CN107976472A - A kind of preparation method of sensor electrode surface stable against biological contamination coating - Google Patents
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Abstract
本发明公开了一种传感器电极表面抗生物污染涂层的制备方法，其特征在于光敏透明质酸的合成、抗生物污染涂层的制备两大步骤。本发明制备的光敏透明质酸具有强亲水性、可光交联性与良好的生物相容性。光敏透明质酸合成简单，原料易得，易于实现工业化生产；光敏单体的引入使得涂层在发挥透明质酸本身优异特性的基础上，可以通过紫外光交联来固定涂层的结构。在应用于原液样品检测时，基于光敏透明质酸涂层修饰的传感器电极具有长期高效的抗生物污染能力。光交联技术、功能涂层技术与生物传感技术的结合，可广泛应用于食品安全、生物医药、环保监测与生命安全等领域。
The invention discloses a preparation method of an anti-biological pollution coating on the surface of a sensor electrode, which is characterized in two steps of synthesis of photosensitive hyaluronic acid and preparation of an anti-biological pollution coating. The photosensitive hyaluronic acid prepared by the invention has strong hydrophilicity, photocrosslinkability and good biocompatibility. The synthesis of photosensitive hyaluronic acid is simple, the raw materials are readily available, and it is easy to realize industrial production; the introduction of photosensitive monomers enables the coating to fix the structure of the coating through ultraviolet light crosslinking on the basis of exerting the excellent properties of hyaluronic acid itself. When applied to the detection of raw liquid samples, the sensor electrode modified based on the photosensitive hyaluronic acid coating has long-term and efficient anti-biological contamination ability. The combination of photocrosslinking technology, functional coating technology and biosensing technology can be widely used in food safety, biomedicine, environmental monitoring and life safety and other fields.
Description
技术领域technical field
本发明涉及高分子材料、生物功能涂层与电化学传感器领域，尤其是涉及一种采用光敏透明质酸修饰传感器电极表面制备抗生物污染涂层的方法。The invention relates to the fields of polymer materials, biological functional coatings and electrochemical sensors, in particular to a method for preparing an anti-biological pollution coating by modifying the surface of sensor electrodes with photosensitive hyaluronic acid.
背景技术Background technique
当传统的生物传感器直接与原生实际样品(如血液、牛奶、果汁、生活污水)接触以检测其中成分时，会因蛋白质、血细胞及微生物黏附等引起电极表面的污染，从而降低检测灵敏度和使用寿命，甚至导致完全失效。以血液分析为例，目前对实际血液进行分析时，首先需要采用离心分离技术来处理血液，然后利用生化分析仪在分离所得到的血清中进行目标物的检测。这样的处理方式存在一些明显的缺陷：生化分析过程需要离心设施，这对小型诊所、家庭或者在线快速检测都有很大的不便之处；血液样品离心处理过程中可能受到污染；由于较长时间的血液处理和分析过程，因此会造成不可忽视的血液成分新陈代谢所带来检测误差，此外血清中目标物浓度完成检测后，还需要经过经验公式转换，但最终结果也并不能完全表达全血中的真实状况。而血液中目标物的准确检测，对于疾病的诊断和治疗来说，其重要性无需赘言。因此，对于血液等实际样品直接检测中的传感器电极表面生物污染己成为制约生物、医学、环境检测的关键科学问题之一。When traditional biosensors are directly in contact with raw actual samples (such as blood, milk, juice, and domestic sewage) to detect the components, the surface of the electrode will be polluted due to the adhesion of proteins, blood cells, and microorganisms, which will reduce the detection sensitivity and service life. , or even lead to complete failure. Take blood analysis as an example. At present, when analyzing actual blood, it is first necessary to use centrifugal separation technology to process the blood, and then use a biochemical analyzer to detect the target substance in the separated serum. There are some obvious defects in this processing method: the biochemical analysis process requires centrifugation facilities, which is very inconvenient for small clinics, families or online rapid testing; blood samples may be contaminated during centrifugation; due to the long time Therefore, it will cause detection errors caused by the metabolism of blood components that cannot be ignored. In addition, after the concentration of the target substance in the serum is detected, it needs to be converted by an empirical formula, but the final result cannot fully express the concentration of the whole blood. real situation. The importance of accurate detection of target substances in blood for the diagnosis and treatment of diseases goes without saying. Therefore, the biocontamination on the surface of sensor electrodes in the direct detection of actual samples such as blood has become one of the key scientific issues restricting biological, medical and environmental detection.
为了解决传感器电极表面的生物污染问题，科学家们开发出一系列抗生物粘附的材料与方法。例如，将抗炎药物如氟美松被包埋在电极表面的涂层内，以通过抗炎药物的缓慢释放抑制蛋白在电极表面的非特异性吸附。这种方法虽然体现很好的抗生物粘附特性，然而这种抗粘附只能在短期内有效，随着药物的消耗，这种涂层的抗蛋白污染效果会逐渐降低。近年来，一些两性离子型聚合物与亲水聚合物被广泛应用于抗生物蛋白粘附涂层，然而这些聚合物缺乏合适的结构难以长期锚定在导电基材表面。因此，发展合适的电极表面抗生物污染材料成为生物电极构建中的一大挑战。In order to solve the problem of biological contamination on the sensor electrode surface, scientists have developed a series of anti-bioadhesion materials and methods. For example, anti-inflammatory drugs such as dexamethasone are embedded in the coating on the electrode surface to inhibit the non-specific adsorption of proteins on the electrode surface through the slow release of anti-inflammatory drugs. Although this method exhibits good anti-bioadhesion properties, this anti-adhesion is only effective in a short period of time. As the drug is consumed, the anti-protein contamination effect of this coating will gradually decrease. In recent years, some zwitterionic polymers and hydrophilic polymers have been widely used in avidin adhesion coatings. However, these polymers lack suitable structures and are difficult to anchor on the surface of conductive substrates for a long time. Therefore, the development of suitable anti-biofouling materials on the electrode surface has become a major challenge in the construction of bioelectrodes.
发明内容Contents of the invention
针对现有技术存在的上述问题，本发明提供了一种基于光敏透明质酸的抗生物污染涂层，并将其用于构建抗生物污染传感器电极的制备方法。透明质酸具有无免疫原性、生物相容性好、亲水性强的特性，本发明采用光敏透明质酸制备抗生物污染电极，光敏透明质酸在紫外光照下可以发生交联，从而可以有效增强涂层与电极的结合强度，而透明质酸作为涂层主体材料，可以有效抑制蛋白的非特异性吸附。本发明抗生物污染传感器电极对于实际样品的检测具有信号稳定性好、抗生物污染能力强、长期持续抗生物污染等优点。Aiming at the above-mentioned problems in the prior art, the present invention provides a photosensitive hyaluronic acid-based anti-biofouling coating, which is used in a preparation method for constructing an anti-biofouling sensor electrode. Hyaluronic acid has the characteristics of non-immunogenicity, good biocompatibility, and strong hydrophilicity. The present invention uses photosensitive hyaluronic acid to prepare an anti-biological contamination electrode. Photosensitive hyaluronic acid can be cross-linked under ultraviolet light, so that it can Effectively enhance the bonding strength between the coating and the electrode, and hyaluronic acid, as the main material of the coating, can effectively inhibit the non-specific adsorption of proteins. The anti-biological pollution sensor electrode of the invention has the advantages of good signal stability, strong anti-biological pollution ability, long-term continuous anti-biological pollution and the like for the detection of actual samples.
本发明的技术方案如下：Technical scheme of the present invention is as follows:
一种传感器电极表面抗生物污染涂层的制备方法，光敏透明质酸的合成、抗生物污染涂层的制备的具体步骤如下：A method for preparing an anti-biological pollution coating on the surface of a sensor electrode, the specific steps of the synthesis of photosensitive hyaluronic acid and the preparation of the anti-biological pollution coating are as follows:
(1)光敏透明质酸的合成(1) Synthesis of photosensitive hyaluronic acid
将透明质酸(HA)溶解在水中，搅拌至完全溶解，形成透明质酸水溶液；顺次加入4-二甲氨基吡啶(DMAP)、光敏单体、四丁基溴化铵(TBAB)，室温反应12h～48h；将反应液用丙酮沉淀后，转入透析袋隔水透析；将透析好的溶液冷冻干燥，得到光敏透明质酸，避光储存备用；Dissolve hyaluronic acid (HA) in water and stir until completely dissolved to form a hyaluronic acid aqueous solution; add 4-dimethylaminopyridine (DMAP), photosensitive monomer, tetrabutylammonium bromide (TBAB) sequentially, and keep at room temperature React for 12h to 48h; Precipitate the reaction solution with acetone, transfer it to a dialysis bag for water-proof dialysis; freeze-dry the dialyzed solution to obtain photosensitive hyaluronic acid, and store it away from light for later use;
(2)抗生物污染涂层的制备(2) Preparation of anti-biological contamination coating
将步骤(1)中光敏透明质酸溶于水中，加入水溶性光引发剂，避光搅拌至形成澄清透明的光敏透明质酸水溶液；将光敏透明质酸涂覆在传感器电极表面，室温干燥后用紫外光照射使涂层发生交联，即在电极表面修饰抗生物污染涂层；将涂层修饰后的传感器电极在干燥环境下储存备用。Dissolving the photosensitive hyaluronic acid in step (1) in water, adding a water-soluble photoinitiator, and stirring in the dark until a clear and transparent photosensitive hyaluronic acid aqueous solution is formed; coating the photosensitive hyaluronic acid on the surface of the sensor electrode, and drying at room temperature The coating is irradiated with ultraviolet light to cross-link, that is, the anti-biological contamination coating is modified on the surface of the electrode; the sensor electrode modified by the coating is stored in a dry environment for future use.
所述步骤(1)中光敏单体为甲基丙烯酸缩水甘油酯、甲基丙烯酸羟乙酯、甲基丙烯酸酐、甲基丙烯酸缩水甘油醚中的一种；所述光敏单体与透明质酸中羧基含量的物质的量之比为0.2～100。In the step (1), the photosensitive monomer is one of glycidyl methacrylate, hydroxyethyl methacrylate, methacrylic anhydride, and glycidyl methacrylate; the photosensitive monomer and hyaluronic acid The amount ratio of the carboxyl group content is 0.2-100.
所述步骤(2)中光敏透明质酸水溶液的浓度为0.5mg/mL～10mg/mL；所述光引发剂为水溶性光引发剂；光敏透明质酸涂覆方式为旋涂、刮涂、电沉积中的一种；The concentration of the photosensitive hyaluronic acid aqueous solution in the step (2) is 0.5mg/mL～10mg/mL; the photoinitiator is a water-soluble photoinitiator; the photosensitive hyaluronic acid coating method is spin coating, scraping coating, One of electrodeposition;
所述步骤(2)中传感器电极基材可以为金、玻碳、碳浆、石墨烯、碳纳米管、碳纤维、ITO中的一种；传感器电极涂覆前需要预处理，预处理方法为：依次用丙酮、乙醇、去离子水超声清洗；In the step (2), the sensor electrode substrate can be one of gold, glassy carbon, carbon paste, graphene, carbon nanotubes, carbon fibers, and ITO; the sensor electrode needs pretreatment before coating, and the pretreatment method is: Ultrasonic cleaning with acetone, ethanol, and deionized water in sequence;
所述步骤(2)中紫外光照射条件为：紫外光源功率为10W～1000W，光源与涂层距离为1cm～10cm，照射时间为5min～60min。The ultraviolet light irradiation conditions in the step (2) are as follows: the power of the ultraviolet light source is 10W-1000W, the distance between the light source and the coating is 1cm-10cm, and the irradiation time is 5min-60min.
所述涂层修饰后的传感器电极需保存在干燥环境下。The sensor electrode modified by the coating needs to be stored in a dry environment.
本发明有益的技术效果在于：The beneficial technical effects of the present invention are:
1、本发明通过光敏单体改性透明质酸，并将这种光敏透明质酸修饰于传感器电极表面，以解决直接应用于原生样品检测时生物传感器表面生物污染的问题。光敏透明质酸合成简单，原料易得，易于实现工业化生产；光敏透明质酸保持了透明质酸本身的生物相容性与超亲水性，而光敏单体的引入使得其在制备涂层时可以通过光交联来稳定涂层的结构，从而使得制备的传感器电极具有长期稳定的抗生物污染能力。1. The present invention modifies hyaluronic acid with a photosensitive monomer, and modifies the photosensitive hyaluronic acid on the surface of the sensor electrode to solve the problem of biological contamination on the surface of the biosensor when it is directly applied to the detection of native samples. The synthesis of photosensitive hyaluronic acid is simple, the raw materials are easy to obtain, and it is easy to realize industrial production; photosensitive hyaluronic acid maintains the biocompatibility and superhydrophilicity of hyaluronic acid itself, and the introduction of photosensitive monomers makes it easier to The structure of the coating can be stabilized by photocrosslinking, so that the prepared sensor electrode has long-term stable anti-biological contamination ability.
2、本发明在传感器电极表面制备的抗生物污染涂层具有稳定性好、抗生物污染能力强、不影响传感器的检测性能等优点。2. The anti-biological pollution coating prepared on the sensor electrode surface of the present invention has the advantages of good stability, strong anti-biological pollution ability, and does not affect the detection performance of the sensor.
3、本发明制备的抗生物污染涂层适用性强，可广泛修饰于各类传感器电极表面，以解决传感器在实际样品检测中的生物蛋白与微生物的非特异性污染问题。3. The anti-biological pollution coating prepared by the present invention has strong applicability and can be widely modified on the surface of various sensor electrodes to solve the problem of non-specific pollution of biological proteins and microorganisms in the actual sample detection of sensors.
4、本发明将高分子材料技术、生物涂层技术与电化学传感技术可构建新型、多样化电化学传感器，有望广泛应用于食品安全、生物医药、环保监测与生命安全等领域。4. The present invention combines polymer material technology, biological coating technology and electrochemical sensing technology to construct new and diversified electrochemical sensors, which are expected to be widely used in food safety, biomedicine, environmental monitoring and life safety and other fields.
附图说明Description of drawings
图1：本发明的传感器电极表面抗生物污染涂层的示意图；Fig. 1: the schematic diagram of anti-biological contamination coating on sensor electrode surface of the present invention;
图2：本发明实施例1中光敏透明质酸HA-GMA的结构示意图；Figure 2: Schematic diagram of the structure of photosensitive hyaluronic acid HA-GMA in Example 1 of the present invention;
图3：本发明实施例1中抗生物污染涂层的水接触角测试图；Fig. 3: the water contact angle test figure of anti-biological contamination coating in the embodiment of the present invention 1;
图4：本发明实施例1中抗生物污染涂层的体外细胞实验图。Figure 4: In vitro cell experiment diagram of the anti-biological contamination coating in Example 1 of the present invention.
具体实施方式Detailed ways
下面结合附图和实施例，对本发明进行具体描述。The present invention will be specifically described below in conjunction with the accompanying drawings and embodiments.
本发明的抗生物污染涂层的制备方法如图1所示，由图中可以看出，光敏透明质酸修饰传感器电极后，由于透明质酸的强亲水性，可以在电极表面形成水化层，从而可以抑制蛋白、微生物等生物污染物在电极表面的非特异性粘附。The preparation method of the anti-biological pollution coating of the present invention is shown in Figure 1, as can be seen from the figure, after photosensitive hyaluronic acid is modified sensor electrodes, due to the strong hydrophilicity of hyaluronic acid, hydration can be formed on the electrode surface. layer, which can inhibit the non-specific adhesion of biological pollutants such as proteins and microorganisms on the electrode surface.
实施例1Example 1
一种传感器电极表面抗生物污染涂层的制备方法，包括如下具体步骤：A preparation method for an anti-biological contamination coating on the surface of a sensor electrode, comprising the following specific steps:
(1)光敏透明质酸的合成(1) Synthesis of photosensitive hyaluronic acid
将1.0g透明质酸(HA)溶解在水中，搅拌至完全溶解，形成透明质酸水溶液；顺次加入4-二甲氨基吡啶(DMAP)(40％mol.相对于HA重复单元数)、甲基丙烯酸缩水甘油酯(GMA)(10倍于HA重复单元数)、四丁基溴化铵(TBAB)(80％mol.相对于HA重复单元数)，室温反应48h；将反应液用丙酮沉淀，转入透析袋隔水透析4d；将透析好的溶液冷冻干燥，得到光敏透明质酸HA-GMA，避光储存备用；Dissolve 1.0g of hyaluronic acid (HA) in water and stir until completely dissolved to form an aqueous solution of hyaluronic acid; add 4-dimethylaminopyridine (DMAP) (40% mol. relative to the number of HA repeating units), formazan Glycidyl acrylate (GMA) (10 times the number of HA repeating units), tetrabutylammonium bromide (TBAB) (80% mol. relative to the number of HA repeating units), react at room temperature for 48h; precipitate the reaction solution with acetone , transferred to a dialysis bag for dialysis against water for 4 days; the dialysis solution was freeze-dried to obtain photosensitive hyaluronic acid HA-GMA, which was stored away from light for later use;
(2)抗生物污染涂层的制备(2) Preparation of anti-biological contamination coating
将步骤(1)中50mg光敏透明质酸HA-GMA溶于10mL水中，加入水溶性光引发剂Irgacure 2959(0.5wt％相对于HA-GMA)，避光搅拌至形成澄清透明的光敏透明质酸水溶液；施加2.0V电压下电沉积180s使光敏透明质酸沉积在ITO电极表面，室温干燥后用60W紫外光照射30min使涂层发生交联(光源与涂层距离1cm)，即在ITO电极表面修饰抗生物污染涂层；将涂层修饰后的ITO电极在干燥环境下储存备用。Dissolve 50mg of photosensitive hyaluronic acid HA-GMA in step (1) in 10mL of water, add water-soluble photoinitiator Irgacure 2959 (0.5wt% relative to HA-GMA), and stir in the dark until a clear and transparent photosensitive hyaluronic acid is formed Aqueous solution; apply 2.0V voltage for 180s to deposit photosensitive hyaluronic acid on the surface of the ITO electrode, and after drying at room temperature, irradiate the coating with 60W ultraviolet light for 30 minutes to cross-link the coating (the distance between the light source and the coating is 1cm), that is, on the surface of the ITO electrode Modify the anti-biological fouling coating; store the ITO electrode modified by the coating in a dry environment for future use.
实施例2Example 2
一种传感器电极表面抗生物污染涂层的制备方法，包括如下具体步骤：A preparation method for an anti-biological contamination coating on the surface of a sensor electrode, comprising the following specific steps:
(1)光敏透明质酸的合成(1) Synthesis of photosensitive hyaluronic acid
将0.5g透明质酸(HA)溶解在水中，搅拌至完全溶解，形成透明质酸水溶液；顺次加入4-二甲氨基吡啶(DMAP)(100％mol.相对于HA重复单元数)、甲基丙烯酸缩水甘油酯(GMA)(5倍于HA重复单元数)、四丁基溴化铵(TBAB)(100％mol.相对于HA重复单元数)，室温反应24h；将反应液用丙酮沉淀，转入透析袋隔水透析7d；将透析好的溶液冷冻干燥，得到光敏透明质酸HA-GMA，避光储存备用；Dissolve 0.5g of hyaluronic acid (HA) in water and stir until completely dissolved to form an aqueous solution of hyaluronic acid; sequentially add 4-dimethylaminopyridine (DMAP) (100% mol. relative to the number of HA repeating units), formazan Glycidyl acrylate (GMA) (5 times the number of HA repeating units), tetrabutylammonium bromide (TBAB) (100% mol. relative to the number of HA repeating units), react at room temperature for 24h; precipitate the reaction solution with acetone , transferred to a dialysis bag for dialysis against water for 7 days; freeze-dried the dialyzed solution to obtain photosensitive hyaluronic acid HA-GMA, and stored it away from light for later use;
(2)抗生物污染涂层的制备(2) Preparation of anti-biological contamination coating
将步骤(1)中20mg光敏透明质酸HA-GMA溶于10mL水中，加入水溶性光引发剂Irgacure 2959(1.0wt％相对于HA-GMA)，避光搅拌至形成澄清透明的光敏透明质酸水溶液；取20μL光敏透明质酸溶液积在玻碳电极表面(直径3mm)，室温干燥后用1000W紫外光照射5min使涂层发生交联(光源与涂层距离5cm)，即在玻碳电极表面修饰抗生物污染涂层；将涂层修饰后的玻碳电极在干燥环境下储存备用。Dissolve 20 mg of photosensitive hyaluronic acid HA-GMA in step (1) in 10 mL of water, add water-soluble photoinitiator Irgacure 2959 (1.0 wt % relative to HA-GMA), and stir in the dark until a clear and transparent photosensitive hyaluronic acid is formed Aqueous solution; take 20 μL of photosensitive hyaluronic acid solution and deposit it on the surface of the glassy carbon electrode (diameter 3mm), and after drying at room temperature, irradiate the coating with 1000W ultraviolet light for 5 minutes to crosslink the coating (the distance between the light source and the coating is 5cm), that is, on the surface of the glassy carbon electrode Modify the anti-biological fouling coating; store the modified glassy carbon electrode in a dry environment for future use.
实施例3Example 3
一种传感器电极表面抗生物污染涂层的制备方法，包括如下具体步骤：A preparation method for an anti-biological contamination coating on the surface of a sensor electrode, comprising the following specific steps:
(1)光敏透明质酸的合成(1) Synthesis of photosensitive hyaluronic acid
将1g透明质酸(HA)溶解在水中，搅拌至完全溶解，形成透明质酸水溶液；顺次加入4-二甲氨基吡啶(DMAP)(200％mol.相对于HA重复单元数)、甲基丙烯酸羟乙酯(HEMA)(20倍于HA重复单元数)、四丁基溴化铵(TBAB)(400％mol.相对于HA重复单元数)，室温反应12h；将反应液用丙酮沉淀，转入透析袋隔水透析7d；将透析好的溶液冷冻干燥，得到光敏透明质酸HA-HEMA，避光储存备用；Dissolve 1g of hyaluronic acid (HA) in water and stir until completely dissolved to form an aqueous solution of hyaluronic acid; add 4-dimethylaminopyridine (DMAP) (200% mol. relative to the number of HA repeating units), methyl Hydroxyethyl acrylate (HEMA) (20 times the number of HA repeating units), tetrabutylammonium bromide (TBAB) (400% mol. relative to the number of HA repeating units), reacted at room temperature for 12h; the reaction solution was precipitated with acetone, Transfer to a dialysis bag for dialysis against water for 7 days; freeze-dry the dialyzed solution to obtain photosensitive hyaluronic acid HA-HEMA, and store it away from light for later use;
(2)抗生物污染涂层的制备(2) Preparation of anti-biological contamination coating
将步骤(1)中100mg光敏透明质酸HA-HEMA溶于10mL水中，加入水溶性光引发剂Irgacure 2959(0.5wt％相对于HA-HEMA)，避光搅拌至形成澄清透明的光敏透明质酸水溶液；取0.5mL光敏透明质酸溶液滴在ITO电极表面(长4cm*宽4cm)，随后利用匀胶机通过旋涂将HA-HEMA修饰在ITO电极表面；室温干燥后用300W紫外光照射20min使涂层发生交联(光源与涂层距离5cm)，即在玻碳电极表面修饰抗生物污染涂层；将涂层修饰后的玻碳电极在干燥环境下储存备用。Dissolve 100 mg of photosensitive hyaluronic acid HA-HEMA in step (1) in 10 mL of water, add water-soluble photoinitiator Irgacure 2959 (0.5 wt % relative to HA-HEMA), and stir in the dark until a clear and transparent photosensitive hyaluronic acid is formed Aqueous solution; take 0.5mL photosensitive hyaluronic acid solution and drop it on the surface of ITO electrode (length 4cm*width 4cm), and then use a homogenizer to modify HA-HEMA on the surface of ITO electrode by spin coating; after drying at room temperature, irradiate with 300W ultraviolet light for 20min The coating is cross-linked (the distance between the light source and the coating is 5 cm), that is, the anti-biological pollution coating is modified on the surface of the glassy carbon electrode; the glassy carbon electrode modified by the coating is stored in a dry environment for later use.
测试例：Test case:
抗生物污染涂层修饰电极的体外细胞实验In Vitro Cell Experiment of Electrode Modified by Anti-Biofouling Coating
按照实施例1中操作方法，将同一只ITO电极一部分表面涂覆光敏透明质酸HA-GMA，另一部分表面不涂覆任何材料。将该ITO用紫外光照消毒后，放入24孔培养版，每孔加入500μL含6×103个细胞的细胞悬液，将培养电极置于37℃、含5％CO2、相对湿度95％的烘箱中培养24h。接着取出后用无菌PBS冲洗两次，随后用戊二醛进行交联，干燥后使用扫描显微镜观察L929细胞在样品表面的粘附情况。According to the operation method in Example 1, a part of the surface of the same ITO electrode was coated with photosensitive hyaluronic acid HA-GMA, and the other part was not coated with any material. After sterilizing the ITO with ultraviolet light, put it into a 24-well culture plate, add 500 μL of cell suspension containing 6×10 3 cells to each well, and place the culture electrode at 37°C with 5% CO 2 and a relative humidity of 95%. Incubate in an oven for 24 hours. After taking it out, it was rinsed twice with sterile PBS, then cross-linked with glutaraldehyde, and after drying, the adhesion of L929 cells on the surface of the sample was observed using a scanning microscope.
结果如图4所示，由图4可以看出，未涂覆HA-GMA的ITO区域粘附大量L929细胞，而对HA-GMA涂覆ITO区域没有细胞粘附，由此证明，该抗生物污染涂层修饰的ITO电极表面具有优异的抗生物体非特异性吸附的能力。The results are shown in Figure 4. It can be seen from Figure 4 that a large number of L929 cells adhered to the ITO area not coated with HA-GMA, but there was no cell adhesion to the ITO area coated with HA-GMA, thus proving that the antibiotic The surface of the ITO electrode modified by the fouling coating has excellent ability to resist non-specific adsorption of organisms.
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