A method of depositing hydroxyapatite in a three-dimensional porous sample

Abstract

This invention discloses a method for depositing hydroxyapatite in a three-dimensional porous sample, comprising: S1, pretreatment of the three-dimensional porous sample; S2, placing the three-dimensional porous sample in an O... 2 In a plasma spraying apparatus; S3, prepare SBF1, immerse the sample treated in step S2 into SBF1, seal the container tightly, and place it in an incubator. After immersion in SBF1, remove the sample, wash and dry it. SBF1 contains the following components: CaCl2, MgCl2·6H2O, NaHCO3, K2HPO4·3H2O, Na2SO4, KCl, and NaCl; S4, prepare SBF2, immerse the sample treated in step S3 into SBF2, seal the container tightly, and place it in an incubator at 37°C. SBF2 contains the following components: NaCl, CaCl2, and K2HPO4·3H2O; S5, remove the sample, wash and dry it to obtain a three-dimensional porous sample with deposited hydroxyapatite. The method provided by this invention uniformly deposits HA in the porous three-dimensional void surface, and the composite interface can accelerate early bone healing, increase the integration of the bone implant surface with the bone, and promote the early stability of the implant.

Description

Technical Field

[0001] This invention relates to the field of biomedical materials technology, specifically a method for depositing hydroxyapatite in a three-dimensional porous sample. Background Technology

[0002] For successful implantation of various bone implants, establishing a well-integrated osseointegrated internal surface structure between the implant and the recipient bone is a crucial prerequisite. A poorly integrated osseointegrated internal surface can lead to insufficient adhesion between the implant and the bone interface, causing the implant to loosen and resulting in implantation failure. Furthermore, a poorly integrated osseointegrated internal surface can create a "stress shielding" effect between the implant and the recipient bone, leading to bone atrophy in the recipient area over long-term implantation and affecting the lifespan of the bone implant. Therefore, optimizing the internal surface structure of bone implants is a vital step in bone implant development.

[0003] Porous structures possess a three-dimensional, interconnected open pore structure, facilitating the transport of body fluids and nutrients. This enables the establishment of effective connections between the bone tissue within the pores and the external bone tissue, enhancing the bone intercalation strength between the bone tissue and the implant. However, the ability of porous structures alone to induce new bone ingrowth is limited. Some researchers have modified the osteogenic activity of porous structures by filling them with hydrogels or spraying bioactive materials onto their surfaces. Therefore, enhancing the osteogenic activity of porous structures is an important clinical problem that needs to be addressed.

[0004] Hydroxyapatite (HA) possesses excellent biocompatibility and osteoconductive properties, allowing it to form coatings on various materials. These coatings chemically bond with human bone tissue, inducing the differentiation of bone progenitor cells into new bone and accelerating bone healing during early implantation. Therefore, simultaneously depositing HA coatings on porous structures can significantly increase the integration of the implant's inner surface with the bone, promoting early healing and stability. Uniform and effective deposition of HA on porous surfaces is crucial not only for the success of material construction but also for its osteogenic properties. Currently, mainstream methods for HA preparation include plasma spraying, laser deposition, and electrophoretic deposition. However, due to the complex microscopic three-dimensional voids in porous structures, traditional methods suffer from poor preparation strength and inhomogeneity. Summary of the Invention

[0005] This invention overcomes these difficulties and provides a method for depositing hydroxyapatite in a three-dimensional porous sample. The method of this invention can uniformly deposit HA in the porous three-dimensional void surface, and the composite interface can accelerate early bone healing, increase the integration of the bone implant surface with the bone, and promote the early stability of the implant.

[0006] To achieve this objective, the present invention provides the following technical solution:

[0007] In a first aspect, this invention provides a method for depositing hydroxyapatite in a three-dimensional porous sample, comprising the following steps: S1, pretreatment of the three-dimensional porous sample; S2, placing the pretreated three-dimensional porous sample in an O2 plasma sprayer for surface treatment under an O2 environment; S3, preparing a No. 1 high-magnification simulated body fluid SBF1, immersing the sample treated in step S2 in SBF1, sealing the container tightly, and placing it in an incubator; after immersion in SBF1, removing the sample, washing, and drying it; the No. 1 high-magnification simulated body fluid SBF1 contains the following components: CaCl2, MgCl2·6H2O, NaHCO3, K2HPO4·3H2O, Na2SO4, KCl, and NaCl; S4, preparing a No. 2 high-magnification simulated body fluid SBF2, immersing the sample treated in step S3 in SBF2, sealing the container tightly, and placing it in an incubator; the No. 2 high-magnification simulated body fluid SBF2 contains the following components: NaCl and CaCl2. K2HPO4·3H2O; S5. Take out the sample, clean and dry it to obtain a three-dimensional porous sample of deposited hydroxyapatite.

[0008] In this invention, compounds were deposited in three-dimensional porous samples using the method of this invention through electron microscopy and EDS energy dispersive spectroscopy. The Ca-P ratio and basic elements of the compounds were the same as those of HA.

[0009] Preferably, step S1 specifically includes the following steps: immersing the three-dimensional porous sample in acetone, 100% ethanol and dd H2O for 0.5-1 hours respectively, and then air-drying it.

[0010] The preferred method for preparing No. 1 high-concentration simulated body fluid SBF1 is as follows: R1, mix dd H2O, CaCl2, MgCl2·6H2O, NaHCO3, and K2HPO4·3H2O separately and adjust the pH to 6.0; R2, add Na2SO4, KCl, and NaCl, and adjust the pH to 6.5.

[0011] Preferably, the weight parts of each component of No. 1 high-concentration simulated body fluid SBF1 are: CaCl2 0.347 parts, MgCl2·6H2O 0.380 parts, NaHCO3 0.441 parts, K2HPO4·3H2O 0.285 parts, Na2SO4 0.089 parts, KCl 0.280 parts, and NaCl 10.01 parts.

[0012] Preferably, the preparation method of No. 2 high-concentration simulated body fluid SBF2 is as follows: dd H2O, NaCl, CaCl2, and K2HPO4·3H2O are thoroughly mixed, and the pH value is adjusted to 6.0; wherein, the weight parts of each component of SBF1 are: NaCl 10.41 parts, CaCl2 0.347 parts, and K2HPO4·3H2O 0.57 parts.

[0013] Preferably, the soaking time in SBF1 is 24-48 hours, and the soaking time in SBF2 is twice the soaking time in SBF1.

[0014] Preferably, in steps S3 and S4, the sample surface area / volume ratio of the immersion solution (high-magnification simulated body fluid SBF1 or high-magnification simulated body fluid SBF2) does not exceed 1.95 cm². 2 / mL.

[0015] Preferably, three-dimensional porous samples include porous metals and their alloys, and porous polymer materials. For example, porous Ta, porous CaP, porous polyethylene, porous Ti, etc.

[0016] Preferably, the porous structure includes biomimetic porous, diamond-shaped, cubic, honeycomb, grid-shaped, dodecahedral, and foam-shaped porous structures.

[0017] In this invention, biomimetic porous structures refer to structures that mimic the porous structure of a compound itself, typically containing layered macropores and micropores. Diamond-type porous structures refer to structures that are diamond-shaped and contain both macropores and micropores.

[0018] A second aspect of the present invention provides a method for preparing an implant, comprising the following steps:

[0019] A1. Obtain biomimetic porous Ti / diamond-type porous Ti;

[0020] A2. First, immerse the biomimetic porous Ti / diamond porous Ti in acetone, 100% ethanol and dd H2O for 0.5 hours each, and then air dry.

[0021] A3. Place the biomimetic porous Ti / diamond porous Ti in an O2 plasma sprayer and perform surface treatment for 2 minutes in an O2 environment.

[0022] A4. Prepare high-concentration simulated body fluid SBF1: 250mL dd H2O; 0.347g CaCl2; 0.380g MgCl2·6H2O; 0.441g NaHCO3; 0.285g K2HPO4·3H2O. Mix thoroughly and adjust the pH to 6.0. Then add 0.089g Na2SO4; 0.280g KCl; 10.01g NaCl, and adjust the pH to 6.5. Filter using a 0.22mm filter.

[0023] A5. Immerse biomimetic porous Ti / diamond porous Ti in SBF1, ensuring the surface area of ​​the biomimetic porous Ti or diamond porous Ti / volume ratio of the immersion solution (No. 1 high-concentration simulated body fluid SBF1) does not exceed 1.95 cm². 2 / mL; Seal the container tightly and place it in an incubator at 37°C. After soaking in SBF1 for 24 hours, remove the sample, wash it three times with dd H2O, and air dry it.

[0024] A6. Prepare No. 2 high-concentration simulated body fluid SBF2: dd H2O 250mL; NaCl 10.41g; CaCl2 0.347g; K2HPO4·3H2O 0.57g. Mix thoroughly, adjust the pH to 6.0, and filter with a 0.22mm filter.

[0025] A7. Soak the previously dried biomimetic porous Ti / diamond porous Ti in SBF2 again, keeping the surface area of ​​biomimetic porous Ti or diamond porous Ti / volume ratio of soaking solution (No. 2 high-concentration simulated body fluid SBF2) not exceeding 1.95 cm2 / mL; after sealing the container, place it in an incubator at 37°C, and soak for twice the time it was in SBF1.

[0026] Compared with existing technologies, the beneficial effects and significant advancements of this invention are as follows: By configuring a high-concentration simulated solution and controlling the pH value and deposition soaking time, this invention ultimately constructs a method for HA deposition in a three-dimensional porous structure. The constructed material forms a Ca and P-containing apatite layer within the interface, similar to the calcium-phosphorus deposition process. The constructed material can rapidly promote new bone formation and exhibits stronger bone integration. Attached Figure Description

[0027] To more clearly illustrate the technical solution of the present invention, the accompanying drawings used in the embodiments of the present invention will be briefly introduced below.

[0028] Obviously, the accompanying drawings described below are only some of the drawings of the embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort, but these other drawings are also within the scope of the drawings required for the embodiments of the present invention.

[0029] Figure 1 This is a flowchart of the method in Embodiment 1 of the present invention;

[0030] Figure 2 The figures shown are from Example 2 of this invention, where AE represents a diamond-type porous Ti-HA sample and its electron microscopy results; FJ represents a biomimetic porous Ti-HA sample and its electron microscopy results; K represents the EDS energy spectrum of the diamond-type porous Ti-HA sample; Q represents the EDS energy spectrum of the biomimetic porous Ti-HA sample; LP represents the layered EDS image of the biomimetic porous Ti-HA sample; and RV represents the layered EDS image of the diamond-type porous Ti-HA sample.

[0031] Figure 3 The figure shows the results of Example 3 of the present invention, where AD represents the appearance of each group of samples; EP represents the surgical implantation process of each group; QT represents the Micro-CT scan results of each group 8 weeks after surgery; and UX represents the hard tissue sections and HE staining results of each group of samples 8 weeks after surgery. Detailed Implementation

[0032] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the present invention is further described below in conjunction with specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and parts by weight. Unless otherwise specified, the experimental materials and reagents used in the following embodiments are commercially available.

[0033] Unless otherwise specified, the 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 pertains. It should be noted that the terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the exemplary embodiments of this application.

[0034] It should be noted that the terms "first," "second," and "third" (if present), etc., in the specification, claims, and accompanying drawings of the embodiments of this invention, are only used to distinguish different objects and not to describe a specific order. Furthermore, the term "comprising" and any variations thereof are intended to cover non-exclusive inclusion. For example, a process, method, system, product, or device that includes a series of steps or units is not limited to the listed steps or units, but may optionally include steps or units not listed, or may optionally include other steps or units inherent to these processes, methods, products, or devices.

[0035] The technical solution of the present invention will now be described in detail with reference to specific embodiments.

[0036] Example 1

[0037] The method for preparing biomimetic porous Ti-HA and diamond-type porous Ti-HA in this embodiment is as follows: Figure 1 As shown.

[0038] Preparation of biomimetic porous Ti-HA

[0039] 1. Obtain biomimetic porous Ti.

[0040] 2. First, immerse the porous sample to be deposited with HA in acetone, 100% ethanol and dd H2O for 0.5 h each, and then air dry.

[0041] 3. Then place the biomimetic porous Ti in an O2 plasma sprayer and perform surface treatment for 2 minutes in an O2 environment;

[0042] 4. Prepare high-concentration simulated body fluid No. 1 (SBF1): 250mL dd H2O; 0.347g CaCl2; 0.380g MgCl2·6H2O; 0.441g NaHCO3; 0.285g K2HPO4·3H2O. Mix thoroughly and adjust the pH to 6.0. Then add 0.089g Na2SO4; 0.280g KCl; 10.01g NaCl, and adjust the pH to 6.5. Filter using a 0.22mm filter.

[0043] 5. Immerse the biomimetic porous Ti in SBF1 (keeping the ratio of biomimetic porous Ti surface area to immersion liquid volume not exceeding 1.95 cm2 / mL); seal the container tightly and place it in an incubator at 37°C.

[0044] 6. After soaking in SBF1 for 24 hours, remove the sample, wash it three times with dd H2O, and air dry it.

[0045] 7. Prepare high-concentration simulated body fluid No. 2 (SBF2): dd H2O 250mL; NaCl 10.41g; CaCl2 0.347g; K2HPO4·3H2O 0.57g. Mix thoroughly and adjust the pH to 6.0. Filter with a 0.22mm filter.

[0046] 8. Immerse the previously dried biomimetic porous Ti in SBF2 again (keeping the ratio of biomimetic porous Ti surface area to immersion liquid volume no more than 1.95 cm²). 2 / mL); after sealing the container, place it in an incubator at 37°C for twice the soaking time in SBF1.

[0047] 9. Remove the porous sample, wash it three times with dd H2O and air dry it. The relevant preparation process is now complete.

[0048] Preparation of diamond-type porous Ti-HA

[0049] 1. Obtain diamond-shaped porous Ti.

[0050] 2. First, immerse the porous sample to be deposited with HA in acetone, 100% ethanol and dd H2O for 0.5 h each, and then air dry.

[0051] 3. Then place the diamond-shaped porous Ti in an O2 plasma sprayer and perform surface treatment for 2 minutes in an O2 environment;

[0052] 4. Prepare high-concentration simulated body fluid No. 1 (SBF1): 250mL dd H2O; 0.347g CaCl2; 0.380g MgCl2·6H2O; 0.441g NaHCO3; 0.285g K2HPO4·3H2O. Mix thoroughly and adjust the pH to 6.0. Then add 0.089g Na2SO4; 0.280g KCl; 10.01g NaCl, and adjust the pH to 6.5. Filter using a 0.22mm filter.

[0053] 5. Immerse the diamond-type porous Ti in SBF1 (keeping the ratio of diamond-type porous Ti surface area to immersion liquid volume not exceeding 1.95 cm2 / mL); seal the container tightly and place it in an incubator at 37°C.

[0054] 6. After soaking in SBF1 for 24 hours, remove the sample, wash it three times with dd H2O, and air dry it.

[0055] 7. Prepare high-concentration simulated body fluid No. 2 (SBF2): dd H2O 250mL; NaCl 10.41g; CaCl2 0.347g; K2HPO4·3H2O 0.57g. Mix thoroughly and adjust the pH to 6.0. Filter with a 0.22mm filter.

[0056] 8. Soak the previously dried diamond porous Ti in SBF2 again (keeping the diamond porous Ti surface area / soaking liquid volume ratio not exceeding 1.95 cm2 / mL); place it in a sealed container in an incubator at 37°C for twice the soaking time in SBF1.

[0057] 9. Remove the porous sample, wash it three times with dd H2O and air dry it. The relevant preparation process is now complete.

[0058] Example 2

[0059] The biomimetic porous Ti-HA and diamond-type porous Ti-HA prepared in Example 1 were observed using scanning electron microscopy. Further EDS energy dispersive spectroscopy analysis was performed.

[0060] The results are as follows Figure 2 As shown, AE represents the diamond-type porous Ti-HA sample and its electron microscopy (EM) results; FJ represents the biomimetic porous Ti-HA sample and its EEM results; K represents the diamond-type porous Ti-HA sample's EDS energy spectrum; Q represents the biomimetic porous Ti-HA sample's EDS energy spectrum; LP represents the layered EDS image of the biomimetic porous Ti-HA sample; and RV represents the layered EDS image of the diamond-type porous Ti-HA sample. It is evident that SEM observation reveals the formation of sheet-like HA deposits on the surface of both the biomimetic and diamond-type porous Ti-HA samples, with well-defined deposition within the porous interior. Furthermore, EDS energy spectrum analysis shows that the Ca / P ratio and basic elements are identical to those of HA, indicating a stable chemically synthesized structure.

[0061] This embodiment fully verifies that the method described in this embodiment can deposit hydroxyapatite (HA) in biomimetic porous Ti and diamond-type porous Ti, and the chemically synthesized HA has a stable structure.

[0062] Example 3: Canine mandibular surface implantation experiment

[0063] Experimental groups: biomimetic porous Ti, diamond porous Ti, biomimetic porous Ti-HA and diamond porous Ti-HA prepared in Example 1.

[0064] Experimental Methods: Under general anesthesia, an incision was made under the canine jaw to separate the muscle and periosteum, exposing the lateral surface of the mandible. Several bone holes were drilled using a drill bit. The porous Ti-HA sample to be implanted was then placed against the drilled area, fixed with titanium screws, and then sutured in layers. Samples were harvested 8 weeks postoperatively, and each group of materials underwent Micro-CT scanning and histological examination.

[0065] The results are as follows Figure 3 As shown, AD represents the appearance of each group of samples; EP represents the surgical implantation process of each group; QT represents the Micro-CT scan results of each group at 8 weeks post-operation; and UX represents the hard tissue sections and HE staining results of each group of samples at 8 weeks post-operation. The results showed that the porous interiors of the diamond-shaped porous Ti and biomimetic porous Ti samples were empty with no new bone ingrowth, while new bone ingrowth was observed in the biomimetic porous Ti-HA and diamond-shaped porous Ti-HA samples. Hard tissue sections and HE staining results of each group of samples at 8 weeks post-operation showed that a large amount of fibrous tissue edged between the surface of the diamond-shaped porous Ti and biomimetic porous Ti samples and the newly formed bone tissue, while the biomimetic porous Ti-HA and diamond-shaped porous Ti-HA samples were in direct contact with the newly formed bone tissue.

[0066] This embodiment fully verifies that the method of the present invention, through the uniform deposition of HA in a porous three-dimensional void surface, enables the composite interface to accelerate early bone healing, increase the integration of the bone implant surface with the bone, and promote the early stability of the implant.

[0067] In the description process of the above instruction manual:

[0068] The terms "this embodiment," "an embodiment of the present invention," "as shown," "further," and "further improved technical solutions," etc., indicate that the specific features, structures, materials, or characteristics described in the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the illustrative expressions of the above terms are not necessarily directed at the same embodiment or example, and the specific features, structures, materials, or characteristics described can be combined or combined in any suitable manner in one or more embodiments or examples. Furthermore, without causing contradiction, those skilled in the art can combine or combine the different embodiments or examples described in this specification and the features of the different embodiments or examples.

[0069] Finally, it should be noted that:

[0070] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them;

[0071] Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention. Non-essential improvements, adjustments or substitutions made by those skilled in the art based on the content of this specification are all within the scope of protection claimed by the present invention.

Claims

1. A method for depositing hydroxyapatite in a three-dimensional porous sample, characterized in that, Includes the following steps: S1. Preprocessing of three-dimensional porous samples; S2. Place the pretreated three-dimensional porous sample in an O2 plasma sprayer and perform surface treatment in an O2 environment. S3. Prepare high-magnification simulated body fluid SBF1 (No. 1). Immerse the sample treated in step S2 in SBF1, seal the container tightly, and place it in an incubator. After immersion in SBF1, remove the sample, wash it, and air dry it. High-magnification simulated body fluid SBF1 contains the following components: CaCl2, MgCl2·6H2O, NaHCO3, K2HPO4·3H2O, Na2SO4, KCl, and NaCl. S4. Prepare high-magnification simulated body fluid SBF2 (No. 2). Immerse the sample treated in step S3 in SBF2, seal the container tightly, and place it in an incubator. High-magnification simulated body fluid SBF2 (No. 2) contains the following components: NaCl, CaCl2, and K2HPO4·3H2O. S5. Remove the sample, clean and dry it to obtain a three-dimensional porous sample of deposited hydroxyapatite.

2. The method for depositing hydroxyapatite in a three-dimensional porous sample as described in claim 1, characterized in that, Step S1 specifically includes the following steps: immersing the three-dimensional porous sample in acetone, 100% ethanol and dd H2O for 0.5-1 hours respectively, and then air-drying it.

3. The method for depositing hydroxyapatite in a three-dimensional porous sample as described in claim 1, characterized in that, The preparation method of No. 1 high-magnification simulated body fluid SBF1 is as follows: R1. Mix dd H2O, CaCl2, MgCl2·6H2O, NaHCO3, and K2HPO4·3H2O, and adjust the pH to 6.0; R2, then add Na2SO4, KCl, and NaCl, and adjust the pH to 6.

5.

4. A method for depositing hydroxyapatite in a three-dimensional porous sample as described in claim 1 or 3, characterized in that, The weight parts of each component of No. 1 high-concentration simulated body fluid SBF1 are as follows: CaCl2 0.347 parts, MgCl2·6H2O 0.380 parts, NaHCO3 0.441 parts, K2HPO4·3H2O 0.285 parts, Na2SO4 0.089 parts, KCl 0.280 parts, and NaCl 10.01 parts.

5. The method for depositing hydroxyapatite in a three-dimensional porous sample as described in claim 1, characterized in that, The preparation method of No. 2 high-concentration simulated body fluid SBF2 is as follows: dd H2O, NaCl, CaCl2, and K2HPO4·3H2O are thoroughly mixed, and the pH value is adjusted to 6.0; the weight parts of each component of SBF1 are: NaCl 10.41 parts, CaCl2 0.347 parts, and K2HPO4·3H2O 0.57 parts.

6. The method for depositing hydroxyapatite in a three-dimensional porous sample as described in claim 1, characterized in that, The soaking time in SBF1 is 24-48 hours, and the soaking time in SBF2 is twice that in SBF1.

7. The method for depositing hydroxyapatite in a three-dimensional porous sample as described in claim 1, characterized in that, In steps S3 and S4, the ratio of the surface area of ​​the three-dimensional porous sample to the volume of either high-magnification simulated body fluid SBF1 (No. 1) or SBF2 (No. 2) does not exceed 1.95 cm². 2 / mL.

8. The method for depositing hydroxyapatite in a three-dimensional porous sample as described in claim 1, characterized in that, Three-dimensional porous samples include porous metals and their alloys, and porous polymer materials.

9. The method for depositing hydroxyapatite in a three-dimensional porous sample as described in claim 8, characterized in that, Porous structures include biomimetic porous structures, diamond-shaped porous structures, cubic porous structures, honeycomb porous structures, grid-shaped porous structures, dodecahedral porous structures, and foam porous structures.

10. A method for preparing an implant, characterized in that, Includes the following steps: A1. Obtain biomimetic porous Ti / diamond-type porous Ti; A2. First, immerse the biomimetic porous Ti / diamond porous Ti in acetone, 100% ethanol and dd H2O for 0.5 hours each, and then air dry. A3. Place the biomimetic porous Ti / diamond porous Ti in an O2 plasma sprayer and perform surface treatment for 2 minutes in an O2 environment. A4. Prepare high-concentration simulated body fluid SBF1: 250mL dd H2O, 0.347g CaCl2, 0.380g MgCl2·6H2O, 0.441g NaHCO3, 0.285g K2HPO4·3H2O. Mix thoroughly and adjust the pH to 6.

0. Then add 0.089g Na2SO4, 0.280g KCl, and 10.01g NaCl, and adjust the pH to 6.

5. Filter using a 0.22mm filter. A5. Immerse biomimetic porous Ti / diamond porous Ti in SBF1, ensuring the surface area of ​​the biomimetic porous Ti or diamond porous Ti / the volume ratio of No. 1 high-magnification simulated body fluid SBF1 does not exceed 1.95 cm². 2 / mL; Seal the container tightly and place it in an incubator at 37°C. After soaking in SBF1 for 24 hours, remove the sample, wash it three times with dd H2O, and air dry it. A6. Prepare No. 2 high-concentration simulated body fluid SBF2: dd H2O 250mL, NaCl 10.41g, CaCl2 0.347g, K2HPO4·3H2O 0.57g, mix thoroughly, adjust the pH value to 6.0, and filter with a 0.22mm filter; A7. Immerse the previously dried biomimetic porous Ti / diamond porous Ti in SBF2 again, ensuring that the surface area of ​​the biomimetic porous Ti or diamond porous Ti / the volume of No. 2 high-concentration simulated body fluid SBF2 does not exceed 1.95 cm². 2 / mL; after sealing the container, place it in an incubator at 37°C for twice the soaking time in SBF1.

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