CN114574434A - Engineered magnetic control cell assembly and preparation method and application thereof - Google Patents

Engineered magnetic control cell assembly and preparation method and application thereof Download PDF

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CN114574434A
CN114574434A CN202210167246.2A CN202210167246A CN114574434A CN 114574434 A CN114574434 A CN 114574434A CN 202210167246 A CN202210167246 A CN 202210167246A CN 114574434 A CN114574434 A CN 114574434A
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magnetic
antioxidant
cells
cell assembly
assembly
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成昱
邓翠君
李珍光
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Tongji University
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Tongji University
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Abstract

The invention relates to an engineered magnetic control cell assembly and a preparation method and application thereof. The magnetic cells can form three-dimensional cell assemblies with different shapes under the control of the variable-frequency rotating magnetic field. The cell assembly can enhance the mechanical transduction in and among cells under the action of micro-magnetic force, has obvious in-vivo osteoarthritis treatment effect, and can achieve the effect of integrating anti-inflammation and tissue repair. Therefore, the engineered magnetic control cell assembly has wide application prospect in the field of inflammatory disease tissue repair and treatment.

Description

Engineered magnetic control cell assembly and preparation method and application thereof
Technical Field
The invention relates to the field, in particular to an engineered magnetic control cell assembly and a preparation method and application thereof.
Background
The assembly of cells is actually a manual assembly life. Since most scientists define life as a cell that can survive, grow and replicate independently of other cells. With the rapid development of cell molecular biology, human research on stem cells has been achieved with tremendous success. Stem cell research subjects include embryonic stem cells and adult stem cells. The stem cells with strong differentiation potential provide abundant cell sources for the research of cell assembly.
On the other hand, with the development of physical technology and micro-fabrication technology, some enabling technologies for precisely manipulating micro-and even nano-particles and micro-droplets, such as optical tweezers, Laser Guided Direct Writing (LGDW), micro-droplet jetting, micro-pen, dip-pen nanotechnology, etc., have been developed. These techniques use physical effects such as light, electricity, heat, and piezoelectricity to manipulate particles for assembly in three dimensions. Some of these enabling techniques have been used to assemble cells because of less physiological damage to the cells.
It can be seen that the conventional cell assembly preparation method comprises: spontaneous cell aggregation, suspension culture, 3D printing scaffold culture, overlay liquid culture, spin cell culture, porous microsphere carriers, and the like, and a large number of cell assemblies have been used in the field of tissue repair. However, the above methods require relatively long time for preparing 3D cell assemblies, are inefficient, have uncontrollable dimensions, and have poor disease applicability.
Therefore, in order to achieve the goal of rapidly preparing a 3D cell assembly with controllable morphology and disease universality, it is necessary to develop a new efficient and controllable cell assembly strategy.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide an engineering magnetic control cell assembly which is controllable in size height, adjustable in appearance, short in preparation process time, simple in process, controllable in condition and low in cost, and a preparation method and application thereof.
The purpose of the invention can be realized by the following technical scheme:
an engineered magnetically controlled cell assembly comprising cells and antioxidant magnetic nanomaterials (MFNPs) distributed within the cells.
Further, the cells include, but are not limited to, stem cells, liver cells, cartilage cells, osteoblasts, or cardiac muscle cells, among other somatic cells.
Furthermore, the antioxidant magnetic nano material is a magnetic induction substance modified by an antioxidant substance. The antioxidant magnetic nano material has a wide function of eliminating free radicals, including superoxide anion free radicals, hydroxyl free radicals, hydrogen peroxide, nitrogen free radicals and the like.
Further, the magnetic induction substances include but are not limited to magnetic nano materials such as ferroferric oxide, cobalt ferrite, nickel ferrite or neodymium iron boron;
the antioxidant substance includes, but is not limited to, an antioxidant material having a radical scavenging activity, such as melanin, lignin, ferulic acid, tannic acid, or N-acetyl-L-cysteine.
A method of preparing an engineered magnetic controlled cell assembly as described above, comprising the steps of:
(1) preparing an antioxidant magnetic nano material: preparing an antioxidant magnetic nanomaterial by taking an antioxidant substance as a ligand by adopting a typical method for preparing the magnetic nanomaterial such as a coprecipitation method or a hydrothermal method; such as: under the inert gas atmosphere, dissolving ferric salt and ferrous salt in deionized water, then adding an antioxidant, adding inorganic alkali liquor after mixing uniformly to adjust the pH of the solution to be more than 10, heating for reaction to obtain an antioxidant magnetic nano material, and fully washing and purifying the antioxidant magnetic nano material by using deionized water through magnetic separation;
(2) co-culturing the antioxidant magnetic nano material and cells: preparing a dispersion solution from an antioxidant magnetic nano material by using a cell complete culture medium, adding the dispersion solution into a culture dish containing cells, placing the culture dish into an incubator for continuous culture, and washing an obtained cell compound by using a PBS phosphate buffer solution to obtain magnetic cells;
(3) and (3) placing the magnetic cells in a magnetic field, and acting for a period of time to obtain the engineered magnetic control cell assembly. The stem cells are regulated and differentiated in specific directions including but not limited to chondroblasts, osteogenesis and the like by generating micro-magnetic force through magnetic induction media in the magnetic cells.
Further, the concentration of the antioxidant substance is 0.5-1mg/mL, the concentration of ferric salt is 40-100mM, and the molar ratio of ferric salt to ferrous salt is 2: 1; the temperature of the temperature rise reaction is 70-85 ℃, and the time is 1-3 h.
Further, the inorganic alkali solution comprises ammonia water, potassium hydroxide or sodium hydroxide, and preferably ammonia water.
Further, the concentration of the antioxidant magnetic nano material in the dispersion liquid is 10-500 mu g/mL; the temperature in the incubator is 35-38 ℃, preferably 36.5-37.3 ℃, and the volume concentration of carbon dioxide is 0-6%, preferably 4-6%; the continuous culture time is 12-72 h. It is not difficult to presume that the content of the magnetic substance in the magnetic cell is 10 to 500. mu.g/mL.
Further, the intensity of the magnetic field is 0-260mT, the frequency is 0-30Hz, preferably 0-20 Hz; the action time is 0-60min, preferably 0-10 min. Can assist cells to resist the oxidative stress physiopathological microenvironment by adjusting the strength of the micro-magnetic force.
Further, the magnetic field is a rotating magnetic field or a pulse magnetic field.
Further, the cell assembly includes, but is not limited to, a two-dimensional sheet assembly and a three-dimensional spherical assembly.
The application of the engineered magnetic control cell assembly is used for preparing an implant material with anti-inflammatory and tissue repair functions.
Compared with the prior art, the invention has the following advantages:
(1) the invention endows the cells with oxidation resistance and magnetic control property by the oxidation-resistant magnetic nano material for the first time, and realizes the rapid cell ball preparation technology under the control of the variable-frequency rotating magnetic field for the first time, and the obtained cell assembly has the advantages of highly controllable size, adjustable appearance, short preparation process time, simple process, controllable conditions, low cost and the like;
(2) meanwhile, the cell assembly can respond to a magnetic field, generates mild magnetic mechanical force stimulation inside and among cells, enhances mechanical transduction inside and among the cells, promotes the directional differentiation of the stem cells, and can adjust the magnitude of micro magnetic force generated by a magnetic induction medium by adjusting parameters of the magnetic field so as to adjust and control the differentiation direction of the stem cells;
(3) in addition, the antioxidant magnetic nano material of the invention endows stem cells with antioxidant performance, and can help the stem cells to be prevented from oxidative stress damage at in-vivo inflammation parts. Under the control of a rotating magnetic field, magnetic stem cells are firstly found to form a cell assembly in a joint cavity and promote the cell assembly to be differentiated into chondroblasts, have remarkable osteoarthritis treatment and cartilage repair biological functional characteristics, and can achieve the effect of integration of anti-inflammation and cartilage repair;
(4) therefore, the engineered magnetic control cell assembly has wide application prospect in the field of inflammatory disease tissue repair treatment.
Drawings
FIG. 1 is a graph of MFNP performance characterization from example 1;
FIG. 2 shows the biological activity and ROS-scavenging effect of MFNP in example 1;
FIG. 3 shows that the MFNPs and micromagnetic forces of example 1 assist in enhancing the anti-inflammatory ability of stem cells;
FIG. 4 is a diagram of stem cell assemblies obtained by different magnetic field parameters in example 1
FIG. 5 is the assembly of stem cells in the joint cavity by micromagnetic manipulation in example 1;
FIG. 6 is the micro-magnetic force regulating stem cell chondrogenic differentiation of example 1;
FIG. 7 is a graph showing the effect of the stem cell assembly on the in vivo treatment of arthritis in example 1.
Detailed Description
The invention is described in detail below with reference to the figures and specific embodiments. The present embodiment is implemented on the premise of the technical solution of the present invention, and a detailed implementation manner and a specific operation process are given, but the protection scope of the present invention is not limited to the following embodiments.
The specific embodiment of the invention is briefly described as follows:
1. the preparation and performance characterization of the antioxidant magnetic nano material are as follows: taking the synthesis of antioxidant magnetic nano ferroferric oxide as an example
(1) Preparing an antioxidant magnetic nano material: under the inert gas atmosphere, ferric salt and ferrous salt are dissolved in deionized water, then antioxidant substances are added, inorganic alkali liquor is added after uniform mixing to adjust the pH of the solution to be more than 10, the temperature is raised to 80 ℃ for reaction for at least 1h, and the obtained antioxidant magnetic nano material is fully washed and purified by using the deionized water through magnetic separation.
(2) The physicochemical property characterization of the antioxidant magnetic nano material: characterizing the appearance and size of the antioxidant magnetic nanomaterial by a Transmission Electron Microscope (TEM); the saturation magnetization of the oxidation-resistant magnetic nanomaterial was characterized by a Vibrating Sample Magnetometer (VSM). The activity of the antioxidant magnetic nano material for eliminating free radicals is tested by using 1, 1-diphenyl-2-trinitrophenylhydrazine free radical (DPPH).
(3) The free radical scavenging effect of the antioxidant magnetic nano material is characterized in that: ROS and RNS eliminating efficiency of MFNP is characterized by an electron paramagnetic resonance spectrometer (EPR), a fluorescence imaging system, a fluorescence spectrometer, an ultraviolet visible spectrophotometer and a response free radical detection kit (DPPH, ABTS, MDA, PTIO and the like).
(4) And (3) biological activity characterization of the antioxidant magnetic nano material: the MFNP is loaded into the cell by utilizing the active uptake function of the cell, the feeding amount and the feeding time of different MFNPs are changed, the cells with different MFNP contents are obtained, and the influence of the antioxidant magnetic nano material on the cell activity of the cell is researched by utilizing reagents such as CCK8, MTT, Live/Dead, AM/PI and the like.
2. Research on life activity and tissue repair performance of cell assembly
2.1 Life viability Studies of cell assemblies
Assembling cells into a 3D assembly by using a rotating magnetic field, and observing the morphology of the cell assembly by using an optical microscope, an SEM (scanning electron microscope), a laser confocal microscope (CLSM) and the like; performing real-time dynamic observation on stem cells through a living cell workstation, and evaluating the behaviors of cell assemblies such as adhesion and proliferation by using reagents such as CCK 8; whether the MFNP can cause autophagy, apoptosis or necrosis of a cell assembly is characterized by adopting an autophagy and apoptosis necrosis kit; and (3) representing whether the antioxidant magnetic nano material can induce the cell assembly to generate Iron death by using MDA (multidose Measure) kits, Iron Assay kits, BODIPY C-11 kits and the like.
2.2 study of cell Assembly differentiation Capacity
Placing the assembly in an ultra-low adhesion culture dish for continuous culture, applying micro-magnetic stimulation every day, and researching the expression of genes related to proliferation, adhesion and differentiation of the cell assembly by the micro-magnetic stimulation; characterizing the acid protein differentiated from the osteogenic cartilage by using alisnew blue, and carrying out qualitative and quantitative analysis on markers of the osteogenic early stage and the osteogenic final stage by using alkaline phosphatase and alizarin red dye liquor; the adipogenic differentiation capacity of the cell assemblies was characterized by oil red staining.
2.3 in vivo therapeutic efficacy Studies of cell assemblies
An SD rat osteoarthritis model was constructed by a surgical modeling method, and magnetic cells were injected to the joint cavity site while micro-magnetic treatment was given every day. The treatment is carried out once a day for 2 weeks, the materials are respectively taken 2 weeks and 6 weeks after the treatment is finished, the treatment effect is evaluated by living body imaging, tissue section and the like, and then the repair effect is evaluated by the international association for cartilage repair (ICRS) scoring standard. The invention proves that the micro-magnetic force can promote cells to form an assembly in a joint cavity for the first time, and the cell assembly is found to have efficient anti-inflammatory and cartilage repair functions for the first time. The fluorescence in vivo imaging result shows that the monodisperse cells can be aggregated to form an assembly body in the joint cavity under the influence of micro-magnetic force, and the micro-magnetic force can effectively promote the formation of the aggregation body compared with the situation that the micro-magnetic force group is not applied. Histochemical staining analysis shows that after 2 weeks of micro-magnetic treatment, inflammation can be obviously reduced and the cartilage tissue repair can be promoted, while the blank control group (CTR), the pure cell group, the pure magnetic field group and the material group have no obvious effect on the cartilage tissue repair.
The present invention will be described in further detail with reference to examples. It is also to be understood that the following examples are illustrative of the present invention and are not to be construed as limiting the scope of the invention, and that certain insubstantial modifications and adaptations of the invention by those skilled in the art may be made in light of the above teachings. The specific process parameters and the like of the following examples are also only one example of suitable ranges, i.e., those skilled in the art can select the appropriate ranges through the description herein, and are not limited to the specific values exemplified below.
Example 1
An engineered magnetic control cell assembly and a preparation method and application thereof, specifically comprising the following steps:
(1) preparing and characterizing the antioxidant magnetic nano material: weighing ferric chloride hexahydrate and ferrous chloride tetrahydrate, dissolving in deionized water under inert gas atmosphere to form 40mM/20mM Fe3+/Fe2+Adding melanin 0.5mg/mL, mixing, adding ammonia water to adjust pH>And 10, heating to 80 ℃ for reaction for 1 hour, and fully washing and purifying the obtained antioxidant magnetic nano material by deionized water through magnetic separation. The characterization result shows that the antioxidant magnetic nano material is irregular spherical particles with the particle size of 10-20nm (figure 1A), the saturation magnetization is about 75emu/g (figure 1B), the DPPH & scavenging activity of the antioxidant magnetic nano material is increased along with the increase of the concentration (figures 1C and D), and the scavenging rate can reach more than 95% at 200 mug/mL (figure 1D).
Fig. 1 is a MFNP performance characterization graph. (A) Transmission electron micrograph of MFNP, (B) saturation magnetization of MFNP, (C) uv-visible spectrum of DPPH radical scavenging by MFNP, and (D) DPPH radical scavenging rate by MFNP. The results show that MFNP has a dimensional morphology, excellent saturation magnetization, and excellent DPPH radical scavenging effect.
FIG. 2 shows the biological activity of MFNP and ROS scavenging effect. (A) Proliferation activity of MFNP-endocytosed stem cells before and after micromagnetic treatment, (B) cell viability of MFNP-endocytosed stem cells in a hydrogen peroxide environment, and (C) ability of MFNP-endocytosed stem cells to scavenge ROS before and after micromagnetic treatment. It is shown that MFNP has good biological activity and micromagnetic forces can promote cell proliferation and assist stem cells against oxidative stress microenvironments.
(2) Preparation and viability characterization of cell assemblies: digesting cells cultured by an antioxidant magnetic nano material of 12.5 mu g/mL by pancreatin, and diluting a cell suspension into 500 w/mL; ② adding 3 ten thousand cells into a 96-well plate, and adding PBS for example to make up to 100 μ L; thirdly, adjusting the magnetic field intensity to 260mT, acting on the magnetic field for 20min at the frequency of 10Hz, and then taking a picture of the assembled body by using a microscope; and fourthly, putting the assembly back to the incubator for continuous culture, observing and recording the morphological change of the assembly by using a microscope every day, and simultaneously evaluating the life activity of the assembly by using CCK8 and a Live/Dead kit.
FIG. 3 shows that MFNP and micromagnetic force assist in enhancing anti-inflammatory ability of stem cells. (A) MMP3 gene expression, (B) Adamts-5 gene expression, (C) COX-2 gene expression, (D) IL-4 gene expression, (E) IL-10 gene expression, and (F) IL-13 gene expression. The figure shows that MFNP material and micromagnetic treatment can down-regulate the expression of stem cell inflammatory factors and promote the expression of anti-inflammatory related genes.
FIG. 4 shows stem cell assemblies obtained with different magnetic field parameters. (A) The cell assemblies are obtained under different magnetic field strengths, (B) the cell assemblies are formed under different frequencies, (C) the cell assemblies are formed at different times, (D) the assemblies are formed by stem cells with different magnetic contents, and (E) the cell assemblies with different shapes are obtained by changing the parameters of the magnetic field. The figure shows that assemblies with different shapes and sizes can be obtained in vitro by regulating and controlling different magnetic field strengths, frequencies and action times.
Fig. 5 micromagnetic force is used for controlling the aggregation and assembly of stem cells in a joint cavity. (A) The cell aggregation condition before and after the magnetic force after the injection of 25 ten thousand stem cells, and the cell aggregation condition of the stem cells whether the magnetic force of 100 ten thousand stem cells acts or not is (B) injected. The figure shows that the micro-magnetic force can enhance the fluorescence intensity of the joint part after the stem cells are injected, and the micro-magnetic force can promote the stem cells to gather in the joint cavity.
(3) Characterization of osteogenic differentiation properties of cell assemblies: firstly, after the preparation of the assembly is finished, replacing an osteogenesis induction culture medium to continuously culture the assembly, and carrying out micro-magnetic treatment (260mT, 5Hz, 10min) every day; ② cell RNA is extracted 7 days later and the expression of osteogenesis related genes (COL1, BMP2, OCN, OPN, RUNX2 and the like) is detected; thirdly, after 14 days, the synthesis and secretion of the alkaline phosphatase are detected by using an alkaline phosphatase qualitative and quantitative detection kit; and fourthly, after 21 days, characterizing the calcium nodules synthesized by the assembly by using an alizarin red staining kit.
FIG. 6 micromagnetic force regulates chondrogenic differentiation of stem cells. (A) COL2 gene expression, (B) Aggrecan gene expression, (C) SOX9 gene expression, and (D) NACD gene expression. (E-H) when TGF-beta is added, the micro-magnetic force is adjusted up to the expression of the cartilage related gene. (E) COL2 gene expression, (F) Aggrecan gene expression, (G) SOX9 gene expression, and (H) NACD gene expression. The figure shows that the micro-magnetic force can be used for up-regulating the expression of genes related to the chondrocytes, and the chondrogenic differentiation effect is more remarkable after the micro-magnetic force is cooperated with TGF-beta.
FIG. 7 Effect of stem cell assemblies on in vivo treatment of arthritis. (A) Macroscopic photograph of joint sample, (B) SO staining of joint sample, (C) SO staining amplification, (D) Alisin blue staining, and (E) toluidine blue staining. The figure shows that micromagnetic treatment can effectively treat osteoarthritis.
Example 2
An engineered magnetic control cell assembly and a preparation method and application thereof, specifically comprising the following steps:
(1) preparing and characterizing the antioxidant magnetic nano material: weighing ferric nitrate nonahydrate and ferrous chloride tetrahydrate and dissolving in deionized water under the inert gas atmosphere to form 70mM/35mM Fe3+/Fe2+Adding 0.75mg/mL tannic acid, mixing, adding sodium hydroxide solution to adjust pH>And 10, heating to 80 ℃ for reaction for 1h, and fully washing and purifying the obtained antioxidant magnetic nano material by using deionized water through magnetic separation.
(2) Preparation and viability characterization of cell assemblies: digesting cells cultured by an antioxidant magnetic nano material of 25 mu g/mL by pancreatin, and diluting a cell suspension into 500 w/mL; ② adding 3 ten thousand cells into a 96-well plate, and adding PBS for supplementing to 100 mu L; thirdly, adjusting the magnetic field intensity to 200mT, the frequency to 5Hz, acting on the magnetic field for 5min, and then taking a picture of the assembly by using a microscope; and fourthly, putting the assembly back to the incubator for continuous culture, observing and recording the morphological change of the assembly every day by using a microscope, and simultaneously evaluating the life activity of the assembly by using a CCK8 and a Live/Dead kit.
(3) Chondrogenic differentiation performance characterization of cell assemblies: firstly, after the assembly is prepared, the assembly is replaced by a cartilage induction culture medium to be continuously cultured, and micro-magnetic treatment (200mT, 2Hz, 5min) is carried out every day; ② 7 days later, extracting cell RNA and detecting the expression of osteogenesis related genes (COL2, Aggrecan, SOX9, NCAD, etc.); ③ detecting the synthesis and secretion of acidic protein related to chondrogenesis secreted by cells by using alisnew blue staining solution after 14 days; after 28 days, the cell pellet was embedded and sectioned, and the degree of chondrogenic differentiation of the cells was evaluated with safranin O.
Example 3
An engineered magnetic control cell assembly and a preparation method and application thereof, specifically comprising the following steps:
(1) preparing and characterizing the antioxidant magnetic nano material: weighing ferric chloride hexahydrate and ferrous sulfate heptahydrate in deionized water under the inert gas atmosphere to form 100mM/50mM Fe3+/Fe2+Adding 1mg/mL lignin, mixing, adding potassium hydroxide to adjust pH>And 10, heating to 80 ℃ for reaction for 1h, and fully washing and purifying the obtained antioxidant magnetic nano material by using deionized water through magnetic separation.
(2) Preparation and viability characterization of cell assemblies: digesting cells cultured by using an antioxidant magnetic nano material of 50 mu g/mL by using pancreatin, and diluting a cell suspension into 500 w/mL; ② adding 3 ten thousand cells into a 96-well plate, and adding PBS for example to make up to 100 μ L; thirdly, adjusting the magnetic field intensity to 150mT, the frequency to 1Hz, acting on the magnetic field for 10min, and then taking a picture of the assembly by using a microscope; and fourthly, putting the assembly back to the incubator for continuous culture, observing and recording the morphological change of the assembly every day by using a microscope, and simultaneously evaluating the life activity of the assembly by using a CCK8 and a Live/Dead kit.
(3) Characterization of adipogenic differentiation properties of cell assemblies: firstly, after the preparation of the assembly is finished, a fat induction culture medium is used for continuously culturing the assembly, and micro-magnetic treatment (150mT, 5Hz, 3min) is carried out every day; ② 7 days later, extracting cell RNA and detecting the expression of fat differentiation related genes (PPAR-gamma, C/EBP-alpha, FAS, ACC and the like); measuring the content of triglyceride and total cholesterol after 14 days; fourthly, after 21 days, the cell balls were embedded and sectioned, and the degree of adipogenic differentiation of the cells was evaluated with oil red O.
The foregoing is directed to preferred embodiments of the present invention, other and further embodiments of the invention may be devised without departing from the basic scope thereof, and the scope thereof is determined by the claims that follow. However, any simple modification, equivalent change and modification of the above embodiments according to the technical essence of the present invention will still fall within the protection scope of the technical solution of the present invention.

Claims (10)

1. An engineered magnetically controlled cell assembly, comprising cells and antioxidant magnetic nanomaterials distributed within the cells.
2. The engineered magnetron cell assembly of claim 1, wherein the cells include, but are not limited to, stem cells, hepatocytes, chondrocytes, osteoblasts, or cardiomyocytes.
3. The engineered magnetically controlled cell assembly of claim 1, wherein the antioxidant magnetic nanomaterial is a magnetically responsive substance modified with an antioxidant substance.
4. The assembly of claim 3, wherein said magnetically responsive substance comprises but is not limited to ferroferric oxide, cobalt ferrite, nickel ferrite, or neodymium iron boron;
the antioxidant substances include, but are not limited to, melanin, lignin, ferulic acid, tannic acid, or N-acetyl-L-cysteine.
5. A method for preparing an engineered magnetron cell assembly as claimed in any one of claims 1 to 4, comprising the steps of:
(1) preparing an antioxidant magnetic nano material: preparing an antioxidant magnetic nano material by adopting a coprecipitation method or a hydrothermal method and taking an antioxidant substance as a ligand;
(2) co-culturing the antioxidant magnetic nano material and cells: preparing the antioxidant magnetic nano material into dispersion liquid by using a cell complete culture medium, then adding the dispersion liquid into a culture dish containing cells, placing the culture dish into an incubator for continuous culture, and washing an obtained cell compound by using PBS phosphate buffer solution to obtain magnetic cells;
(3) and (3) placing the magnetic cells in a magnetic field, and acting for a period of time to obtain the engineered magnetic control cell assembly.
6. The method of claim 5, wherein the concentration of the anti-oxidizing magnetic nanomaterial in the dispersion is 10-500 μ g/mL; the temperature in the incubator is 35-38 ℃, and the volume concentration of carbon dioxide is 0-6%; the continuous culture time is 12-72 h.
7. The method of claim 5, wherein the magnetic field has a strength of 0-260mT and a frequency of 0-30 Hz; the action time is 0-60 min.
8. The method of claim 5, wherein the magnetic field is a rotating magnetic field or a pulsed magnetic field.
9. The method of claim 5, wherein the cell assembly includes, but is not limited to, two-dimensional sheet assembly and three-dimensional spherical assembly.
10. Use of an engineered magnetron cell assembly as claimed in any one of claims 1 to 4 in the preparation of an implant material with anti-inflammatory and tissue repair properties.
CN202210167246.2A 2022-02-23 2022-02-23 Engineered magnetic control cell assembly and preparation method and application thereof Pending CN114574434A (en)

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