CN116570761A - Preparation and application of icariin-carrying porous magnesium alloy - Google Patents
Preparation and application of icariin-carrying porous magnesium alloy Download PDFInfo
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- CN116570761A CN116570761A CN202310561250.1A CN202310561250A CN116570761A CN 116570761 A CN116570761 A CN 116570761A CN 202310561250 A CN202310561250 A CN 202310561250A CN 116570761 A CN116570761 A CN 116570761A
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- magnesium alloy
- icariin
- porous magnesium
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- porous
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- 238000002360 preparation method Methods 0.000 title claims abstract description 11
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Classifications
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
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- A61L27/02—Inorganic materials
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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- A61L27/54—Biologically active materials, e.g. therapeutic substances
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/10—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing inorganic materials
- A61L2300/102—Metals or metal compounds, e.g. salts such as bicarbonates, carbonates, oxides, zeolites, silicates
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- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/216—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials with other specific functional groups, e.g. aldehydes, ketones, phenols, quaternary phosphonium groups
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
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- Engineering & Computer Science (AREA)
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- Molecular Biology (AREA)
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Abstract
The invention discloses a preparation method and application of a porous magnesium alloy carrying icariin, belonging to the technical field of biomedicine, and comprising the following steps: sintering titanium particles into a porous template, preparing a composite material of magnesium alloy and titanium particles by using percolation casting, and finally dissolving titanium filling particles by using hydrofluoric acid to obtain the porous magnesium alloy. The icariin is loaded on the magnesium alloy, long-term icariin stimulation can be provided along with degradation of the magnesium alloy into cartilage defect parts, and the icariin stimulation and released magnesium ions play a synergistic effect to jointly accelerate the repair of the cartilage defect.
Description
Technical Field
The invention belongs to the technical field of biomedicine, and particularly relates to preparation and application of a porous magnesium alloy carrying icariin.
Background
With the development of material science, various cartilage repair materials have been developed, which can be classified into metallic materials and nonmetallic materials, and magnesium alloy materials are commonly used in the metallic materials. Magnesium ions are indispensable elements in a body and participate in various metabolism processes, magnesium ions generated after the magnesium alloy is degraded can supplement magnesium elements in the body, promote metabolism acceleration, and accelerate angiogenesis and cartilage repair.
The traditional Chinese medicine composition has long been applied in orthopaedics diseases, and icariin is used as the main component of epimedium, and can be used for repairing cartilage defect, bone defect and peripheral nerve defect. As an accelerant for cartilage formation, icariin has the functions of anti-inflammatory, anti-aging, immunoregulation and the like, has wide physiological activity and has higher medicinal and health care values.
Cartilage defects are a difficult clinical treatment point, and are mainly caused by wounds, inflammations and the like, and are difficult to repair by themselves after being damaged due to separation from a circulatory system. In clinic, cartilage defects are usually repaired by taking autologous cartilage, but the curative effect is often poor due to the fact that the autologous cartilage is not enough in source, the pain of a patient is high due to secondary operation, the operation is complex, the cost is high, complications such as infection can be caused in a donor area, and the like. Development of new methods to repair cartilage defects is an urgent endeavor.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a preparation method and application of a porous magnesium alloy carrying icariin.
The aim of the invention can be achieved by the following technical scheme:
an alloy material comprises a porous magnesium alloy, wherein icariin is carried on the porous magnesium alloy.
The preparation of the icariin-carrying porous magnesium alloy comprises the following steps:
sintering titanium particles into a porous template, preparing a composite material of magnesium alloy and titanium particles by using percolation casting, and finally dissolving titanium filling particles by using hydrofluoric acid to obtain a porous magnesium alloy;
dissolving 10 mu mol/L icariin in absolute ethyl alcohol, soaking the porous magnesium alloy in the icariin solution, taking out and drying, and adsorbing the icariin on the porous magnesium alloy to obtain the icariin-carrying porous magnesium alloy.
Further, the titanium particles are prepared into a porous template by sintering.
Further, the diameter of the cylindrical material is 2mm, and the height is 2mm.
Further, the time for immersing the porous magnesium alloy in the icariin solution is 1 day.
An application of porous magnesium alloy carrying icariin in preparing cartilage defect repairing medicine is provided.
The invention has the beneficial effects that:
the invention provides a preliminary application of icariin-carrying porous magnesium alloy in cartilage defect repair, the obtained icariin-carrying porous magnesium alloy can provide icariin and magnesium ions released along with the degradation of a bracket for cartilage tissues at defect positions, the porous magnesium alloy can be used as a filling material at cartilage defect positions, and can provide climbing materials and pores for nascent chondrocytes, blood vessels and the like, and the released icariin and magnesium ions can be used as stimulating substances to act on surrounding bone marrow mesenchymal stem cells (BMSCs) and chondrocytes, so that the former is continuously differentiated into chondrocytes and the latter are continuously proliferated, and finally the cartilage defect repair is completed from bottom to top, thereby reducing donor deficiency, patient pain, complex operation, cost increase and the like caused by secondary operation of repairing the defect positions by taking autologous cartilage.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described, and it will be obvious to those skilled in the art that other drawings can be obtained according to these drawings without inventive effort.
FIG. 1 is a drawing of a rat model of the present invention, wherein a cartilage defect of 2mm diameter is drilled in the non-load area of the medial femoral condyle of the knee joint of the rat, and then a porous magnesium alloy material is implanted;
FIG. 2 is a macroscopic view of the porous magnesium alloy stent of the present invention;
FIG. 3 is a scanning electron microscope image of the stent of the present invention. A is a scanning electron microscope image of a simple porous magnesium alloy; the diagram B is a scanning electron microscope diagram (SEM MAG:10.0 kx) of the icariin/porous magnesium alloy novel bracket;
FIG. 4 is a release profile of icariin of the present invention;
fig. 5 is a macroscopic view of the repair of cartilage defects in the knee of the present invention. Graph a/E is group a: icariin/porous magnesium alloy novel bracket group, B/F is group B: pure porous magnesium alloy group, C/G is C group: no scaffold group, D/H is D group: a sham surgical team. a-D are general specimen figures; E-H is a micro-CT image;
FIG. 6 is a graph of data from micro-CT analysis of cartilage defect repair in each group according to the present invention. A is icariin/porous magnesium alloy novel bracket group, B is simple porous magnesium alloy group, C is bracket-free group, and D is artificial operation group. BV/TV is relative bone volume, tb.N is bone trabecular Liang Shuliang, tb.Sp is bone trabecular separation, tb.Th is bone trabecular thickness. * P <0.05;
FIG. 7 is a microscopic view of the repair of cartilage defects in the knee of the present invention. HE dyeing, safranine O-fast green dyeing and toluidine blue dyeing are sequentially carried out from top to bottom, and icariine/porous magnesium alloy novel bracket group, simple porous magnesium alloy group, bracket-free group and artificial operation group are sequentially carried out from left to right;
fig. 8 is an improved Mankin score evaluation of the invention for knee cartilage defect repair. If the score at the defect is high, cartilage defect repair is poor. A. B, C, D the new type of bracket group of icariin/porous magnesium alloy, the pure porous magnesium alloy group, the no bracket group and the artificial operation group are in turn. * P <0.05; * P <0.01; * P <0.001;
FIG. 9 shows the expression of the factor protein of interest (X200) detected by immunohistochemical staining of the present invention. Aggrecan, collagen II, sox9, beta-catenin, wnt5a, wnt1 and sFRP1 proteins are sequentially arranged from top to bottom, and icariin/porous magnesium alloy novel bracket groups, pure porous magnesium alloy groups, bracket-free groups and artificial operation groups are sequentially arranged from left to right;
FIG. 10 shows the expression of the factor protein of the present invention. A. B, C, D the new type of bracket group of icariin/porous magnesium alloy, the pure porous magnesium alloy group, the no bracket group and the artificial operation group are in turn. * P <0.05; * P <0.01; * P <0.001; * P <0.0001.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Example 1
1. Construction of novel icariin/porous magnesium alloy bracket
1.1 preparation of icariin/porous magnesium alloy
The required materials are mainly provided by Shanghai university light alloy precision molding national engineering research center (refer to porous magnesium-based materials described in CN 105039771A), magnesium is mainly used, the main component is Mg-3.1Nd-0.2Zn-0.4Zr, a small amount of Nd, zn and Zr are added for alloying, titanium particles are adopted for sintering into a porous template, then a composite material of magnesium alloy and titanium particles is prepared by using percolation casting, finally titanium filling particles are dissolved by using hydrofluoric acid, porous magnesium alloy is obtained, and the porous magnesium alloy is processed into a cylindrical material with the diameter of 2mm and the height of 2mm. ICA (physical adsorption) of 10 mu mol/L was adsorbed on a porous magnesium alloy by dissolving ICA in absolute ethyl alcohol, immersing the porous magnesium alloy in ICA solution for 1d, taking out, drying in a vacuum oven.
1.2 evaluation of morphology characterization of novel icariin/porous magnesium alloy stent
And photographing by using a camera to observe the surface morphology of the novel porous magnesium alloy and icariin/porous magnesium alloy bracket. The dried porous magnesium alloy and icariin/porous magnesium alloy novel stent are fixed by spraying metal, and the microstructure is observed by using a field emission scanning electron microscope (SEM, TESCAN, mira3 XMH), and the accelerating voltage is 5kV.
1.3 icariin standard curve and Release curve
The relevant data were measured using a High Performance Liquid Chromatograph (HPLC) and standard icariin concentration curves were made. Icariin with different concentrations was dissolved in PBS to prepare 0mg/L, 2.5mg/L, 5mg/L, 10mg/L, 20mg/L, 30mg/L, 40mg/L solutions. Absorbance was monitored at wavelength 270nm using an ultraviolet light detector (UV 230 ii), flow rate: 1ml/min. Finally, the absorbance is plotted against the concentration to obtain a standard curve. Experiments were performed using three independently prepared icariin solutions. The novel icariin/porous magnesium alloy scaffold material having biological activity was weighed to 180mg and immersed in 20ml of PBS solution at 37℃to release icariin. At each time point, 100. Mu.l of PBS solution was taken out and their ultraviolet absorption spectrum at an absorption wavelength of 270nm was examined, and compared with a standard curve prepared, thereby determining the concentration of icariin in PBS. Measurements were made using three separate sample materials.
2. Experimental animal
3 female 10 week old Sprague Dawley (SD) rats (weight 220-240 g) and 24 female 6 week old SD rats (weight 250-350 g) were purchased from the Guangdong province laboratory animal center. SD rats used for the experiments were kept in an environmental control room of Specific Pathogen Free (SPF) barrier region of Shenzhen Hospital laboratory animal center, beijing university, 12h light/12 h dark cycle, relative humidity of 50% -55%, room temperature of 24+ -2 ℃. All rats were free to access food and sterile water. The study was approved by the Shenzhen hospital ethics committee and the animal research committee at the university of Beijing. All experimental procedures and treatments were conducted under the relevant regulations of the national institutes of health "laboratory animal care and use guidelines".
3. Biological property evaluation of icariin/magnesium alloy novel stent material
Artificial scaffold block according to 1g:100ml of the solution was completely immersed in a volume of physiological saline and allowed to stand at room temperature for 48 hours to prepare a physiological saline extract. Taking 3 SD rats, removing back hair after anesthesia, selecting 1 injection point on each side of the spine, respectively injecting leaching solution and 0.9% physiological saline by using a sterile injector to form a hillock, placing the hillock under the same condition for culture after injection, and observing the injection sites and the occurrence of subcutaneous stimulation reactions such as congestion, swelling and the like of peripheral skin at 1h, 12h, 24h and 48h respectively.
4. Modeling and grouping of rats
SD rats were anesthetized using sodium pentobarbital intraperitoneal injection (anesthetic dose calculated as 30 mg/kg). After the rat is in an anesthetic state, skin preparation is carried out on the right knee joint, then 0.5% iodophor is used for disinfecting the skin of the right knee joint for 3 times, the inner side edge of the patella ligament of the right knee joint is cut, the patella is led in from the inner side of the patella ligament, the knee joint is bent to dislocate the patella, so that the femoral pulley is exposed, an electric drill with a 2mm diameter Kirschner wire is used for drilling holes in a non-load area in the middle of the femoral condyle of the right knee joint, the drilling holes reach the cartilage and the subchondral bone full layer until blood in a bone marrow cavity infiltrates into the joint cartilage defect to provide BMSCs in the bone marrow cavity. SD rats were randomly divided into 4 groups: group A: the novel bracket carrying 10 mu mol/L icariin/porous magnesium alloy is placed into the cartilage defect of knee joint (n=8); group B: the simple porous magnesium alloy bracket material is placed in the cartilage defect part (blank group) of the knee joint (n=8); group C: no biological material is placed in the cartilage defect of the knee joint (n=8); group D: only the joint capsule was dissected on the contralateral side of each rat and untreated articular cartilage was used as a normal control (n=24).
5. Observation of general specimens of articular cartilage
Normal feeding of rats to 12 weeks after surgery, killing each group of SD rats by using a cervical dislocation method, sequentially cutting the skin, muscle and joint capsule of the knee joint according to the original operation incision, observing the repair condition of cartilage defects of the knee joint, observing the appearance of synovitis such as the presence or absence of joint effusion and synovial swelling of the knee joint, taking out the distal femur ends of the two sides of the rats completely, and soaking the distal femur ends into prepared paraformaldehyde for tissue fixation.
6. micro-CT macroscopic observation of rat knee joint cartilage repair condition
The surface and internal structure of subchondral bone at the repair site are observed by using a distal femur specimen on both sides of a rat sacrificed by micro-CT scanning, and the voltage of an X-ray tube is 80kV and the current is 450 muA. The number of views is 400, the exposure time is 400ms, and the lens scanning technique is selected. The effective pixel size is 0.046mm.
7. Conventional and special staining observation of rat knee joint cartilage repair condition
Tissue specimens were fixed with 4% paraformaldehyde, decalcified and embedded in paraffin, and sectioned with a paraffin microtome to a thickness of 5-6 μm. The tissue slice for HE staining is firstly placed in hematoxylin staining solution for 2min, and then is placed in running water to be washed for bluing; then placing the mixture in eosin staining solution for 2min, and washing the mixture with water again. The tissue slice for safranin O solid-green staining is placed in Weibert dye liquor for 5min, differentiated for 5s by acid differentiation liquor, then placed in solid-green dye liquor for 20min, washed with weak acid for 5s, placed in safranin dye liquor for 30min, and finally washed with water for superfluous flooding. The tissue slice for toluidine blue staining is placed in toluidine blue staining solution for 5min, then placed in 95% ethanol for 1min, and finally the slice is washed.
8. Assessment of rat knee cartilage repair by improved Mankin score
We scored and statistically analyzed cartilage defect repair by cartilage structure, chondrocyte number and distribution, matrix staining and wet line integrity according to the modified Mankin scoring criteria.
9. Immunohistochemical staining for detecting protein expression condition of Wnt/beta-catenin signal channel related factor and chondrogenic differentiation related factor
Tissue specimens were fixed with 4% paraformaldehyde, decalcified and embedded in paraffin, and sectioned with a paraffin microtome to a thickness of 5-6 μm. The tissue sections were dewaxed and washed with running water, subjected to antigen retrieval using pepsin, washed with PBS solution, treated with endogenous peroxidase blocker, washed with PBS solution, and then washed with PBS after treatment with non-specific staining blocker, and the sections were incubated with Sox9, collagen ii, aggrecan, β -catenin, wnt1, wnt5a, sFRP1 antibodies (dilution 1:400; bioss, beijing, china). Followed by washing with PBS solution, incubation with secondary antibody, treatment with freshly prepared DAB solution. The sites where Sox9, collagen ii, aggrecan, β -catenin, wnt1, wnt5a, sFRP1 proteins were highly expressed were found to be brown-stained under a microscope. Positive areas were photographed using a positive fluorescence microscope and areas of the same size were cut out. Specific values of the brown region were calculated using Image Pro 6.0.
10. Statistical analysis
All metrology data are expressed as mean ± standard deviation (mean ± SD), and independent sample t-test and one-way analysis of variance (ANOVA) are performed using SPSS 26.0 software. P <0.05 indicates that the inter-group differences are statistically significant.
Experimental results
1. Porous magnesium alloy stent morphology and characterization
As shown in FIG. 2 and FIG. 3, we constructed a porous magnesium alloy with a diameter of 2mm and a height of 2mm, and macroscopic observation can show the surface microporous structure. Scanning electron microscope observation further shows that the surfaces of the novel brackets of the pure porous magnesium alloy and the icariin/porous magnesium alloy are respectively provided with a micropore structure, and the surface of the bracket carrying the medicine is coarser than the surface of the bracket of the pure porous magnesium alloy, so that the bracket is ICA loaded on the bracket.
2. Medicine release condition of icariin/porous magnesium alloy biological scaffold
As shown in FIG. 4, when icariin is released from the icariin/porous magnesium alloy biological scaffold material, the release concentration is high in the first 7 days, and reaches a peak value of 23.730. Mu. Mol/L at the 5 th day, after which the drug release concentration gradually decreases during the 7 th to 14 th days, and a small amount of icariin is still released at the 21 st to 28 th days. Icariin/porous magnesium alloy biological material can slowly release icariin.
3. Biological safety of icariin/porous magnesium alloy stent
After the icariin/porous magnesium alloy biological scaffold leaching solution and 0.9% physiological saline are injected into the skin at two sides of the spine, congestion and swelling of the skin at two sides are not seen within 48 hours, and skin swelling disappears. The subcutaneous stimulation test is negative, and proves that the icariin/porous magnesium alloy stent has better in-vitro biological safety and is suitable for subsequent experiments.
4. Rat observations after modeling
All groups of SD rats survived 12 weeks post-surgery. In the observation period, the method finds that all rats have good states, the general vital signs such as food intake, hair condition, stool and urine are not obviously different, the operation wounds are well healed, and the conditions such as wound breakage, blood seepage and liquid seepage, infection purulence and the like are avoided.
5. Macroscopic observation of repair of cartilage defects in knee joints
As shown in fig. 5, the general sample observation shows that the repair condition of the cartilage defect in the group A is the best in three groups, the cartilage defect is filled by milky cartilage tissue, the cartilage surface is smooth, the integrity of intercondylar fossa is basically restored, the color is the same as that of the surrounding cartilage tissue, and the overall cartilage shape is more similar to that of the group D; the defect part of the group B is filled with cartilage-like tissue, but the smoothness and the integrity of the surface of the repaired tissue are lower than those of the group A; the C group cartilage defect is poor in repair, the defect area is large, obvious pits exist on the surface, and the defect is filled with a large amount of similar fibrous tissues.
As shown in fig. 6 and table 1, in the swept and reconstructed 3D image, group a had subchondral bone repair similar to natural bone tissue in the defect area, morphologically closer to group D; the subchondral bone defect of the B group is obvious, but is better than that of the C group, and partial bone repair is still carried out; the subchondral bone defect area in group C is the largest, and there is a significant depression, indicating that the rats themselves have limited intrinsic repairability. We calculated and analyzed the relative bone volume (BV/TV), number of trabeculae (Tb.N), degree of trabecular separation (Tb.Sp), and trabecular thickness (Tb.Th) for each group of defect repair sites. The tb.n values were found to be minimal for group C compared to both A, B groups, and the differences were statistically significant (P < 0.005). The values BV/TV, tb.Sp and Tb.Th were not statistically significant for each group of differences analyzed.
Table 1micro CT detection of the values of the amount of cartilage to repair cartilage defects in each group
Note that: a, group A and group B are different; b, group B and group C are different;
6. microscopic observations of repair of cartilage defects in knee joints
As shown in FIG. 7, we performed decalcification treatment on each group of specimens, and conventional and special staining was performed after tissue sections. The method is used for observing the structure and cell change of each layer of rat femoral cartilage tissue through conventional HE staining, and safranine O solid-green staining and toluidine blue staining are used for observing cartilage defect repair and change of damp line. We find that the cartilage defect in group A is basically repaired, each layer of cartilage has clear structure, smooth surface, normal distribution of chondrocytes, and compact and regular arrangement of subchondral bones. Safranin O and toluidine blue are deeply and uniformly dyed, a tide line is complete and has no blood vessel passing, and the tissue morphology and the dyeing degree are more similar to those of the group D; the cartilage defect in the B group is not completely repaired, part of fibrous tissue remains in subchondral bone, the cartilage surface is slightly rough, chondrocytes are reduced and disordered, subchondral bone is relatively regular, part of the subchondral bone grows at the defect, safranine O and toluidine blue are deeply dyed but unevenly distributed, the dyeing at the middle of the defect is shallow, the joint surface is uneven, and the wet line is discontinuous; the cartilage defect in the group C is obvious, the surface is uneven, the morphology is uneven, each layer of structure of cartilage is indistinguishable, the defect part is mainly covered by a large amount of fibrous tissues, the chondrocytes are few and distributed in clusters, the subchondral bone hardly grows into the defect part, safranine O and toluidine blue are dyed shallowest in the four groups, the number of the chondrocytes is obviously reduced, and the cartilage defect repair level is lowest in the four groups. Each layer of cartilage in the group D has clear structure, smooth cartilage surface and uniform distribution of cartilage cells. Safranin O and toluidine blue are deeply and uniformly dyed, and the tidal line is neat and complete.
As shown in fig. 8, the results of the modified Mankin scores of four knee joints in the A, B, C, D group are respectively 2.400±0.548, 5.375 ±1.109, 10.750 ±0.957 and 0.750±0.500, and the difference is found to have statistical significance (P < 0.0001) compared with the Mankin score of the B, C group in the a group through statistical analysis. The man kin score differed significantly (P < 0.0001) in group B compared to group C. Therefore, the icariin/porous magnesium alloy novel scaffold has the best cartilage defect repairing condition, and the simple porous magnesium alloy has the following composition.
7. Mechanism for detecting knee joint cartilage defect repair by immunohistochemical staining
As shown in fig. 9 and 10, in the immunohistochemical results of Aggrecan, collagen ii and Sox9 chondrogenic related factor, the three protein expression levels of group a were all up-regulated compared to B, C, the difference was statistically significant (P < 0.05), and the three protein expression levels of group a were all up-regulated compared to group B, the difference was statistically significant (P < 0.05); in the result of immunohistochemical of the related factors beta-catenin, wnt5a and Wnt1 of the Wnt/beta-catenin signal pathway, the expression quantity of three proteins in the A group is up-regulated compared with the expression quantity of three proteins in the B, C group, the difference is statistically significant (P is less than 0.05), the expression quantity of three proteins in the A group is up-regulated compared with the expression quantity of three proteins in the B group, and the difference is statistically significant (P is less than 0.05). The novel icariin/porous magnesium alloy scaffold is shown to be capable of realizing cartilage defect repair by activating related factors in Wnt/beta-catenin channels.
TABLE 2 immunohistochemical staining score for each protein in each group of rat knee joints
Note that: a, group A and group B are different; b, group a and group C differ; c, group A and group D differ; d, group B and group C are different; e, group B and group D differ; f, group C and group D differ.
In the description of the present specification, the descriptions of the terms "one embodiment," "example," "specific example," and the like, mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The foregoing has shown and described the basic principles, principal features and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, and that the above embodiments and descriptions are merely illustrative of the principles of the present invention, and various changes and modifications may be made without departing from the spirit and scope of the invention, which is defined in the appended claims.
Claims (6)
1. The alloy material is characterized by comprising a porous magnesium alloy, wherein icariin is carried on the porous magnesium alloy.
2. The preparation method of the icariin-carrying porous magnesium alloy is characterized by comprising the following steps of:
preparing titanium particles into a porous template, preparing a composite material of magnesium alloy and titanium particles by using percolation casting, and finally dissolving titanium filling particles by using hydrofluoric acid to obtain a porous magnesium alloy;
dissolving 10 mu mol/L icariin in absolute ethyl alcohol, soaking the porous magnesium alloy in the icariin solution, taking out and drying, and adsorbing the icariin on the porous magnesium alloy to obtain the icariin-carrying porous magnesium alloy.
3. The method for preparing icariin-carrying porous magnesium alloy and application thereof according to claim 2, wherein the porous template is made of titanium particles by sintering.
4. The preparation and application of icariin-carrying porous magnesium alloy according to claim 2, wherein the diameter of the cylindrical material is 2mm and the height is 2mm.
5. The method for preparing icariin-carrying porous magnesium alloy and application thereof according to claim 2, wherein the time for immersing the porous magnesium alloy in the icariin solution is 1 day.
6. The use of a icariin-carrying porous magnesium alloy as claimed in claim 1 in the preparation of a cartilage defect repair medicament.
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