observed at the ends of the permanent magnet. Such their behavior is understandable in comparison with the topography of the magnetic field of a permanent magnet (Fig. 7B).

Thus, the proposed group of inventions allows us to study the process of magnetically controlled delivery of MNPs in vitro, depending on the volumetric flow rate, salt composition and viscosity of the model fluid.

Claims (3)

1. A stand for studying the process of magnetically controlled delivery of nanoparticles into the vascular system, including a U-shaped tube, a single end of which, located between the magnet and the recorder, is connected to the storage tank, the output of the latter is connected through a pump and flow meter to one of the forked ends of the U-shaped tube forming a closed loop simulating the circulatory system, and the aforementioned bifurcated end of the U-shaped tube is also connected to a pressure sensor and a nanoparticle delivery element, which is a joint eggs. 2. A stand for studying the process of magnetically controlled delivery of nanoparticles to the vascular system, including a U-shaped tube, a single end of which, located between the magnet and the recorder, is connected to the storage tank, the output of the latter is connected through a pump and flow meter to one of the forked ends of the U-shaped tube forming a closed loop simulating the circulatory system, and the aforementioned bifurcated end of the tube is also connected to a pressure sensor, and an element of access is placed in the other bifurcated end of the U-shaped tube Application of nanoparticles representing an electromagnetic probe connected to a power source. 3. The stand according to claim 2, characterized in that the electromagnetic probe is made in the form of a solenoid with a length significantly exceeding its diameter.

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