CN111150884A - Magnetic induction coil-type vertebral body cage with superparamagnetic iron oxide magnetic nanocoating - Google Patents

Abstract

The invention discloses a magnetic induction coil type vertebral body fusion cage with a superparamagnetic iron oxide magnetic nano coating, which is used for reinforcing stable support between vertebral bodies in cervical vertebra surgery, the fusion cage is of a magnetic induction coil type structure, the surface of the fusion cage is provided with a superparamagnetic iron oxide magnetic nano coating, the superparamagnetic iron oxide magnetic nano coating is gamma-Fe 2O3 magnetic nano particles coated with polysaccharide quaternary ammonium salt on the surface, and under the action of an external magnetic field, the vertebral body fusion cage with the superparamagnetic iron oxide magnetic nano coating generates corresponding electromagnetic effect and other indirect effect to promote the repair of bone tissue injury. The specific magnetic induction coil structure and the external magnetic field are used for the cervical vertebra fusion operation, so that the growth and the repair of bone tissues can be promoted under the action of the external specific magnetic field after the operation, and the effect of accelerating the recovery is achieved.

Description

Magnetic induction coil type vertebral body fusion cage with superparamagnetic iron oxide magnetic nano coating

Technical Field

The invention relates to preparation and processing of a magnetic induction coil type vertebral fusion cage with a superparamagnetic iron oxide magnetic nano coating, in particular to surface modification and coating preparation of the vertebral fusion cage, and an effect of promoting vertebral fusion and accelerating recovery under the action of a magnetic field in and after a cervical vertebra fusion operation.

Background

At present, cervical spondylosis becomes a common disease and a frequently encountered disease, the morbidity of the cervical spondylosis is gradually increased in a young population under the background of the high-speed development of the current society, the cervical spondylosis is also called cervical syndrome, is a general name of cervical osteoarthritis, proliferative cervical spondylitis, cervical radicular syndrome and cervical disc herniation, and is a disease based on the change of a degenerative pathology. Mainly, due to long-term strain of cervical vertebrae, hyperosteogeny, or prolapse of intervertebral disc and ligament thickening, the spinal cord, nerve root or vertebral artery of cervical vertebrae are pressed, and a series of clinical syndromes of dysfunction appear. When the disease condition is treated by the vertebral body fusion cage, the postoperative rapid rehabilitation is particularly important.

Titanium metal has very strong fatigue resistance and corrosion resistance compared to other metals. Titanium metal also has better biocompatibility, and the rejection reaction of titanium metal is smaller compared with other metals when the titanium metal is implanted into an animal body.

The magnetic field is a non-invasive physical factor and can directly act on organisms to generate corresponding biological effects, and when the magnetic field of the organisms is changed, the activity of cell molecules and the activity of the organisms are influenced. A tissue of a living body contains a large number of free electrons, and these free electrons are stationary in a natural state. However, when the organism tissue is in an electromagnetic environment, the free electrons in the organism tissue are excited to move continuously, so that the organism in the changing electromagnetic field can generate a very strong response under very small stimulation. At present, the research on treating bone diseases by utilizing the electromagnetic effect is carried out in China, and the feasibility of the electromagnetic effect for exerting the treatment effect is proved.

The magnetic nanoparticles are a chemical compound formed by combining nanoparticles and magnetic elements such as iron, nickel and the like, and have the advantages of special magnetic effect, magnetic guidance, good biocompatibility, low toxicity, long blood circulation time, specific advantages due to small size and the like. On the one hand, the gamma-Fe 2O3 iron oxide particles used in the present invention have been certified for safety by ISO International quality control System with low cytotoxicity to organisms; on the other hand, the residual magnetism Mr and the coercive force Hc of the superparamagnetic nano-particles basically tend to be zero, so that only weak magnetic dipole effect exists between the particles, and stable magnetic fluid can be formed. At present, the iron oxide nanoparticles are widely applied biomedical nano materials, have good biocompatibility, in-vivo stability and special electromagnetic properties, can generate a plurality of unique effects under an external electromagnetic field, and are increasingly researched and paid attention to magnetic drug carriers, tumor thermotherapy and brain deep magnetic stimulation.

Disclosure of Invention

The purpose of the invention is as follows: the invention designs and manufactures a magnetic induction coil type vertebral body fusion device with a superparamagnetic iron oxide magnetic nano coating, which plays roles of providing stable support between vertebral bodies and maintaining the height of a vertebral body gap in a cervical vertebra fusion operation, and can promote bone repair and accelerate the time of vertebral body fusion under the action of a magnetic field.

The technical scheme is as follows: the invention relates to a magnetic induction coil type vertebral body fusion cage with a superparamagnetic iron oxide magnetic nano coating.

Wherein:

the magnetic induction coil type structure main body component is metal pure titanium, the chemical composition meets the regulation of Grade2 in GB/T3810, and the grain size is not coarser than Grade 5.

The magnetic induction coil type structure is a single-wire spiral structure, a multi-wire side-by-side spiral structure or a multi-wire twisted spiral structure.

The magnetic induction coil type structure is integrally cylindrical, the diameter range of the bottom is 8mm-22mm, the height range is 8mm-28mm, the height of a single-layer spiral cone of the magnetic induction coil type cone fusion device is 1.5mm-2mm, and the thickness is 0.8mm-1.2 mm.

The superparamagnetic iron oxide magnetic nano coating is gamma-Fe 2O3 magnetic nano particles coated with polysaccharide quaternary ammonium salt on the surface, and under the action of an external magnetic field, the vertebral body fusion cage of the superparamagnetic iron oxide magnetic nano coating generates corresponding electromagnetic effect and other indirect effect to promote the repair of bone tissue damage.

The coating thickness of the gamma-Fe 2O3 magnetic nano-particles coated with the surface polysaccharide quaternary ammonium salt on the magnetic induction coil type structure is 200-400 mu m.

The preparation method of the magnetic induction coil type vertebral fusion cage with the superparamagnetic iron oxide magnetic nano coating comprises the following steps:

step 1: before the preparation of the superparamagnetic iron oxide magnetic nano coating, washing the processed vertebral body fusion cage with a neutral detergent for 20-40 min, then washing with deionized water at normal temperature for 15-20 min, and then washing in distilled water for 20-30 min;

step 2: after heat treatment is carried out for 2 hours at 400-500 ℃ in a drying furnace, the mixture is cooled to room temperature along with the furnace;

and step 3: placing the treated vertebral body fusion device in NaOH (10 mol. L) at the temperature of 40-50 DEG C^-1) Soaking in the solution for 24-36 h, taking out the vertebral fusion cage, placing in distilled water, cleaning for 15-30 min, and drying in a drying furnace at 40-50 ℃ for 24-36 h;

and 4, step 4: placing the dried vertebral body fusion device in a resistance furnace at 5-15 ℃ for min^-1Heating to 500-600 deg.c at the heating rate, maintaining the temperature for 1-1.5 hr, and cooling to room temperature;

and 5: after the steps are completed, the vertebral body fusion cage is firstly placed in a quaternary ammonium salt colloidal solution for 15min to 20min, the temperature of the solution is 40 ℃ to 50 ℃, the vertebral body fusion cage is taken out and dried in a drying furnace for 1h to 1.5h at the temperature of 40 ℃ to 50 ℃, then the vertebral body fusion cage is placed in a gamma-Fe 2O3 colloidal solution for 15min to 20min at the temperature of 40 ℃ to 50 ℃, the vertebral body fusion cage is taken out and dried in the drying furnace for 1h to 1.5h at the temperature of 40 ℃ to 50 ℃;

step 6: after the magnetic induction coil type vertebral body fusion cage is alternately placed for 4 times, the preparation of the superparamagnetic iron oxide nanoparticle gamma-Fe 2O3 composite film is completed on the surface of the magnetic induction coil type vertebral body fusion cage.

Has the advantages that: the superparamagnetic iron oxide nanoparticles are assembled on the surface of a magnetic inductive coil type vertebral body fusion cage and implanted into a damaged part of a vertebral body, and the specific action is carried out on bone tissues by utilizing the electromagnetic effect and other indirect effects of the magnetic nanoparticles under an external electromagnetic field, such as the magnetocaloric effect, the magnetoelastic effect and the like. Because the magnetic material can gather the electromagnetic field, the action intensity of the electromagnetic field is locally enhanced in the bone tissue, the electromagnetic action energy at the interface of the magnetic nanoparticle assembly structure and the tissue cells is greatly increased, and the point can ensure that a stronger electromagnetic effect can be generated at the interface of the nano material even if only the electromagnetic field with lower intensity exists in the external environment. Meanwhile, the electromagnetic field generated by the polarization or magnetization of the nanoparticle interface is quickly attenuated along with the increase of the distance, so that the electromagnetic effect is a local electromagnetic effect, the in-vivo electromagnetic effect is only acted on the local part of the bone injury tissue, and the simultaneous influence on multiple parts of the whole body is avoided.

Drawings

Fig. 1 is a mechanical structure diagram of the present invention.

Detailed Description

The invention relates to a magnetic induction coil type vertebral body fusion cage with a superparamagnetic iron oxide magnetic nano coating.

Wherein:

the magnetic induction coil type structure is a single-wire spiral structure, a multi-wire side-by-side spiral structure or a multi-wire twisted spiral structure.

The magnetic induction coil type structure is integrally cylindrical, the diameter range of the bottom is 8mm-22mm, the height range is 8mm-28mm, the height of a single-layer spiral cone of the magnetic induction coil type cone fusion device is 1.5mm-2mm, and the thickness is 0.8mm-1.2 mm.

The superparamagnetic iron oxide magnetic nano coating is gamma-Fe 2O3 magnetic nano particles coated with polysaccharide quaternary ammonium salt on the surface, and under the action of an external magnetic field, the vertebral body fusion cage of the superparamagnetic iron oxide magnetic nano coating generates corresponding electromagnetic effect and other indirect effect to promote the repair of bone tissue damage.

The coating thickness of the gamma-Fe 2O3 magnetic nano-particles coated with the surface polysaccharide quaternary ammonium salt on the magnetic induction coil type structure is 200-400 mu m.

The invention relates to the manufacture of a magnetic induction coil type vertebral body fusion cage with a superparamagnetic iron oxide magnetic nano coating, which is characterized in that the method for manufacturing the magnetic induction coil type vertebral body fusion cage utilizes commercially available metal pure titanium, has chemical components meeting the requirements of Grade2 in GB/T3810, has the grain size not larger than 5 Grade, and is processed into the size and the shape of a magnetic induction coil type by numerical control processing or turning processing.

The preparation method comprises the following steps: before the preparation of the superparamagnetic iron oxide magnetic nano coating, the processed vertebral body fusion cage is washed for 20min by using a neutral detergent, then washed for 15min by using deionized water at normal temperature, then placed in distilled water for washing for 20min, finally treated for 2 hours in a drying furnace by heat treatment at 400 ℃, and cooled to room temperature along with the furnace. After the treatment, the treated vertebral body fusion device is placed in NaOH (10 mol. L) at 50 DEG C^-1) Soaking in the solution for 24h, taking out the vertebral fusion cage, washing in distilled water for 15min, and drying in a drying furnace at 40 ℃ for 24 h. Finally, the dried vertebral body fusion cage is placed in a resistance furnace at 15 ℃ for min^-1The temperature is raised to 600 ℃ at the heating rate, the temperature is kept for 1h, and then the furnace is cooled to the room temperature. After the steps are finished, the vertebral body fusion cage is firstly placed in a quaternary ammonium salt colloidal solution for 15min, the temperature of the solution is 40 ℃, the vertebral body fusion cage is taken out and dried in a drying furnace for 1h at 40 ℃, then the vertebral body fusion cage is placed in a gamma-Fe 2O3 colloidal solution for 15min at the temperature of 40 ℃, the vertebral body fusion cage is taken out and dried in the drying furnace for 1h at 40 ℃, and after the vertebral body fusion cage is placed alternately for 4 times, superparamagnetic iron oxide nano-particle gamma is finished on the surface of the magnetic induction coil type vertebral body fusion cagePreparation of Fe2O3 composite film.

Experimental example 1 influence of magnetic induction coil type vertebral fusion cage with superparamagnetic iron oxide magnetic nano coating on rat vertebral lesion under action of external magnetic field

① A female C57BL/6 mouse (C57BL/6J JAXTMmic strain) of 5-6 weeks old is taken, the weight is about 15-18g, bilateral ovaries are cut off, a bone injury model is established at 19 th, 20 th and 21 st spines of the rat after 3 months, a superparamagnetic iron oxide magnetic nano-coating vertebral fusion device (the bottom diameter is 2mm, the height is 6mm) is implanted into a spine injury part of the rat, different types of electromagnetic fields are applied regularly, the external magnetic field is a rotating magnetic field, the rotating magnetic field is generated by uniformly rotating a pair of symmetrically arranged conical magnets at 1-20Hz, the waveform of the magnetic field generated at the stimulation part is in a sequence pulse shape, and the field intensity of the rotating magnetic field is less than 0.5T.

② the experiment is divided into a control group, a common vertebral body fusion device, a magnetic induction coil type vertebral body fusion device implanted with a superparamagnetic iron oxide nano coating, an electromagnetic field group externally applied to the magnetic induction coil type vertebral body fusion device implanted with the common vertebral body fusion device and an electromagnetic field group externally applied to the magnetic induction coil type vertebral body fusion device implanted with the superparamagnetic iron oxide nano coating.

③ separating and purifying marrow mesenchymal stem cells from the vertebra of healthy rats of the same strain, taking the marrow mesenchymal stem cells as seed cells for bone tissue repair, inoculating the seed cells on the prepared superparamagnetic iron oxide nano-coating vertebral body fusion device, and implanting the seed cells into the bone defect of the rats;

④ at 4, 8, 12 weeks after the stent is implanted, taking X-ray pictures to observe the bone healing condition, analyzing parameters such as bone density by combining bone quantitative analysis software, simultaneously extracting blood from the bone repair part, separating serum, detecting biochemical indexes such as Ca2+, alkaline phosphatase (ALP), alanine aminotransferase, creatinine kinase and the like, detecting the levels of several important inflammatory factors by using a high-flux protein chip, and comprehensively analyzing the bone metabolism level and the magnetic induction coil type vertebral body fusion device with the superparamagnetic iron oxide nano coating in biocompatibility and the effectiveness of bone repair.

⑤ the rats are killed at 12 weeks after the stent is implanted, HE staining of tissues at the bone defect position is adopted to observe the generation condition of the bone trabecula, VonKossa staining is adopted to observe the bone mineralization condition, tartrate-resistant acid phosphatase (TRAP) staining is adopted to observe the activity and quantity of osteoclasts, Prussian blue staining is adopted to identify the phagocytosis condition of the cells to the nanoparticles, Micro-CT is adopted to measure the bone volume, the expression of callus osteoprotegerin is quantitatively analyzed, and the like, and the magnetic induction coil type vertebral body fusion device of the superparamagnetic iron oxide nano coating and the capability of an external electromagnetic field for promoting the healing of the bone defect are comprehensively evaluated.

⑥ separating the cells at the bone repairing position, detecting the apoptosis and proliferation activity of the cells by apoptosis immunoassay kit (TUNEL kit) and flow cytometry, identifying the phagocytosis of the cells to nanoparticles by tissue slice Prussian blue staining, analyzing the adhesion, morphological change and the like of the cells by scanning electron microscope and cytoskeleton fluorescent staining, and detecting the transcription of various related mRNAs of the new bone cells on the vertebral fusion cage by quantitative PCR as the evaluation index of the performance of the fusion cage.

⑦ femoral bone marrow cells are separated and induced to differentiate into osteoblasts and osteoclasts in vitro, the early differentiation of osteoblasts is identified by ALP staining, the mineralization ability of mature osteoblasts in vitro is identified by Von Kossa staining, osteoblast differentiation markers (osteocalcin, osteopontin, etc.) are detected by quantitative PCR and Western Blot, the differentiation of osteoclasts is identified by TRAP staining, the bone resorption ability of osteoclasts is identified by toluidine blue staining, and the mRNA expression levels of osteoclast differentiation markers (cFms, RANK, Cath K, Integrin β 3, MMP9, IkB α, NFATc1, etc.) are detected by quantitative PCR and Western Blot.

⑧ data analysis, using SPSS 176 statistic software package to make statistic analysis, the data is represented by average + -standard error, the comparison of the averages among groups is variance analysis (ANOVA), the comparison of the averages in the groups is pair t test, the result shows that the collective magnetism property of the iron oxide nanometer particle assembly structure is changed from superparamagnetism to weak ferromagnetism, the assembly structure generates remanence, using magnetic induction protein as indicator to prove that remanence promotes the differentiation of mouse primary marrow cells to osteogenesis direction, the magnetic induction coil type centrum fusion device of superparamagnetism iron oxide nanometer coating promotes the growth and differentiation of marrow cells by magnetism.

Claims (7)

1. A magnetic induction coil type vertebral body fusion cage with a superparamagnetic iron oxide magnetic nano coating is characterized in that: the fusion cage is of a magnetic induction coil structure, and the surface of the fusion cage is provided with a superparamagnetic iron oxide magnetic nano coating.

2. The magnetic induction coil-type vertebral fusion cage with superparamagnetic iron oxide magnetic nano-coating according to claim 1, characterized in that: the magnetic induction coil type structure main body component is metal pure titanium, the chemical composition meets the regulation of Grade2 in GB/T3810, and the grain size is not coarser than Grade 5.

3. The magnetic induction coil-type vertebral fusion cage with superparamagnetic iron oxide nano-coating according to claim 1 or 2, characterized in that: the magnetic induction coil type structure is a single-wire spiral structure, a multi-wire side-by-side spiral structure or a multi-wire twisted spiral structure.

4. The magnetic induction coil-type vertebral fusion cage with superparamagnetic iron oxide magnetic nano-coating according to claim 3, characterized in that: the magnetic induction coil type structure is integrally cylindrical, the diameter range of the bottom is 8mm-22mm, the height range is 8mm-28mm, the height of a single-layer spiral cone of the magnetic induction coil type cone fusion device is 1.5mm-2mm, and the thickness is 0.8mm-1.2 mm.

5. The magnetic induction coil-type vertebral fusion cage with superparamagnetic iron oxide magnetic nano-coating according to claim 1, characterized in that: the superparamagnetic iron oxide magnetic nano coating is gamma-Fe 2O3 magnetic nano particles coated with polysaccharide quaternary ammonium salt on the surface, and under the action of an external magnetic field, the vertebral body fusion cage of the superparamagnetic iron oxide magnetic nano coating generates corresponding electromagnetic effect and other indirect effect to promote the repair of bone tissue damage.

6. The magnetic induction coil-type vertebral fusion cage with superparamagnetic iron oxide magnetic nano-coating according to claim 5, wherein: the coating thickness of the gamma-Fe 2O3 magnetic nano-particles coated with the surface polysaccharide quaternary ammonium salt on the magnetic induction coil type structure is 200-400 mu m.

7. A method for preparing the magnetic induction coil type vertebral body fusion cage with the superparamagnetic iron oxide magnetic nano-coating according to claim 5, wherein the method for preparing comprises the following steps:

step 1: before the preparation of the superparamagnetic iron oxide magnetic nano coating, washing the processed vertebral body fusion cage with a neutral detergent for 20-40 min, then washing with deionized water at normal temperature for 15-20 min, and then washing in distilled water for 20-30 min;

step 2: after heat treatment is carried out for 2 hours at 400-500 ℃ in a drying furnace, the mixture is cooled to room temperature along with the furnace;

and step 3: placing the treated vertebral body fusion device in NaOH (10 mol. L) at the temperature of 40-50 DEG C^-1) Soaking in the solution for 24-36 h, taking out the vertebral fusion cage, placing in distilled water, cleaning for 15-30 min, and drying in a drying furnace at 40-50 ℃ for 24-36 h;

and 4, step 4: placing the dried vertebral body fusion device in a resistance furnace at 5-15 ℃ for min^-1Heating to 500-600 ℃ at the heating rate, keeping the temperature for 1-2 h, and cooling to room temperature along with the furnace;

and 5: after the steps are completed, the vertebral body fusion cage is firstly placed in a quaternary ammonium salt colloidal solution for 15min to 20min, the temperature of the solution is 40 ℃ to 50 ℃, the vertebral body fusion cage is taken out and dried in a drying furnace for 1h to 1.5h at the temperature of 40 ℃ to 50 ℃, then the vertebral body fusion cage is placed in a gamma-Fe 2O3 colloidal solution for 15min to 20min at the temperature of 40 ℃ to 50 ℃, the vertebral body fusion cage is taken out and dried in the drying furnace for 1h to 1.5h at the temperature of 40 ℃ to 50 ℃;

step 6: after the magnetic induction coil type vertebral body fusion cage is alternately placed for 4 times, the preparation of the superparamagnetic iron oxide nanoparticle gamma-Fe 2O3 composite film is completed on the surface of the magnetic induction coil type vertebral body fusion cage.

Images