辣根过氧化物酶在壳聚糖和无掺杂金刚石纳米粒子共沉积膜上的固定及其直接电化学

无掺杂的金刚石纳米粒子(UND)可以和壳聚糖共沉积到玻碳电极表面,形成壳聚糖-UND复合膜.此复合膜可以通过吸附的方法,固定辣根过氧化物酶(HRP),并且能够实现HRP的直接电化学,保持HRP对过氧化氢的良好催化能力.为了进一步研究此复合膜及HRP在此复合膜上的电化学活性,运用扫描电子显微镜(SEM)、全反射红外(ATR)、电化学交流阻抗、循环伏安等技术来跟踪各步修饰过程.结果表明,此复合膜有良好的生物相容性,能够很好的保持HRP的生物活性.     

疏水疏油纳米银修饰不锈钢材料的制备及生物相容性

采用恒电位电化学沉积法在316L不锈钢表面沉积银纳米修饰层,并置于全氟硅烷溶液中进行疏水修饰,通过控制沉积时间和修饰过程,制备了一系列具有不同界面亲疏水特性的修饰材料.采用扫描电子显微镜(SEM)、能量散射X射线光谱(EDX)仪及接触角测量仪等对材料性能进行了表征;利用四甲基偶氮唑盐(MTT)法和流式细胞仪等检验了材料的血液相容性;并将修饰后的316L不锈钢植入实验动物体内,检验其组织相容性.实验结果表明,该纳米银全氟硅烷修饰的316L不锈钢材料比316L裸不锈钢具有更好的血液及组织相容性,有望成为一种理想的血管支架应用材料.     

以金纳米颗粒-二氧化钛纳米线阵列为支架的电化学免疫传感的构建及其应用

首先以掺杂氟的二氧化锡导电玻璃为基片,采用水热法制备出二氧化钛纳米线阵列,然后使用电化学沉积法将金纳米颗粒修饰于二氧化钛纳米线阵列表面以增强其导电性和生物相容性.将纳米金-二氧化钛纳米线阵列用作电化学免疫传感器支架,利用纳米金与癌胚抗原抗体之间的静电吸附作用,将包被抗体负载于电极表面制得检测癌胚抗原的电流型免疫传感器.同样,水热法还被用于制备圆柱形二氧化钛纳米棒,以辛二酸双(3-磺基-N-羟基琥珀酰亚胺酯)钠盐为双氨基交联剂,将辣根过氧化物酶和信号抗体一起固定于二氧化钛纳米棒表面作为示踪标记物.扫描电子显微镜、透射电子显微镜和X射线衍射分析仪均被用于分析上述材料的结构、形貌和组成.通过夹心型免疫反应,示踪标记物上信号抗体被定量负载于免疫传感器表面.通过一个以过氧化氢为媒介体的辣根过氧化物酶催化反应,差示脉冲伏安法被用于癌胚抗原的定量测定.结果表明,该癌胚抗原免疫传感器线性范围为0.01~120μg/L,检测限为6 ng/L.     

316L钢表面纳米银镀层的制备、性质及血液相容性

采用恒电位电沉积方法在316L不锈钢表面沉积银纳米镀层, 并将钢板置于全氟硅烷溶液中浸泡, 通过动态凝血实验、 抗凝血时间测定、 血小板黏附实验、 溶血实验和蛋白吸附实验等手段, 测试材料的血液相容性. 结果表明, 通过上述方法可明显改善316L不锈钢的血液相容性, 而抗凝血性能、 溶血率及纤维蛋白吸附量不亚于裸钢板. 与316L裸不锈钢相比, 银镀膜全氟硅烷浸泡316L不锈钢具有良好的血液相容性, 是一种比较理想的冠状动脉支架材料, 具有良好的应用前景.     

纳米二氧化钛对辣根过氧化酶催化作用

将辣根过氧化物酶(HRP)固定在二氧化钛(TiO2)纳米颗粒或纳米管修饰玻碳电极(GC)上,形成纳米TiO2微粒/HRP修饰GC电极和TiO2纳米管/HRP修饰GC电极.比较了HRP在纳米TiO2微粒、TiO2纳米管电极上的直接电子转移反应.实验表明,HRP在TiO2纳米管电极表面更能有效地促进它的电活性中心发生电子交换反应.此外,还测定了HRP标记抗体的电化学性能,为抗原抗体免疫反应信号的选择提供了参考依据.     

酶功能化聚多巴胺包覆纳米普鲁士蓝的制备及电化学分析应用

在纳米金表面原位沉积普鲁士蓝,然后在核壳结构纳米金-普鲁士蓝的表面包覆一层易氧化聚合的多巴胺保护膜,利用多巴胺聚合表面残留的大量氨基和羟基进一步将纳米铂粒子修饰于聚多巴胺膜表面制得普鲁士蓝-聚多巴胺-纳米铂多层纳米复合材料。将此复合材料修饰于金电极表面,协同使用辣根过氧化物酶用于H_2O_2浓度的检测。结果表明:聚多巴胺的引入有效增加了普鲁士蓝的稳定性,增大了纳米铂的负载量以及辣根过氧化物酶的生物活性;由于普鲁士蓝、纳米铂和辣根过氧化物酶的多重信号放大作用,酶功能化纳米复合材料修饰电极对H_2O_2表现出良好的电还原活性。优化条件下,对H_2O_2的检测范围为2.0×10~(-7)~1.0×10~(-3)mol·L~(-1),检出限(S/N=3)为1.2×10~(-7)mol·L~(-1)。     

表面等离子体子共振技术实时研究蛋白质在修饰表面的静电吸附行为

研究蛋白质在固相表面的静电吸附特性,进而控制蛋白质在修饰表面的静电吸附尤为重要,表面等离子体子共振可以检测金属表面吸附物质厚度和折射率的变化^[1]。这种技术已在研究生物分子相互作用^[2]和考察自组装单层的形成^[3]及蛋白质在固体表面吸附行为^[9-11]等方面得到广泛的应用。对蛋白质在固体表面吸附行为的研究多为考察不同的蛋白质在不同的修饰表面的吸附行为。然而,对蛋白质在修饰表面静电吸附的本质影响因素的研究却少有报道^[4]。本文使用表面等离子体子共振技术实时研究了蛋白质在甲羧基化葡聚糖修饰表面的静电吸附与溶液pH值及离子强度的依赖关系。     

纳米孔技术对PD-1抗原抗体结合检测的实验研究

肿瘤的免疫疗法是利用人体自身的免疫系统去治疗肿瘤的一种方法，程序性死亡受体1（PD-1）是肿瘤免疫疗法中的一种免疫检查点，利用PD-1或PD-L1的单克隆抗体，可以阻断PD-1/PD-L1信号通路，恢复T细胞的免疫杀伤功能，实现肿瘤的免疫治疗.为了研究PD-1抗体药物与人体PD-1抗原的结合有效性，以PD-1蛋白质分子为实验对象，用纳米孔传感器件开展了PD-1、PD-1抗体及其结合物的辨识.分别对纯PD-1蛋白质分子和其抗体分子进行了纳米孔过孔实验研究，发现PD-1及其抗体的相对堵塞电流幅值比分别为0.00404和0.01297，实现了抗原抗体分子的区分.并对Si₃N₄纳米孔内壁进行了PD-1抗体修饰，再将PD-1蛋白质分子通过经抗体修饰后的纳米孔，以期实现结合物的检测，实验结果显示部分抗原抗体实现了特异性结合，剩下部分呈游离状，所以纳米孔技术可实现抗原抗体结合物的区分.未来，纳米孔有望成为无标志检测药物有效性的一种新手段.     

固态纳米孔对蛋白质易位的实验研究

蛋白质因其多样性和功能性,是生物体内一类非常重要的分子.通常蛋白质的表征需要借助荧光或者酶的标记.而纳米孔技术,得益于免标记、单分子检测等优势,为蛋白质的表征提供了新方向.我们使用固态纳米孔完成了单个蛋白质分子及蛋白质-蛋白质结合物的检测.可以发现,外部电压和电解质溶液的酸碱度会直接影响蛋白质分子表面带电量,从而加快或延迟其在孔内的易位时间.抗原、抗体本质上都是蛋白质,两者之间具有高度特异性.通过比较抗体溶液在添加特异性抗原前后的易位事件,实现了单个蛋白质分子和蛋白质-蛋白质结合物的区分.未来,纳米孔技术有望应用于多蛋白质分子的辨识、蛋白质分子相互作用机制等方面的研究.     

生物医用纳米颗粒表面的两性离子化设计

生物医用纳米颗粒的表面设计对维持纳米颗粒稳定性和抑制蛋白质非特异性吸附从而实现体内长效循环等具有重要意义.具有细胞膜仿生结构的两性离子界面能通过离子静电作用形成高效水合层,不仅可有效增强纳米颗粒的稳定性和抗免疫清除能力,通过提高体内循环时间增强其"被动"靶向能力,而且当与环境响应性或生物活性分子复合后,还可有效实现纳米颗粒的"主动"靶向功能,因此"两性离子化"已经发展为纳米颗粒表面设计的新策略.本文主要概述了两性离子材料在生物医用纳米表面设计中的应用进展,包括小分子和聚合物两性离子对无机纳米颗粒的表面修饰、聚合物两性离子组装体用于抗肿瘤药物传递等,同时也介绍了混合电荷材料的一些特殊性质和应用.     

Adsorption and protein-induced metal release from chromium metal and stainless steel

A research effort is undertaken to understand the mechanism of metal release from, e.g., inhaled metal particles or metal implants in the presence of proteins. The effect of protein adsorption on the metal release process from oxidized chromium metal surfaces and stainless steel surfaces was therefore examined by quartz crystal microbalance with energy dissipation monitoring (QCM-D) and graphite furnace atomic absorption spectroscopy (GFAAS). Differently charged and sized proteins, relevant for the inhalation and dermal exposure route were chosen including human and bovine serum albumin (HSA, BSA), mucin (BSM), and lysozyme (LYS). The results show that all proteins have high affinities for chromium and stainless steel (AISI 316) when deposited from solutions at pH 4 and at pH 7.4 where the protein adsorbed amount was very similar. Adsorption of albumin and mucin was substantially higher at pH 4 compared to pH 7.4 with approximately monolayer coverage at pH 7.4, whereas lysozyme adsorbed in multilayers at both investigated pH. The protein-surface interaction was strong since proteins were irreversibly adsorbed with respect to rinsing. Due to the passive nature of chromium and stainless steel (AISI 316) surfaces, very low metal release concentrations from the QCM metal surfaces in the presence of proteins were obtained on the time scale of the adsorption experiment. Therefore, metal release studies from massive metal sheets in contact with protein solutions were carried out in parallel. The presence of proteins increased the extent of metals released for chromium metal and stainless steel grades of different microstructure and alloy content, all with passive chromium(III)-rich surface oxides, such as QCM (AISI 316), ferritic (AISI 430), austentic (AISI 304, 316L), and duplex (LDX 2205).     

A Quartz Crystal Microbalance Characterization of Metal‐Oil Interfaces and Interactions with Wax Molecules

The solid‐liquid interface between stainless steel and model petroleum fluids is investigated at isothermal conditions using a quartz crystal microbalance. AISI 316 (Fe/Cr18/Ni10/Mo3) stainless steel is chosen to represent the metal surface. Paraffin components dissolved in dodecane constitute the petroleum fluid phase. Commercial macro‐crystalline and micro‐crystalline waxes provide primarily linear and branched paraffin components, respectively. Paraffin solubility conditions are established through a van't Hoff relationship. Model fluids prepared with the single‐component alkanes n‐C₃₆ or n‐C₃₀ paraffin provide well‐defined solubility conditions. Monitored changes in resonance frequency and energy dissipation of the quartz crystal resonator immersed in the model fluids confirm that no continual deposition of paraffin components occurs at isothermal conditions. Solid paraffin crystals dispersed in solution show no adherence to the stainless steel surface. The absence of attractive interactions between the stainless steel surface and the dispersed paraffin crystals suggests that a surface adsorption and/or surface nucleation mechanism is responsible for the formation of incipient paraffin wax deposits under nonquiescent conditions.     

Protein-associated water and secondary structure effect removal of blood proteins from metallic substrates

Removing adsorbed protein from metals has significant health and industrial consequences. There are numerous protein-adsorption studies using model self-assembled monolayers or polymeric substrates but hardly any high-resolution measurements of adsorption and removal of proteins on industrially relevant transition metals. Surgeons and ship owners desire clean metal surfaces to reduce transmission of disease via surgical instruments and minimize surface fouling (to reduce friction and corrosion), respectively. A major finding of this work is that, besides hydrophobic interaction adhesion energy, water content in an adsorbed protein layer and secondary structure of proteins determined the access and hence ability to remove adsorbed proteins from metal surfaces with a strong alkaline-surfactant solution (NaOH and 5 mg/mL SDS in PBS at pH 11). This is demonstrated with three blood proteins (bovine serum albumin, immunoglobulin, and fibrinogen) and four transition metal substrates and stainless steel (platinum (Pt), gold (Au), tungsten (W), titanium (Ti), and 316 grade stainless steel (SS)). All the metallic substrates were checked for chemical contaminations like carbon and sulfur and were characterized using X-ray photoelectron spectroscopy (XPS). While Pt and Au surfaces were oxide-free (fairly inert elements), W, Ti, and SS substrates were associated with native oxide. Difference measurements between a quartz crystal microbalance with dissipation (QCM-D) and surface plasmon resonance spectroscopy (SPR) provided a measure of the water content in the protein-adsorbed layers. Hydrophobic adhesion forces, obtained with atomic force microscopy, between the proteins and the metals correlated with the amount of the adsorbed protein-water complex. Thus, the amount of protein adsorbed decreased with Pt, Au, W, Ti and SS, in this order. Neither sessile contact angle nor surface roughness of the metal substrates was useful as predictors here. All three globular proteins behaved similarly on addition of the alkaline-surfactant cleaning solution, in that platinum and gold exhibited an increase, while tungsten, titanium, and stainless steel showed a decrease in weight. According to dissipation measurements with the QCM-D, the adsorbed layer for platinum and gold was rigid, while that for the tungsten, titanium, and stainless steel was much more flexible. The removal efficiency of adsorbed-protein by alkaline solution of SDS depended on the water content of the adsorbed layers for W, Ti, and SS, while for Pt and Au, it depended on secondary structural content. When protein adsorption was high (Pt, Au), protein-protein interactions and protein-surface interactions were dominant and the removal of protein layers was limited. Water content of the adsorbed protein layer was the determining factor for how efficiently the layer was removed by alkaline SDS when protein adsorption was low. Hence, protein-protein and protein-surface interactions were minimal and protein structure was less perturbed in comparison with those for high protein adsorption. Secondary structural content determined the efficient removal of adsorbed protein for high adsorbed amount.     

表面改性SUS316L不锈钢的电化学行为研究

分别以表面镀Rh,表面离子束增强沉积Ta2O5膜及溶胶凝胶法沉积TiO2膜对冠状动脉支架用材料SUS316L不锈钢进行表面改性.采用电化学方法研究了该表面改性试样在Tyrode's模拟人工体液中的电化学行为.结果表明,上述3种表面改性方法均可提高SUS316L不锈钢在模拟人工体液中的阳极极化性能.其中对于采用离子束增强法沉积的Ta2O5膜和溶胶凝胶法沉积的TiO2膜,因Ta和Ti上的d轨道空位已被氧的电子占据,不利于氢吸附,从而抑制了阴极的析氢过程.X射线衍射分析发现,3种改性方法在SUS316L不锈钢表面依次形成均匀而致密的Rh金属层,Ta2O5的无序膜层和TiO2晶态膜层,阻止了合金元素的溶解,改善不锈钢的电化学性能.     

Effects of cyclonic plasma deposited organosilicon nano-coating on 316 stainless steel and its surface characterization

Cyclonic atmospheric pressure plasma is developed to synthesize the organosilicon nano-coating on 316 L stainless steel surface with hexamethyldisilazane (HMDSN) and HMDSN/N₂ monomers. The modified 316 L stainless steel surface characteristics of cyclonic plasma deposited organosilicon nano-coating were evaluated by the static contact angle measurement, FTIR, SEM, AFM, and XPS detections. The chemical analysis with FTIR and XPS depicts that cyclonic plasma deposited nano-coating obtains the relatively inorganic characteristics. The surface morphological determination with SEM and AFM refers cyclonic plasma deposited 316 L stainless steel surface roughness alteration with switching monomer inputs. This study shows the potential of chamber-less deposition to create the plasma deposited organosilicon nano-coating for 316 L stainless.     

TiO_2/316L不锈钢薄膜电极在NaCl溶液中的耐腐蚀性能

应用sol gel法和提拉技术于 316L不锈钢表面构筑纳米TiO2薄膜,再经水热后处理以消除膜中的细小龟裂.SEM和XRD技术表征膜的形貌和厚度,线性极化法分别考察膜厚度、pH、和Cl浓度对纳米膜电极耐腐蚀性能影响.电化学交流阻抗检测纳米TiO2膜在 0. 5mol/LNaCl溶液中的阻抗随浸泡时间的变化,光电子能谱技术测定了经浸泡 1008h后的纳米膜中各元素相对百分含量和价态.结果表明:在中性或碱性条件下,厚度为 375~464nm的纳米膜其耐腐蚀性随浸泡时间的延长呈现初期增加而后稳定,浸泡 48h后腐蚀电流较之浸泡初期降低 2个数量级,耐腐蚀电阻增加 2个数量级,在浸泡 1 008h内没有发现腐蚀的产物,Fe是以原子态扩散到膜中.     

磁场对316L不锈钢微生物腐蚀行为的影响

采用循环极化、微生物分析法、扫描电镜及表面能谱分析等方法,研究了磁场对316L不锈钢在含硫酸盐还原菌(SRB)的土壤模拟溶液中的腐蚀行为。结果表明,磁场可以抑制SRB的生长;未外加磁场时316L不锈钢表面膜层以局部堆积为主,加有磁场时,局部堆积明显减小,膜层均匀致密的排列于基体表面;无论有或没有外加磁场,316L不锈钢表面均发生钝化膜破裂型点蚀,未外加磁场时的点蚀电位低于加有磁场时的。在相同的浸泡时间,未外加磁场时循环极化滞后环面积明显比加有磁场时的大,说明磁场可以有效抑制316L不锈钢点蚀的形成与发展,降低316L不锈钢的点蚀诱发能力。     

A porous silicon immunoassay platform for fluorometric determination of α-synuclein in human cerebrospinal fluid

Levels of total and/or oligomeric α-synuclein may be used as a biomarker tool to aid in the diagnosis and development of new disease-modifying therapies. We report here on a porous silicon antibody microarray for the fluorimetric determination of cerebrospinal fluid levels of total α-synuclein, a protein involved the pathology of Parkinson’s disease. The surface of porous silicon has a 3-dimensional macro- and nanoporous structure, and this offers a large binding capacity for capturing probe molecules. Porous silicon also warrants efficient immobilization of antibodies by surface adsorption, and does not require chemical immobilization. The platform requires 10 μL of cerebrospinal fluid, and each test requires 4 h for assay only (including immobilization of capturing antibody). The limit of detection is 35 pg mL^?1 of α-synuclein in cerebrospinal fluid, and the dynamic analytical range extends from 0.01 to 100 ng·mL^?1.

Perfluorocarbon thin films and polymer brushes on stainless steel 316 L for the control of interfacial properties

Perfluorocarbon thin films and polymer brushes were formed on stainless steel 316 L (SS316L) to control the surface properties of the metal oxide. Substrates modified with the films were characterized using diffuse reflectance infrared Fourier transform spectroscopy (DRIFT), contact angle analysis, atomic force microscopy (AFM), and cyclic voltammetry (CV). Perfluorooctadecanoic acid (PFOA) was used to form thin films by self-assembly on the surface of SS316L. Polypentafluorostyrene (PFS) polymer brushes were formed by surface-initiated polymerization using SAMs of 16-phosphonohexadecanoic acid (COOH-PA) as the base. PFOA and PFS were effective in significantly reducing the surface energy and thus the interfacial wetting properties of SS316L. The SS316L control exhibited a surface energy of 38 mN/m compared to PFOA and PFS modifications, which had surface energies of 22 and 24 mN/m, respectively. PFOA thin films were more effective in reducing the surface energy of the SS316L compared to PFS polymer brushes. This is attributed to the ordered PFOA film presenting aligned CF(3) terminal groups. However, PFS polymer brushes were more effective in providing corrosion protection. These low-energy surfaces could be used to provide a hydrophobic barrier that inhibits the corrosion of the SS316L metal oxide surface.