Magnesium metal-guided bone regeneration membrane and preparation method thereof, product prepared from magnesium metal-guided bone regeneration membrane, and bone repair combination product comprising magnesium metal-guided bone regeneration membrane

Abstract

The invention belongs to the technical field of oral implant materials, and relates to a magnesium metal-guided bone regeneration membrane, a preparation method thereof and a product for preparing the magnesium metal-guided bone regeneration membrane. The magnesium metal-guided bone regeneration membrane comprises a bone repair combination product of the magnesium metal-guided bone regeneration membrane. The preparation method comprises the following steps: respectively providing an electrochemical deposition solution and a polysaccharide-containing solution; the electrochemical deposition liquid comprises a calcium ion donor, a phosphate ion donor and mussel mucin; the method comprises the following steps: placing a magnesium metal matrix in an electrochemical deposition solution, and carrying out first-stage electrochemical deposition under a first electrochemical deposition condition to prepare an intermediate 1; placing the intermediate 1 in an electrochemical deposition solution, performing second-stage electrochemical deposition under a second electrochemical deposition condition, cleaning, and drying to prepare an intermediate 2; placing the intermediate 2 in a solution containing polysaccharide, dip-coating the polysaccharide, and drying; the first electrochemical deposition condition and the second electrochemical deposition condition are respectively as follows: the voltage is 1-5V and 10-60V; the membrane has the biological functions of low corrosion rate, high bonding strength, inflammation resistance, bacteria resistance, osteogenesis promotion and the like.

Description

Technical Field

[0001] This application belongs to the field of oral implant materials technology, specifically relating to magnesium metal guided bone regeneration membranes, their preparation methods, products made therefrom, and bone repair combination products including magnesium metal guided bone regeneration membranes. Background Technology

[0002] Biodegradable magnesium metal has achieved groundbreaking applications in the biomedical field due to its unique biodegradability, biocompatibility, and mechanical adaptability. Implants made from biodegradable magnesium metal, such as cardiovascular stents, orthopedic fixation devices, and guided bone regeneration membranes, have been successfully applied in cardiovascular intervention, bone trauma repair, and oral and maxillofacial regeneration.

[0003] Compared to traditional titanium alloys or polymer materials, biodegradable magnesium metal avoids the trauma of secondary surgery, significantly shortens the patient's recovery period, and reduces the burden on the healthcare system. In particular, biodegradable magnesium metal-guided bone regeneration membranes, with their dynamic matching characteristics between degradation rate and bone tissue regeneration process, have shown potential to replace collagen membranes and titanium meshes, becoming a research hotspot in the field of oral bone defect repair.

[0004] However, magnesium degrades relatively quickly in physiological environments, especially in the high-humidity, high-microbial-load environment of the oral cavity. This rapid degradation can lead to premature loss of mechanical support, failing to maintain spatial stability during the initial stages of bone defect repair. For example, in weight-bearing areas (such as the mandible), excessively rapid degradation may cause premature failure of fixation devices, affecting bone healing. Furthermore, the mismatch between the degradation rate and bone regeneration dynamics can trigger local pH increases and hydrogen accumulation, necessitating surface treatment to reduce the magnesium degradation rate.

[0005] To reduce the degradation rate of magnesium metal, surface treatments (acid / alkali surface treatment, micro-arc oxidation surface treatment, etc.) and surface coatings (inorganic coatings, organic coatings) are commonly used for surface modification. Specifically: hydrofluoric acid surface treatment: forms a magnesium fluoride (MgF2) conversion layer on the magnesium surface through acid etching, significantly improving corrosion resistance and biocompatibility; sodium hydroxide surface treatment: cleans the magnesium metal surface with sodium hydroxide solution, removing oxides and forming a uniform alkaline passivation layer, reducing interference from impurities in subsequent treatments; micro-arc oxidation surface treatment: applies high voltage in an electrolyte, generating a porous magnesium oxide (MgO) coating on the magnesium surface through plasma discharge, improving the corrosion resistance and mechanical properties of magnesium metal; inorganic coatings: such as calcium phosphate coatings prepared by depositing calcium phosphate compounds on the magnesium metal surface through chemical conversion, hydrothermal methods, and electrochemical deposition methods, to reduce the degradation rate of magnesium metal and simulate bone tissue components, promoting osteoblast proliferation and enhancing bone integration performance.

[0006] CN118121773A describes an ultrapure magnesium-guided bone regeneration membrane and its preparation method, which involves immersing the magnesium membrane in a hydrofluoric acid solution with a concentration of 15-29 mol / L for 2-30 hours to prepare a single-layer magnesium fluoride coating with a thickness of 1-3 μm on the surface of the magnesium membrane.

[0007] CN118531384A describes a method for preparing a fine needle-like micro / nano structured calcium phosphate coating on the surface of a biodegradable magnesium alloy. The method involves cleaning the surface of a magnesium film, immersing it in a pre-prepared calcium phosphate solution, and heat-treating it at 92-96℃ for 1-9 hours to form a fine needle-like micro / nano structured calcium phosphate coating with a thickness of 1-20 μm on the surface of the magnesium alloy substrate.

[0008] CN106757251A describes a method for preparing a composite coating on a magnesium alloy surface, wherein a magnesium hydroxide coating with a thickness of 5-50 μm is prepared on the magnesium alloy surface by cathodic electrodeposition. Subsequently, the electrodeposited sample is repeatedly immersed in a polylactic acid solution and subjected to lifting, drying, and coating with an organic coating.

[0009] CN113730655 A describes a medical magnesium alloy barrier membrane for alveolar bone defect repair and its preparation method, including the following steps:

[0010] Step 1: Obtain the pretreated surface by mechanical grinding and chemical polishing.

[0011] For ease of testing, medical magnesium alloy plates or rods were cut into 100mm×100mm×1mm square pieces or Φ100×1mm round pieces using a wire cutting machine. The surfaces to be coated were pretreated by mechanical grinding with sandpaper to remove surface oxide scale and impurities. Then, they were washed with 20g / L NaOH to remove surface oil. Finally, they were ultrasonically cleaned in acetone and anhydrous ethanol for 15 minutes each and dried with cold air.

[0012] Step 2: Laser remelting to prepare the pre-treated surface

[0013] A patterned pretreatment surface was prepared on the medical magnesium alloy pretreatment surface obtained in the first step using a low-power CO2 laser. The laser power was 10-50W, the scanning rate was 200-600mm / s, the single-pass scanning interval was 0.005-0.035mm, and the scanning path was a series of concentric circles or concentric squares.

[0014] Step 3: Micro-arc oxidation to prepare calcium phosphate coating

[0015] A calcium-phosphorus coating was prepared on the pretreated surface of the medical magnesium alloy obtained in the second step using a micro-arc oxidation device. The electrolyte was composed of...

[0016] The electrolyte is composed of Ca(NO3)2, Na3PO4·12H2O, and NaOH. The proportions of each component are adjusted to make the Ca / P ratio of the electrolyte 1.2-1.8 and the pH value 8-11. The electrolyte is kept at 15-30℃ by a circulating water cooling device. A two-stage constant voltage mode is selected. The voltage of the first stage is 250-400V and the treatment time is 10-20min. The voltage of the second stage is 400-500V and the treatment time is 4-8min.

[0017] Step 4: Preparation of chitosan coating by vacuum impregnation and lifting

[0018] A chitosan coating was prepared on the surface of the medical magnesium alloy calcium phosphate coating obtained in the third step using a vacuum impregnation and lifting machine, with a vacuum degree of 10. -2 -10 -3 Pa, chitosan with a purity of 99.5-99.9% and an average molecular weight of 50,000-150,000, is dissolved in a 1:1 mass ratio of dimethylformamide and tetrahydrofuran solution. The solution is magnetically stirred at room temperature for 24 hours until uniformly mixed to prepare a 5-20 wt.% coating solution. The lifting rate is 5-30 cm / min, and the number of lifting cycles is 2-10. After each lifting, the solution is dried in an oven at 80℃ for 20 minutes.

[0019] However, the coating adhesion strength of surface-modified magnesium metal guided bone regeneration membranes prepared using traditional processes needs to be improved. Summary of the Invention

[0020] Based on this, one or more embodiments of this application provide a magnesium metal guided bone regeneration membrane, a method for preparing the same, a product thereof, and a bone repair combination product including the magnesium metal guided bone regeneration membrane. The technical solutions include the following:

[0021] One or more embodiments of this application provide a method for preparing a magnesium metal guided bone regeneration membrane, the preparation method comprising the following steps:

[0022] An electrochemical deposition solution and a polysaccharide-containing solution are provided respectively; the electrochemical deposition solution includes a calcium ion donor, a phosphate ion donor, and mussel adhesive protein;

[0023] A magnesium metal substrate is placed in the electrochemical deposition solution, and a first-stage electrochemical deposition is performed under the first electrochemical deposition conditions to prepare intermediate 1.

[0024] The membrane intermediate 1 was placed in the electrochemical deposition solution, and a second-stage electrochemical deposition was performed under the second electrochemical deposition conditions. After cleaning and drying, intermediate 2 was prepared.

[0025] The intermediate 2 was placed in the polysaccharide-containing solution, coated with polysaccharide, and dried to prepare a magnesium metal guiding bone regeneration membrane;

[0026] in,

[0027] The first electrochemical deposition conditions include: a voltage of 1-5V;

[0028] The first electrochemical deposition conditions include a voltage of 10-60V.

[0029] In some embodiments of this application, the preparation method satisfies one or more of the following conditions:

[0030] (1) The first electrochemical deposition conditions include: keeping the electrochemical deposition solution static, a temperature of 10-60°C, a voltage of 1-5V, and a time of 1-10 min; and,

[0031] (2) The second electrochemical deposition conditions include: stirring the electrochemical deposition solution, with a temperature of 10-60℃, a voltage of 10-60V, and a time of 5-60min;

[0032] Optionally, the stirring speed is 200-400 rpm.

[0033] In some embodiments of this application, the electrochemical deposition solution satisfies one or more of the following conditions:

[0034] 1) The calcium ion donor includes calcium nitrate;

[0035] 2) The phosphate ion donor includes dihydrogen phosphate, optionally calcium dihydrogen phosphate or / and ammonium dihydrogen phosphate;

[0036] 3) The electrochemical deposition solution comprises 0.01M-0.08M of the calcium ion donor, 0.005M-0.08M of the phosphate ion donor, and 0.5mg / mL-2.5mg / mL of mussel adhesive protein;

[0037] 4) The pH of the electrochemical deposition solution is 1-6;

[0038] Optionally, the mussel adhesive protein includes recombinant humanized mussel adhesive protein;

[0039] Optionally, the molecular weight of the mussel adhesive protein is 10-100 kDa.

[0040] In some embodiments of this application, the polysaccharide-containing solution includes one or more of hyaluronic acid, chitosan, sodium alginate, chondroitin sulfate, and heparin.

[0041] Optionally, the polysaccharide-containing solution satisfies one or more of the following conditions:

[0042] (A) The molecular weight of the hyaluronic acid is 10-2000 kDa;

[0043] (B) The molecular weight of the chitosan is 10-300 kDa;

[0044] (C) The molecular weight of the sodium alginate is 50-200 kDa;

[0045] (D) The polysaccharide-containing solution comprises 0.1-5 wt% of the hyaluronic acid;

[0046] (E) The polysaccharide-containing solution comprises 0.1-2 wt% of the chitosan; and,

[0047] (F) The polysaccharide-containing solution comprises 0.1-8 wt% of the sodium alginate.

[0048] In some embodiments of this application, the number of dip coatings is 1-10, the time for each dip coating is 1-15 minutes, and after each dip coating is completed, it is heated to set before proceeding to the next dip coating.

[0049] Optionally, the heat setting conditions include a temperature of 55-65℃ and a time of 3-8 minutes.

[0050] In some embodiments of this application, if the polysaccharide-containing solution satisfies the conditions shown in (A) and (D), then the drying temperature is 20-100°C and the drying time is 10-480 min;

[0051] If the polysaccharide-containing solution meets the conditions shown in (B) and (E), then the drying temperature is 20-60℃ and the drying time is 6-12h.

[0052] If the polysaccharide-containing solution meets the conditions shown in (C) and (F), the drying temperature is 30-60℃ and the drying time is 3-12h. Optionally, the preparation method further includes: after coating the polysaccharide, the obtained intermediate 3 is immersed in a crosslinking agent; optionally, the crosslinking agent includes calcium salt; optionally, the immersion conditions include: the concentration of calcium salt in the immersion system is 4-5wt% and the immersion time is 1-3h.

[0053] In some embodiments of this application, the preparation method satisfies one or more of the following conditions:

[0054] A) The magnesium metal matrix is ​​pretreated as follows: polishing, cleaning and drying;

[0055] Optionally, sanding can be done with 2000-8000 grit sandpaper;

[0056] Optionally, during the pretreatment process, the cleaning includes washing with acetone, ethanol and water in sequence;

[0057] Optionally, the drying temperature is 40-80℃ and the time is 10-60 minutes;

[0058] B) During the preparation of intermediate 2, the cleaning process includes washing with water; and,

[0059] C) During the preparation of intermediate 2, the drying temperature is 40-80℃ and the time is 10-60min.

[0060] One or more embodiments of this application provide a magnesium metal guided bone regeneration membrane, prepared by the preparation method described above.

[0061] One or more embodiments of this application provide a bone repair combination product, the bone repair combination product comprising the magnesium metal guided bone regeneration membrane.

[0062] One or more embodiments of this application provide a product for preparing a magnesium metal guided bone regeneration membrane, the product comprising an electrochemical deposition solution as defined above;

[0063] Optionally, the product may also include a polysaccharide-containing solution as defined above.

[0064] Details of one or more embodiments of this application are set forth in the following description, and other features, objects, and advantages of this application will become apparent from the specification and its claims. Attached Figure Description

[0065] To more clearly illustrate the technical solutions in the embodiments of this application and to more completely understand this application and its beneficial effects, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0066] Figure 1 This is a scanning electron microscope image of the sample surface in Example 1.

[0067] Figure 2 The image shows a scanning electron microscope (SEM) image of the sample surface in Comparative Example 2.

[0068] Figure 3 The image shows a scanning electron microscope (SEM) image of the sample surface in Comparative Example 3.

[0069] Figure 4 This is a scanning electron microscope image of the sample surface in Comparative Example 4.

[0070] Figure 5 Images of the cell morphology of the sample from Example 1, which is magnesium metal.

[0071] Figure 6Images of cell migration in the sample from Example 1, which is magnesium metal.

[0072] Figure 7 Images of stained tissue samples of magnesium metal and coated magnesium metal.

[0073] Figure 8 The graphs show the degradation of hyaluronic acid in samples from Example 1, Comparative Example 1, and Comparative Example 2. Detailed Implementation

[0074] The present application will be further described in detail below with reference to the accompanying drawings, embodiments, and examples. It should be understood that these embodiments and examples are for illustrative purposes only and are not intended to limit the scope of the present application. The purpose of providing these embodiments and examples is to enable a more thorough and comprehensive understanding of the disclosure of the present application. It should also be understood that the present application can be implemented in many different forms and is not limited to the embodiments and examples described herein. Those skilled in the art can make various modifications or alterations without departing from the spirit of the present application, and the equivalent forms obtained also fall within the protection scope of the present application. Furthermore, numerous specific details are set forth in the following description to provide a fuller understanding of the present application. It should be understood that the present application can be implemented without one or more of these details.

[0075] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the specification of this application is for descriptive purposes only and is not intended to be limiting of the application.

[0076] the term

[0077] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0078] The terms "and / or," "or / and," and "and / or" as used herein include any one of two or more of the related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical AND," and also undoubtedly includes technical solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0079] In this application, the terms "multiple", "various", "multiple times", "multi-dimensional", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0080] The terms “combinations of,” “any combination of,” and “any combination of” used in this article include all suitable combinations of any two or more of the listed items.

[0081] In this document, the term "suitable" as used in phrases such as "suitable combination," "suitable method," and "any suitable method" refers to the ability to implement the technical solution of this application, solve the technical problem of this application, and achieve the expected technical effect of this application.

[0082] In this document, terms such as “preferred,” “better,” “more suitable,” and “ideal” are merely used to describe implementation methods or examples that achieve better results, and should be understood not to limit the scope of protection of this application.

[0083] In this application, terms such as "further," "even further," and "particularly" are used to describe purposes and indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0084] In this application, "optionally," "optionally," and "optional" mean that something is optional, that is, it means that it is selected from either "with" or "without." If there are multiple "optional" entries in a technical solution, unless otherwise specified, and there are no contradictions or mutual constraints, each "optional" entry shall be independent.

[0085] In this application, the terms "first aspect," "second aspect," "third aspect," "fourth aspect," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first," "second," "third," "fourth," etc., serve only as a non-exhaustive enumeration and should be understood not to constitute a closed limitation on quantity.

[0086] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0087] In this application, numerical intervals (i.e., numerical ranges) are involved. Unless otherwise specified, the selected numerical distributions within the aforementioned numerical intervals are considered continuous and include the two endpoints (i.e., the minimum and maximum values) of the numerical range, as well as every value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints. In this document, this is equivalent to directly listing every integer. For example, if t is an integer selected from 1 to 10, it means that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. Furthermore, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0088] Unless otherwise specified, the temperature parameters in this application are permitted to be either constant-temperature treatment or variations within a certain temperature range. It should be understood that the constant-temperature treatment allows temperature fluctuations within the precision range of the instrument control, such as ±5℃, ±4℃, ±3℃, ±2℃, or ±1℃.

[0089] In this application, % (w / w) and wt% both represent weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass-volume percentage.

[0090] All references to this application are incorporated herein by reference as if each document were individually incorporated herein by reference. Unless they conflict with the purpose and / or technical solution of this application, all cited references are incorporated herein by reference in their entirety and for all purposes. When references are cited in this application, the definitions of relevant technical features, terms, nouns, phrases, etc., are also incorporated herein by reference. Examples and preferred embodiments of the cited technical features may also be incorporated herein by reference, but only to the extent that they enable the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, this application shall prevail or modifications shall be made adaptably to the description in this application.

[0091] A first aspect of this application provides a method for preparing a magnesium metal guided bone regeneration membrane, the method comprising the following steps:

[0092] An electrochemical deposition solution and a polysaccharide-containing solution are provided respectively; the electrochemical deposition solution includes a calcium ion donor, a phosphate ion donor, and mussel adhesive protein;

[0093] A magnesium metal substrate is placed in the electrochemical deposition solution, and a first-stage electrochemical deposition is performed under the first electrochemical deposition conditions to prepare intermediate 1.

[0094] The membrane intermediate 1 was placed in the electrochemical deposition solution, and a second-stage electrochemical deposition was performed under the second electrochemical deposition conditions. After cleaning and drying, intermediate 2 was prepared.

[0095] The intermediate 2 was placed in the polysaccharide-containing solution, coated with polysaccharide, and dried to prepare a magnesium metal guiding bone regeneration membrane;

[0096] in,

[0097] The first electrochemical deposition conditions include: a voltage of 1-5V (e.g., 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5V);

[0098] The first electrochemical deposition conditions include a voltage of 10-60V (e.g., 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60V).

[0099] In some examples of this application, the preparation method satisfies one or more of the following conditions:

[0100] (1) The first electrochemical deposition conditions include: keeping the electrochemical deposition solution static, a temperature of 10-60°C (e.g., 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60V), a voltage of 1-5V (e.g., 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5V), and a time of 1-10min (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10min); and,

[0101] (2) The second electrochemical deposition conditions include: stirring the electrochemical deposition solution, maintaining a temperature of 10-60℃ (e.g., 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60℃), and a voltage of 10-60V (e.g., 10, 12, 14, 16, 18, 20, 22, 24℃). 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60V), for a time of 5-60 minutes (e.g., 5, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 minutes);

[0102] Optionally, the stirring speed is 200-400 rpm (e.g., 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400 rpm).

[0103] In some examples of this application, the electrochemical deposition solution satisfies one or more of the following conditions:

[0104] 1) The calcium ion donor includes calcium nitrate;

[0105] 2) The phosphate ion donor includes dihydrogen phosphate, optionally calcium dihydrogen phosphate or / and ammonium dihydrogen phosphate;

[0106] 3) The electrochemical deposition solution comprises 0.01M-0.08M (e.g., 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08M) of calcium ion donor, 0.005M-0.08M (e.g., 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08M) of phosphate ion donor, and 0.5-2.5 mg / mL (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5 mg / mL) of mussel adhesive protein;

[0107] 4) The pH of the electrochemical deposition solution is 1-6 (e.g., 1, 2, 3, 4, 5, 6);

[0108] Optionally, the mussel adhesive protein includes recombinant humanized mussel adhesive protein;

[0109] Optionally, the molecular weight of the mussel agaric is 10-100 kDa (e.g., 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 kDa).

[0110] In some examples of this application, the polysaccharide-containing solution includes one or more of hyaluronic acid, chitosan, sodium alginate, chondroitin sulfate, and heparin.

[0111] Optionally, the polysaccharide-containing solution satisfies one or more of the following conditions:

[0112] (A) The molecular weight of the hyaluronic acid is 10-2000 kDa (e.g., 10, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000 kDa);

[0113] (B) The molecular weight of the chitosan is 10-300 kDa (e.g., 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300 kDa);

[0114] (C) The molecular weight of the sodium alginate is 50-200 kDa (e.g., 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200 kDa);

[0115] (D) The polysaccharide-containing solution comprises 0.1-5 wt% (e.g., 0.1 wt%, 0.5 wt%, 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, 5 wt%) of the hyaluronic acid;

[0116] (E) The polysaccharide-containing solution comprises 0.1-2 wt% (e.g., 0.1 wt%, 0.5 wt%, 1 wt%, 1.5 wt%, 2 wt%) of the chitosan; and,

[0117] (F) The polysaccharide-containing solution comprises 0.1-8 wt% (e.g., 0.1 wt%, 0.5 wt%, 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, 3 wt%, 3.5 wt%, 4 wt%, 4.5 wt%, 5 wt%, 5.5 wt%, 6 wt%, 6.5 wt%, 7 wt%, 7.5 wt%, 8 wt%) of the sodium alginate.

[0118] In some examples of this application, the number of dip coatings is 1-10 times (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 times), and the time for each dip coating is 1-15 minutes (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 minutes). After each dip coating is completed, it is heated to set the shape before proceeding to the next dip coating.

[0119] Optionally, the heat setting conditions include: a temperature of 55-65℃ (e.g., 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65℃) and a time of 3-8 min (e.g., 3, 4, 5, 6, 7, 8 min).

[0120] In some examples of this application, the polysaccharide-containing solution satisfies the conditions shown in (A) and (D), and the drying temperature is 20-100℃ (e.g., 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100℃), and the drying time is 10-480 min (e.g., 10, 30, 60, 90, 120, 150, 180, 210, 240, 270, 300, 330, 360, 390, 420, 450, 480 min);

[0121] If the polysaccharide-containing solution satisfies the conditions shown in (B) and (E), then the drying temperature is 20-60℃ (e.g., 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60℃), and the drying time is 6-12h (e.g., 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12h).

[0122] If the polysaccharide-containing solution meets the conditions shown in (C) and (F), then the drying temperature is 30-60℃ (e.g., 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60℃), and the drying time is 3-12h (e.g., 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12h). Optionally, The preparation method further includes: after impregnating the polysaccharide, the obtained intermediate 3 is immersed in a crosslinking agent; optionally, the crosslinking agent includes calcium salt; optionally, the immersion conditions include: the concentration of calcium salt in the immersion system is 4-5 wt% (e.g., 4 wt%, 4.2 wt%, 4.4 wt%, 4.6 wt%, 4.8 wt%, 5 wt%), and the immersion time is 1-3 h (e.g., 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, 3 h).

[0123] In some examples of this application, the preparation method satisfies one or more of the following conditions:

[0124] A) The magnesium metal matrix is ​​pretreated as follows: polishing, cleaning and drying;

[0125] Optionally, sanding can be done with 2000-8000 grit sandpaper;

[0126] Optionally, during the pretreatment process, the cleaning includes washing with acetone, ethanol and water in sequence;

[0127] Optionally, the drying temperature is 40-80℃ (e.g., 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80℃), and the time is 10-60 min (e.g., 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 min);

[0128] B) During the preparation of intermediate 2, the cleaning process includes washing with water; and,

[0129] C) During the preparation of intermediate 2, the drying temperature is 40-80℃ (e.g., 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80℃) and the time is 10-60 min (e.g., 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 min).

[0130] A second aspect of this application provides a magnesium metal guided bone regeneration membrane, prepared by the aforementioned preparation method.

[0131] A third aspect of this application provides a bone repair combination product, the bone repair combination product comprising the aforementioned magnesium metal guided bone regeneration membrane.

[0132] A fourth aspect of this application is a product for preparing a magnesium metal guided bone regeneration membrane, the product comprising an electrochemical deposition solution as defined in the first aspect;

[0133] Optionally, the product may also include a polysaccharide-containing solution as defined in the first aspect.

[0134] A two-step process is used to construct a coating on the surface of magnesium metal that enhances the bioactivity of magnesium metal and promotes the adhesion and growth of osteoblasts, possessing anti-inflammatory, antibacterial, and osteogenic biological functions.

[0135] The biodegradable calcium phosphate-polysaccharide coating for magnesium metal surface with multiple biological functions and its preparation method utilize electrochemical deposition technology. By adjusting the composition of the electrochemical deposition solution and optimizing parameters such as electrochemical deposition voltage and deposition time, a calcium phosphate coating with uniform thickness and good adhesion is prepared on the magnesium metal surface.

[0136] In this application, polysaccharides are coated onto the calcium phosphate coating by dip coating. The polysaccharides used can be hyaluronic acid, chitosan, sodium alginate, heparin, chondroitin sulfate, and other biocompatible polysaccharides and their derivatives that have anti-inflammatory, antibacterial, and osteopromoting biological functions.

[0137] The embodiments of this application will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this application, or follow experimental manuals or conventional conditions in the art, or follow the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0138] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0139] This application employs electrochemical deposition technology. By optimizing the composition of the calcium phosphate deposition solution, deposition voltage, and deposition time, a calcium phosphate coating with uniform thickness, good adhesion, and the ability to extend the retention time of the outer layer material is stably prepared on the magnesium metal surface.

[0140] Polysaccharide substances with antibacterial, anti-inflammatory, and osteogenic biological functions are selected. By using a dip-coating method, the concentration, composition, and number of dip-coating solutions are optimized to coat the calcium-phosphorus coating with a polysaccharide coating. This enhances the bioactivity of magnesium metal and promotes the adhesion and growth of osteoblasts, thus endowing it with antibacterial, anti-inflammatory, and osteogenic biological functions.

[0141] Example 1

[0142] This embodiment provides a biodegradable calcium phosphorus-hyaluronic acid coating for magnesium metal surfaces with anti-inflammatory and bone-promoting functions, and its preparation method, mainly including the following steps:

[0143] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0144] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.016mol / L, Ca(H2PO4)2·H2O is 0.0067mol / L, and mussel adhesive protein is 1mg / mL.

[0145] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 4.5 and 6.

[0146] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 3V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, complete the first stage of electrochemical deposition, and prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal, that is, prepare intermediate 1.

[0147] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0148] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample, which is the preparation intermediate 2.

[0149] Step 7: Prepare a hyaluronic acid solution with a mass concentration of 0.5% using hyaluronic acid with a molecular weight of 150kDa (Bloomage Biotechnology Co., Ltd., HA-EP2).

[0150] Step 8: Immerse the calcium-phosphorus coated magnesium metal sample prepared in Step 6 in the hyaluronic acid solution prepared in Step 7. After soaking for 60 seconds, slowly remove the sample and treat it at a drying temperature of 60°C for 5 minutes to complete one dip-coating. Repeat the above dip-coating operation 3 times, and then dry the obtained sample at a temperature of 60°C for 6 hours to obtain a calcium-phosphorus-hyaluronic acid coated biodegradable magnesium metal product with anti-inflammatory and bone-promoting functions.

[0151] Example 2

[0152] This embodiment provides a biodegradable calcium phosphorus-hyaluronic acid coating for magnesium metal surfaces with anti-inflammatory and bone-promoting functions, and its preparation method, mainly including the following steps:

[0153] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0154] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.016mol / L, Ca(H2PO4)2·H2O is 0.0067mol / L, and mussel adhesive protein is 1mg / mL.

[0155] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 4.5 and 6.

[0156] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 3V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0157] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0158] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0159] Step 7: Prepare a hyaluronic acid solution with a mass concentration of 0.5% using hyaluronic acid with a molecular weight of 2000kDa (Bloomage Biotechnology Co., Ltd., HA-EP2).

[0160] Step 8: Immerse the calcium-phosphorus coated magnesium metal sample prepared in Step 6 in the hyaluronic acid solution prepared in Step 7. After immersion for 60 seconds, slowly remove the sample and treat it at a drying temperature of 60°C for 5 minutes to complete one dip-coating. Repeat the above dip-coating operation 3 times, and then dry the sample at a temperature of 60 minutes for 6 hours to obtain a calcium-phosphorus-hyaluronic acid coated biodegradable magnesium metal product with anti-inflammatory and bone-promoting functions.

[0161] Example 3

[0162] This embodiment provides a biodegradable calcium phosphorus-hyaluronic acid coating for magnesium metal surfaces with anti-inflammatory and bone-promoting functions, and its preparation method, mainly including the following steps:

[0163] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0164] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.016mol / L, Ca(H2PO4)2·H2O is 0.0067mol / L, and mussel adhesive protein is 1mg / mL.

[0165] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 4.5 and 6.

[0166] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 3V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0167] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0168] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0169] Step 7: Prepare a 5% hyaluronic acid solution using hyaluronic acid with a molecular weight of 150 kDa (Bloomage Biotechnology Co., Ltd., HA-EP2).

[0170] Step 8: Immerse the calcium-phosphorus coated magnesium metal sample prepared in Step 6 in the prepared hyaluronic acid solution. After soaking for 60 seconds, slowly remove the sample and treat it at a drying temperature of 60°C for 5 minutes to complete one dip-coating. Repeat the above dip-coating operation 3 times, and then dry the sample at 60°C for 6 hours to obtain a calcium-phosphorus-hyaluronic acid coated biodegradable magnesium metal product with anti-inflammatory and bone-promoting functions.

[0171] Example 4

[0172] This embodiment provides a biodegradable calcium phosphate-chitosan coating for magnesium metal surfaces with antibacterial and osteopromoting functions, and its preparation method, mainly including the following steps:

[0173] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0174] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.016mol / L, Ca(H2PO4)2·H2O is 0.0067mol / L, and mussel adhesive protein is 1mg / mL.

[0175] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 4.5 and 6.

[0176] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 3V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0177] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0178] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0179] Step 7: Prepare a 0.5% chitosan solution using chitosan with a molecular weight of 150 kDa (Macklin, C804726).

[0180] Step 8: Immerse the calcium-phosphorus coated magnesium metal sample prepared in Step 6 in the prepared chitosan solution, soak for 60 seconds, then slowly remove it and treat it at a drying temperature of 60℃ for 5 minutes to complete one dip coating.

[0181] After repeating the above dip-coating operation three times, the sample was dried at 60°C for 6 hours to obtain a calcium phosphorus-chitosan coating biodegradable magnesium metal product with antibacterial and bone-promoting functions.

[0182] Example 5

[0183] This embodiment provides a biodegradable calcium phosphate-sodium alginate coating for magnesium metal surfaces and its preparation method, mainly including the following steps:

[0184] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0185] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.016mol / L, Ca(H2PO4)2·H2O is 0.0067mol / L, and mussel adhesive protein is 1mg / mL.

[0186] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 4.5 and 6.

[0187] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 3V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0188] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0189] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0190] Step 7: Prepare a sodium alginate solution with a mass concentration of 0.5% using sodium alginate with a molecular weight of 150 kDa (Macklin, S817373).

[0191] Step 8: Immerse the calcium phosphate coated magnesium metal sample in the prepared sodium alginate solution, soak for 60 seconds, then slowly remove it and treat it at a drying temperature of 60°C for 5 minutes to complete one dip coating. Repeat the above dip coating operation 3 times.

[0192] Step 9: Prepare a 5% (w / w) CaCl2 solution using anhydrous calcium chloride (CaCl2, Macklin, C885902). Immerse the sample in the CaCl2 solution for 2 hours, then rinse the sample with purified water. Finally, dry the sample at 60°C for 6 hours to obtain a calcium phosphate-sodium alginate coated biodegradable magnesium metal product.

[0193] Example 6

[0194] This embodiment provides a biodegradable calcium phosphorus-hyaluronic acid coating for magnesium metal surfaces with anti-inflammatory and bone-promoting functions, and its preparation method, mainly including the following steps:

[0195] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0196] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.032mol / L, Ca(H2PO4)2·H2O is 0.0134mol / L, and mussel adhesive protein is 0.5mg / mL.

[0197] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 1 and 3.

[0198] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 10±1℃, fix the deposition voltage to 5V and the deposition time to 1min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0199] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 60±1℃, fix the deposition voltage at 30V and the deposition time at 30min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0200] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0201] Step 7: Prepare a hyaluronic acid solution with a mass concentration of 0.1% using hyaluronic acid with a molecular weight of 10 kDa (Bloomage Biotechnology Co., Ltd., HA-EP2).

[0202] Step 8: Immerse the calcium-phosphorus coated magnesium metal sample prepared in Step 6 in the hyaluronic acid solution prepared in Step 7. After soaking for 10 minutes, slowly remove the sample and treat it at a drying temperature of 55°C for 3 minutes to complete one dip-coating. After dip-coating, dry the obtained sample at a temperature of 20°C for 8 hours to obtain a calcium-phosphorus-hyaluronic acid coated biodegradable magnesium metal product with anti-inflammatory and bone-promoting functions.

[0203] Example 7

[0204] This embodiment provides a biodegradable calcium phosphorus-hyaluronic acid coating for magnesium metal surfaces with anti-inflammatory and bone-promoting functions, and its preparation method, mainly including the following steps:

[0205] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0206] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein are 0.08 mol / L and 2.5 mg / mL, respectively.

[0207] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 3 and 4.5.

[0208] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 60±1℃, fix the deposition voltage to 1V and the deposition time to 10min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0209] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 10±1℃, fix the deposition voltage at 60V and the deposition time at 60min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0210] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0211] Step 7: Prepare a 5% hyaluronic acid solution using hyaluronic acid with a molecular weight of 1000 kDa (Bloomage Biotechnology Co., Ltd., HA-EP2).

[0212] Step 8: Immerse the calcium-phosphorus coated magnesium metal sample prepared in Step 6 in the hyaluronic acid solution prepared in Step 7, soak for 15 minutes, then slowly remove it and treat it at a drying temperature of 65°C for 8 minutes to complete one dip coating.

[0213] After repeating the above dip-coating operation 9 times, the obtained sample was dried at 100°C for 10 minutes to obtain a calcium phosphorus-hyaluronic acid coated biodegradable magnesium metal product with anti-inflammatory and bone-promoting functions.

[0214] Example 8

[0215] This embodiment provides a biodegradable calcium phosphate-chitosan coating for magnesium metal surfaces with antibacterial and osteopromoting functions, and its preparation method, mainly including the following steps:

[0216] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0217] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.032mol / L, Ca(H2PO4)2·H2O is 0.0134mol / L, and mussel adhesive protein is 0.5mg / mL.

[0218] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 1 and 3.

[0219] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 10±1℃, fix the deposition voltage to 5V and the deposition time to 1min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0220] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 60±1℃, fix the deposition voltage at 30V and the deposition time at 30min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0221] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0222] Step 7: Prepare a 0.1% chitosan solution using chitosan with a molecular weight of 10 kDa (Macklin, C804726).

[0223] Step 8: Immerse the calcium-phosphorus coated magnesium metal sample prepared in Step 6 in the prepared chitosan solution. After soaking for 10 minutes, slowly remove the sample and treat it at a drying temperature of 55℃ for 3 minutes to complete one dip-coating. After dip-coating, dry the sample at 20℃ for 8 hours to obtain a calcium-phosphorus-chitosan coated biodegradable magnesium metal product with antibacterial and osteopromoting functions.

[0224] Example 9

[0225] This embodiment provides a biodegradable calcium phosphate-chitosan coating for magnesium metal surfaces with antibacterial and osteopromoting functions, and its preparation method, mainly including the following steps:

[0226] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0227] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein are 0.08 mol / L and 2.5 mg / mL, respectively.

[0228] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 3 and 4.5.

[0229] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 60±1℃, fix the deposition voltage to 1V and the deposition time to 10min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0230] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 10±1℃, fix the deposition voltage at 60V and the deposition time at 60min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0231] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0232] Step 7: Prepare a 2% chitosan solution using chitosan (Macklin, C804726) with a molecular weight of 300 kDa.

[0233] Step 8: Immerse the calcium-phosphorus coated magnesium metal sample prepared in Step 6 in the prepared chitosan solution, soak for 15 minutes, then slowly remove it and treat it at a drying temperature of 65°C for 8 minutes to complete one dip coating.

[0234] After repeating the above dip-coating operation 9 times, the sample was dried at 60°C for 12 hours to obtain a calcium phosphate-chitosan coating biodegradable magnesium metal product with antibacterial and bone-promoting functions.

[0235] Example 10

[0236] This embodiment provides a biodegradable calcium phosphate-sodium alginate coating for magnesium metal surfaces and its preparation method, mainly including the following steps:

[0237] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0238] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.032mol / L, Ca(H2PO4)2·H2O is 0.0134mol / L, and mussel adhesive protein is 0.5mg / mL.

[0239] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 1 and 3.

[0240] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 10±1℃, fix the deposition voltage to 5V and the deposition time to 1min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0241] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 60±1℃, fix the deposition voltage at 30V and the deposition time at 30min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0242] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0243] Step 7: Prepare a sodium alginate solution with a mass concentration of 0.1% using sodium alginate with a molecular weight of 50 kDa (Macklin, S817373).

[0244] Step 8: Immerse the calcium phosphate coated magnesium metal sample in the prepared sodium alginate solution, soak for 10 minutes, then slowly remove it and treat it at a drying temperature of 55°C for 3 minutes to complete one dip coating.

[0245] Step 9: Prepare a 5% (w / w) CaCl2 solution using anhydrous calcium chloride (CaCl2, Macklin, C885902). Immerse the sample in the CaCl2 solution for 2 hours, then rinse the sample with purified water. Finally, dry the sample at 30°C for 3 hours to obtain a calcium phosphate-sodium alginate coated biodegradable magnesium metal product.

[0246] Example 11

[0247] This embodiment provides a biodegradable calcium phosphate-sodium alginate coating for magnesium metal surfaces and its preparation method, mainly including the following steps:

[0248] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0249] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein are 0.08 mol / L and 2.5 mg / mL, respectively.

[0250] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 3 and 4.5.

[0251] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 60±1℃, fix the deposition voltage to 1V and the deposition time to 10min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0252] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 10±1℃, fix the deposition voltage at 60V and the deposition time at 60min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0253] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0254] Step 7: Prepare a 5% sodium alginate solution using sodium alginate with a molecular weight of 200 kDa (Macklin, S817373).

[0255] Step 8: Immerse the calcium phosphate coated magnesium metal sample in the prepared sodium alginate solution, soak for 15 minutes, then slowly remove it and treat it at a drying temperature of 65°C for 8 minutes to complete one dip coating. Repeat the above dip coating operation 9 times.

[0256] Step 9: Prepare a 5% (w / w) CaCl2 solution using anhydrous calcium chloride (CaCl2, Macklin, C885902). Immerse the sample in the CaCl2 solution for 2 hours, then rinse the sample with purified water. Finally, dry the sample at 50°C for 12 hours to obtain a calcium phosphate-sodium alginate coated biodegradable magnesium metal product.

[0257] Example 12

[0258] This embodiment provides a biodegradable calcium phosphorus-tea polyphenol coating on a magnesium metal surface and its preparation method, mainly including the following steps:

[0259] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0260] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.016mol / L, Ca(H2PO4)2·H2O is 0.0067mol / L, and mussel adhesive protein is 1mg / mL.

[0261] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 4.5 and 6.

[0262] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 3V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0263] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0264] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0265] Step 7: Prepare a 0.25 mg / L tea polyphenol solution using tea polyphenols (Macklin, T821916).

[0266] Step 8: Immerse the calcium-phosphorus coated magnesium metal sample prepared in Step 6 in the tea polyphenol solution prepared in Step 7. After soaking for 60 seconds, slowly remove the sample and treat it at a drying temperature of 60°C for 5 minutes to complete one dip coating.

[0267] After repeating the above dip-coating operation three times, the resulting sample was dried at 60°C for 6 hours to obtain a calcium phosphorus-tea polyphenol-coated biodegradable magnesium metal product.

[0268] Comparative Example 1

[0269] This comparative example provides a biodegradable calcium phosphorus-hyaluronic acid coating for magnesium metal surfaces with anti-inflammatory and osteopromoting functions, and its preparation method. Compared with Example 1, step 4 is omitted in this comparative example, and the main steps are as follows:

[0270] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0271] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.016mol / L, Ca(H2PO4)2·H2O is 0.0067mol / L, and mussel adhesive protein is 1mg / mL.

[0272] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 4.5 and 6.

[0273] Step 4: Omitted.

[0274] Step 5: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 10min, and stir at a stirring speed of about 300rpm during the deposition process to keep the calcium-phosphorus deposition solution uniform.

[0275] Step 6: Place the magnesium metal with the electrochemically deposited calcium-phosphorus coating in purified water and wash it 5 times to remove the poorly adhered calcium-phosphorus compounds. Dry it at 60°C for 30 minutes to obtain the calcium-phosphorus coated magnesium metal sample.

[0276] Step 7: Prepare a hyaluronic acid solution with a mass concentration of 0.5% using hyaluronic acid with a molecular weight of 150kDa (Bloomage Biotechnology Co., Ltd., HA-EP2).

[0277] Step 8: Immerse the calcium phosphate coated magnesium metal sample in the hyaluronic acid solution prepared in Step 7, soak for 60 seconds, then slowly remove it and treat it at a drying temperature of 60°C for 5 minutes to complete one dip coating.

[0278] After repeating the above dip-coating operation three times, the sample was dried at 60°C for 6 hours to obtain a calcium phosphorus-hyaluronic acid coating biodegradable magnesium metal product with anti-inflammatory and bone-promoting functions.

[0279] Comparative Example 2

[0280] This comparative example prepares a calcium-phosphorus coating on a magnesium metal surface by electrochemical deposition. Compared with Example 1, this comparative example omits mussel adhesive protein in step 2 and mainly includes the following steps:

[0281] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0282] Step 2: Prepare a calcium-phosphorus precipitation solution using Ca(NO3)2·4H2O and Ca(H2PO4)2·H2O, wherein the concentration of Ca(NO3)2·4H2O is 0.016 mol / L and the concentration of Ca(H2PO4)2·H2O is 0.0067 mol / L.

[0283] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 4.5 and 6.

[0284] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 3V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition.

[0285] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0286] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0287] Step 7: Prepare a hyaluronic acid solution with a mass concentration of 0.5% using hyaluronic acid with a molecular weight of 150kDa (Bloomage Biotechnology Co., Ltd., HA-EP2).

[0288] Step 8: Immerse the calcium-phosphorus coated magnesium metal sample prepared in Step 6 in the prepared hyaluronic acid solution, soak for 60 seconds, then slowly remove it and treat it at a drying temperature of 60℃ for 5 minutes to complete one dip coating.

[0289] After repeating the above dip-coating operation three times, the sample was dried at 60°C for 6 hours to obtain a calcium phosphorus-hyaluronic acid coating biodegradable magnesium metal product with anti-inflammatory and bone-promoting functions.

[0290] Comparative Example 3

[0291] This comparative example prepares a calcium phosphate coating on a magnesium metal surface by electrochemical deposition. Compared with Example 1, steps 7 and 8 are omitted in this comparative example, and the main steps are as follows:

[0292] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0293] Step 2: Prepare a calcium-phosphorus precipitation solution containing mussel adhesive protein using Ca(NO3)2·4H2O, Ca(H2PO4)2·H2O, and mussel adhesive protein (Jinpu Nuoan, GPE009001, molecular weight 16.06kDa). The concentrations of Ca(NO3)2·4H2O are 0.016mol / L, Ca(H2PO4)2·H2O is 0.0067mol / L, and mussel adhesive protein is 1mg / mL.

[0294] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 4.5 and 6.

[0295] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 3V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0296] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0297] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0298] Steps 7 and 8 are omitted.

[0299] Comparative Example 4

[0300] This comparative example prepares a calcium-phosphorus coating on a magnesium metal surface using an electrochemical deposition method. Compared to Example 1, this comparative example omits mussel adhesive protein in step 2, and also omits steps 7 and 8. The main steps are as follows:

[0301] Step 1: Perform surface pretreatment on magnesium metal. The surface pretreatment steps are as follows: grinding (using 2000-8000# sandpaper), cleaning (using acetone, ethanol, and purified water in sequence), and drying at 60℃ for 30 minutes.

[0302] Step 2: Prepare a calcium-phosphorus precipitation solution using Ca(NO3)2·4H2O and Ca(H2PO4)2·H2O, wherein the concentration of Ca(NO3)2·4H2O is 0.016 mol / L and the concentration of Ca(H2PO4)2·H2O is 0.0067 mol / L.

[0303] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid to bring the pH of the calcium phosphate deposition solution to between 4.5 and 6.

[0304] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 3V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0305] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0306] Step 6: Place the magnesium metal after the second-stage electrochemical deposition of calcium phosphate coating in purified water and wash it 5 times to remove the poorly adhered calcium phosphate compounds. Dry it at 60°C for 30 minutes to obtain the calcium phosphate coated magnesium metal sample.

[0307] Steps 7 and 8 are omitted.

[0308] Comparative Example 5

[0309] This embodiment is a comparative example of Example 1. The only difference from Example 1 is that the fixed deposition voltage is the same for the first stage electrochemical deposition and the second stage electrochemical deposition. Specifically:

[0310] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 3V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0311] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 3V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0312] Comparative Example 6

[0313] This embodiment is a comparative example of Example 1. The only difference from Example 1 is that the fixed deposition voltage is the same for the first stage electrochemical deposition and the second stage electrochemical deposition. Specifically:

[0314] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, fix the deposition voltage to 10V and the deposition time to 3min, keep the calcium-phosphorus deposition solution still during the deposition process, and complete the first stage of electrochemical deposition to prepare a uniform adsorption layer of mussel adhesive protein on the surface of magnesium metal.

[0315] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0316] Comparative Example 7

[0317] This embodiment is a comparative example of Example 1. The only difference from Example 1 is that step 4 uses a dip-coating method instead of an electrochemical deposition method. Specifically:

[0318] Step 4: Place the surface-pretreated magnesium metal into the calcium-phosphorus deposition solution prepared in Step 3, adjust the temperature of the calcium-phosphorus deposition solution to 15±1℃, and immerse for 120 minutes. Keep the calcium-phosphorus deposition solution still during the deposition process to prepare a mussel adhesive protein adsorption layer on the surface of the magnesium metal.

[0319] Comparative Example 8

[0320] This embodiment is a comparative example of Embodiment 1, and the only difference from Embodiment 1 is:

[0321] Step 2: Prepare a calcium-phosphorus precipitation solution using Ca(NO3)2·4H2O and Ca(H2PO4)2·H2O, wherein the concentration of Ca(NO3)2·4H2O is 0.016 mol / L and the concentration of Ca(H2PO4)2·H2O is 0.0067 mol / L; at the same time, prepare a protein solution with a mussel adhesive protein concentration of 1 mg / mL.

[0322] Step 3: Adjust the pH of the calcium phosphate deposition solution using acetic acid, so that the pH of the calcium phosphate deposition solution and the protein solution is between 4.5 and 6.

[0323] In step 4, the pretreated magnesium metal is placed into the protein solution prepared in step 3. The temperature of the protein solution is adjusted to 15±1℃, the deposition voltage is fixed at 3V and the deposition time is 3min. The solution is kept still during the deposition process to complete the first stage of electrochemical deposition and prepare a uniform adsorption layer of mussel adhesive protein on the surface of the magnesium metal.

[0324] Step 5: Place the magnesium metal that has completed the first stage of electrochemical deposition into the calcium phosphorus deposition solution prepared in Step 3. Adjust the temperature of the calcium phosphorus deposition solution to 25±1℃, fix the deposition voltage at 10V and the deposition time at 7min. During the deposition process, stir at a stirring speed of about 300rpm to keep the calcium phosphorus deposition solution uniform. This completes the second stage of electrochemical deposition.

[0325] I. Testing Method:

[0326] 1. Scanning electron microscopy (SEM) detection method

[0327] Cut the sample to an appropriate size, remove contaminants, and ensure it is completely dry. Sputter-coat the sample surface (gold spray) to avoid the charging effect. Securely mount the sample on the sample stage and place it in the SEM sample chamber. After evacuating the chamber, set the accelerating voltage to 3kV and use a secondary electron detector to observe the surface morphology of the coating, noting its uniformity, density, and the presence of pores, cracks, etc.

[0328] 2. Electrochemical corrosion performance testing methods

[0329] The corrosion of magnesium metal samples in biological simulated fluid (SBF) was analyzed using an electrochemical workstation with a three-electrode system. The sample was used as the working electrode, a platinum sheet as the counter electrode, and a saturated calomel electrode as the reference electrode. Potentiodynamic planning was performed at a potential scan rate of 1 mV / s and a relative open-circuit voltage of ±0.4 V. The corrosion current density, corrosion voltage, and corrosion rate of different samples were estimated using the Tafel method.

[0330] 3. Cytotoxicity detection methods

[0331] Cell preparation: Mouse fibroblasts were revived in MEM medium containing 10% FBS and cultured at 37°C, 5% CO2, and saturated humidity. After 2-3 passages until the cells reached the logarithmic growth phase, the cells were digested with trypsin, collected, and the cell concentration was adjusted to 1×10⁶ cells / year. 5 The following experiments were conducted using cells / mL.

[0332] Preparation of extraction solution: Using MEM medium as the extraction medium, the samples were placed at 6cm intervals. 2 The extraction medium was added at a ratio of / mL, and the extraction was carried out at 37℃ and 125rpm for 72h.

[0333] Preparation of control samples: Negative samples were prepared by extracting MEM medium containing 10% FBS into a 50 mL sterile centrifuge tube at 37°C with shaking for 72 h. Positive samples were prepared using MEM complete medium containing 10% DMSO. Blank samples were prepared using MEM medium containing 10% FBS.

[0334] Experimental method: 100 μL of cell suspension was added to each well of a 96-well plate, for a total of 1 × 10⁶ cells / well. 4 Cells per well were cultured at 37°C and 5% CO2 for 24 hours. After culture, the culture medium was discarded, and sample extract, negative control, blank control, and positive control were added to each well, with two parallel controls per group and eight wells per control. Cells were then incubated at 37°C, 5% CO2, and saturated humidity for 24 hours, followed by microscopic examination and photographic recording of cell morphology. The culture medium was then discarded, and MTT staining agent was added to each well. Cells were then incubated at 37°C, 5% CO2, and saturated humidity for 3 hours. After culture, the liquid in the culture plate was discarded, and isopropanol was added to each well. The plates were mixed by shaking in the dark for 30 minutes and then analyzed using a microplate reader at a detection wavelength of 570 nm and a reference wavelength of 650 nm.

[0335] 4. In vitro cell migration assay methods:

[0336] Preparation of extraction solution: Using MEM medium as the extraction medium, the samples were placed at 6cm intervals. 2 The extraction medium was added at a ratio of / mL, and the sample was extracted for 72 hours under the conditions of 37℃ and 125rpm oscillation to obtain the sample extract.

[0337] Experimental Methods: Cell suspension was added to 6-well plates and cultured at 37°C, 5% CO2, and saturated humidity until cell confluence reached 90%. The culture medium was removed, creating a slit. Exfoliated cells were washed three times with PBS, and cell morphology was recorded under a microscope. 3 mL of sample extract was added to each well, and the plates were incubated at 37°C, 5% CO2, and saturated humidity for 20 h. Cell morphology was then recorded under a microscope to compare the effects of different samples on cell migration ability.

[0338] 5. Histological staining experimental methods

[0339] Sample preparation: After cleaning with ethanol for 10 min, the sample was cleaned three times with water for injection (5 min each time), dried, and then sterilized by irradiation with a 25 KGy electron beam.

[0340] Experimental Methods: SD rats were anesthetized by intraperitoneal injection of sodium pentobarbital solution. Before surgery, the top of the skull was prepared and disinfected using an electric razor. A sagittal incision was made in the skin of the left skull using ophthalmic scissors. The subcutaneous fascia and periosteum were bluntly dissected using a periosteal separator to fully expose the bone surface. A bone defect was then created in the top of the skull using a trephine. The sample was implanted into the defect, and the incision was sutured. Postoperatively, the rats were kept in solitary confinement and treated with anti-infection measures. The rats were sacrificed 28 days postoperatively, and skull samples were collected. The skull samples were fixed at the implantation site with 4% paraformaldehyde, decalcified, stained with hematoxylin and eosin (HE), and examined under a microscope for photographic morphology recording.

[0341] 6. Detection method for degradation of hyaluronic acid coating

[0342] Sample preparation: After cleaning the sample with ethanol for 10 min, it was cleaned 3 times with water for injection (5 min / time), dried, and then sterilized by irradiation with a 25 KGy electron beam.

[0343] Experimental method: with 5 mL / cm 2 The sample was immersed in simulated saliva at a specific ratio, and the system was placed in a constant-temperature shaker set at 37℃ and a shaking speed of 70 rpm. At 0.5h, 2h, 4h, 8h, 12h, and 24h, 1 mL of the extract was collected and 1 mL of fresh extract was added. Finally, the concentration of hyaluronic acid in the extract was measured, and a line graph of concentration change was plotted.

[0344] 7. Coating Adhesion Strength Testing Method

[0345] Sample preparation: Prepare a coating on the magnesium film surface, with a coating length of not less than 15 mm; measure and record the coating thickness using a micrometer; apply an adhesive to the uncoated magnesium film surface; in accordance with GB / T 23101.4-2023 Surgical Implants Hydroxyapatite Part 4: Determination of Coating Bond Strength and ISO 4587-2003 standards, bond the coated magnesium film to the magnesium film coated with adhesive, ensuring an overlap length of 12.5 ± 0.5 mm.

[0346] Experimental method: A tensile load was applied at a constant loading rate of 2.5 ± 0.5 mm / min until the parts were completely separated, and the maximum applied load was recorded; Coating adhesion strength calculation: σ = F / A

[0347] σ: Coating adhesion strength, measured in megapascals (MPa)

[0348] F: Maximum load, in Newtons (N).

[0349] A: Area of ​​the overlapping portion, in square millimeters (mm) 2 )

[0350] II. Test Results

[0351] like Figure 1 As shown in the scanning electron microscope image of the sample surface in Example 1, the rough surface structure of the calcium phosphate coating was covered after coating with hyaluronic acid.

[0352] like Figure 2 As shown in Comparative Example 2, after coating the calcium phosphate coating with hyaluronic acid using the same process, the hyaluronic acid also covered the rough surface of the calcium phosphate coating, but the exposed portion of the calcium phosphate coating was larger. This indicates that whether the calcium phosphate deposition solution contains mussel adhesive protein has a certain impact on the subsequent preparation of the hyaluronic acid layer. The presence of mussel adhesive protein in the calcium phosphate coating can enhance the binding of the hyaluronic acid layer.

[0353] like Figure 3 As shown, in Comparative Example 3, a dense and rough calcium phosphate coating was uniformly deposited on the surface of magnesium metal by electrochemical deposition.

[0354] like Figure 4 As shown, the sample surface in Comparative Example 4 also has a rough calcium-phosphorus coating, but compared with the sample in Comparative Example 3 with added mussel adhesive protein, the calcium-phosphorus coating on the surface of the sample in Comparative Example 4 has a sheet-like structure and the coating density is significantly reduced.

[0355] Table 1. Electrochemical corrosion performance data of samples

[0356] Sample <![CDATA[Corrosion current density (mA / cm 2 )]]> Corrosion rate (mm / year) Bare magnesium metal <![CDATA[0.832×10 -2 ]]> 0.0214 Example 1 <![CDATA[6.48×10 -4 ]]> 0.0017 Example 2 <![CDATA[1.37×10 -4 ]]> 0.0014 Example 3 <![CDATA[8.71×10 -4 ]]> 0.0022 Example 4 <![CDATA[8.87×10 -5 ]]> 0.0012 Example 5 <![CDATA[2.58×10 -4 ]]> 0.0014 Example 6 <![CDATA[8.48×10 -4 ]]> 0.0027 Example 7 <![CDATA[7.51×10 -4 ]]> 0.0021 Example 8 <![CDATA[8.91×10 -5 ]]> 0.0013 Example 9 <![CDATA[8.88×10 -5 ]]> 0.0012 Example 10 <![CDATA[3.51×10 -4 ]]> 0.0014 Example 11 <![CDATA[4.23×10 -4 ]]> 0.0015 Example 12 <![CDATA[8.14×10 -4 ]]> 0.0031 Comparative Example 1 <![CDATA[3.09×10 -3 ]]> 0.0058 Comparative Example 2 <![CDATA[6.89×10 -3 ]]> 0.0091 Comparative Example 3 <![CDATA[9.82×10 -4 ]]> 0.0035 Comparative Example 4 <![CDATA[7.42×10 -3 ]]> 0.0121 Comparative Example 5 <![CDATA[1.73×10 -3 ]]> 0.0044 Comparative Example 6 <![CDATA[1.72×10 -3 ]]> 0.0044 Comparative Example 7 <![CDATA[1.96×10 -3 ]]> 0.0078 Comparative Example 8 <![CDATA[1.09×10 -3 ]]> 0.0066

[0357] As shown in Table 1, the corrosion rate of Example 1 was reduced by 92.05% compared to bare magnesium metal, and the corrosion resistance of the coated sample was significantly improved; the corrosion rate of Example 1 was reduced by 82.35% compared to Example 12; the corrosion rate of Example 1 was reduced by 241.18% compared to Comparative Example 1; the corrosion rate of Example 1 was reduced by 435.29% compared to Comparative Example 2; the corrosion rate of Example 1 was reduced by 105.88% compared to Comparative Example 3; the corrosion rate of Example 1 was reduced by 611.76% compared to Comparative Example 4; the corrosion rate of Example 1 was reduced by 158.82% compared to Comparative Example 5; the corrosion rate of Example 1 was reduced by 158.82% compared to Comparative Example 6; the corrosion rates of Examples 2-11 were all significantly reduced compared to Example 12, and the corrosion resistance was significantly improved.

[0358] The above results indicate that: 1) The coating sample obtained through two-stage electrochemical deposition has the lowest corrosion degradation rate, meaning that Example 1 exhibits the best corrosion resistance. 2) The sample with added mussel adhesive protein in the calcium-phosphorus deposition solution has an even lower corrosion degradation rate due to the enhanced adhesion and density of the calcium-phosphorus coating; the addition of mussel adhesive protein can improve its corrosion resistance.

[0359] like Figure 5 As shown, the magnesium metal sample cells were shrunken and detached from their cell walls, with a relative cell proliferation rate of only 19% and a cytotoxicity grade of 4. The cells in Example 1 showed no morphological changes, but the relative cell proliferation rate reached 87%, and the cytotoxicity grade was 1. After surface coating modification, the sample exhibited good biocompatibility.

[0360] like Figure 6 As shown, in Example 1, the number of migrating cells was significantly increased compared to the magnesium metal sample, indicating that the coating modification treatment can enhance the cell migration ability of magnesium metal, giving it a stronger tissue repair ability.

[0361] like Figure 7 As shown, the sample in Example 1 showed a significant amount of new bone formation, which was fully integrated with the tissue and showed no adverse reactions such as inflammation. This indicates that the coating modification treatment can enhance the bone formation capacity of magnesium metal, giving it a stronger bone repair ability.

[0362] like Figure 8 As shown, the degradation rate of hyaluronic acid in Example 1 and Comparative Example 1 was significantly lower than that in Comparative Example 2, indicating that the mussel adhesive protein component in the calcium phosphorus coating can increase the retention time of hyaluronic acid and prolong its action time. Moreover, the degradation rate of hyaluronic acid in Example 1 was also significantly lower than that in Comparative Example 1, indicating that staged electrochemical deposition can increase the retention time of hyaluronic acid and prolong its action time by optimizing the structure of the calcium phosphorus layer.

[0363] Table 2. Adhesion strength data of sample coatings

[0364] Sample Bond strength (MPa) Example 1 18.5 Example 2 17.8 Example 3 17.5 Example 4 19.7 Example 5 20.6 Example 6 18.1 Example 7 17.4 Example 8 18.3 Example 9 18.1 Example 10 18.7 Example 11 18.0 Example 12 16.1 Comparative Example 1 6.8 Comparative Example 2 7.3 Comparative Example 3 8.8 Comparative Example 4 6.4 Comparative Example 5 9.3 Comparative Example 6 9.2 Comparative Example 7 8.7 Comparative Example 8 8.8

[0365] As shown in Table 2, Example 1 exhibits the highest coating adhesion strength. These results indicate that: 1) the mussel adhesive protein component in the calcium-phosphorus coating can increase the coating adhesion strength; 2) the coating obtained by staged electrochemical deposition has even higher adhesion strength. This is because during staged electrochemical deposition, the low-temperature static environment in the first stage protects protein activity, allowing the DOPA groups of the mussel adhesive protein to form chelate bonds with the magnesium surface, creating a pre-adsorption layer. In the second stage, the calcium-phosphorus crystals undergo in-situ mineralization, thereby enhancing interfacial bonding.

[0366] In summary, this application constructs a mussel adhesive protein and calcium-phosphorus-polysaccharide coating on the surface of biodegradable magnesium metal through a continuous two-step process. This not only reduces the corrosion rate of magnesium metal to match the bone repair rate, but also endows it with anti-inflammatory, antibacterial and osteogenic biological functions through the introduction of calcium-phosphorus and polysaccharide (hyaluronic acid, chitosan, sodium alginate, heparin, chondroitin sulfate, etc.) components.

[0367] The technical features of the above-described embodiments and examples can be combined in any suitable manner. For the sake of brevity, not all possible combinations of the technical features in the above-described embodiments and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0368] The embodiments described above are merely illustrative of several implementation methods of this application, intended to facilitate a detailed understanding of the technical solutions of this application, but should not be construed as limiting the scope of protection of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the scope of protection of this application. Furthermore, it should be understood that after reading the above teachings of this application, those skilled in the art can make various alterations or modifications to this application, and the equivalent forms obtained also fall within the scope of protection of this application. It should also be understood that technical solutions obtained by those skilled in the art based on the technical solutions provided in this application through logical analysis, reasoning, or limited experimentation are all within the scope of protection of the appended claims. Therefore, the scope of protection of this patent application should be determined by the content of the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. A method for preparing a magnesium metal-guided bone regeneration membrane, characterized in that, The preparation method includes the following steps: An electrochemical deposition solution and a polysaccharide-containing solution are provided respectively; the electrochemical deposition solution includes a calcium ion donor, a phosphate ion donor, and mussel adhesive protein; A magnesium metal substrate is placed in the electrochemical deposition solution, and a first-stage electrochemical deposition is performed under the first electrochemical deposition conditions to prepare intermediate 1. The membrane intermediate 1 was placed in the electrochemical deposition solution, and a second-stage electrochemical deposition was performed under the second electrochemical deposition conditions. After cleaning and drying, intermediate 2 was prepared. The intermediate 2 was placed in the polysaccharide-containing solution, coated with polysaccharide, and dried to prepare a magnesium metal guiding bone regeneration membrane; in, The first electrochemical deposition conditions include: a voltage of 1-5V; The first electrochemical deposition conditions include a voltage of 10-60V.

2. The method for preparing the magnesium metal guided bone regeneration membrane according to claim 1, characterized in that, The preparation method satisfies one or more of the following conditions: (1) The first electrochemical deposition conditions include: keeping the electrochemical deposition solution static, a temperature of 10-60°C, a voltage of 1-5V, and a time of 1-10 min; and, (2) The second electrochemical deposition conditions include: stirring the electrochemical deposition solution, with a temperature of 10-60℃, a voltage of 10-60V, and a time of 5-60min; Optionally, the stirring speed is 200-400 rpm.

3. The method for preparing the magnesium metal guided bone regeneration membrane according to claim 1, characterized in that, The electrochemical deposition solution satisfies one or more of the following conditions: 1) The calcium ion donor includes calcium nitrate; 2) The phosphate ion donor includes dihydrogen phosphate, optionally calcium dihydrogen phosphate or / and ammonium dihydrogen phosphate; 3) The electrochemical deposition solution comprises 0.01M-0.08M of the calcium ion donor, 0.005M-0.08M of the phosphate ion donor, and 0.5mg / mL-2.5mg / mL of mussel adhesive protein; 4) The pH of the electrochemical deposition solution is 1-6; Optionally, the mussel adhesive protein includes recombinant humanized mussel adhesive protein; Optionally, the molecular weight of the mussel adhesive protein is 10-100 kDa.

4. The method for preparing the magnesium metal guided bone regeneration membrane according to any one of claims 1 to 3, characterized in that, The polysaccharide-containing solution includes one or more of the following: hyaluronic acid, chitosan, sodium alginate, chondroitin sulfate, and heparin. Optionally, the polysaccharide-containing solution satisfies one or more of the following conditions: (A) The molecular weight of the hyaluronic acid is 10-2000 kDa; (B) The molecular weight of the chitosan is 10-300 kDa; (C) The molecular weight of the sodium alginate is 50-200 kDa; (D) The polysaccharide-containing solution comprises 0.1-5 wt% of the hyaluronic acid; (E) The polysaccharide-containing solution comprises 0.1-2 wt% of the chitosan; and, (F) The polysaccharide-containing solution comprises 0.1-8 wt% of the sodium alginate.

5. The method for preparing the magnesium metal guided bone regeneration membrane according to claim 4, characterized in that, The number of dip coatings is 1-10, and the time for each dip coating is 1-15 minutes. After each dip coating is completed, heat it to set the shape before starting the next dip coating. Optionally, the heat setting conditions include a temperature of 55-65℃ and a time of 3-8 minutes.

6. The method for preparing the magnesium metal guided bone regeneration membrane according to claim 4, characterized in that, If the polysaccharide-containing solution meets the conditions shown in (A) and (D), then the drying temperature is 20-100℃ and the drying time is 10-480 min; If the polysaccharide-containing solution meets the conditions shown in (B) and (E), then the drying temperature is 20-60℃ and the drying time is 6-12h. If the polysaccharide-containing solution meets the conditions shown in (C) and (F), the drying temperature is 30-60℃ and the drying time is 3-12h. Optionally, the preparation method further includes: after coating the polysaccharide, the obtained intermediate 3 is immersed in a crosslinking agent; optionally, the crosslinking agent includes calcium salt; optionally, the immersion conditions include: the concentration of calcium salt in the immersion system is 4-5wt% and the immersion time is 1-3h.

7. The method for preparing the magnesium metal guided bone regeneration membrane according to any one of claims 1 to 3 and 5 to 6, characterized in that, The preparation method satisfies one or more of the following conditions: A) The magnesium metal matrix is ​​pretreated as follows: polishing, cleaning and drying; Optionally, sanding can be done with 2000-8000 grit sandpaper; Optionally, during the pretreatment process, the cleaning includes washing with acetone, ethanol and water in sequence; Optionally, the drying temperature is 40-80℃ and the time is 10-60 minutes; B) During the preparation of intermediate 2, the cleaning process includes washing with water; and, C) During the preparation of intermediate 2, the drying temperature is 40-80℃ and the time is 10-60min.

8. A magnesium metal guided bone regeneration membrane, characterized in that, Prepared by the preparation method according to any one of claims 1 to 7.

9. A bone repair combination product, characterized in that, The bone repair combination product includes the magnesium metal guided bone regeneration membrane as described in claim 8.

10. A product for preparing a magnesium metal guided bone regeneration membrane, characterized in that, The product includes an electrochemical deposition solution as defined in any one of claims 1 to 7; Optionally, the product further includes a polysaccharide-containing solution as defined in any one of claims 1 to 7.

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