CN110227195B - Method for removing virus and pathological cells in blood - Google Patents
Method for removing virus and pathological cells in blood Download PDFInfo
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- CN110227195B CN110227195B CN201910368818.1A CN201910368818A CN110227195B CN 110227195 B CN110227195 B CN 110227195B CN 201910368818 A CN201910368818 A CN 201910368818A CN 110227195 B CN110227195 B CN 110227195B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
- A61M1/3618—Magnetic separation
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
- A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
- A61M1/362—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits changing physical properties of target cells by binding them to added particles to facilitate their subsequent separation from other cells, e.g. immunoaffinity
Abstract
The invention relates to the field of blood, and provides a method for removing viruses and pathological cells in blood, which is used for improving the removal efficiency of the pathological cells or the viruses in the blood. The invention provides a method for removing viruses and pathological cells in blood, which comprises the following steps: combining blood with the magnetic composite particles, and fully contacting the blood with the magnetic composite particles to combine viruses or pathological cells in the blood with the magnetic composite particles; separating the magnetic composite particles combined with the virus or the pathological cells from the blood by using a magnetic field; the magnetic composite particles are modified nano ferroferric oxide, and the modified nano ferroferric oxide is prepared by compounding nano ferroferric oxide modified by silicon with biological macromolecular particles. Can separate the diseased cells or viruses from the blood with high efficiency and delay the migration and the diffusion of the viruses or the diseased cells in the body.
Description
Technical Field
The invention relates to the field of blood, in particular to a method for removing viruses and pathological cells in blood.
Background
Blood is the root of human life, and the invasion of some pathogenic substances to blood can directly threaten the life and health of human body. Therefore, it is of great importance to safely control blood products and to purify blood of patients with diseases. For viral infections, most of them can heal themselves within several weeks, some viral infections can remove viruses such as measles virus, hepatitis B virus and the like by the action of their own immune system after several months, but some viruses are difficult to remove after chronic disease infection, mainly including herpes virus, retrovirus, hepatitis c virus, echinovirus, polyoma virus and the like, and these difficult-to-remove viruses infect blood and further cause infection of blood cells, plasma and plasma derivatives, such as acquired immunodeficiency syndrome virus (HIV) is a causative agent of aids, which infects specific cells of the immune system and endangers the immune response of the infected individual. Therefore, the removal of viral particles, viral nucleic acid fragments and infected blood cells from blood products or human blood is of great importance to the safety of blood.
The treatment of the virus contained in the blood products at present mainly comprises two forms of inactivation and removal, wherein the inactivation method comprises a pasteurization method, an organic solvent combined surfactant (S/D) method, a dry heat inactivation method, a photochemical inactivation method and the like, and the methods have respective defects, such as that the pasteurization method is not ideal for inactivating some heat-resistant viruses, the S/D method cannot effectively inactivate non-lipid enveloped viruses, and the photochemical method has larger activity damage to some plasma proteins.
The composition of the polymer or organic reagent and the magnetic particles can introduce functional groups such as amino, carboxyl, sulfydryl, hydroxyl and the like on the surfaces of the particles, and then biological molecules such as enzyme, antibody, cell, nucleic acid, oligonucleotide and the like are fixed on the surfaces through covalent interaction, so that the composition is applied to the biological and medical fields such as cell sorting, enzyme immobilization, immunodetection, purification and separation of drug carriers and nucleic acid, targeted treatment research of tumors and the like.
The application aims to provide the magnetic composite particles which can improve the magnetic sorting effect, and further remove the virus or pathological cells in blood.
Disclosure of Invention
The invention provides a method for removing virus and diseased cells in blood, aiming at improving the removal efficiency of diseased cells or virus in blood.
In order to solve the technical problems, the technical scheme provided by the invention is as follows:
a method for removing viruses and diseased cells from blood, comprising: s1, combining blood with magnetic composite particles, and fully contacting the blood with the magnetic composite particles to combine viruses or pathological cells in the blood with the magnetic composite particles; s2, separating the magnetic composite particles combined with the viruses or the pathological cells from blood by using a magnetic field; the magnetic composite particles are modified nano ferroferric oxide, and the modified nano ferroferric oxide is formed by compounding nano ferroferric oxide modified by silicon with biological macromolecular particles.
The pathological cells or viruses are combined with the magnetic composite particles, so that the pathological cells or viruses are separated from the blood, the separation efficiency is high, and the influence on normal cells in the blood is small.
Can separate the diseased cells or viruses from the blood with high efficiency and delay the migration and the diffusion of the viruses or the diseased cells in the body.
Preferably, the method for modifying the nano ferroferric oxide by silicon comprises the following steps: s21, adding 40-50 parts by mass of nano ferroferric oxide into a hydrolysate of ethyl orthosilicate, and performing ultrasonic dispersion, wherein the hydrolysate of the ethyl orthosilicate comprises 5-10 parts by mass of ethyl orthosilicate, 500-600 parts by mass of deionized water, 400-500 parts by mass of ethanol and 1-2 parts by mass of concentrated ammonia water; s22, adding 3-5 parts by mass of octadecyl trimethyl ammonium bromide, uniformly stirring, adding 1-2 parts by mass of methyl orthosilicate, uniformly stirring, collecting a product, washing with a mixed solution of ethanol and water, drying, and roasting at 500-600 ℃ for 8 hours to obtain the modified nano ferroferric oxide. The magnetic nano ferroferric oxide is combined with silicon dioxide, so that the particle size of the nano ferroferric oxide is more uniform, the nano ferroferric oxide can be more fully mixed with cells, the sorting efficiency is improved, and the effect of removing viruses or pathological cells is better.
Preferably, the method for modifying the nano ferroferric oxide by silicon comprises the following steps: s21, adding 45-50 parts by mass of nano ferroferric oxide into a hydrolysate of ethyl orthosilicate, and performing ultrasonic dispersion, wherein the hydrolysate of the ethyl orthosilicate comprises 6-10 parts by mass of ethyl orthosilicate, 550-600 parts by mass of deionized water, 450-500 parts by mass of ethanol and 1.5-2 parts by mass of concentrated ammonia water; s22, adding 4-5 parts by mass of octadecyl trimethyl ammonium bromide, uniformly stirring, adding 1.5-2 parts by mass of methyl orthosilicate, uniformly stirring, collecting a product, washing with a mixed solution of ethanol and water, drying, and roasting at 550-600 ℃ for 8 hours to obtain the modified nano ferroferric oxide.
Preferably, the method for modifying the nano ferroferric oxide by silicon comprises the following steps: s21, adding 45 parts by mass of nano ferroferric oxide into a hydrolysate of ethyl orthosilicate, and performing ultrasonic dispersion, wherein the hydrolysate of the ethyl orthosilicate comprises 6 parts by mass of ethyl orthosilicate, 550 parts by mass of deionized water, 450 parts by mass of ethanol and 1.5 parts by mass of concentrated ammonia water; s22, adding 4 parts by mass of octadecyl trimethyl ammonium bromide, uniformly stirring, adding 1.5 parts by mass of methyl orthosilicate, uniformly stirring, collecting a product, washing with a mixed solution of ethanol and water, drying, and roasting at 550 ℃ for 8 hours to obtain the modified nano ferroferric oxide.
Preferably, the nano ferroferric oxide is modified by nano layered double hydroxide before being modified by silicon, and the modification method comprises the following steps: dispersing 30-50 parts by mass of nano ferroferric oxide in 800-1000 parts by mass of deionized water, adding 5-10 parts by mass of nano layer-transition double hydroxide, performing ultrasonic dispersion, and drying to obtain nano layered hydroxide modified nano ferroferric oxide. The nano ferroferric oxide modified by the nano layered double hydroxide can have better biocompatibility, can be quickly combined with cells, cannot affect normal cells, and avoids the nano particles from damaging certain normal cells.
Preferably, the method for preparing the nano-layered hydroxide comprises the following steps: taking 1-3 parts by mass of magnesium nitrate, 0.5-2 parts by mass of aluminum nitrate and 40-60 parts by mass of deionized water; dissolving nitric acid and aluminum nitrate in deionized water, adding the solution into a sodium hydroxide solution which is stirred vigorously, carrying out hydrothermal reaction, drying a reaction product, and crushing to obtain the nano layered hydroxide. The nano layered hydroxide prepared by the method can better improve the cell compatibility of the nano ferroferric oxide.
Preferably, the sodium hydroxide solution contains 0.5-0.8 parts by mass of sodium hydroxide and 40-60 parts by mass of deionized water. Sodium hydroxide provides an alkaline environment.
Preferably, the conditions of the hydrothermal reaction are: reacting for 24-36 h at 80-120 ℃. The particle size of the layered nano hydroxide prepared by the hydrothermal reaction is more consistent.
Preferably, the nano layered hydroxide is used for modifying nano ferroferric oxide after being compounded with nano titanium dioxide, and the compounding method of the nano layered hydroxide and the nano titanium dioxide comprises the following steps: dispersing 10-20 parts by mass of nano layered hydroxide into 300-400 parts by mass of deionized water, adding 1-2 parts by mass of nano titanium dioxide, and performing ultrasonic dispersion uniformly to obtain nano layered hydroxide compounded with the nano titanium dioxide. The titanium modified nano layered hydroxide can further improve the biocompatibility of nano ferroferric oxide, and meanwhile, the nano ferroferric oxide is combined with biological macromolecules more tightly, so that the nano ferroferric oxide can be effectively combined with pathological cells, and the removal effect of the pathological cells is improved.
Preferably, the nano titanium dioxide is modified nano titanium dioxide, and the preparation method of the modified nano titanium dioxide comprises the following steps: adding ethyl titanate and molybdenum acetylacetonate into cyclohexane to obtain precursor solution, wherein the concentration of the ethyl titanate is 0.4mol/L, and the acetylacetoneThe concentration of the ketone molybdenum is 0.012 mol/L; the precursor solution is subjected to external oxygen-assisted shearing atomization burner at a feeding speed of 5ml/min to form fine atomized liquid drops, and the micro liquid drops are in H2/O2The mixed gas is converted into nano powder under the assistance of diffusion flame, the shearing pressure at the shearing atomizing nozzle is 0.28MPa, and H in the diffusion flame2The flow rate of (2) is 240L/h, O2The flow rate of (2) is 950L/h; and calcining the nano powder for 8 hours in an air atmosphere at the calcining temperature of 400 ℃, and cooling to obtain the modified titanium dioxide. The modified titanium dioxide, especially the modified titanium dioxide prepared by adopting the spray combustion technology, can better improve the biocompatibility of the nano ferroferric oxide.
Compared with the prior art, the invention has the beneficial effects that: the pathological cells or viruses can be efficiently separated from blood, and the migration and diffusion of the viruses or the pathological cells in vivo can be delayed; the biocompatibility of the nano ferroferric oxide can be further improved, and meanwhile, the nano ferroferric oxide is combined with biological macromolecules more tightly, so that the nano ferroferric oxide can be effectively combined with pathological cells, and the removal effect of the pathological cells is improved.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Example 1
A method for removing viruses and diseased cells from blood, comprising: s1, combining blood with magnetic composite particles, and fully contacting the blood with the magnetic composite particles to combine viruses or pathological cells in the blood with the magnetic composite particles; s2, separating the magnetic composite particles combined with the viruses or the pathological cells from blood by using a magnetic field; the magnetic composite particles are modified nano ferroferric oxide, and the modified nano ferroferric oxide is formed by compounding nano ferroferric oxide modified by silicon with biological macromolecular particles. The method for modifying the nano ferroferric oxide by silicon comprises the following steps: s21, adding 45 parts by mass of nano ferroferric oxide into a hydrolysate of ethyl orthosilicate, and performing ultrasonic dispersion, wherein the hydrolysate of ethyl orthosilicate comprises 6 parts by mass of ethyl orthosilicate, 550 parts by mass of deionized water and ethyl acetate450 parts of alcohol and 1.5 parts of concentrated ammonia water; s22, adding 4 parts by mass of octadecyl trimethyl ammonium bromide, uniformly stirring, adding 1.5 parts by mass of methyl orthosilicate, uniformly stirring, collecting a product, washing with a mixed solution of ethanol and water, drying, and roasting at 550 ℃ for 8 hours to obtain the modified nano ferroferric oxide. The nano ferroferric oxide is modified by nano layered double hydroxide before being modified by silicon, and the modification method comprises the following steps: dispersing 40 parts by mass of nano ferroferric oxide in 900 parts by mass of deionized water, adding 7 parts by mass of nano layer-transition double hydroxide, performing ultrasonic dispersion, and drying to obtain nano layered hydroxide modified nano ferroferric oxide. The preparation method of the nano layered hydroxide comprises the following steps: taking 2 parts by mass of magnesium nitrate, 1 part by mass of aluminum nitrate and 50 parts by mass of deionized water; dissolving nitric acid and aluminum nitrate in deionized water, adding the solution into a sodium hydroxide solution which is stirred vigorously, carrying out hydrothermal reaction, drying a reaction product, and crushing to obtain the nano layered hydroxide. The sodium hydroxide solution contains 0.6 part by mass of sodium hydroxide and 50 parts by mass of deionized water. The conditions of the hydrothermal reaction are as follows: the reaction was carried out at 100 ℃ for 27 h. The nano layered hydroxide is compounded with nano titanium dioxide and then used for modifying nano ferroferric oxide, and the compounding method of the nano layered hydroxide and the nano titanium dioxide comprises the following steps: dispersing 15 parts by mass of nano layered hydroxide into 350 parts by mass of deionized water, adding 1.5 parts by mass of nano titanium dioxide, and performing ultrasonic dispersion uniformly to obtain nano layered hydroxide compounded with the nano titanium dioxide. The nano titanium dioxide is modified nano titanium dioxide, and the preparation method of the modified nano titanium dioxide comprises the following steps: adding ethyl titanate and molybdenum acetylacetonate into cyclohexane to obtain a precursor solution, wherein the concentration of the ethyl titanate is 0.4mol/L, and the concentration of the molybdenum acetylacetonate is 0.012 mol/L; the precursor solution is subjected to external oxygen-assisted shearing atomization burner at a feeding speed of 5ml/min to form fine atomized liquid drops, and the micro liquid drops are in H2/O2The mixed gas is converted into nano powder under the assistance of diffusion flame, the shearing pressure at the shearing atomizing nozzle is 0.28MPa, and H in the diffusion flame2The flow rate of (2) is 240L/h, O2At a flow rate of 950L/h; and calcining the nano powder for 8 hours in an air atmosphere at the calcining temperature of 400 ℃, and cooling to obtain the modified titanium dioxide.
Can separate the diseased cells or viruses from the blood with high efficiency and delay the migration and the diffusion of the viruses or the diseased cells in the body. The magnetic nano ferroferric oxide is combined with silicon dioxide, so that the particle size of the nano ferroferric oxide is more uniform, the nano ferroferric oxide can be more fully mixed with cells, the sorting efficiency is improved, and the effect of removing viruses or pathological cells is better. The nano ferroferric oxide modified by the nano layered double hydroxide can have better biocompatibility, can be quickly combined with cells, cannot affect normal cells, and avoids the nano particles from damaging certain normal cells. The nano layered hydroxide prepared by the method can better improve the cell compatibility of the nano ferroferric oxide. Sodium hydroxide provides an alkaline environment. The particle size of the layered nano hydroxide prepared by the hydrothermal reaction is more consistent. The titanium modified nano layered hydroxide can further improve the biocompatibility of nano ferroferric oxide, and meanwhile, the nano ferroferric oxide is combined with biological macromolecules more tightly, so that the nano ferroferric oxide can be effectively combined with pathological cells, and the removal effect of the pathological cells is improved. The modified titanium dioxide, especially the modified titanium dioxide prepared by adopting the spray combustion technology, can better improve the biocompatibility of the nano ferroferric oxide.
Example 2
A method for removing viruses and diseased cells from blood, comprising: s1, combining blood with magnetic composite particles, and fully contacting the blood with the magnetic composite particles to combine viruses or pathological cells in the blood with the magnetic composite particles; s2, separating the magnetic composite particles combined with the viruses or the pathological cells from blood by using a magnetic field; the magnetic composite particles are modified nano ferroferric oxide, and the modified nano ferroferric oxide is formed by compounding nano ferroferric oxide modified by silicon with biological macromolecular particles. The method for modifying the nano ferroferric oxide by silicon comprises the following steps: s21, adding 40 parts by mass of nano ferroferric oxide into water of ethyl orthosilicatePerforming ultrasonic dispersion on hydrolysate, wherein the hydrolysate of the ethyl orthosilicate comprises 5 parts by mass of ethyl orthosilicate, 500 parts by mass of deionized water, 400 parts by mass of ethanol and 1 part by mass of concentrated ammonia water; s22, adding 3 parts by mass of octadecyl trimethyl ammonium bromide, uniformly stirring, adding 1 part by mass of methyl orthosilicate, uniformly stirring, collecting a product, washing with a mixed solution of ethanol and water, drying, and roasting at 500 ℃ for 8 hours to obtain the modified nano ferroferric oxide. The nano ferroferric oxide is modified by nano layered double hydroxide before being modified by silicon, and the modification method comprises the following steps: dispersing 30 parts by mass of nano ferroferric oxide in 800 parts by mass of deionized water, adding 5 parts by mass of nano-layer-transition double hydroxide, performing ultrasonic dispersion, and drying to obtain nano-layered hydroxide modified nano ferroferric oxide. The preparation method of the nano layered hydroxide comprises the following steps: taking 1 part by mass of magnesium nitrate, 0.5 part by mass of aluminum nitrate and 40 parts by mass of deionized water; dissolving nitric acid and aluminum nitrate in deionized water, adding the solution into a sodium hydroxide solution which is stirred vigorously, carrying out hydrothermal reaction, drying a reaction product, and crushing to obtain the nano layered hydroxide. The sodium hydroxide solution contains 0.5 part by mass of sodium hydroxide and 40 parts by mass of deionized water. The conditions of the hydrothermal reaction are as follows: the reaction was carried out at 80 ℃ for 24 h. The nano layered hydroxide is compounded with nano titanium dioxide and then used for modifying nano ferroferric oxide, and the compounding method of the nano layered hydroxide and the nano titanium dioxide comprises the following steps: dispersing 10 parts by mass of nano layered hydroxide into 300 parts by mass of deionized water, adding 1 part by mass of nano titanium dioxide, and uniformly dispersing by ultrasonic to obtain the nano layered hydroxide compounded with the nano titanium dioxide. The nano titanium dioxide is modified nano titanium dioxide, and the preparation method of the modified nano titanium dioxide comprises the following steps: adding ethyl titanate and molybdenum acetylacetonate into cyclohexane to obtain a precursor solution, wherein the concentration of the ethyl titanate is 0.4mol/L, and the concentration of the molybdenum acetylacetonate is 0.012 mol/L; the precursor solution is subjected to external oxygen-assisted shearing atomization burner at a feeding speed of 5ml/min to form fine atomized liquid drops, and the micro liquid drops are in H2/O2Down-conversion into nanopowders with the aid of diffusion flame, shear atomizing nozzleThe shear pressure at the opening is 0.28MPa, and H in the diffusion flame2The flow rate of (2) is 240L/h, O2The flow rate of (2) is 950L/h; and calcining the nano powder for 8 hours in an air atmosphere at the calcining temperature of 400 ℃, and cooling to obtain the modified titanium dioxide.
Example 3
A method for removing viruses and diseased cells from blood, comprising: s1, combining blood with magnetic composite particles, and fully contacting the blood with the magnetic composite particles to combine viruses or pathological cells in the blood with the magnetic composite particles; s2, separating the magnetic composite particles combined with the viruses or the pathological cells from blood by using a magnetic field; the magnetic composite particles are modified nano ferroferric oxide, and the modified nano ferroferric oxide is formed by compounding nano ferroferric oxide modified by silicon with biological macromolecular particles. The method for modifying the nano ferroferric oxide by silicon comprises the following steps: s21, adding 40-50 parts by mass of nano ferroferric oxide into a hydrolysate of ethyl orthosilicate, and performing ultrasonic dispersion, wherein the hydrolysate of the ethyl orthosilicate comprises 10 parts by mass of ethyl orthosilicate, 600 parts by mass of deionized water, 500 parts by mass of ethanol and 2 parts by mass of concentrated ammonia water; s22, adding 5 parts by mass of octadecyl trimethyl ammonium bromide, uniformly stirring, adding 2 parts by mass of methyl orthosilicate, uniformly stirring, collecting a product, washing with a mixed solution of ethanol and water, drying, and roasting at 600 ℃ for 8 hours to obtain the modified nano ferroferric oxide. The nano ferroferric oxide is modified by nano layered double hydroxide before being modified by silicon, and the modification method comprises the following steps: dispersing 50 parts by mass of nano ferroferric oxide in 1000 parts by mass of deionized water, adding 10 parts by mass of nano layer-transition double hydroxide, performing ultrasonic dispersion, and drying to obtain nano layered hydroxide modified nano ferroferric oxide. The preparation method of the nano layered hydroxide comprises the following steps: taking 3 parts by mass of magnesium nitrate, 2 parts by mass of aluminum nitrate and 60 parts by mass of deionized water; dissolving nitric acid and aluminum nitrate in deionized water, adding the solution into a sodium hydroxide solution which is stirred vigorously, carrying out hydrothermal reaction, drying a reaction product, and crushing to obtain the nano layered hydroxide. The sodium hydroxide solution contains 0.8 part by mass of sodium hydroxide and 40 parts by mass of deionized water. The water heatThe reaction conditions were: the reaction was carried out at 120 ℃ for 36 h. The nano layered hydroxide is compounded with nano titanium dioxide and then used for modifying nano ferroferric oxide, and the compounding method of the nano layered hydroxide and the nano titanium dioxide comprises the following steps: dispersing 20 parts by mass of nano layered hydroxide into 400 parts by mass of deionized water, adding 2 parts by mass of nano titanium dioxide, and uniformly dispersing by ultrasonic to obtain the nano layered hydroxide compounded with the nano titanium dioxide. The nano titanium dioxide is modified nano titanium dioxide, and the preparation method of the modified nano titanium dioxide comprises the following steps: adding ethyl titanate and molybdenum acetylacetonate into cyclohexane to obtain a precursor solution, wherein the concentration of the ethyl titanate is 0.4mol/L, and the concentration of the molybdenum acetylacetonate is 0.012 mol/L; the precursor solution is subjected to external oxygen-assisted shearing atomization burner at a feeding speed of 5ml/min to form fine atomized liquid drops, and the micro liquid drops are in H2/O2The mixed gas is converted into nano powder under the assistance of diffusion flame, the shearing pressure at the shearing atomizing nozzle is 0.28MPa, and H in the diffusion flame2The flow rate of (2) is 240L/h, O2The flow rate of (2) is 950L/h; and calcining the nano powder for 8 hours in an air atmosphere at the calcining temperature of 400 ℃, and cooling to obtain the modified titanium dioxide.
Example 4
Example 4 is different from example 1 in that the nano ferroferric oxide is not modified by nano layered hydroxide.
Example 5
Example 5 differs from example 1 in that the nano-layered hydroxide is not modified with titanium dioxide.
Example 6
Example 6 differs from example 1 in that the nano titanium dioxide is not modified.
Comparative example 1
Comparative example 1 is different from example 1 in that the magnetic composite particles are nano ferroferric oxide.
Examples of the experiments
(one) incubation of cells with immunomagnetic microspheres: adding 1mL of cultured MCF-7 cells into a 50mL human blood sample, adding 50 mu L of immunomagnetic microspheres, standing and incubating at room temperature for 1h, and counting the cells.
(II) magnetic separation: the incubated sample is placed on a magnetic separation rack for magnetic separation to remove diseased cells, and the cells obtained by separation are counted.
TABLE 1 Capture Effect of magnetic separation on cancer cells
| Capture rate | |
| Example 1 | 96.53±3.94% |
| Example 2 | 93.42±2.45% |
| Example 3 | 94.29±4.73% |
| Example 4 | 74.56±7.94% |
| Example 5 | 88.93±6.73% |
| Example 6 | 81.53±8.34% |
| Comparative example 1 | 46.21±9.06% |
As can be seen from table 1, the nano ferroferric oxide modified by the nano layered hydroxide modified by the modified titanium dioxide can effectively trap cancer cells.
The capture rate of the silicon modified nano ferroferric oxide compounded with the biomacromolecules in the examples 1-3 modified by the modified titanium dioxide nano layered hydroxide is obviously higher than that of the examples 4-6 and the comparative example, which shows that the capture rate of the multiple modified nano ferroferric oxide to cancer cells is higher, and the capture effect of the magnetic particles with the optimized formula, namely the example 1, to the cancer cells is stronger.
In comparative example 1, the effect is poor when only nano ferroferric oxide is used for capturing, which shows that the effect of effectively capturing cancer cells cannot be generated when only magnetic particles are used.
The above detailed description is specific to possible embodiments of the present invention, and the above embodiments are not intended to limit the scope of the present invention, and all equivalent implementations or modifications that do not depart from the scope of the present invention should be included in the present claims.
Claims (6)
1. A method for removing viruses and diseased cells from blood, comprising: s1, combining blood with magnetic composite particles, and fully contacting the blood with the magnetic composite particles to combine viruses or pathological cells in the blood with the magnetic composite particles; s2, separating the magnetic composite particles combined with the viruses or the pathological cells from blood by using a magnetic field; the magnetic composite particles are modified nano ferroferric oxide, and the modified nano ferroferric oxide is prepared by compounding nano ferroferric oxide modified by silicon with biological macromolecular particles;
the method for modifying the nano ferroferric oxide by silicon comprises the following steps: s21, adding 40-50 parts by mass of nano ferroferric oxide into a hydrolysate of ethyl orthosilicate, and performing ultrasonic dispersion, wherein the hydrolysate of the ethyl orthosilicate comprises 5-10 parts by mass of ethyl orthosilicate, 500-600 parts by mass of deionized water, 400-500 parts by mass of ethanol and 1-2 parts by mass of concentrated ammonia water; s22, adding 3-5 parts by mass of octadecyl trimethyl ammonium bromide, uniformly stirring, adding 1-2 parts by mass of methyl orthosilicate, uniformly stirring, collecting a product, washing with a mixed solution of ethanol and water, drying, and roasting at 500-600 ℃ for 8 hours to obtain modified nano ferroferric oxide;
the nano ferroferric oxide is modified by nano layered double hydroxide before being modified by silicon, and the modification method comprises the following steps: dispersing 30-50 parts by mass of nano ferroferric oxide in 800-1000 parts by mass of deionized water, adding 5-10 parts by mass of nano layer-transition double hydroxide, performing ultrasonic dispersion, and drying to obtain nano layered hydroxide modified nano ferroferric oxide;
the preparation method of the nano layered hydroxide comprises the following steps: taking 1-3 parts by mass of magnesium nitrate, 0.5-2 parts by mass of aluminum nitrate and 40-60 parts by mass of deionized water; dissolving nitric acid and aluminum nitrate in deionized water, adding the solution into a sodium hydroxide solution which is stirred vigorously, carrying out hydrothermal reaction, drying a reaction product, and crushing to obtain a nano layered hydroxide;
the nano layered hydroxide is compounded with nano titanium dioxide and then used for modifying nano ferroferric oxide, and the compounding method of the nano layered hydroxide and the nano titanium dioxide comprises the following steps: dispersing 10-20 parts by mass of nano layered hydroxide into 300-400 parts by mass of deionized water, adding 1-2 parts by mass of nano titanium dioxide, and performing ultrasonic dispersion uniformly to obtain nano layered hydroxide compounded with the nano titanium dioxide.
2. The method for removing viruses and diseased cells in blood according to claim 1, wherein the method for modifying the nano ferroferric oxide by silicon comprises the following steps: s21, adding 45-50 parts by mass of nano ferroferric oxide into a hydrolysate of ethyl orthosilicate, and performing ultrasonic dispersion, wherein the hydrolysate of the ethyl orthosilicate comprises 6-10 parts by mass of ethyl orthosilicate, 550-600 parts by mass of deionized water, 450-500 parts by mass of ethanol and 1.5-2 parts by mass of concentrated ammonia water; s22, adding 4-5 parts by mass of octadecyl trimethyl ammonium bromide, uniformly stirring, adding 1.5-2 parts by mass of methyl orthosilicate, uniformly stirring, collecting a product, washing with a mixed solution of ethanol and water, drying, and roasting at 550-600 ℃ for 8 hours to obtain the modified nano ferroferric oxide.
3. The method for removing viruses and diseased cells in blood according to claim 2, wherein the method for modifying the nano ferroferric oxide by silicon comprises the following steps: s21, adding 45 parts by mass of nano ferroferric oxide into a hydrolysate of ethyl orthosilicate, and performing ultrasonic dispersion, wherein the hydrolysate of the ethyl orthosilicate comprises 6 parts by mass of ethyl orthosilicate, 550 parts by mass of deionized water, 450 parts by mass of ethanol and 1.5 parts by mass of concentrated ammonia water; s22, adding 4 parts by mass of octadecyl trimethyl ammonium bromide, uniformly stirring, adding 1.5 parts by mass of methyl orthosilicate, uniformly stirring, collecting a product, washing with a mixed solution of ethanol and water, drying, and roasting at 550 ℃ for 8 hours to obtain the modified nano ferroferric oxide.
4. The method of claim 1, wherein the sodium hydroxide solution comprises 0.5 to 0.8 parts by mass of sodium hydroxide and 40 to 60 parts by mass of deionized water.
5. The method according to claim 1, wherein the hydrothermal reaction is carried out under the following conditions: reacting for 24-36 h at 80-120 ℃.
6. The method for removing viruses and diseased cells in blood according to claim 1, wherein the nano titanium dioxide is modified nano titanium dioxide, and the method for preparing the modified nano titanium dioxide comprises: adding ethyl titanate and molybdenum acetylacetonate into cyclohexane to obtain a precursor solution, wherein the concentration of the ethyl titanate is 0.4mol/L, and the concentration of the molybdenum acetylacetonate is 0.012 mol/L; the precursor solution is subjected to external oxygen-assisted shearing atomization burner at a feeding speed of 5ml/min to form fine atomized liquid drops, and the micro liquid drops are in H2/O2The mixed gas is converted into nano powder under the assistance of diffusion flame, the shearing pressure at the shearing atomizing nozzle is 0.28MPa, and H in the diffusion flame2The flow rate of (2) is 240L/h, O2The flow rate of (2) is 950L/h; and calcining the nano powder for 8 hours in an air atmosphere at the calcining temperature of 400 ℃, and cooling to obtain the modified titanium dioxide.
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Address after: Building F2, 39 Ruihe Road, Huangpu District, Guangzhou City, Guangdong Province 510000 Applicant after: Lancy Purcell Biotechnology (Guangzhou) Co.,Ltd. Address before: 510000 Room D09, 4th Floor, 131 Airport Road, Baiyun District, Guangzhou City, Guangdong Province Applicant before: Lancy Purcell Biotechnology (Guangzhou) Co.,Ltd. |
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Denomination of invention: A method for clearing viruses and diseased cells in the blood Effective date of registration: 20230629 Granted publication date: 20210928 Pledgee: Hunan Pingjiang Rural Commercial Bank Co.,Ltd. Pledgor: Chen Zhongping|Lancy Purcell Biotechnology (Guangzhou) Co.,Ltd. Registration number: Y2023980046730 |