Preparation method and application of surface-nanocrystallized TLM titanium alloy material

Abstract

The application provides a preparation method and application of a surface-nanocrystallized TLM titanium alloy material, and the surface-nanocrystallized TLM titanium alloy material is prepared by the following steps: step 1, polishing a TLM titanium alloy ultrafine-grain composite plate by using silicon carbide sandpaper in stages; step 2, using the polished TLM titanium alloy ultrafine-grain composite plate as an anode, using a platinum foil as a cathode, and performing anodic oxidation in an electrolyte containing fluorides; and step 3, performing heat treatment. The application forms nanotubes with different diameters on the surface of the original TLM titanium alloy foil after treatment, so as to construct a three-dimensional structure which is beneficial to cell adhesion and proliferation. The surface-nanocrystallized TLM titanium alloy material has the characteristics of good histocompatibility, suitable hardness and toughness, and easy cutting and shaping, which is very beneficial to serving as a cartilage scaffold material. The surface nanotubes have a suitable three-dimensional space structure and can adsorb and release nutrient molecules, so as to exchange substances with surrounding tissue cells, which is beneficial to cell adhesion, growth and proliferation.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology, and particularly relates to a preparation method and application of a surface-nanocrystallized TLM titanium alloy material. BACKGROUND

[0002] Biomedical metallic material is a metal or alloy used for diagnosing, treating, repairing or replacing damaged tissues or organs of a living body or improving the function thereof. The biomedical metallic material is mainly used for repairing and replacing hard tissues such as bones and teeth, repairing cardiovascular and soft tissues, and manufacturing artificial organs, and is required to have excellent corrosion resistance, biocompatibility and good processing performance in addition to good mechanical and physical properties, and thus can serve in a human body for a long time. Titanium alloy is one of the most promising biomedical metallic materials. In medicine, titanium alloy can be used as an implant device to replace damaged hard tissues, such as artificial hip joints, artificial knee joints, bone plates, bone fracture fixation screws, heart valve prostheses, pacemakers and artificial hearts. Pure titanium and Ti6Al4V have been the most important medical titanium alloys.

[0003] In recent years, based on the high demand for medical devices and the rapid development of titanium alloys, researchers have developed a batch of new titanium alloys, such as Ti6AI7Nb alloy (ASTM F1295), Ti13Nb13Zr alloy (ASTM F1713), Ti25Nb3Mo3Zr2.2Sn alloy (TLM) and Ti12Mo6Zr alloy (ASTM F1813) and other new materials.

[0004] TLM titanium alloy contains a certain proportion of zirconium (Zr), molybdenum (Mo), niobium (Nb) and tin (Sn) elements beneficial to the human body, so that the alloy has a more similar elastic modulus to human bone, can greatly reduce the stress shielding phenomenon of the implanted part and the surrounding tissue, and reduce the risk of secondary surgery. At the same time, it has excellent mechanical properties including low elastic modulus, high strength, high plasticity and toughness, high fatigue limit, and wide and adjustable comprehensive mechanical properties; at the same time, it has excellent cold and hot processing performance, and its comprehensive performance is better than that of various third-generation new medical β-type titanium alloys reported in the world. TLM titanium alloy is a Ti-Zr-Sn-Mo-Nb alloy system, and the added elements zirconium, molybdenum, tin and niobium belong to alloy additive elements with excellent biocompatibility and non-toxicity. The alloy has certain strength, elasticity, toughness and high fatigue resistance, and is not easy to deform under external force, and its shape and size can be shaped and cut at will according to needs, which is very beneficial to serve as a scaffold implant material for cartilage and the like.

[0005] At present, the TLM titanium alloy new material has passed engineering test verification, and the material has developed high-tech products such as vascular stents, artificial joints, dental repair products, orthopedic internal fixation devices and the like which can be used for repair and replacement of human soft and hard tissues. SUMMARY

[0006] The present application aims at solving the problems existing in the prior art, and provides a preparation method and application of a surface-nanocrystallized TLM alloy biomaterial.

[0007] The present application is realized by the following technical solutions:

[0008] In a first aspect, the present application provides a preparation method of a surface-nanocrystallized TLM titanium alloy material, which specifically comprises the following steps:

[0009] The TLM titanium alloy ultra-fine crystal composite plate is polished by using silicon carbide sandpaper in stages, then the polished TLM titanium alloy ultra-fine crystal composite plate is used as an anode, a platinum foil is used as a cathode, anodic oxidation is carried out in an electrolyte containing fluoride, and finally heat treatment is carried out to obtain the surface-nanocrystallized TLM titanium alloy material.

[0010] Further improvement of the present application is that:

[0011] The electrolyte containing fluoride is an aqueous solution of HF and HNO3, wherein the volume concentration of HF is 1%-5%, and the volume concentration of HNO3 is 1%-10%.

[0012] Further improvement of the present application is that:

[0013] During the anodic oxidation process, the voltage is maintained at 10-120V, and the time is 30-120min.

[0014] Further improvement of the present application is that:

[0015] The heat treatment is heat preservation at 500-600℃ for 30-60min in an atmospheric atmosphere, and then the furnace is cooled.

[0016] Further improvement of the present application is that:

[0017] The preparation method of the TLM titanium alloy ultra-fine crystal composite plate is that: after heat treatment and cooling of the TLM titanium alloy hot-rolled plate, the plate is sequentially subjected to pickling and shot blasting treatment, and then is subjected to one-time stacking of multiple layers of plates for cladding, and cold rolling to obtain the TLM titanium alloy ultra-fine crystal composite plate.

[0018] Further improvement of the present application is that:

[0019] The TLM titanium alloy hot-rolled plate is selected from a TLM titanium alloy hot-rolled plate with a thickness of 1.0 mm, and the phase transition point is 710 DEG C.

[0020] The further improvement of the present application is that:

[0021] The temperature of the heat treatment is 610-810 DEG C, the time is 30 min, and the cooling mode is water cooling.

[0022] The further improvement of the present application is that:

[0023] The pickling is carried out in a mixed solution composed of HF, HNO3 and H2O for 2-7 min, and the volume ratio of HF, HNO3 and H2O in the mixed solution is 1:5:7.

[0024] The sand blasting treatment is selected from white corundum coarse blasting, the pressure of sand blasting is 0-0.45 Mpa, and the duration is 10-30 s.

[0025] The further improvement of the present application is that:

[0026] The intermediate stress relief annealing is carried out in the rolling process, the annealing temperature is 300-500 DEG C, and the holding time is 2-7 min.

[0027] In the second aspect of the present application, the surface nanocrystallization TLM titanium alloy material prepared by the preparation method is applied to a cartilage scaffold material.

[0028] Compared with the prior art, the present application has the following beneficial effects:

[0029] The surface nanocrystallization TLM titanium alloy material provided by the present application is formed into nanotubes with different diameters on the surface of the original TLM titanium alloy foil, so as to construct a three-dimensional structure conducive to cell adhesion and proliferation.

[0030] The surface nanocrystallization TLM titanium alloy material has the characteristics of good tissue compatibility, suitable hardness and toughness, easy cutting and shaping, etc., which is very beneficial to serving as a cartilage scaffold material, the surface nanotube has a suitable three-dimensional space structure and can absorb and release nutrient molecules and exchange materials with surrounding tissue cells, which is conducive to cell adhesion, growth and proliferation, and the 3D printing technology can be used to accurately construct a repair material according to tissue defects. BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1 The microstructure diagrams of the TLM titanium alloy hot-rolled plates after heat treatment of example 1, example 2 and example 3, wherein (a) 610, 30 min; (b) 710 DEG C, 30 min; (c) 810, 30 min;

[0032] Figure 2Microstructure images of TLM titanium alloy ultrafine grain composite plates prepared for example 1, example 4, example 5 and example 6, wherein (a) single layer; (b) 2 layers; (c) 4 layers; (d) 8 layers;

[0033] Figure 3 XRD spectrum of 8-layer TLM titanium alloy ultrafine grain composite plate prepared for example 6;

[0034] Figure 4 Transmission electron microscope and diffraction spectrum of 8-layer TLM titanium alloy ultrafine grain composite plate prepared for example 6;

[0035] Figure 5 Scanning electron microscope image of surface nanocrystallized TLM titanium alloy material prepared for example 7;

[0036] Figure 6 Scanning electron microscope image of surface nanocrystallized TLM titanium alloy material prepared for example 8;

[0037] Figure 7 Scanning electron microscope image of surface nanocrystallized TLM titanium alloy material prepared for example 9;

[0038] Figure 8 Scanning electron microscope image of surface nanocrystallized TLM titanium alloy material prepared for example 10;

[0039] Figure 9 Scanning electron microscope image of surface nanocrystallized TLM titanium alloy material prepared for example 11;

[0040] Figure 10 Scanning electron microscope images of surface nanocrystallized TLM titanium alloy materials with nanotube diameters of 100 nm, 200 nm and 300 nm, wherein (a) 100 nm; (b) 200 nm; (c) 300 nm;

[0041] Figure 11 Electron microscope scanning image of adipose stromal stem cells cultured with nanocrystallized TLM titanium alloy;

[0042] Figure 12 Inverted microscope image of adipose stromal stem cells cultured with nanocrystallized TLM titanium alloy;

[0043] Figure 13 Absorbance values of surface nanocrystallized TLM titanium alloy material of the present application and other materials on the market, wherein A: zinc material; B: nickel-titanium alloy; C: beta-titanium; D: TLM; E: TC4; F: 316L stainless steel; *p<0.05, **p<0.01, ***p<0.001;

[0044] Figure 14The fluorescence staining results of the proliferation and differentiation of adipose-derived stem cells on various alloy scaffolds, wherein Bar = 100 μm. DETAILED DESCRIPTION

[0045] The application will be further described in detail below with reference to the accompanying drawings:

[0046] The application provides a preparation method of a surface-nanocrystallized TLM titanium alloy material, and specifically comprises the following steps:

[0047] Step 1, preparation of a TLM titanium alloy composite plate

[0048] A TLM titanium alloy hot-rolled plate with a thickness of 1.0 mm is selected, and the phase transition point is 710 ℃. After heat treatment, cooling is performed. The heat treatment temperature is 610-810 ℃, the time is 30 min, and the cooling method is water cooling. Then, the surface oxide layer is removed by pickling and sandblasting treatment (sandblasting can also improve the mechanical occlusion degree of the composite plate). After a plurality of TLM titanium alloy hot-rolled plates are stacked layer by layer, they are covered with a cover material. The cover material is selected to be a 304 stainless steel plate with a thickness of 2 mm. Finally, the multi-layer composite plate is rolled by a two / four-roller rolling mill. During the rolling process, intermediate stress relief annealing is required. The annealing temperature is 300-500 ℃, and the holding time is 2-7 min. The multi-layer composite plate is processed by a cold rolling mill and a flat cold rolling mill. A 0.2 mm thick 2-layer, 4-layer or 8-layer TLM titanium alloy ultra-fine grain composite plate is obtained.

[0049] The pickling is performed in a mixed solution composed of HF, HNO3 and H2O for 2-7 min, and the volume ratio of HF, HNO3 and H2O in the mixed solution is 1:5:7.

[0050] The sandblasting treatment selects white corundum coarse spraying, the sandblasting pressure is 0-0.45 Mpa, the sandblasting is uniform, and the duration is 10-30 s.

[0051] Step 2, preparation of a surface-nanocrystallized TLM titanium alloy material

[0052] The TLM titanium alloy ultra-fine grain composite plate prepared in step 1 is polished by using silicon carbide sandpaper in stages. Then, the polished TLM titanium alloy ultra-fine grain composite plate is used as an anode, and a platinum foil is used as a cathode. Anodic oxidation is performed in an electrolyte containing fluoride. The electrolyte containing fluoride is an aqueous solution of HF and HNO3. The volume concentration of HF is 1%-5%, and the volume concentration of HNO3 is 1%-10%. During the anodic oxidation process, the voltage is maintained at 10-120 V, and the time is 30-120 min. After anodic oxidation, heat treatment is performed. In an atmospheric atmosphere, the temperature is maintained at 500-600 ℃ for 30-60 min, and then the furnace is cooled. Thus, a surface-nanocrystallized TLM titanium alloy material is prepared.

[0053] The surface nanotube diameter of the surface nanocrystallized TLM titanium alloy material prepared by the method is 50-300nm.

[0054] The method for preparing the surface nanocrystallized TLM titanium alloy material is further explained below through specific examples.

[0055] Example 1

[0056] Preparation of the TLM titanium alloy composite plate: select a TLM titanium alloy hot-rolled plate with a thickness of 1.0mm, a phase transition point of 710℃, and perform water cooling after heat treatment, the heat treatment temperature is 610℃, and the time is 30min; then remove the surface oxide layer by adopting pickling and shot blasting technology, take a piece of the treated TLM titanium alloy hot-rolled plate for cladding, the cladding material selects a 304 stainless steel plate with a thickness of 2mm, and finally perform cold rolling, the cold rolling adopts a two / four roller mill to roll the multi-layer composite plate, intermediate stress relief annealing needs to be performed during the rolling process, the annealing temperature is 400℃, and the holding time is 5min; the multi-layer composite plate is processed by adopting a cold rolling mill and a flat cold rolling mill, and a single-layer TLM titanium alloy ultra-fine grain composite plate with a thickness of 0.2mm is obtained.

[0057] Example 2

[0058] Preparation of the TLM titanium alloy composite plate: select a TLM titanium alloy hot-rolled plate with a thickness of 1.0mm, a phase transition point of 710℃, and perform water cooling after heat treatment, the heat treatment temperature is 710℃, and the time is 30min; then remove the surface oxide layer by adopting pickling and shot blasting technology, take a piece of the treated TLM titanium alloy hot-rolled plate for cladding, the cladding material selects a 304 stainless steel plate with a thickness of 2mm, and finally perform cold rolling, the cold rolling adopts a two / four roller mill to roll the multi-layer composite plate, intermediate stress relief annealing needs to be performed during the rolling process, the annealing temperature is 400℃, and the holding time is 5min; the multi-layer composite plate is processed by adopting a cold rolling mill and a flat cold rolling mill, and a single-layer TLM titanium alloy ultra-fine grain composite plate with a thickness of 0.2mm is obtained.

[0059] Example 3

[0060] Preparation of TLM titanium alloy composite sheet: select TLM titanium alloy hot-rolled sheet with thickness of 1.0 mm, phase transition point of 710°C, after heat treatment, water cooling, heat treatment temperature of 810°C, time of 30 min; then adopt pickling and shot blasting technology to remove surface oxide layer, take a piece of treated TLM titanium alloy hot-rolled sheet to perform cladding, cladding material selects 304 stainless steel plate with thickness of 2 mm, finally after cold rolling, adopt two / four roller mill to perform rolling of multi-layer composite sheet, need to perform intermediate stress relief annealing during rolling, annealing temperature of 400°C, holding for 5 min; adopt cold rolling mill and flat cold rolling mill to process multi-layer composite sheet, process to obtain 0.2 mm thick single-layer TLM titanium alloy ultra-fine grain composite sheet.

[0061] Figure 1 Microstructure diagrams of TLM titanium alloy hot-rolled sheets after heat treatment of example 1, example 2 and example 3. The alloy sheet treated at 610°C (100°C below the phase transition point) has no recrystallization, and obvious original slab rolling flow lines, as shown in Figure 2 a; the alloy sheet after heat treatment and water cooling at 710°C has no obvious flow lines, and obvious recrystallization, and the grain size is about 10 microns, as shown in Figure 2 b; the alloy sheet treated at 810°C (100°C above the phase transition point) has an average grain size of about 100 microns, as shown in Figure 2 c.

[0062] Table 1 is the mechanical properties of single-layer TLM titanium alloy ultra-fine grain composite sheet after 80% deformation. From table 1, it can be seen that under the same heat treatment temperature and processing deformation, the mechanical properties of the cold-rolled TLM sheet are obviously better than those of the hot-rolled sheet, and the surface of the cold-rolled sheet is bright, the flatness is good, and the elastic modulus reaches 38 GPa, which is very close to the modulus (E=10-30 GPa) of the human natural bone, and the strength also reaches more than 900 MPa. At the same time, it also shows that with the increase of heat treatment temperature, the comprehensive properties such as strength, plasticity and elastic modulus of the sheet processed below the phase transition point of 610°C are the best.

[0063] Table 1 is the mechanical properties of single-layer TLM titanium alloy ultra-fine grain composite sheet after 80% deformation.

[0064]

[0065]

Example 4

[0066] Referring to example 1, the difference between example 1 and example 4 is that two pieces of treated TLM titanium alloy hot-rolled sheets are stacked (one sheet is placed on another sheet) to perform cladding, and finally a 2-layer TLM titanium alloy ultra-fine grain composite sheet with thickness of 0.2 mm is processed.

[0067] Example 5

[0068] Referring to Example 1, the difference between Example 1 is that four pieces of the processed TLM titanium alloy hot-rolled plate are stacked and then covered with a cover, and finally a 4-layer TLM titanium alloy ultra-fine grain composite plate with a thickness of 0.2 mm is obtained.

[0069] Example 6

[0070] Referring to Example 1, the difference between Example 1 is that eight pieces of the processed TLM titanium alloy hot-rolled plate are stacked and then covered with a cover, and finally an 8-layer TLM titanium alloy ultra-fine grain composite plate with a thickness of 0.2 mm is obtained.

[0071] Figure 2 Microstructure diagrams of the TLM titanium alloy ultra-fine grain composite plates prepared in Example 1, Example 4, Example 5 and Example 6. As shown in Figure 2 It can be seen that the average deformation rate of each layer of the TLM titanium alloy ultra-fine grain composite plates prepared in Example 1, Example 4, Example 5 and Example 6 is 80%, 90%, 95% and 97.5%, respectively. Figure 2 The alloy in a is deformed by 80% cold rolling, and it can be seen that the elongated grains are locally broken; as shown in Figure 2 b, with the increase of the deformation rate, the grains are further obviously refined, and the local grain refinement is below 10 μm, the bonding interface between the two layers of thin plates is clear, and no obvious mechanical interlocking is achieved; Figure 2 c, Figure 2 d are the microstructures of the 4-layer and 8-layer composite plates in the rolling direction, respectively, indicating that the microstructure is a typical fibrous structure, and there is no obvious interface between the layers, and good mechanical interlocking is achieved.

[0072] The TLM alloy plate can obtain good mechanical properties matched with human bones by adopting the processing mode of annealing at 610°C and then cold rolling. As shown in the table, under the same heat treatment temperature and processing deformation, the mechanical properties of the TLM plate obtained by cold rolling are obviously better than those of the hot-rolled plate, and the surface of the cold-rolled plate is bright and flat, and the elastic modulus reaches 38 GPa, which is very close to the modulus (E=10-30 GPa) of the natural bone of human body, and the strength also reaches more than 900 MPa. At the same time, it also shows that with the increase of the heat treatment temperature, the plate processed below the phase transition point at 610°C has the best comprehensive properties such as strength, plasticity and elastic modulus.

[0073] Figure 3 XRD pattern of the 8-layer TLM titanium alloy ultra-fine grain composite plate prepared in Example 6. As shown in Figure 3It can be seen that the phase structure of the alloy after large deformation multi-pass cold rolling and intermediate annealing is composed of α, β and a small amount of deformation-induced α" martensite, wherein the percentage of α phase in the structure is 19%, which is increased compared with the percentage of α phase in the alloy plate before rolling, which is 10%. This shows that the deformation temperature rise generated by the cladding cumulative pack rolling can decompose the β phase, and with the increase of the deformation amount, the deformation-induced α" martensite is formed, and the α phase is also increased, so that the microstructure of the deformed material is more complex, thereby improving the strength and plasticity of the material under the interaction of different micro deformation mechanisms.

[0074] Figure 4 The transmission electron microscope image and diffraction pattern of the 8-layer TLM titanium alloy ultra-fine grain composite plate prepared in Example 6, Figure 4 It is illustrated that the uniformity of the grain size of the plate, and the continuous diffraction ring indicates that the grain size in the microstructure is refined to the nanometer scale, and with the increase of the total deformation amount, the nanometer size continues to decrease to the minimum of 80-100 nm, but no deformation twinning is found. Due to the formation of dual-phase structure of the TLM alloy plate under two-phase zone annealing and large deformation, the dislocation movement is restricted by the dual-nanometer phase, thereby realizing the improvement of the strength of the alloy. It is proved that the phase transformation has a certain promoting effect on the refinement of the structure, which is mainly caused by the stress-strain-induced inhomogeneous nucleation martensitic phase transformation. The martensitic phase transformation not only can refine the structure, but also can obtain bulk amorphous by controlling cold deformation.

[0075]

Example 7

[0076] Preparation of surface nanocrystalline TLM titanium alloy material: the 0.2mm thick 8-layer TLM titanium alloy ultra-fine grain composite plate prepared in Example 6 is polished by using silicon carbide sandpaper step by step (120, 320, 800 silicon carbide sandpaper is used step by step), then the polished TLM titanium alloy ultra-fine grain composite plate is used as anode, platinum foil is used as cathode, and anodic oxidation is carried out in an electrolyte containing fluoride. The electrolyte containing fluoride is an aqueous solution of HF and HNO3, wherein the volume concentration of HF is 1% and the volume concentration of HNO3 is 3%; the voltage is maintained at 20V during anodic oxidation, and the time is 120min; after anodic oxidation, heat treatment is carried out: in an atmosphere, 550℃ for 60min, and then cooled in the furnace, to prepare a surface nanocrystalline TLM titanium alloy material with a surface nanotube diameter of 50nm array.

[0077]

Example 8

[0078] Preparation of surface-nanocrystallized TLM titanium alloy material: refer to Example 7, except that the voltage is maintained at 30 V during anodization for 120 min, and the material is heat treated after anodization: in an atmospheric atmosphere, heat treatment at 550 ℃ for 60 min and furnace cooling, to obtain a surface-nanocrystallized TLM titanium alloy material with a surface nanotube array of 100 nm in diameter.

[0079] Example 9

[0080] Preparation of surface-nanocrystallized TLM titanium alloy material: refer to Example 7, except that the voltage is maintained at 40 V during anodization for 120 min, and the material is heat treated after anodization: in an atmospheric atmosphere, heat treatment at 550 ℃ for 60 min and furnace cooling, to obtain a surface-nanocrystallized TLM titanium alloy material with a surface nanotube array of 150 nm in diameter.

[0081] Example 10

[0082] Preparation of surface-nanocrystallized TLM titanium alloy material: refer to Example 7, except that the voltage is maintained at 50 V during anodization for 120 min, and the material is heat treated after anodization: in an atmospheric atmosphere, heat treatment at 550 ℃ for 60 min and furnace cooling, to obtain a surface-nanocrystallized TLM titanium alloy material with a surface nanotube array of 200 nm in diameter.

[0083] Example 11

[0084] Preparation of surface-nanocrystallized TLM titanium alloy material: refer to Example 7, except that the voltage is maintained at 60 V during anodization for 120 min, and the material is heat treated after anodization: in an atmospheric atmosphere, heat treatment at 550 ℃ for 60 min and furnace cooling, to obtain a surface-nanocrystallized TLM titanium alloy material with a surface nanotube array of 300 nm in diameter.

[0085] Figures 5 to 9 are scanning electron microscope images of the surface-nanocrystallized TLM titanium alloy materials prepared in Examples 7 to 11, respectively. Figures 5 to 9 It can be seen that the nanotubes on the surface of the TLM titanium alloy material are arranged in an array and have a suitable three-dimensional spatial structure.

[0086] Figure 10 are scanning electron microscope images of the surface-nanocrystallized TLM titanium alloy materials with nanotube diameters of 100 nm, 200 nm and 300 nm, respectively. Figure 10As can be seen, the nanotubes on the surface of the surface-nanocrystallized TLM titanium alloy material have a structure similar to that of a siphon, which has a function similar to that of a siphon and can adsorb nutrient components such as cytokines, thereby playing a role in providing a three-dimensional space for cell adhesion and releasing the components such as cytokines to surrounding tissue cells, thereby promoting the further proliferation and differentiation of stem cells in vivo and in vitro.

[0087] Example 12

[0088] Preparation of the surface-nanocrystallized TLM titanium alloy material: see Example 7, except that the electrolyte containing fluoride is an aqueous solution of HF and HNO3, wherein the volume concentration of HF is 2% and the volume concentration of HNO3 is 5%; the voltage is maintained at 10 V during the anodization process for 100 min; and the heat treatment after anodization is performed at 500 °C for 40 min in an atmospheric atmosphere and then cooled in the furnace, thereby obtaining the surface-nanocrystallized TLM titanium alloy material.

[0089] Example 13

[0090] Preparation of the surface-nanocrystallized TLM titanium alloy material: see Example 7, except that the electrolyte containing fluoride is an aqueous solution of HF and HNO3, wherein the volume concentration of HF is 3% and the volume concentration of HNO3 is 10%; the voltage is maintained at 80 V during the anodization process for 80 min; and the heat treatment after anodization is performed at 580 °C for 50 min in an atmospheric atmosphere and then cooled in the furnace, thereby obtaining the surface-nanocrystallized TLM titanium alloy material.

[0091] Example 14

[0092] Preparation of the surface-nanocrystallized TLM titanium alloy material: see Example 7, except that the electrolyte containing fluoride is an aqueous solution of HF and HNO3, wherein the volume concentration of HF is 4% and the volume concentration of HNO3 is 7%; the voltage is maintained at 100 V during the anodization process for 50 min; and the heat treatment after anodization is performed at 600 °C for 30 min in an atmospheric atmosphere and then cooled in the furnace, thereby obtaining the surface-nanocrystallized TLM titanium alloy material.

[0093] Example 15

[0094] Preparation of surface nano-crystallized TLM titanium alloy material: See Example 7, except that: the electrolyte containing fluoride is an aqueous solution of HF and HNO3, wherein the volume concentration of HF is 5% and the volume concentration of HNO3 is 1%; the voltage is maintained at 120V for 30 minutes during the anodizing process, and heat treatment is performed after anodizing: in an atmospheric atmosphere, the material is kept at 520°C for 40 minutes and then cooled in the furnace to obtain a surface nano-crystallized TLM titanium alloy material.

[0095] [Example 16]

[0096] Adipose-derived stromal stem cells and surface nano-TLM titanium alloy composite test, specifically:

[0097] A surface-nanostructured TLM titanium alloy sample was treated in a 10% acrylamide solution of nitric acid (0.04 mol / L) and cerium salt (0.002 mol / L) at 70°C for 3 hours under a nitrogen atmosphere. Then, a certain amount of water was added to the reactor, the stirrer was started, and non-ionic polyacrylamide with relative molecular weights of 8 million and 10 million was slowly added to a concentration of about 2%. The temperature was raised to 55°C with ventilation. After the polyacrylamide was dissolved, a certain amount of NaOH was added to reach a pH value of 10-11, and the reaction was carried out for 1 hour. A certain amount of hypochlorite solution was then added, and the pH was maintained at about 10. The temperature was maintained at 55°C for 2 hours. Acid was added to adjust the pH to 7, and after cooling, the surface amination of the material was achieved, increasing its affinity with cytokines, etc.

[0098] The nano-TLM titanium alloy sample after the amino treatment was 60 After irradiation sterilization, the cells were placed in a six-well plate (nanotubes facing upwards), and DMEM culture medium containing epidermal growth factor and ordinary DMEM culture medium were added respectively. The third generation of adipose-derived stromal stem cells were selected to make a culture medium with a concentration of 3.5×10 6 / ml cell suspension was added dropwise to the culture medium of each well and cultured in a carbon dioxide incubator overnight to observe the cell adhesion and proliferation. Figure 11 As shown in the figure, scanning electron microscopy shows that the fat matrix stems and nano-TLM titanium alloy are well attached, and the three-dimensional pore size is suitable for cell attachment and growth; Figure 12 As shown in the figure, under an inverted microscope, adipose-derived stromal stem cells are evenly distributed on the surface of the nano-TLM titanium alloy, and the cell density and growth state are good, indicating that after the adipose-derived stromal stem cells are co-cultured with the nano-TLM titanium alloy, the adipose-derived stromal stem cells are evenly distributed on the surface of the nano-TLM titanium alloy and the growth state is good. The composite effect of the two is good, which is suitable for local continued proliferation, growth and differentiation.

[0099] [Example 17]

[0100] The surface-nanocrystallized TLM titanium alloy material of the present application is compared with other materials on the market in terms of cytotoxicity.

[0101] Other commonly used materials on the market are selected and made into cell scaffolds, including zinc, nickel-titanium alloy, titanium, TC4, TLM, 316L stainless steel, and the diameter is 10mm.

[0102] The alloy material cytotoxicity test is performed, the adipose stromal stem cells are digested into a single cell suspension, dropped onto the alloy material which has been placed in a 24-well plate and sterilized by high temperature and high pressure in advance, and after 6 hours of constant temperature culture, 50ul of cell culture solution is taken from each well and reacted with the LDH (lactate dehydrogenase) reaction solution prepared in advance in a 96-well plate at room temperature for 30min, then the reaction termination solution is added, and then the 96-well plate is placed in an enzyme marker to read the absorbance value at 490nm wavelength, and the statistical results are shown in Figure 13 The cytotoxicity test results show that the TLM titanium alloy material has the smallest cytotoxicity, and in order to detect the growth of adipose stromal stem cells on various alloy materials, a comparison of cell culture on various alloy materials is performed, and then the fluorescence staining results show that the cell morphology on the surface-nanocrystallized TLM titanium alloy material is normal, has no difference with normal cells, and the cell proliferation function is not affected, as shown in Figure 14 .

[0103] The above technical solution is only one embodiment of the present application, and for those skilled in the art, on the basis of the principles disclosed in the present application, various types of improvements or modifications can be easily made, and the technical solution described in the above specific embodiments is not limited to the present application, therefore the above description is only preferred, and does not have the meaning of limitation.

Claims

1. A method for preparing a surface nano-structured TLM titanium alloy material, characterized in that: The following steps are involved: A 1.0 mm thick TLM titanium alloy hot-rolled plate with a phase transition point of 710°C was heat-treated at 610°C for 30 min and then water-cooled. After pickling and sandblasting, eight layers of the plate were stacked at once for sheathing and cold rolling. During the cold rolling process, stress relief annealing was performed at 400°C for 5 min. Finally, an 8-layer TLM titanium alloy ultrafine-grained composite plate with a thickness of 0.2 mm was obtained. The TLM titanium alloy ultrafine-grained composite plate is polished step by step with silicon carbide sandpaper, and then anodized in an electrolyte containing fluoride using the polished TLM titanium alloy ultrafine-grained composite plate as the anode and platinum foil as the cathode. Finally, a heat treatment is performed in an atmospheric atmosphere at 500-600°C for 30-60 minutes and then cooled with the furnace to prepare a surface nano-crystallized TLM titanium alloy material; the fluoride-containing electrolyte is an aqueous solution of HF and HNO3, wherein the volume concentration of HF is 1%-5% and the volume concentration of HNO3 is 1%-10%.

2. The method for preparing the surface nano-structured TLM titanium alloy material according to claim 1, characterized in that: During the anodizing process, the voltage is maintained at 10-120V and the time is 30-120min.

3. The method for preparing the surface nano-structured TLM titanium alloy material according to claim 1, characterized in that: Acid cleaning is carried out in a mixed solution of HF, HNO3 and H2O for 2-7 minutes, and the volume ratio of HF, HNO3 and H2O in the mixed solution is 1:5:7; The sandblasting treatment uses white corundum coarse blasting, the sandblasting pressure is 0-0.45Mpa, and the duration is 10-30s.

4. Application of the surface nano-crystallized TLM titanium alloy material prepared by the preparation method according to any one of claims 1 to 3 in cartilage scaffold materials.

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