CN113368315B

Movatterモバイル変換

Info

Publication number: CN113368315B
Application number: CN202110639873.7A
Authority: CN
Inventors: 王东辉; 郝雪青; 梁春永; 李宝娥; 王洪水
Original assignee: Hebei University of Technology
Current assignee: Hebei University of Technology
Priority date: 2021-06-09
Filing date: 2021-06-09
Publication date: 2022-04-19
Anticipated expiration: 2041-06-09
Also published as: CN113368315A

Abstract

Translated fromChinese

本发明为一种具有温敏性水凝胶涂层的医用镍钛合金材料及其制备方法与应用。该复合材料自下而上为镍钛合金基体、多级多孔的二氧化钛纳米层和聚异丙基丙烯酰胺温敏性水凝胶涂层。其制备方法是首先通过阳极氧化在医用镍钛合金表面制备多级多孔的二氧化钛纳米层，然后通过BIBAPTES溶液的引发剂接枝到表面，随后通过化学接枝将聚异丙基丙烯酰胺温敏性水凝胶负载在样品表面，得到了镍钛合金基体/二氧化钛纳米层/聚异丙基丙烯酰胺温敏性水凝胶涂层的复合材料。本发明样品制备方法简单、成本较低，同时聚异丙基丙烯酰胺温敏性水凝胶具有较好的生物相容性，表面光滑柔软，能够满足医用植入的需求。

The invention relates to a medical nickel-titanium alloy material with a temperature-sensitive hydrogel coating and a preparation method and application thereof. The composite material is composed of a nickel-titanium alloy matrix, a multi-level porous titanium dioxide nanolayer and a polyisopropylacrylamide temperature-sensitive hydrogel coating from bottom to top. The preparation method is to first prepare a multi-level porous titanium dioxide nanolayer on the surface of medical nickel-titanium alloy by anodization, and then graft it to the surface by the initiator of BIBAPTES solution, and then chemically graft the polyisopropylacrylamide thermosensitive. The hydrogel was loaded on the surface of the sample, and the composite material of nickel-titanium alloy matrix/titanium dioxide nanolayer/polyisopropylacrylamide thermosensitive hydrogel coating was obtained. The sample preparation method of the invention is simple and low in cost, and the polyisopropylacrylamide temperature-sensitive hydrogel has good biocompatibility, smooth and soft surface, and can meet the needs of medical implantation.

Description

Translated fromChinese

一种具有温敏性水凝胶涂层的医用镍钛合金材料及其制备方法与应用A kind of medical nickel-titanium alloy material with temperature-sensitive hydrogel coating and its preparation method and application

技术领域technical field

本发明属于生物材料表面改性技术领域，具体涉及一种具有温敏性水凝胶涂层的医用镍钛合金材料及其制备方法和应用，具体说，是涉及一种以生物医用镍钛合金材料为基底，通过电化学阳极氧化在镍钛合金表面制备多级多孔的二氧化钛纳米层，再将温敏性水凝胶化学接枝在样品表面，以此来实现选择性生物学效应，即利于细胞粘附，促进内皮化的同时有着较强的抗菌、抗污性能，避免发生感染。The invention belongs to the technical field of surface modification of biological materials, and in particular relates to a medical nickel-titanium alloy material with a temperature-sensitive hydrogel coating and a preparation method and application thereof, in particular to a biological medical nickel-titanium alloy The material is used as the substrate, and the multi-level porous titanium dioxide nanolayer is prepared on the surface of the nickel-titanium alloy by electrochemical anodization, and then the temperature-sensitive hydrogel is chemically grafted on the surface of the sample to achieve selective biological effects, which is beneficial to It has strong antibacterial and antifouling properties while promoting endothelialization and preventing infection.

背景技术Background technique

镍钛合金凭借其独特的形状记忆功能、优异的机械性能和生物相容性广泛应用于各腔道支架。但高的镍离子含量使人们对镍钛合金作为长期植入材料的安全性产生了怀疑，同时镍钛合金高的表面模量和硬度会对组织造成机械损伤，并且由于镍钛合金表面并不具备抗菌性，因此镍钛合金的应用并不能完全满足临床要求，需要对镍钛合金表面进行改性处理，使其尽可能去除一部分镍离子的同时能满足抗菌、抗污的性能。Nitinol alloys are widely used in various cavity stents due to their unique shape memory function, excellent mechanical properties and biocompatibility. However, the high nickel ion content casts doubt on the safety of Nitinol as a long-term implant material. At the same time, the high surface modulus and hardness of Nitinol can cause mechanical damage to the tissue, and because the surface of Nitinol does not It has antibacterial properties, so the application of nickel-titanium alloys cannot fully meet the clinical requirements. It is necessary to modify the surface of nickel-titanium alloys to remove part of the nickel ions as much as possible while meeting the antibacterial and antifouling properties.

温敏性水凝胶是一种具有三维网络结构的高分子材料，最低临界温度以下有着大量亲水基团，具有良好的吸水性。由于它表面富含水合层，同时具有多孔结构及非常柔软的特性，可以非常接近地模拟正常组织，具有良好生物相容性的同时，其界面自由能较低，从而保持了细菌、细胞及蛋白质在其表面较低的吸附趋势，不易导致结壳或生物膜的产生，具有良好的防污性能；在最低临界温度以上，聚合物与水分子之间的氢键遭到破坏，构象发生转变，水合层消失，聚合物表现为疏水，表面不再具有抗菌性。聚异丙基丙烯酰胺温敏性水凝胶具有优异的生物相容性，利于细胞粘附，促进内皮化，利于伤口愈合。但是，由于其机械性能较差，无法单独作为医用植入材料使用。Thermosensitive hydrogel is a polymer material with a three-dimensional network structure. It has a large number of hydrophilic groups below the lowest critical temperature and has good water absorption. Due to its rich hydration layer on its surface, porous structure and very soft characteristics, it can simulate normal tissue very closely. It has good biocompatibility and low interfacial free energy, thus maintaining bacteria, cells and proteins. It has a low adsorption tendency on its surface, which is not easy to lead to the formation of crusts or biofilms, and has good antifouling performance; above the minimum critical temperature, the hydrogen bond between the polymer and the water molecule is destroyed, and the conformation changes. The hydration layer disappears, the polymer appears hydrophobic, and the surface is no longer antimicrobial. Polyisopropylacrylamide thermosensitive hydrogel has excellent biocompatibility, which facilitates cell adhesion, promotes endothelialization, and facilitates wound healing. However, due to its poor mechanical properties, it cannot be used alone as a medical implant material.

本文基于上述调研，对医用镍钛合金材料进行阳极氧化脱合金，构建多级多孔的二氧化钛纳米层，随后利用化学接枝在其表面负载聚异丙基丙烯酰胺水凝胶涂层，以达到温度响应的选择性生物学效应。Based on the above investigations, this paper conducts anodizing and dealloying of medical nickel-titanium alloy materials to construct multi-level porous titanium dioxide nanolayers. Selective biological effects of response.

发明内容SUMMARY OF THE INVENTION

本发明的目的是为了解决现有技术中的问题，提供了一种具有温敏性水凝胶涂层的医用镍钛合金材料及其制备方法和应用。该复合材料自下而上为镍钛合金基体、多级多孔的二氧化钛纳米层和聚异丙基丙烯酰胺温敏性水凝胶涂层。其制备方法是首先通过阳极氧化在医用镍钛合金表面制备多级多孔的二氧化钛纳米层，然后通过将成分为3～5％v/v的3-(2-溴异丁酰溴)丙基(三乙氧基)硅烷(BIBAPTES)和95～97％v/v的无水乙醇的引发剂接枝到表面，随后通过化学接枝将聚异丙基丙烯酰胺温敏性水凝胶负载在样品表面，得到了镍钛合金基体/二氧化钛纳米层/聚异丙基丙烯酰胺温敏性水凝胶涂层的复合材料。本发明样品制备方法简单、成本较低，同时聚异丙基丙烯酰胺温敏性水凝胶具有较好的生物相容性，表面光滑柔软，能够满足医用植入的需求。The purpose of the present invention is to solve the problems in the prior art, and provide a medical nickel-titanium alloy material with a temperature-sensitive hydrogel coating and a preparation method and application thereof. The composite material is composed of a nickel-titanium alloy matrix, a multi-level porous titanium dioxide nanolayer and a polyisopropylacrylamide temperature-sensitive hydrogel coating from bottom to top. The preparation method is to first prepare a multi-level porous titanium dioxide nanolayer on the surface of medical nickel-titanium alloy by anodizing, and then prepare 3-(2-bromoisobutyryl bromide) propyl ( The initiator of triethoxy)silane (BIBAPTES) and 95-97% v/v absolute ethanol was grafted to the surface, and then the polyisopropylacrylamide thermosensitive hydrogel was loaded on the sample by chemical grafting On the surface, a composite of nickel-titanium alloy matrix/titanium dioxide nanolayer/polyisopropylacrylamide thermosensitive hydrogel coating was obtained. The sample preparation method of the invention is simple and low in cost, and the polyisopropylacrylamide temperature-sensitive hydrogel has good biocompatibility, smooth and soft surface, and can meet the needs of medical implantation.

为实现上述目的，本发明采用如下的技术方案：For achieving the above object, the present invention adopts the following technical scheme:

一种具有温敏性水凝胶涂层的医用镍钛合金材料，该合金材料自下而上为镍钛合金基体、多级多孔的二氧化钛纳米层和聚异丙基丙烯酰胺温敏性水凝胶涂层，其中，多级多孔的二氧化钛纳米层厚度约为3～15μm，其表面的多级多孔结构是由微米级和纳米级两种级别的孔组成的，其中微米孔孔径约1～10μm，纳米孔孔径约200～600nm，纳米孔非均匀分布于微米孔内；聚异丙基丙烯酰胺温敏性水凝胶涂层厚度约为20～300nm。A medical nickel-titanium alloy material with a temperature-sensitive hydrogel coating, the alloy material is a nickel-titanium alloy matrix, a multi-level porous titanium dioxide nanolayer and a polyisopropylacrylamide temperature-sensitive hydrogel from bottom to top Adhesive coating, wherein, the thickness of the multi-level porous titanium dioxide nanolayer is about 3-15 μm, and the multi-level porous structure on the surface is composed of micro-scale and nano-scale pores, wherein the micro-pore diameter is about 1-10 μm , the diameter of the nanopores is about 200-600nm, the nanopores are unevenly distributed in the micropores; the thickness of the polyisopropylacrylamide thermosensitive hydrogel coating is about 20-300nm.

所述的具有温敏性水凝胶涂层的医用镍钛合金材料的制备方法，包括如下步骤：The preparation method of the medical nickel-titanium alloy material with temperature-sensitive hydrogel coating comprises the following steps:

(1)材料表面预处理：对镍钛合金材料进行酸洗，然后清洗，烘干；(1) Material surface pretreatment: pickling the nickel-titanium alloy material, then cleaning and drying;

(2)阳极氧化：将步骤(1)预处理后的镍钛合金作为阳极，石墨片作为阴极，在电解液中进行2～100min的恒电压脱合金处理，在镍钛合金表面会形成多级多孔的二氧化钛纳米层结构，电解后清洗、烘干备用；(2) Anodizing: using the pretreated nickel-titanium alloy in step (1) as the anode, and the graphite sheet as the cathode, a constant voltage dealloying treatment is carried out in the electrolyte for 2 to 100 minutes, and multi-level formation will be formed on the surface of the nickel-titanium alloy. Porous titanium dioxide nano-layer structure, cleaning and drying after electrolysis;

其中，电解液是浓度为1.0～2.5mol/L的HNO₃溶液，阳极氧化脱合金处理的电压为2～6 V，两电极之间距离为30～50mm；Wherein, the electrolyte is an HNO₃ solution with a concentration of 1.0-2.5 mol/L, the voltage of the anodic oxidation dealloying treatment is 2-6 V, and the distance between the two electrodes is 30-50 mm;

(3)热氧化：将步骤(2)处理的镍钛合金进行热氧化，随后浸泡在NaOH溶液中10～30 min，取出后清洗、烘干；(3) Thermal oxidation: thermally oxidize the nickel-titanium alloy treated in step (2), then soak it in NaOH solution for 10-30 min, take it out, clean and dry;

(4)接枝引发剂：将步骤(3)处理好的样品浸泡在3～5％v/v的3-(2-溴异丁酰溴)丙基 (三乙氧基)硅烷(BIBAPTES)的乙醇溶液中40～50min，取出后用无水乙醇冲洗，烘干；(4) Grafting initiator: soak the sample treated in step (3) in 3-5% v/v 3-(2-bromoisobutyryl bromide)propyl(triethoxy)silane (BIBAPTES) in ethanol solution for 40-50min, take out, rinse with absolute ethanol, and dry;

(5)通过原子转移自由基聚合法负载聚异丙基丙烯酰胺温敏性水凝胶涂层：室温下将经过步骤(4)处理的样品浸泡到聚异丙基丙烯酰胺溶液中10～15min，取出后用离子水清洗，再在氮气氛围下干燥，得到具有温敏性水凝胶涂层的医用镍钛合金材料；(5) Loading polyisopropylacrylamide thermosensitive hydrogel coating by atom transfer radical polymerization method: soak the sample treated in step (4) in polyisopropylacrylamide solution for 10-15min at room temperature , washed with ionized water after taking out, and then dried under nitrogen atmosphere to obtain a medical nickel-titanium alloy material with a temperature-sensitive hydrogel coating;

其中，所述的聚异丙基丙烯酰胺溶液中，包含8～10％w/v异丙基丙烯酰胺,0.4～0.5％w/v 溴化亚铜,0.1～0.2％w/v溴化铜,1.3～1.5％v/v五甲基二乙烯三胺组成的溶液中。Wherein, the polyisopropylacrylamide solution contains 8-10%w/v isopropylacrylamide, 0.4-0.5%w/v cuprous bromide, and 0.1-0.2%w/v copper bromide , 1.3 to 1.5% v/v pentamethyl diethylene triamine in a solution.

所述的步骤(1)中，酸洗时，酸洗液由质量分数为40％的氢氟酸(HF)、质量分数为60％的硝酸(HNO₃)和H₂O混合而成，酸洗时间为1～10min；HF、HNO₃和H₂O的体积比为1～5：5～10：30～80。In the step (1), during pickling, the pickling solution is mixed with hydrofluoric acid (HF) with a mass fraction of 40%, nitric acid (HNO₃ ) with a mass fraction of 60% and H₂ O, and the acid is mixed with H2O. The washing time is 1-10 min; the volume ratio of HF, HNO₃ and H₂ O is 1-5:5-10:30-80.

所述的步骤(3)中进行热氧化的温度为350～650℃，升温速率为2～10℃/min，保温0.5～3 h。NaOH溶液的浓度为1.5～2.0mol/L，随后用去离子水冲洗，并在空气中烘干。In the step (3), the temperature for thermal oxidation is 350-650° C., the heating rate is 2-10° C./min, and the temperature is kept for 0.5-3 h. The concentration of the NaOH solution is 1.5-2.0 mol/L, followed by rinsing with deionized water and drying in the air.

所述的步骤(4)中BIBAPTES的乙醇溶液的制备方法为：0℃，氮气条件下，在四氢呋喃中依次加入三乙胺、2-溴异丁酰溴、3-氨基丙基三乙氧基硅烷，随后升至常温下搅拌过夜，然后然后真空抽滤；随后将真空抽滤得到的滤液加入到无水乙醇，得到BIBAPTES的乙醇溶液；In the described step (4), the preparation method of the ethanol solution of BIBAPTES is as follows: at 0° C., under nitrogen conditions, triethylamine, 2-bromoisobutyryl bromide and 3-aminopropyltriethoxy are successively added to tetrahydrofuran. Silane was then raised to room temperature and stirred overnight, then vacuum filtration; then the filtrate obtained by vacuum filtration was added to absolute ethanol to obtain an ethanolic solution of BIBAPTES;

其中，在每20～30ml四氢呋喃中加入2～5ml三乙胺、5.8～6.0g的2-溴异丁酰溴、4.0～4.5 g的3-氨基丙基三乙氧基硅烷；滤液和无水乙醇的体积比3～5：95～97。Wherein, 2-5 ml of triethylamine, 5.8-6.0 g of 2-bromoisobutyryl bromide and 4.0-4.5 g of 3-aminopropyltriethoxysilane are added to every 20-30 ml of tetrahydrofuran; the filtrate and anhydrous The volume ratio of ethanol is 3～5:95～97.

所述的具有温敏性水凝胶涂层的医用镍钛合金材料的应用，用于制备与人体内腔道接触的医用镍钛合金器件中。The application of the medical nickel-titanium alloy material with the temperature-sensitive hydrogel coating is used in the preparation of medical nickel-titanium alloy devices in contact with the human body cavity.

本发明的实质性特点为：The essential features of the present invention are:

本发明提出的技术方案解决了传统医用镍钛合金表面改性方法存在的问题，首先，温敏性水凝胶涂层的负载赋予了医用镍钛合金表面优异的抑菌性能，解决了传统阳极氧化方法处理医用镍钛合金表面抑菌性能缺乏的问题；其次，负载于医用镍钛合金表面的温敏性水凝胶涂层可以有效的抑制镍钛合金基体中镍离子的溶出，消除了医用镍钛合金植入人体后存在的安全隐患；最后，通过阳极氧化制备的多级多孔结构显著的提高了医用镍钛合金基体与温敏性水凝胶涂层之间的结合力，避免了医用镍钛合金在服役过程中涂层脱落的风险。The technical solution proposed by the present invention solves the problems existing in the surface modification method of traditional medical nickel-titanium alloy. First, the load of the temperature-sensitive hydrogel coating endows the surface of medical nickel-titanium alloy with excellent bacteriostatic properties, which solves the problem of traditional anode The oxidation method can solve the problem of lack of bacteriostatic properties on the surface of medical nickel-titanium alloy; secondly, the temperature-sensitive hydrogel coating loaded on the surface of medical nickel-titanium alloy can effectively inhibit the dissolution of nickel ions in the nickel-titanium alloy matrix, eliminating the need for medical nickel-titanium alloys. The safety hazard after Nitinol is implanted into the human body; finally, the multi-level porous structure prepared by anodization significantly improves the bonding force between the medical Nitinol substrate and the temperature-sensitive hydrogel coating, avoiding the need for medical treatment. Risk of coating peeling off Nitinol during service.

得到的医用镍钛合金表面制备多级多孔的微观结构，再将聚异丙基丙烯酰胺温敏性水凝胶涂层负载在其表面，得到的复合材料自下而上依次为镍钛合金基体、多级多孔的二氧化钛纳米层和聚异丙基丙烯酰胺温敏性水凝胶涂层。A multi-level porous microstructure is prepared on the surface of the obtained medical nickel-titanium alloy, and then a polyisopropylacrylamide temperature-sensitive hydrogel coating is loaded on the surface, and the obtained composite material is a nickel-titanium alloy matrix from bottom to top. , Hierarchical porous titanium dioxide nanolayers and polyisopropylacrylamide thermosensitive hydrogel coatings.

与现有技术相比，本发明的有益效果在于：Compared with the prior art, the beneficial effects of the present invention are:

1.在医用镍钛合金表面进行阳极氧化随后通过引发剂接枝聚异丙基丙烯酰胺温敏性水凝胶涂层的实验方案具有创新性，之前尚未报道。1. The experimental protocol of anodizing on the surface of medical nickel-titanium alloy followed by grafting of polyisopropylacrylamide thermosensitive hydrogel coating by an initiator is innovative and has not been reported before.

2.本发明相比现有的表面改性方法有进步，首先，聚异丙基丙烯酰胺温敏性水凝胶涂层的负载赋予了医用镍钛合金表面优异的抑菌性能，解决了传统阳极氧化方法处理医用镍钛合金表面抑菌性能缺乏的问题；其次，负载于医用镍钛合金表面的温敏性水凝胶涂层可以有效的抑制镍钛合金基体中镍离子的溶出，消除了医用镍钛合金植入人体后存在的安全隐患；最后，通过阳极氧化制备的多级多孔结构去除一部分镍离子的同时显著的提高了医用镍钛合金基体与温敏性水凝胶涂层之间的结合力，避免了医用镍钛合金在服役过程中涂层脱落的风险。2. Compared with the existing surface modification methods, the present invention has progress. First, the load of the polyisopropylacrylamide thermosensitive hydrogel coating endows the surface of the medical nickel-titanium alloy with excellent bacteriostatic properties, which solves the problem of traditional methods. The problem of lack of bacteriostatic properties on the surface of medical nickel-titanium alloy was treated by anodizing method; secondly, the temperature-sensitive hydrogel coating loaded on the surface of medical nickel-titanium alloy can effectively inhibit the dissolution of nickel ions in the nickel-titanium alloy matrix, eliminating the need for The potential safety hazard after medical nickel-titanium alloy is implanted into the human body; finally, the multi-level porous structure prepared by anodization removes part of the nickel ions and significantly improves the gap between the medical nickel-titanium alloy substrate and the temperature-sensitive hydrogel coating. The bonding force avoids the risk of coating peeling off of the medical nickel-titanium alloy during service.

3.本技术方案相比之前，镍离子含量减少，见实施例1表1；温敏性水凝胶涂层和医用镍钛合金基体的结合力也有改善，见实施例1图2、图3；抑菌效果有提升，数见实施例6图6；生物相容性提高，促进基因表达，见实施例8图9。3. Compared with the previous technical solution, the nickel ion content is reduced, see Table 1 in Example 1; the bonding force between the temperature-sensitive hydrogel coating and the medical nickel-titanium alloy substrate is also improved, see Figure 2 and Figure 3 in Example 1 ; The bacteriostatic effect is improved, as shown in Figure 6 in Example 6; the biocompatibility is improved, and gene expression is promoted, as shown in Figure 9 in Example 8.

本发明提出的技术方案解决了传统医用镍钛合金表面改性方法存在的问题，通过阳极氧化可以去除一部分镍离子，消除了医用镍钛合金植入人体后存在的安全隐患，同时在镍钛合金表面制备多级多孔的二氧化钛纳米层，可以增强温敏性水凝胶涂层与镍钛合金基体的结合强度,避免了医用镍钛合金在服役过程中涂层脱落的风险(见实施例1)。温敏性水凝胶表面平滑柔软，可以舒缓高表面模量对组织造成的内在机械刺激，在32℃以下，聚异丙基丙烯酰胺水凝胶表面凭借其低的界面能和高的水合率，不易与细菌、细胞或蛋白质等发生粘连，起到抗菌抗污的效果；接近人体温度时，水合层消失，但多孔的二氧化钛纳米层仍可将水分捕捉到纳米结构中，使得表面仍具有优异的抗菌性，解决了传统阳极氧化方法处理医用镍钛合金表面抑菌性能缺乏的问题(见实施例6、实施例7)；同时聚异丙基丙烯酰胺温敏性水凝胶具备的优良生物相容性利于细胞、蛋白质的粘附，促进内皮化，加速伤口愈合，以此可达到选择性生物学效应(见实施例8)。The technical solution proposed by the invention solves the problems existing in the traditional medical nickel-titanium alloy surface modification method. Part of the nickel ions can be removed by anodizing, which eliminates the potential safety hazard after the medical nickel-titanium alloy is implanted into the human body. The preparation of multi-level porous titanium dioxide nanolayers on the surface can enhance the bonding strength of the temperature-sensitive hydrogel coating and the nickel-titanium alloy substrate, and avoid the risk of coating peeling off of the medical nickel-titanium alloy during service (see Example 1) . The surface of the thermosensitive hydrogel is smooth and soft, which can relieve the inherent mechanical stimulation of the tissue caused by the high surface modulus. Below 32 °C, the surface of the polyisopropylacrylamide hydrogel is characterized by its low interfacial energy and high hydration rate. It is not easy to adhere to bacteria, cells or proteins, and has the effect of antibacterial and antifouling; when the temperature is close to the human body, the hydration layer disappears, but the porous titanium dioxide nanolayer can still capture water into the nanostructure, so that the surface still has excellent It solved the problem of lack of antibacterial properties on the surface of medical nickel-titanium alloys treated by traditional anodizing methods (see Example 6 and Example 7); Compatibility is conducive to the adhesion of cells and proteins, promotes endothelialization, and accelerates wound healing, thereby achieving selective biological effects (see Example 8).

附图说明Description of drawings

图1为本发明阳极氧化装置示意图Fig. 1 is the schematic diagram of the anodic oxidation device of the present invention

图2为实施例1中的样品低倍与高倍扫描电镜形貌图Fig. 2 is the low-magnification and high-magnification scanning electron microscope topography of the sample in Example 1

图3为经实施例1处理后得到的样品横截面扫描电镜形貌图；图3a为样品低倍下横截面扫描电镜形貌图，图3b,c分别为图3a中样品横截面局部放大扫描电镜形貌图Fig. 3 is the SEM morphology of the cross-section of the sample obtained after the treatment in Example 1; Fig. 3a is the SEM morphology of the cross-section under low magnification of the sample, and Fig. 3b and c are respectively the magnified scanning of the partial cross-section of the sample in Fig. 3a Electron microscope topography

图4为经实施例1处理后得到样品的XRD图谱Fig. 4 is the XRD pattern of the sample obtained after the treatment of Example 1

图5为经实施例1处理后得到样品的XPS图谱：其中，图5a为表面的宽扫描光谱图，图5b为表面的碳1s核心能级光谱图Fig. 5 is the XPS spectrum of the sample obtained after the treatment in Example 1: wherein, Fig. 5a is the wide scan spectrum of the surface, and Fig. 5b is thecarbon 1s core energy level spectrum of the surface

图6为经实施例6处理后得到样品的琼脂培养板涂布结果图Figure 6 is a graph showing the results of coating on the agar plate of the sample obtained after the treatment in Example 6

图7为经实施例7处理后得到的样品在空气中测得水的接触角图Fig. 7 is the contact angle diagram of water measured in air for the sample obtained after the treatment in Example 7

图8为经实施例7处理后得到的样品在水中测得1,2-二氯乙烷的接触角图Fig. 8 is the contact angle diagram of 1,2-dichloroethane measured in water for the sample obtained after the treatment in Example 7

图9为经实施例8处理后得到的样品的三种基因表达量图；图9a为RunX2蛋白的基因表达量，图9b为ALP蛋白的基因表达量，图9c为OCN蛋白的基因表达量Fig. 9 is a graph showing the expression levels of three genes in the samples treated in Example 8; Fig. 9a is the gene expression level of RunX2 protein, Fig. 9b is the gene expression level of ALP protein, Fig. 9c is the gene expression level of OCN protein

具体实施方式Detailed ways

下面结合附图和具体实施方式对本发明作进一步详细说明，具体步骤如下：The present invention is described in further detail below in conjunction with the accompanying drawings and specific embodiments, and the specific steps are as follows:

(1)材料预处理：将医用镍钛合金(镍、钛原子比为1：1)材料通过线切割加工成

的薄片状样品，进行酸洗，酸洗时，酸洗液由质量分数为40％的HF、质量分数为60％的HNO₃和H₂O混合而成，酸洗时间为1～10min，去除表面油污、杂质等；HF、HNO₃和H₂O的体积比为1～5：5～10：30～80。清洗烘干，并用去离子水中超声清洗10min，在室温下干燥后备用。(1) Material pretreatment: The medical nickel-titanium alloy (the atomic ratio of nickel and titanium is 1:1) is processed by wire cutting into

The flake samples were pickled. During pickling, the pickling solution was mixed with HF with a mass fraction of 40%, HNO₃ and H₂ O with a mass fraction of 60%, and the pickling time was 1 to 10 min. Surface oil, impurities, etc.; the volume ratio of HF, HNO₃ and H₂ O is 1-5:5-10:30-80. After cleaning and drying, ultrasonically cleaned in deionized water for 10 min, and dried at room temperature for later use.

(2)阳极氧化：将浓度为1.0～2.5mol/L的HNO₃溶液作为电解液，以步骤1处理好的镍钛合金作为阳极，石墨片作为阴极，两电极间距设置为40mm，在2～6V恒定电压下氧化 2～100min，反应结束后取出镍钛合金用去离子水多次冲洗后，并在去离子水中超声5min，烘干待用。(2) Anodizing: use_HNO3 solution with a concentration of 1.0-2.5mol/L as the electrolyte, the nickel-titanium alloy treated in step 1 as the anode, the graphite sheet as the cathode, and the distance between the two electrodes is set to 40mm, and the distance between the two electrodes is set to 40mm. Oxidize at a constant voltage of 6V for 2-100min. After the reaction, take out the nickel-titanium alloy and rinse it with deionized water for several times, then ultrasonicate it in deionized water for 5min, and dry it for use.

(3)热氧化：将步骤(2)处理的镍钛合金以2～10℃/min的升温速率升至350～650℃，并保温0.5～3h。随后将样品浸泡在1.5～2.0mol/L NaOH溶液中10～30min，用去离子水清洗并烘干。(3) Thermal oxidation: the nickel-titanium alloy treated in step (2) is raised to 350-650° C. at a heating rate of 2-10° C./min, and kept for 0.5-3 hours. Then the samples were soaked in 1.5-2.0 mol/L NaOH solution for 10-30 min, washed with deionized water and dried.

(4)接枝引发剂：在0℃，氮气条件下，在20～30ml四氢呋喃中依次加入2～5ml三乙胺、5.8～6.0g的2-溴异丁酰溴、4.0～4.5g的3-氨基丙基三乙氧基硅烷，随后升至常温下搅拌过夜。过滤两次，并在真空中蒸发掉多余的四氢呋喃；然后将真空抽滤得到的滤液加入到无水乙醇中(滤液和无水乙醇的体积比3～5：95～97)，得到BIBAPTES的乙醇溶液。(4) Grafting initiator: in 20-30 ml of tetrahydrofuran, 2-5 ml of triethylamine, 5.8-6.0 g of 2-bromoisobutyryl bromide, 4.0-4.5 g of 3 -aminopropyltriethoxysilane, then warmed to ambient temperature and stirred overnight. Filter twice, and evaporate excess tetrahydrofuran in vacuum; then add the filtrate obtained by vacuum filtration into absolute ethanol (the volume ratio of filtrate and absolute ethanol is 3～5: 95～97) to obtain the ethanol of BIBAPTES solution.

随后将第三步得到的样品浸泡在该溶液中40～50min，取出样品用无水乙醇冲洗三次，氮气氛围下干燥。Subsequently, the sample obtained in the third step was soaked in the solution for 40-50 min, the sample was taken out, washed three times with absolute ethanol, and dried under nitrogen atmosphere.

(5)通过原子转移自由基聚合法负载聚异丙基丙烯酰胺温敏性水凝胶涂层：在氮气氛围中，将样品浸入到含有8～10％w/v异丙基丙烯酰胺,0.4～0.5％w/v溴化亚铜,0.1～0.2％w/v 溴化铜,1.3～1.5％v/v五甲基二乙烯三胺组成的水溶液中，在室温下反应12min。完成后用去离子水冲洗三次，并在氮气氛围下干燥，则温敏性水凝胶涂层接枝到了样品表面。(5) Loading the polyisopropylacrylamide thermosensitive hydrogel coating by atom transfer radical polymerization method: in a nitrogen atmosphere, immerse the sample in a solution containing 8-10% w/v isopropylacrylamide, 0.4 ～0.5%w/v cuprous bromide, 0.1～0.2%w/v copper bromide, 1.3～1.5%v/v pentamethyldiethylenetriamine in an aqueous solution, react at room temperature for 12min. After rinsing with deionized water three times and drying under nitrogen atmosphere, the temperature-sensitive hydrogel coating was grafted to the surface of the sample.

下面是本发明的具体实施例The following are specific embodiments of the present invention

实施例1Example 1

的薄片状样品，进行酸洗，酸洗时，酸洗液由质量分数为40％的HF、质量分数为60％的HNO₃和H₂O混合而成，酸洗时间为10min；HF、HNO₃和H₂O的体积比为1:5:35 清洗烘干，并用去离子水中超声清洗10min，在室温下干燥后备用，简写为“NiTi”。(1) Material pretreatment: The medical nickel-titanium alloy (the atomic ratio of nickel and titanium is 1:1) is processed by wire cutting into

The flaky sample was pickled. During pickling, the pickling solution was mixed with HF with a mass fraction of 40%, HNO₃ with a mass fraction of 60% and H₂ O, and the pickling time was 10min; HF, HNO The volume ratio of₃ and H₂ O was 1:5:35, washed and dried, and ultrasonically cleaned with deionized water for 10 min, dried at room temperature and used for later use, abbreviated as "NiTi".

(2)阳极氧化：将浓度为1mol/L的HNO₃溶液作为电解液，以步骤1处理好的镍钛合金作为阳极，石墨片作为阴极，两电极间距设置为40mm，在6V电压下氧化2min，反应结束后取出镍钛合金用去离子水多次冲洗后，并在去离子水中超声5min，烘干待用，命名为“TiNL”。(2) Anodization: use_HNO3 solution with a concentration of 1mol/L as the electrolyte, use the nickel-titanium alloy processed in step 1 as the anode, the graphite sheet as the cathode, the distance between the two electrodes is set to 40mm, and oxidized at 6V for 2min , after the reaction, the nickel-titanium alloy was taken out and washed with deionized water for several times, and then ultrasonicated in deionized water for 5 min, dried for use, and named "TiNL".

(3)热氧化：将步骤(2)处理的镍钛合金以5℃/min的升温速率升至350℃，并保温3h。随后将样品浸泡在1.5mol/L NaOH溶液中20min，用去离子水清洗并烘干。(3) Thermal oxidation: the nickel-titanium alloy treated in step (2) was raised to 350°C at a heating rate of 5°C/min, and kept for 3 hours. The samples were then soaked in 1.5 mol/L NaOH solution for 20 min, washed with deionized water and dried.

(4)接枝引发剂：在0℃，氮气条件下，在20ml四氢呋喃中依次加入3ml三乙胺、5.8g的2-溴异丁酰溴、4.0g的3-氨基丙基三乙氧基硅烷，随后升至常温下搅拌过夜。过滤两次，并在真空中蒸发掉多余的四氢呋喃，配制BIBAPTES溶液(真空过滤后的滤液和无水乙醇的体积比是4：96的比例配置混合溶液)。(4) Grafting initiator: 3 ml of triethylamine, 5.8 g of 2-bromoisobutyryl bromide, and 4.0 g of 3-aminopropyltriethoxy group were successively added to 20 ml of tetrahydrofuran at 0°C under nitrogen. silane, then warmed to ambient temperature and stirred overnight. Filter twice, and evaporate the excess tetrahydrofuran in vacuum to prepare a BIBAPTES solution (the volume ratio of the filtrate after vacuum filtration and absolute ethanol is 4:96 to prepare a mixed solution).

随后将步骤(3)处理的样品浸泡在4％v/v 3-(2-溴异丁酰溴)丙基(三乙氧基)硅烷 (BIBAPTES)的乙醇溶液中50min，取出样品用无水乙醇冲洗三次，氮气氛围下干燥。Then the sample treated in step (3) was soaked in 4% v/v ethanol solution of 3-(2-bromoisobutyryl bromide)propyl(triethoxy)silane (BIBAPTES) for 50min, and the sample was taken out with anhydrous Rinse three times with ethanol and dry under nitrogen atmosphere.

(5)通过原子转移自由基聚合法负载聚异丙基丙烯酰胺温敏性水凝胶涂层：在氮气氛围中，将样品浸入到含有8％w/v(g/mL)异丙基丙烯酰胺,0.4％w/v溴化亚铜(g/mL),0.1％w/v溴化铜(g/mL)，1.3％v/v五甲基二乙烯三胺组成的水溶液中，室温下反应12min。完成后用去离子水冲洗三次，并在氮气氛围下干燥，则温敏性水凝胶涂层接枝到了样品表面，命名为“PNIPAM/TiNL”。(5) Loading of polyisopropylacrylamide thermosensitive hydrogel coating by atom transfer radical polymerization method: under nitrogen atmosphere, the sample was immersed in a solution containing 8% w/v (g/mL) isopropyl propylene Amide, 0.4% w/v cuprous bromide (g/mL), 0.1% w/v copper bromide (g/mL), 1.3% v/v pentamethyldiethylenetriamine in water at room temperature The reaction was carried out for 12 minutes. After washing with deionized water three times and drying under nitrogen atmosphere, the temperature-sensitive hydrogel coating was grafted to the surface of the sample, named "PNIPAM/TiNL".

实施例2Example 2

的薄片状样品，进行酸洗，酸洗时，酸洗液由质量分数为40％的HF、质量分数为60％的HNO₃和H₂O混合而成，酸洗时间5min；HF、HNO₃和H₂O的体积比为5:10:35。清洗烘干，并用去离子水中超声清洗10min，在室温下干燥后备用。(1) Material pretreatment: The medical nickel-titanium alloy (the atomic ratio of nickel and titanium is 1:1) is processed by wire cutting into

The flaky samples were pickled. During pickling, the pickling solution was mixed with HF with a mass fraction of 40%, HNO₃ with a mass fraction of 60% and H₂ O, and the pickling time was 5min; HF, HNO₃ and H₂ O in a volume ratio of 5:10:35. After cleaning and drying, ultrasonically cleaned in deionized water for 10 min, and dried at room temperature for later use.

(2)阳极氧化：将浓度为2.5mol/L的HNO₃溶液作为电解液，以步骤1处理好的镍钛合金作为阳极，石墨片作为阴极，两电极间距设置为40mm，在2V电压下氧化90min，反应结束后取出镍钛合金用去离子水多次冲洗后，并在去离子水中超声5min，烘干待用。(₂ ) Anodic oxidation: use the HNO solution with a concentration of 2.5mol/L as the electrolyte, the nickel-titanium alloy processed in step 1 as the anode, the graphite sheet as the cathode, the distance between the two electrodes is set to 40mm, and the oxidation is carried out at a voltage of 2V. 90min, after the reaction, the nickel-titanium alloy was taken out and washed with deionized water for several times, and then ultrasonicated in deionized water for 5min, and dried for later use.

(3)热氧化：将步骤(2)处理的镍钛合金以5℃/min的升温速率升至350℃，并保温3h。随后将样品浸泡在1.5mol/L NaOH溶液中30min，用去离子水清洗并烘干。(3) Thermal oxidation: the nickel-titanium alloy treated in step (2) was raised to 350°C at a heating rate of 5°C/min, and kept for 3 hours. The samples were then soaked in 1.5 mol/L NaOH solution for 30 min, washed with deionized water and dried.

(4)接枝引发剂：在0℃，氮气条件下，在30ml四氢呋喃中依次加入5ml三乙胺、 6g的2-溴异丁酰溴、4.5g的3-氨基丙基三乙氧基硅烷，随后升至常温下搅拌过夜。过滤两次，并在真空中蒸发掉多余的四氢呋喃，配制BIBAPTES溶液(真空过滤后的滤液和无水乙醇的体积比是3：97的比例配置混合溶液)。(4) Grafting initiator: 5ml of triethylamine, 6g of 2-bromoisobutyryl bromide and 4.5g of 3-aminopropyltriethoxysilane were successively added to 30ml of tetrahydrofuran at 0°C under nitrogen conditions , then warmed to room temperature and stirred overnight. Filter twice, and evaporate the excess tetrahydrofuran in vacuum to prepare a BIBAPTES solution (the volume ratio of the filtrate after vacuum filtration and absolute ethanol is 3:97 to prepare a mixed solution).

随后将步骤(3)处理的样品浸泡在3％v/v BIBAPTES的乙醇溶液中45min，取出样品用无水乙醇冲洗三次，氮气氛围下干燥。Subsequently, the sample treated in step (3) was soaked in 3% v/v ethanol solution of BIBAPTES for 45 min, the sample was taken out, washed three times with absolute ethanol, and dried under nitrogen atmosphere.

(5)通过原子转移自由基聚合法负载聚异丙基丙烯酰胺温敏性水凝胶涂层：在氮气氛围中，将样品浸入到含有10％w/v异丙基丙烯酰胺,0.5％w/v溴化亚铜,0.2％w/v溴化铜, 1.5％v/v五甲基二乙烯三胺组成的水溶液中，室温下反应12min。完成后用去离子水冲洗三次，并在氮气氛围下干燥，则温敏性水凝胶涂层接枝到了样品表面。(5) Loading polyisopropylacrylamide thermosensitive hydrogel coating by atom transfer radical polymerization method: in nitrogen atmosphere, immerse the sample in a solution containing 10%w/v isopropylacrylamide, 0.5%w /v cuprous bromide, 0.2% w/v cupric bromide, 1.5% v/v pentamethyldiethylenetriamine in an aqueous solution, react at room temperature for 12 min. After rinsing with deionized water three times and drying under nitrogen atmosphere, the temperature-sensitive hydrogel coating was grafted to the surface of the sample.

实施例3Example 3

的薄片状样品，进行酸洗，酸洗时，酸洗液由质量分数为40％的HF、质量分数为60％的HNO₃和H₂O混合而成，酸洗时间为5min；HF、HNO₃和H₂O的体积比为3:5:60。清洗烘干，并用去离子水中超声清洗10min，在室温下干燥后备用。(1) Material pretreatment: The medical nickel-titanium alloy (the atomic ratio of nickel and titanium is 1:1) is processed by wire cutting into

The flaky samples were pickled. During pickling, the pickling solution was mixed with HF with a mass fraction of 40%, HNO₃ and H₂ O with a mass fraction of 60%, and the pickling time was 5min; HF, HNO The volume ratio of₃ and_H2O was 3:5:60. After cleaning and drying, ultrasonically cleaned in deionized water for 10 min, and dried at room temperature for later use.

(2)阳极氧化：将浓度为2.5mol/L的HNO₃溶液作为电解液，以步骤1处理好的镍钛合金作为阳极，石墨片作为阴极，两电极间距设置为40mm，在5V电压下氧化20min，反应结束后取出镍钛合金用去离子水多次冲洗后，并在去离子水中超声5min，烘干待用。(2) Anodic oxidation:_HNO3 solution with a concentration of 2.5mol/L was used as the electrolyte, the nickel-titanium alloy treated in step 1 was used as the anode, the graphite sheet was used as the cathode, the distance between the two electrodes was set to 40mm, and the oxidation was carried out at a voltage of 5V. 20min, after the reaction, the nickel-titanium alloy was taken out and washed with deionized water for several times, and then ultrasonicated in deionized water for 5min, and dried for use.

(3)热氧化：将步骤(2)处理的镍钛合金以8℃/min的升温速率升至650℃，并保温2h。随后将样品浸泡在1.5mol/L NaOH溶液中30min，用去离子水清洗并烘干。(3) Thermal oxidation: the nickel-titanium alloy treated in step (2) was raised to 650°C at a heating rate of 8°C/min, and kept for 2 hours. The samples were then soaked in 1.5 mol/L NaOH solution for 30 min, washed with deionized water and dried.

(4)接枝引发剂：在0℃，氮气条件下，在25ml四氢呋喃中依次加入5ml三乙胺、 6g的2-溴异丁酰溴、4.5g的3-氨基丙基三乙氧基硅烷，随后升至常温下搅拌过夜。过滤两次，并在真空中蒸发掉多余的四氢呋喃，配制BIBAPTES溶液(真空过滤后的滤液和无水乙醇的体积比是4：96的比例配置混合溶液)。(4) Grafting initiator: 5 ml of triethylamine, 6 g of 2-bromoisobutyryl bromide, and 4.5 g of 3-aminopropyltriethoxysilane were successively added to 25 ml of tetrahydrofuran at 0° C. under nitrogen conditions. , then warmed to room temperature and stirred overnight. Filter twice, and evaporate the excess tetrahydrofuran in vacuum to prepare a BIBAPTES solution (the volume ratio of the filtrate after vacuum filtration and absolute ethanol is 4:96 to prepare a mixed solution).

随后将步骤(3)处理的样品浸泡在4％v/v BIBAPTES的乙醇溶液中45min，取出样品用无水乙醇冲洗三次，氮气氛围下干燥。Subsequently, the sample treated in step (3) was soaked in 4% v/v ethanol solution of BIBAPTES for 45 min, the sample was taken out, washed three times with absolute ethanol, and dried under nitrogen atmosphere.

实施例4Example 4

的薄片状样品，对样品进行酸洗，酸洗时，酸洗液由质量分数为40％的HF、质量分数为60％的HNO₃和H₂O混合而成，酸洗时间为3min；HF、HNO₃和H₂O的体积比为 5:10:80。清洗烘干，并用去离子水中超声清洗10min，在室温下干燥后备用。(1) Material pretreatment: The medical nickel-titanium alloy (the atomic ratio of nickel and titanium is 1:1) is processed by wire cutting into

The flaky sample was pickled. During pickling, the pickling solution was mixed with HF with a mass fraction of 40%, HNO₃ and H₂ O with a mass fraction of 60%, and the pickling time was 3min; HF The volume ratio of , HNO₃ and H₂ O is 5:10:80. After cleaning and drying, ultrasonically cleaned in deionized water for 10 min, and dried at room temperature for later use.

(2)阳极氧化：将浓度为2mol/L的HNO₃溶液作为电解液，以步骤1处理好的镍钛合金作为阳极，石墨片作为阴极，两电极间距设置为30mm，在6V电压下氧化20min，反应结束后取出镍钛合金用去离子水多次冲洗后，并在去离子水中超声5min，烘干待用。(₂ ) anodic oxidation: the HNO solution with a concentration of 2mol/L was used as the electrolyte, the nickel-titanium alloy processed in step 1 was used as the anode, the graphite sheet was used as the cathode, the distance between the two electrodes was set to 30mm, and the oxidation was carried out for 20min at a voltage of 6V. , after the reaction, the nickel-titanium alloy was taken out and washed with deionized water for several times, and then ultrasonicated in deionized water for 5 min, and dried for use.

(3)热氧化：将步骤(2)处理的镍钛合金以2℃/min的升温速率升至400℃，并保温0.5h。随后将样品浸泡在1.8mol/L NaOH溶液中20min，用去离子水清洗并烘干。(3) Thermal oxidation: the nickel-titanium alloy treated in step (2) was raised to 400°C at a heating rate of 2°C/min, and kept for 0.5h. The samples were then soaked in 1.8 mol/L NaOH solution for 20 min, washed with deionized water and dried.

(4)接枝引发剂：在0℃，氮气条件下，在25ml四氢呋喃中依次加入2ml三乙胺、5.9g的2-溴异丁酰溴、4.5g的3-氨基丙基三乙氧基硅烷，随后升至常温下搅拌过夜。过滤两次，并在真空中蒸发掉多余的四氢呋喃，配制BIBAPTES溶液(真空过滤后的滤液和无水乙醇的体积比是5：95的比例配置混合溶液)。(4) Grafting initiator: 2 ml of triethylamine, 5.9 g of 2-bromoisobutyryl bromide and 4.5 g of 3-aminopropyltriethoxy were added to 25 ml of tetrahydrofuran at 0° C. under nitrogen conditions. silane, then warmed to ambient temperature and stirred overnight. Filter twice, and evaporate the excess tetrahydrofuran in vacuum to prepare a BIBAPTES solution (the volume ratio of the filtrate after vacuum filtration and absolute ethanol is 5:95 to prepare a mixed solution).

随后将步骤(3)处理的样品浸泡在5％v/v BIBAPTES的乙醇溶液中40min，取出样品用无水乙醇冲洗三次，氮气氛围下干燥。Subsequently, the sample treated in step (3) was soaked in 5% v/v ethanol solution of BIBAPTES for 40 min, the sample was taken out, washed three times with absolute ethanol, and dried under nitrogen atmosphere.

(5)通过原子转移自由基聚合法负载聚异丙基丙烯酰胺温敏性水凝胶涂层：在氮气氛围中，将样品浸入到含有10％w/v异丙基丙烯酰胺,0.5％w/v溴化亚铜,0.15％w/v溴化铜, 1.4％v/v五甲基二乙烯三胺组成的水溶液中，室温下反应15min。完成后用去离子水冲洗三次，并在氮气氛围下干燥，则温敏性水凝胶涂层接枝到了样品表面。(5) Loading polyisopropylacrylamide thermosensitive hydrogel coating by atom transfer radical polymerization method: in nitrogen atmosphere, immerse the sample in a solution containing 10%w/v isopropylacrylamide, 0.5%w /v cuprous bromide, 0.15% w/v cupric bromide, 1.4% v/v pentamethyldiethylenetriamine in an aqueous solution, react at room temperature for 15 min. After rinsing with deionized water three times and drying under nitrogen atmosphere, the temperature-sensitive hydrogel coating was grafted to the surface of the sample.

实施例5Example 5

的薄片状样品，对样品进行酸洗，酸洗时，酸洗液由质量分数为40％的HF、质量分数为60％的HNO₃和H₂O混合而成，酸洗时间为6min；HF、HNO₃和H₂O的体积比为1:5:40。清洗烘干，并用去离子水中超声清洗10min，在室温下干燥后备用。(1) Material pretreatment: The medical nickel-titanium alloy (the atomic ratio of nickel and titanium is 1:1) is processed by wire cutting into

The flaky sample was pickled. During pickling, the pickling solution was mixed with HF with a mass fraction of 40%, HNO₃ and H₂ O with a mass fraction of 60%, and the pickling time was 6min; HF The volume ratio of , HNO₃ and H₂ O is 1:5:40. After cleaning and drying, ultrasonically cleaned in deionized water for 10 min, and dried at room temperature for later use.

(2)阳极氧化：将浓度为2mol/L的HNO₃溶液作为电解液，以步骤1处理好的镍钛合金作为阳极，石墨片作为阴极，两电极间距设置为50mm，在4V电压下氧化10min，反应结束后取出镍钛合金用去离子水多次冲洗后，并在去离子水中超声5min，烘干待用。(₂ ) Anodic oxidation: use the HNO solution with a concentration of 2mol/L as the electrolyte, the nickel-titanium alloy processed in step 1 as the anode, the graphite sheet as the cathode, the distance between the two electrodes is set to 50mm, and oxidized at 4V for 10min , after the reaction, the nickel-titanium alloy was taken out and washed with deionized water for several times, and then ultrasonicated in deionized water for 5 min, and dried for use.

(3)热氧化：将步骤(2)处理的镍钛合金以10℃/min的升温速率升至650℃，并保温1h。随后将样品浸泡在2mol/L NaOH溶液中15min，用去离子水清洗并烘干。(3) Thermal oxidation: the nickel-titanium alloy treated in step (2) was raised to 650°C at a heating rate of 10°C/min, and kept for 1 hour. The samples were then soaked in 2 mol/L NaOH solution for 15 min, washed with deionized water and dried.

(4)接枝引发剂：在0℃，氮气条件下，在25ml四氢呋喃中依次加入5ml三乙胺、5.9g的2-溴异丁酰溴、4.3g的3-氨基丙基三乙氧基硅烷，随后升至常温下搅拌过夜。过滤两次，并在真空中蒸发掉多余的四氢呋喃，配制BIBAPTES溶液(真空过滤后的滤液和无水乙醇的体积比是5：95的比例配置混合溶液)。(4) Grafting initiator: 5 ml of triethylamine, 5.9 g of 2-bromoisobutyryl bromide, and 4.3 g of 3-aminopropyltriethoxy group were successively added to 25 ml of tetrahydrofuran at 0° C. under nitrogen conditions. silane, then warmed to ambient temperature and stirred overnight. Filter twice, and evaporate the excess tetrahydrofuran in vacuum to prepare a BIBAPTES solution (the volume ratio of the filtrate after vacuum filtration and absolute ethanol is 5:95 to prepare a mixed solution).

随后将步骤(3)处理的样品浸泡在5％v/v BIBAPTES的乙醇溶液中48min，取出样品用无水乙醇冲洗三次，氮气氛围下干燥。Subsequently, the sample treated in step (3) was soaked in 5% v/v ethanol solution of BIBAPTES for 48 min, the sample was taken out, washed with absolute ethanol three times, and dried under nitrogen atmosphere.

(5)通过原子转移自由基聚合法负载聚异丙基丙烯酰胺温敏性水凝胶涂层：在氮气氛围中，将样品浸入到含有9％w/v异丙基丙烯酰胺,0.5％w/v溴化亚铜,0.15％w/v溴化铜, 1.4％v/v五甲基二乙烯三胺组成的水溶液中，室温下反应12min。完成后用去离子水冲洗三次，并在氮气氛围下干燥，则温敏性水凝胶涂层接枝到了样品表面。(5) Loading polyisopropylacrylamide thermosensitive hydrogel coating by atom transfer radical polymerization method: in nitrogen atmosphere, the sample was immersed in 0.5%w isopropylacrylamide containing 9%w/v isopropylacrylamide, 0.5%w /v cuprous bromide, 0.15% w/v cupric bromide, 1.4% v/v pentamethyldiethylenetriamine in an aqueous solution, react at room temperature for 12 min. After rinsing with deionized water three times and drying under nitrogen atmosphere, the temperature-sensitive hydrogel coating was grafted to the surface of the sample.

实施例6Example 6

对实施例1处理的样品表面进行了不同温度下的细菌实验，其步骤如下：用75v/v％的乙醇对样品进行灭菌，置于超净台上通风晾干转移到24孔板。将复苏后保存在4℃冰箱中的大肠杆菌原液震30s，取200μl的原菌液注入15ml离心管，加入1.8ml的生理盐水稀释原菌液(10⁷CFU/ml)，振荡30s后置于37℃培养箱中预热30min。将预热好的菌液振荡 30s后均匀接种到样品表面，每个样品表面60μl稀释菌液，并在细菌培养板孔洞周围补充适量的生理盐水以防菌液蒸干。随后将培养板分别平稳放置在25℃和37℃恒温培养箱中培养。12h后将样品和菌液共同转移到含有4ml生理盐水的离心管中，振荡30s脱附菌液。将100μl上述菌液接种到LB琼脂培养板，并用无菌玻璃棒涂布均匀。将涂好细菌的培养板放入到37℃培养箱中继续培养16h，进行拍照观察。Bacterial experiments at different temperatures were carried out on the surface of the sample treated in Example 1, and the steps were as follows: sterilize the sample with 75v/v% ethanol, place it on an ultra-clean bench to ventilate and dry, and transfer it to a 24-well plate. The E. coli stock solution stored in the refrigerator at 4°C after resuscitation was shaken for 30s, 200μl of the stock solution was injected into a 15ml centrifuge tube, 1.8ml of normal saline was added to dilute the stock solution (10⁷ CFU/ml), and the stock solution was shaken for 30s and then placed in a 15ml centrifuge tube. Preheat in a 37°C incubator for 30 min. The preheated bacterial solution was shaken for 30 s and then evenly inoculated onto the surface of the samples, with 60 μl of diluted bacterial solution on each sample surface, and an appropriate amount of physiological saline was supplemented around the holes of the bacterial culture plate to prevent the bacterial solution from evaporating to dryness. The culture plates were then placed stably in a constant temperature incubator at 25°C and 37°C, respectively. After 12 hours, the sample and the bacterial solution were transferred to a centrifuge tube containing 4 ml of normal saline, and the bacterial solution was desorbed by shaking for 30 s. 100 μl of the above bacterial solution was inoculated onto the LB agar plate and spread evenly with a sterile glass rod. The culture plate coated with bacteria was placed in a 37°C incubator for 16 hours, and photographed for observation.

实施例7Example 7

对实施例1处理的样品进行接触角的测试，以表征材料表面润湿性，其步骤如下：分别在25℃和37℃条件下，将3μl的水垂直悬滴在滴在样品表面，待其稳定后，通过接触角测试仪进行拍照及测量其接触角具体数值；分别在25℃和37℃条件下，将1μl的1,2-二氯乙烷液滴滴在浸泡在水里的样品表面，待其稳定后，通过接触角测试仪进行拍照及测量其接触角具体数值。The contact angle test was carried out on the sample treated in Example 1 to characterize the surface wettability of the material. The steps were as follows: under the conditions of 25 ° C and 37 ° C, drop 3 μl of water vertically on the surface of the sample, and wait for After stabilization, the contact angle tester was used to take pictures and measure the specific value of the contact angle; at 25 °C and 37 °C, drop 1 μl of 1,2-dichloroethane on the surface of the sample immersed in water. , and after it is stabilized, take pictures and measure the specific value of its contact angle with a contact angle tester.

实施例8Example 8

应用聚合酶链反应(PCR)方法检测方法进行样品表面细胞ALP、OCN、RunX2蛋白的表达量检测，其中，样品的制备方法与实施例1中相同。将成骨细胞转移到离心管中，1000rmp，离心5min，将上清液倒掉后加入1ml trizol，吹打均匀至液体澄清且无细胞团块后，室温静止5min，然后把所有液体转移到1.5ml EP管中。每1ml trizol加入0.2ml氯仿，向EP管加入氯仿，盖紧盖子，剧烈上下摇晃10s后，室温静止2min。放入已预冷的离心机，4℃，12000 rmp，离心10min。将上层水层转移到另一个1.5ml EP管中(约450ul)，加入0.5ml异丙醇，混匀，静止10min。然后4℃，12000rmp，离心10min。小心倒掉液体，加入1ml预冷的75％乙醇，轻轻摇晃。然后4℃，12000rmp，离心10min。用枪头尽可能除去液体，超净台内静在置干燥，待白色沉淀变为半透明状，立即加入RNA free水。放入-80℃冰箱保存。用反转录试剂盒将总RNA反转录为cDNA，反转录反应条件为37℃水浴15min，85℃水浴10s。将7.7μl的H₂O、0.4ul Primer F、0.4ul Primer R、10ul EX Taq、1.5μl模板cDNA 混合装到EP管中，放入PCR机中进行扩增。把0.6g琼脂糖溶液放入微波炉加热至完全透明，待琼脂糖溶液冷却至温热时再加入1μl EB替代染料，摇匀并倒入带有梳子的电泳板中冷却凝固。凝固后的琼脂糖块放入电泳槽中，并倒入适量1×TEA溶液。接上电源，电压150V，电流100A进行电泳。用紫外灯进行检测。The polymerase chain reaction (PCR) method was used to detect the expression levels of ALP, OCN and RunX2 proteins in the surface cells of the sample, wherein the sample preparation method was the same as that in Example 1. Transfer the osteoblasts to a centrifuge tube, centrifuge at 1000 rmp for 5 min, pour off the supernatant, add 1 ml of trizol, pipette evenly until the liquid is clear and no cell clumps, rest at room temperature for 5 min, and then transfer all the liquid to 1.5 ml of EP in the tube. Add 0.2 ml of chloroform per 1 ml of trizol, add chloroform to the EP tube, close the lid tightly, shake vigorously up and down for 10 s, and then stand at room temperature for 2 min. Put into a pre-cooled centrifuge, 4 ℃, 12000 rmp, centrifuge for 10min. Transfer the upper aqueous layer to another 1.5ml EP tube (about 450ul), add 0.5ml isopropanol, mix well, and stand for 10min. Then, centrifuge at 12000rmp for 10min at 4°C. Carefully pour off the liquid, add 1 ml of pre-chilled 75% ethanol, and shake gently. Then, centrifuge at 12000rmp for 10min at 4°C. Use a pipette tip to remove the liquid as much as possible, and let it dry in the ultra-clean bench. When the white precipitate becomes translucent, add RNA free water immediately. Store in -80°C refrigerator. The total RNA was reverse transcribed into cDNA using a reverse transcription kit, and the reverse transcription reaction conditions were 37 °C water bath for 15 min and 85 °C water bath for 10 s. Mix 7.7 μl of H₂ O, 0.4 μl Primer F, 0.4 μl Primer R, 10 μl EX Taq, and 1.5 μl template cDNA into an EP tube and put it into a PCR machine for amplification. Put 0.6 g of agarose solution in a microwave oven and heat it until it is completely transparent. When the agarose solution is cooled to warm, add 1 μl of EB substitute dye, shake well and pour it into an electrophoresis plate with a comb to cool and solidify. The solidified agarose block was put into the electrophoresis tank, and an appropriate amount of 1×TEA solution was poured into it. Connect the power supply, the voltage is 150V, and the current is 100A for electrophoresis. Check with UV light.

表1为实施例1的样品的X射线能谱Table 1 is the X-ray energy spectrum of the sample of Example 1

表1Table 1

参见图1，本发明阳极氧化的装置如图1所，用处理过的医用镍钛合金作为阳极，石墨片作为阴极，用硝酸溶液作为电解液进行脱合金处理。Referring to Fig. 1, the anodizing device of the present invention is as shown in Fig. 1, using treated medical nickel-titanium alloy as anode, graphite sheet as cathode, and nitric acid solution as electrolyte for dealloying treatment.

参见图2，图2为实施例1经过步骤(2)处理的样品低倍和高倍的扫描电镜形貌图。图 (a)～图(c)为NiTi样品的不同倍数下的扫描电镜形貌图；图(d)～图(f)为TiNL样品的不同倍数下的扫描电镜形貌图；从图(d)～图(f)可以看到，样品表面形成了多级多孔结构，图(e)中能明显看到其形成的微米级别的孔，孔径约为1～10μm；图(f)中能看到形成的纳米孔，其孔径约为200～600nm。Referring to FIG. 2 , FIG. 2 is a scanning electron microscope topography of the sample at low magnification and high magnification of the sample processed in step (2) of Example 1. Figures (a) to (c) are the SEM images of the NiTi samples at different magnifications; Figures (d) to (f) are the SEM images of the TiNL samples at different magnifications; from Figure (d) ) ~ figure (f), it can be seen that a multi-level porous structure is formed on the surface of the sample. In figure (e), the micron-scale pores formed can be clearly seen, and the pore size is about 1-10 μm; in figure (f), it can be seen that The formed nanopores have a pore diameter of about 200 to 600 nm.

参见图3，图3为经过实施例1处理的样品的横截面低倍和高倍的扫描电镜形貌图。多级多孔的的纳米层厚度约为3～15μm，接枝的聚异丙基丙烯酰胺温敏性水凝胶厚度约为20～300 nm，该纳米结构大大增强了接枝的聚异丙基丙烯酰胺温敏性水凝胶涂层与基体的结合力，可防止该材料在服役过程中表面涂层脱落。Referring to FIG. 3 , FIG. 3 is a low-magnification and high-magnification scanning electron microscope topography of the cross-section of the sample processed in Example 1. The thickness of the multi-level porous nanolayer is about 3-15 μm, and the thickness of the grafted polyisopropylacrylamide thermosensitive hydrogel is about 20-300 nm. The nanostructure greatly enhances the grafted polyisopropylacrylamide. The bonding force between the acrylamide thermosensitive hydrogel coating and the substrate can prevent the surface coating from peeling off during service.

参见图4，图4为经实施例1处理后得到样品的XRD图谱。从图4可以看出, PNIPAM/TiNL的峰比NiTi多了两个Ti的氧化物的峰，这是由于阳极氧化和高温氧化带来的，使得镍钛合金表面富含氧化层。Referring to FIG. 4 , FIG. 4 is the XRD pattern of the sample obtained after the treatment in Example 1. It can be seen from Figure 4 that the peaks of PNIPAM/TiNL are more than that of NiTi by two peaks of Ti oxides, which are caused by anodization and high temperature oxidation, which make the surface of NiTi alloy rich in oxide layer.

参见图5，从图5可以看出，表面的宽扫描光谱显示表面元素的组成主要是碳、氮和氧(图5a)。表面的碳1s核心能级光谱(图5b)可以被曲线化为三个结合能分别为284.8、285.5和288.0 电子伏的峰成分，分别归因于碳氢、碳氮和碳氮氧化合物，碳氮氧化合物是由表面温敏性聚合物带来的，XPS结果证明材料表面成功接枝了聚异丙基丙烯酰胺。Referring to Fig. 5, it can be seen from Fig. 5 that the broad scan spectrum of the surface shows that the composition of surface elements is mainly carbon, nitrogen and oxygen (Fig. 5a). Thecarbon 1s core energy level spectrum of the surface (Fig. 5b) can be plotted into three peak components with binding energies of 284.8, 285.5 and 288.0 eV, respectively, ascribed to hydrocarbon, carbon nitrogen and carbon nitrogen oxides, carbon Nitrogen oxides are brought by surface temperature-sensitive polymers, and XPS results show that polyisopropylacrylamide is successfully grafted on the surface of the material.

参见图6，图6为样品分别在25℃和37℃条件下培养大肠杆菌稀释1000倍后的涂板结果，可以从图(a)和图(d)观察到，无论在25℃还是在37℃下，大肠杆菌在NiTi的琼脂板表面都能形成了密集的菌落；从图(b)和图(e)观察到，TiNL结构的存在几乎对细菌的附着不会有任何抑制作用，由于表面粗糙度的增加会利于细菌粘附；从图(c)和图(f) 观察到，无论在临界温度以下还是在临界温度以上，PNIPAM/TiNL对细菌始终有着较强的抗吸附效果，这是样品表面接枝的水凝胶高水合率和低表面能的功劳。这一结果显示，本发明提出的具有温敏性水凝胶涂层的医用镍钛合金材料表面可以有效抑制细菌粘附，具有优异的抑菌性能，有效避免了医用镍钛合金植入物植入人体后的炎症风险。Referring to Figure 6, Figure 6 shows the plating results after the samples were cultured at 25°C and 37°C after 1000-fold dilution of E. coli. It can be observed from Figures (a) and (d) that no matter at 25°C or at 37°C At ℃, E. coli can form dense colonies on the surface of NiTi agar plates; it can be observed from Figure (b) and Figure (e) that the existence of TiNL structure has almost no inhibitory effect on bacterial attachment, because the surface The increase of roughness will facilitate bacterial adhesion; it is observed from Figures (c) and (f) that PNIPAM/TiNL always has a strong anti-adsorption effect on bacteria no matter below or above the critical temperature, which is The high hydration rate and low surface energy of the hydrogels grafted on the surface of the samples are the credit. This result shows that the surface of the medical nickel-titanium alloy material with the temperature-sensitive hydrogel coating proposed by the present invention can effectively inhibit bacterial adhesion, has excellent antibacterial properties, and effectively avoids the implantation of medical nickel-titanium alloy implants. Inflammation risk in humans.

参见图7，NiTi表面呈现亲水性，且并不受温度影响，而TiNL由于其表面呈现多孔的结构，表面粗糙度增加，因此较NiTi表面更为亲水，也不受温度影响；PNIPAM/TiNL在室温时，由于其表面富含亲水的酰胺基，因此十分亲水，接触角很小，表面水合率非常高，起到抗菌抗污的作用；而当温度升至37℃时，聚合物表面酰胺基和水分子的氢键遭到破坏，疏水的异丙基占据主导地位，因此材料表面接触角骤然增大。Referring to Figure 7, the surface of NiTi is hydrophilic and not affected by temperature, while TiNL is more hydrophilic than the surface of NiTi due to its porous structure and increased surface roughness, and is not affected by temperature; PNIPAM/ At room temperature, TiNL is very hydrophilic because its surface is rich in hydrophilic amide groups, the contact angle is very small, and the surface hydration rate is very high, which plays an antibacterial and antifouling role; when the temperature rises to 37 °C, the polymerization The hydrogen bond between the amide group and the water molecule on the surface of the material is destroyed, and the hydrophobic isopropyl group dominates, so the surface contact angle of the material suddenly increases.

参见图8，在水中测试材料表面的亲疏油(1,2-二氯乙烷)性，NiTi表面具有一定的亲油性，且不受温度变化的影响，TiNL表面多孔结构将水分子捕获在其中，使得其在水中显示出了超疏油性；PNIPAM/TiNL在室温时表面高的水合率使得其表面显示出了超疏油性，温度升高时，多孔结构中依旧时材料表面超疏油性的关键因素。Referring to Figure 8, the oleophobic (1,2-dichloroethane) property of the surface of the material was tested in water. The NiTi surface has a certain lipophilicity and is not affected by temperature changes. The porous structure of the TiNL surface traps water molecules in it. , which makes it show superoleophobicity in water; the high hydration rate of PNIPAM/TiNL surface at room temperature makes its surface show superoleophobicity. factor.

参见图9，ALP、OCN、RunX2三种蛋白的表达在三种样品表面短时间内没有太大差别，表明TiNL和PNIPAM/TiNL并不会对细胞产生毒害作用，具有安全性；而在14天时，PNIPAM/TiNL样品表面的基因表达检测量远远大于了NiTi和TiNL，这印证了聚异丙基丙烯酰胺温敏性水凝胶具有良好的生物相容性，在较长时间内可以利于细胞粘附，促进基因表达，利于伤口恢复。Referring to Figure 9, the expressions of ALP, OCN and RunX2 did not differ much on the surface of the three samples in a short period of time, indicating that TiNL and PNIPAM/TiNL did not have toxic effects on cells and were safe; , the detection amount of gene expression on the surface of PNIPAM/TiNL samples is much greater than that of NiTi and TiNL, which confirms that the polyisopropylacrylamide thermosensitive hydrogel has good biocompatibility and can benefit cells for a long time. Adhesion, promote gene expression, and facilitate wound recovery.

参见表1，表1为经过步骤(2)与未经过步骤(2)的样品成分分析。可以观察到，经过步骤(2)阳极氧化脱合金处理后的镍原子相对占比大大减少，这一结果表明，阳极氧化处理有效的减少了医用镍钛合金表面镍离子相对含量可以有效的避免镍钛合金在植入人体后由于镍离子溶出带来的安全风险。See Table 1. Table 1 shows the composition analysis of the samples after step (2) and without step (2). It can be observed that the relative proportion of nickel atoms after the anodic oxidation dealloying treatment in step (2) is greatly reduced. This result shows that the anodic oxidation treatment can effectively reduce the relative content of nickel ions on the surface of medical nickel-titanium alloy and can effectively avoid nickel. The safety risk of titanium alloys due to the dissolution of nickel ions after implantation in the human body.

本发明未尽事宜为公知技术。Matters not addressed in the present invention are known in the art.

Claims

Translated fromChinese

1.一种具有温敏性水凝胶涂层的医用镍钛合金材料，其特征为该合金材料自下而上为镍钛合金基体、多级多孔的二氧化钛纳米层和聚异丙基丙烯酰胺温敏性水凝胶涂层，其中，多级多孔的二氧化钛纳米层厚度为3~15 μm，其表面的多级多孔结构是由微米级和纳米级两种级别的孔组成的，其中微米孔孔径1~10 μm，纳米孔孔径200~600 nm，纳米孔非均匀分布于微米孔内；聚异丙基丙烯酰胺温敏性水凝胶涂层厚度为20~300 nm。1. a kind of medical nickel-titanium alloy material with thermosensitive hydrogel coating, it is characterized in that this alloy material is nickel-titanium alloy matrix, multi-level porous titanium dioxide nano-layer and polyisopropylacrylamide from bottom to top The temperature-sensitive hydrogel coating, in which the thickness of the multi-level porous titanium dioxide nanolayer is 3-15 μm, and the multi-level porous structure on the surface is composed of micro-scale and nano-scale pores, among which the micro-scale pores The pore size is 1~10 μm, the diameter of nanopores is 200~600 nm, and the nanopores are unevenly distributed in the micropores; the thickness of the polyisopropylacrylamide thermosensitive hydrogel coating is 20~300 nm.

2.如权利要求1所述的具有温敏性水凝胶涂层的医用镍钛合金材料的制备方法，其特征为包括如下步骤：2. the preparation method of the medical nickel-titanium alloy material with thermosensitive hydrogel coating as claimed in claim 1, is characterized in that comprising the steps:

（1）材料表面预处理：对镍钛合金材料进行酸洗，然后清洗，烘干；(1) Material surface pretreatment: pickling the nickel-titanium alloy material, then cleaning and drying;

（2）阳极氧化：将步骤（1）预处理后的镍钛合金作为阳极，石墨片作为阴极，在电解液中进行2~100 min的恒电压脱合金处理，在镍钛合金表面会形成多级多孔的二氧化钛纳米层结构，电解后清洗、烘干备用；(2) Anodizing: Using the pretreated nickel-titanium alloy in step (1) as the anode, and the graphite sheet as the cathode, a constant-voltage dealloying treatment is performed in the electrolyte for 2-100 min, and many formations are formed on the surface of the nickel-titanium alloy. Grade porous titanium dioxide nano-layer structure, cleaned and dried after electrolysis;

其中，电解液是浓度为1.0~2.5 mol/L的HNO₃溶液，阳极氧化脱合金处理的电压为2~6V，两电极之间距离为30~50 mm；Among them, the electrolyte is_HNO3 solution with a concentration of 1.0~2.5 mol/L, the voltage of anodic oxidation dealloying treatment is 2~6V, and the distance between the two electrodes is 30~50 mm;

（3）热氧化：将步骤（2）处理的镍钛合金进行热氧化，随后浸泡在NaOH溶液中10~30min，取出后清洗、烘干；(3) Thermal oxidation: thermally oxidize the nickel-titanium alloy treated in step (2), then soak it in NaOH solution for 10-30 minutes, take it out, wash and dry;

（4）接枝引发剂：将步骤（3）处理好的样品浸泡在3~5% v/v 的3-(2-溴异丁酰溴)丙基(三乙氧基)硅烷(BIBAPTES)的乙醇溶液中40~50 min，取出后用无水乙醇冲洗，烘干；(4) Grafting initiator: soak the sample treated in step (3) in 3-(2-bromoisobutyryl bromide) propyl (triethoxy) silane (BIBAPTES) at 3~5% v/v in ethanol solution for 40-50 min, take out, rinse with absolute ethanol, and dry;

（5）通过原子转移自由基聚合法负载聚异丙基丙烯酰胺温敏性水凝胶涂层：室温下将经过步骤（4）处理的样品浸泡到聚异丙基丙烯酰胺溶液中10~15min，取出后用去离子水清洗，再在氮气氛围下干燥，得到具有温敏性水凝胶涂层的医用镍钛合金材料；(5) Loading polyisopropylacrylamide thermosensitive hydrogel coating by atom transfer radical polymerization method: soak the sample treated in step (4) in polyisopropylacrylamide solution for 10~15min at room temperature , washed with deionized water after taking out, and then dried under nitrogen atmosphere to obtain a medical nickel-titanium alloy material with a temperature-sensitive hydrogel coating;

其中，所述的聚异丙基丙烯酰胺溶液中，包含8~10 % w/v异丙基丙烯酰胺, 0.4~0.5 %w/v溴化亚铜, 0.1~0.2 % w/v 溴化铜, 1.3~1.5 % v/v 五甲基二乙烯三胺。Wherein, in the described polyisopropylacrylamide solution, comprising 8~10% w/v isopropylacrylamide, 0.4~0.5% w/v cuprous bromide, 0.1~0.2% w/v cupric bromide , 1.3~1.5 % v/v pentamethyldiethylenetriamine.

3.如权利要求2所述的具有温敏性水凝胶涂层的医用镍钛合金材料的制备方法，其特征为所述的步骤（1）中，酸洗时，酸洗液由质量分数为40% 的氢氟酸（HF）、质量分数为60%的硝酸（HNO₃）和H₂O混合而成，酸洗时间为1~10 min；HF、HNO₃和H₂O的体积比为1~5：5~10：30~80。3. The method for preparing a medical nickel-titanium alloy material with a temperature-sensitive hydrogel coating as claimed in claim 2, wherein in the step (1), during pickling, the pickling solution is composed of a mass fraction of It is a mixture of 40% hydrofluoric acid (HF), 60% nitric acid (HNO₃ ) and H₂ O, and the pickling time is 1~10 min; the volume ratio of HF, HNO₃ and H₂ O 1~5:5~10:30~80.

4.如权利要求2所述的具有温敏性水凝胶涂层的医用镍钛合金材料的制备方法，其特征为所述的步骤（3）中进行热氧化的温度为350~650 ℃，升温速率为2~10 ℃/min，保温0.5~3 h，NaOH溶液的浓度为1.5~2.0 mol/L。4. The method for preparing a medical nickel-titanium alloy material with a temperature-sensitive hydrogel coating as claimed in claim 2, wherein the temperature for thermal oxidation in the step (3) is 350-650°C, The heating rate was 2~10 ℃/min, the temperature was kept for 0.5~3 h, and the concentration of NaOH solution was 1.5~2.0 mol/L.

5.如权利要求2所述的具有温敏性水凝胶涂层的医用镍钛合金材料的制备方法，其特征为所述的步骤（4）中BIBAPTES的乙醇溶液的制备方法为：0 ℃，氮气条件下，在四氢呋喃中依次加入三乙胺、2-溴异丁酰溴、3-氨基丙基三乙氧基硅烷，随后升至常温下搅拌过夜，然后真空抽滤；随后将真空抽滤得到的滤液加入到无水乙醇，得到BIBAPTES的乙醇溶液；5. The method for preparing a medical nickel-titanium alloy material with a temperature-sensitive hydrogel coating according to claim 2, wherein the method for preparing the ethanol solution of BIBAPTES in the step (4) is: 0 °C , under nitrogen, add triethylamine, 2-bromoisobutyryl bromide, and 3-aminopropyltriethoxysilane to tetrahydrofuran in turn, then raise to room temperature and stir overnight, then vacuum filter; then vacuum The filtrate obtained by filtration is added to absolute ethanol to obtain the ethanolic solution of BIBAPTES;

其中，在每20~30 mL 四氢呋喃中加入2~5 mL三乙胺、5.8~6.0 g的2-溴异丁酰溴、4.0~4.5 g的3-氨基丙基三乙氧基硅烷；滤液和无水乙醇的体积比3~5：95~97。Wherein, 2-5 mL of triethylamine, 5.8-6.0 g of 2-bromoisobutyryl bromide, and 4.0-4.5 g of 3-aminopropyltriethoxysilane were added to every 20-30 mL of tetrahydrofuran; the filtrate and The volume ratio of absolute ethanol is 3~5:95~97.

6.如权利要求1所述的具有温敏性水凝胶涂层的医用镍钛合金材料的应用，其特征为用于制备与人体内腔道接触的医用镍钛合金器件中。6 . The application of the medical nickel-titanium alloy material with a temperature-sensitive hydrogel coating according to claim 1 , wherein it is used in the preparation of a medical nickel-titanium alloy device in contact with the human body cavity. 7 .