HOPG片电化学沉积羟基磷灰石初探

为探索羟基磷灰石(HA)在含碳基体上电化学沉积,采用了高定向裂解石墨(HOPG)片这一具有原子级平整表面的导电材料作为阴极材料,探索了电沉积时间、溶液pH值及溶液浓度对沉积层成分和形貌的影响,利用TEM、SEM、AFM、EDS及XRD等进行了表征分析,并初步探讨了羟基磷灰石电沉积机理。结果表明电解液初始pH值为5和钙离子浓度低于0.007mol/L时有利于促进HA的生长,并且可以形成单层片状结构,得到比较规整完善的多孔状单层HA涂层。而延长电沉积和陈化时间则有助于缺钙型HA垂直于基底表面生长,并促进钙磷比增加,形成缺钙性HA。研究将为电沉积羟基磷灰石的应用提供必要的理论和实验支持。     

电流密度对水热电化学沉积HA涂层性能的影响

在含0.15mol/L HF,2mol/L H_3PO_4的水溶液中对Ti6Al4V基体进行阳极氧化处理,然后在0.02mol/L CaCl_2,0.012mol/L K_2HPO_4·3H_2O,0.139mol/L NaCl的电解液中采用水热电化学沉积方法在预处理的Ti_6Al_4V基体表面制备羟基磷灰石(HA)涂层。采用X射线衍射仪(XRD)、扫描电子显微镜(SEM)、能谱仪(EDS)、台阶仪和万能材料试验机等研究沉积过程中电流密度大小对HA涂层物相、微观形貌、厚度、生物活性及结合强度的影响。结果表明:采用水热电化学沉积方法在不同电流密度下均制备出了HA涂层,涂层表现为分层生长,部分晶体呈花簇状。HA的厚度随电流密度的增加先增大后减小,在1.25mA/cm~2时达到最大为26.4μm,此时涂层最为致密,与基体的结合强度最高,约为20.0MPa。模拟体液(SBF)浸泡实验能较快诱导类骨磷灰石(CHA)的生成,最大直径达到7~8μm,表明涂层具有较好的生物活性。     

静电自组装表面修饰HA/PA6膜:表征及其细胞相容性研究

通过表面改性引入活性生物分子可以用来增强材料表面的生物相容性。采用静电自组装技术将明胶和聚乙烯亚胺引入羟基磷灰石/聚酰胺6(HA/PA6)材料表面,最终获得大分子修饰的复合材料。利用水接触角测量(WCA)、X射线光电子能谱(XPS)和原子力显微镜(AFM)测试手段对表面改性前后HA/PA6膜的表面化学、亲水性和表面形貌进行表征。并研究了其对细胞活性的影响。结果表明,改性后膜表面的亲水性增强,粗糙度增大。体外细胞实验表明,明胶涂层后的HA/PA6表面细胞更利粘附、铺展和生长。该固定方法模拟细胞外基质组成制备出的生物活性膜能进一步满足生物医学工程需求。     

Improvement in the dispersion stability of iron oxide nanoparticles in highly concentrated brine solution using encapsulation with polymer-polymer crosslinked shells

In this study, the dispersion stability of superparamagnetic iron oxide nanoparticles (IONPs) under high salinity was investigated. In our previous study, the stable dispersion of IONPs in 5 wt% NaCl solution was achieved under neutral and basic conditions through the functionalization of IONPs using sulfonated phenolic resin (SPR). However, under acidic conditions, the SPR shell dissociated from the IONP surface, resulting in the loss of dispersion stability. To overcome this limitation, crosslinking with polyvinyl alcohol (PVA) was implemented. The sulfonate groups in the SPR and hydroxyl groups in PVA can form a bond by removing water and consequently a crosslinked structure is formed between SPR and PVA. The stability of the dispersion was evaluated using visual observation of sedimentation and size measurements of the sample for a week. It was found that the dispersion was highly stable in brine, containing up to 10 wt% NaCl, at pH 4. The shell/core structure of the highly crosslinked SPR-PVA was verified using EDS mapping of the S elements in the SPR-IONP sample. The change in the shell structure through the crosslink reaction was characterized using thermogravimetric analysis and Fourier transform infrared spectroscopy.     

金膜表面聚丙烯酰吡咯膜的结构和表面形貌研究

研究了一种拟用于SPR生物传感器的生物材料.为此,首先在金膜表面制备聚丙烯酰吡咯膜,然后对膜的结构和表面形貌进行了研究.其中,制备聚丙烯酰吡咯(PAP)的方法是以丙烯酰氯和吡咯钾盐合成丙烯酰吡咯,然后以偶氮二异丁腈(AIBN)为引发剂,进行自由基聚合;金膜表面制备PAP膜的方法是:PAP溶于N,N-二甲基甲酰胺(DMF),然后旋涂到金膜上;最后,涂膜表面与吡咯在三氯化铁溶液中化学法聚合,形成聚合吡咯层.红外光谱可以看出,所得聚合物物质就是需要的产品;AFM非原位表面形貌图像和三维立体形貌图上显示涂膜覆盖了金膜表面原有的小孔,表面粗糙度比纯金膜增大.     

Morphology of multilayers assembled by electrostatic attraction of oppositely charged model polyelectrolytes

Alternating multilayers assembled stepwise with two prototype polyelectrolytes, i.e. poly(allylamine) hydrochloride (PAA) and poly(vinyl sulfate) potassium salt (PVS), have been examined by means of reflection–absorption infrared (RAIR) spectroscopy and atomic force microscopy (AFM). The growth step revealed by the RAIR spectroscopy was in good agreement with the ellipsometric data. The RAIR spectral data revealed that both macromolecules were randomly oriented in the (PVS/PAA) composite system without any preferential alignment. The atomic force microscopy measurements showed that the morphology of the (PVS/PAA) multilayers prepared in the absence of NaCl were quite homogeneous in agreement with a recent report. However, the morphology of the multilayers prepared in the presence of NaCl was observed to be significantly heterogeneous; the morphology was gradually roughened upon an increase in the number of bilayers. The highly coiled, compact macromolecules present in a NaCl solution seemed preferentially to anchor on the oppositely charged, aggregated surface sites without spreading over the interfacial region. Nonetheless, the morphology of the surface of the (PVS/PAA) system was hardly subjected to change once a stable film had been assembled, regardless of the presence of NaCl.     

两步电沉积法制备HA-Ti/HA复合涂层的研究

为了提高金属基羟基磷灰石（HA）涂层的结合强度,采用复合电沉积一电沉积两步法在含Ti粉的钙磷电解液中制备HA—Ti／HA复合涂层,对涂层的组分结构、表面形貌、热稳定性、结合强度和生物活性进行了研究．实验结果表明：两步法制备的底层为HA—Ti复合涂层,外层为纯HA涂层的HA—Ti／HA复合涂层既提高了涂层的结合强度,又保证了涂层的生物活性．当涂层中Ti粉的质量分数为51．2wt％时,涂层与基体的结合强度达到21．2MPa,约为纯HA涂层的3倍．模拟体液浸泡7天后,涂层表面即被一层球状碳磷灰石覆盖,具有良好的生物活性,与纯HA涂层相比,复合涂层具有更好的耐蚀性能．     

Alternate drop coating for forming dual biointerfaces composed of polyelectrolyte multilayers

Two types of polyelectrolyte multilayers were formed on both sides of a quartz crystal microbalance (QCM) substrate by a novel alternate drop coating process. In this study, poly(diallyldimethylammonium chloride) (PDDA) and poly(sodium 4-styrene sulfonate) (PSS) were used as strong-strong polyelectrolytes. On the other hand, PDDA and poly(acrylic acid) (PAA) were used as strong-weak polyelectrolytes. The novel alternate drop coating process can separately fabricate each polyelectrolyte multilayer on both sides of the substrate. The substrate provides dual biointerfaces, both sides of which comprise different multilayers, by employing a combination of polymers. The formation of the multilayer by alternate drop coating was evaluated in terms of changes in the frequency of the QCM and model protein adsorption for proteins such as bovine serum albumin, and their characteristics were investigated with those of the conventional alternate adsorption process by performing dip coating. There was no significant difference between the surface properties resulting from the two formation conditions. This result strongly supported the fact that the multilayers fabricated by alternate drop coating were similar in quality to those fabricated by conventional dip coating. The resulting dual biointerfaces with polyelectrolyte multilayers provide alternative biofunctions in terms of individual protein loading. In summary, the novel alternate drop coating process for substrates is a good candidate for the preparation of dual biointerfaces in the biomedical field.     

Carbohydrate-protein interactions by "clicked" carbohydrate self-assembled monolayers

A Huisgen 1,3-dipolar cycloaddition "click chemistry" was employed to immobilize azido sugars (mannose, lactose, alpha-Gal) to fabricate carbohydrate self-assembled monolayers (SAMs) on gold. This fabrication was based on preformed SAM templates incorporated with alkyne terminal groups, which could further anchor the azido sugars to form well-packed, stable, and rigid sugar SAMs. The clicked mannose, lactose, and alpha-Gal trisaccharide SAMs were used in the analysis of specific carbohydrate-protein interactions (i.e., mannose-Con A; ECL-lactose, alpha-Gal-anti-Gal). The apparent affinity constant of Con A binding to mannose was (8.7 +/- 2.8) x 10(5) and (3.9 +/- 0.2) x 10(6) M(-1) measured by QCM and SPR, respectively. The apparent affinity constants of lactose binding with ECL and alpha-Gal binding with polyclonal anti-Gal antibody were determined to be (4.6 +/- 2.4) x 10(6) and (6.7 +/- 3.3) x 10(6) M(-1), respectively by QCM. SPR, QCM, AFM, and electrochemistry studies confirmed that the carbohydrate SAM sensors maintained the specificity to their corresponding lectins and nonspecific adsorption on the clicked carbohydrate surface was negligible. This study showed that the clicked carbohydrate SAMs in concert with nonlabel QCM or SPR offered a potent platform for high-throughput characterization of carbohydrate-protein interactions. Such a combination should complement other methods such as ITC and ELISA in a favorable manner and provide insightful knowledge for the corresponding complex glycobiological processes.     

Determination of the ionization state of 11-thioundecyl-1-phosphonic acid in self-assembled monolayers by chemical force microscopy

The present article describes the preparation and preliminary characterization of a novel phosphate-functionalized self-assembled monolayer (SAM) and the determination of the surface ionization states of the phosphate headgroup in aqueous solutions by chemical force microscopy (CFM). The phosphate headgroup used was PO(OH)2, a diprotic acid. The adhesion force between an AFM probe and a flat substrate, both of which were chemically modified with the same phosphate SAM, was also measured as a function of pH and ionic strength. At low ionic strength (10(-4) M), two peaks were observed in the force titration curve (adhesion force versus pH) at pH 4.5 and 8.4. The two peaks are positioned 2.4 and 1.2 pH units higher, respectively, than the acid dissociation constants obtained for the phosphate group free in aqueous solution. At high ionic strength (10(-1) M), the adhesion forces were reduced by 1 order of magnitude and the peaks were replaced by shoulders similar to those previously reported for acid force titrations. On the basis of JKR theory, the surface pKa values of the phosphate group in high ionic strength solutions were found to be 4.5 and 7.7, respectively. However, in light of the effects of ionic strength on the force titration curves, we discuss the applicability of JKR theory to nanoscopic measurements of adhesion force and surface pKa.     

4-Aminophenyl boronic acid modified gold platforms for influenza diagnosis

As a potential pandemic threat to human health, there has been an urgent need for rapid, sensitive, simpler and less expensive detection method for the highly pathogenic influenza A virus. For this purpose, Quartz Crystal Microbalance (QCM) and Surface Plasmon Resonance (SPR) sensors have been developed for the recognition of hemagglutinin (HA) which is a major protein of influenza A virus. 4-Aminophenyl boronic acid (4-APBA) has been synthesized and used as a new ligand for binding of sialic acid (SA) via boronic acid–sugar interaction. SA has an important role in binding of HA. QCM and SPR sensor surfaces have been modified with thiol groups and then 4-APBA and SA have been immobilized on sensor surfaces, respectively. Sensor surfaces have been screened with AFM and used for the determination of HA from aqueous solution. The selective recognition of the QCM and SPR sensors toward Concanavalin A has been reported in this work. Also, the binding capacity and detection limits of QCM and SPR sensors have been calculated and detection limits were found to be 4.7 × 10^? 2 μM, (0.26 μg ml^? 1) and 1.28 × 10^? 1 μM, (0.72 μg ml^? 1) in the 95% confidence interval, respectively.     

Selectively Promoting or Preventing Osteoblast Growth on Titanium Functionalized with Polyelectrolyte Multilayers

Titanium plays an important role in medical applications, such as hip joint implants or fixation plates. These implants must perform differently depending on their clinical application. In particular, the osseointegrative properties required of the implant vary with clinical application. The present work is aimed at the functionalization of titanium surface using polyelectrolyte multilayers consisting of natural biopolymers and testing their cell adhesive properties with respect to the osseointegration capacity. Multilayered coatings were created from chitosan (Chi), hyaluronic acid (HA), and gelatine (Gel) through layer‐by‐layer deposition. Cell adhesion, proliferation, and viability were tested in vitro with the human osteoblast cell line CAL‐72 at timescales up to 7 d. Two multilayer coatings consisting of alternated chitosan/gelatin or chitosan/hyaluronic acid layers with the outmost layer of gelatin (Chi/Gel) or hyaluronic acid (Chi/HA), respectively, were tested. The experimental results showed that surfaces functionalized with Chi/Gel and Chi/HA multilayers demonstrated a good initial adhesion of osteoblasts. After 4 d culturing, osteoblast cells were almost completely detached from the substrates functionalized with Chi/HA multilayers. In contrast to Chi/HA, the proliferation of osteoblasts on substrates with Chi/Gel multilayer coatings was statistically significantly higher compared to the control titanium. We have shown that the growth of osteoblasts can be enhanced or completely prevented on a titanium surface functionalized with polyelectrolyte multilayers consisting of natural biopolymers, as desired. Both multilayer coatings, Chi/Gel and Chi/HA, have potential for applications in the field of titanium implants, where rapid osseointegration is essential, and/or where no ingrowth of the implant is desired, respectively.     

真空和大气等离子喷涂镁黄长石生物活性涂层的对比研究

大气喷涂法制备的镁黄长石涂层是一种具有优良生物活性的新型骨科移植体涂层材料, 但其结晶度较低, 影响涂层的化学稳定性。本研究采用真空等离子喷涂法在钛合金表面制备了高结晶度的镁黄长石涂层。与大气喷涂镁黄长石涂层相比, 真空喷涂镁黄长石涂层具有更高的磷灰石矿化能力, 在SBF中浸泡6 d后表面即沉积了一层类骨磷灰石层, 浸泡14 d后表面沉积的磷灰石层的厚度约为大气涂层的4倍。真空喷涂镁黄长石涂层的离子释放明显低于大气涂层, 显示出更高的化学稳定性。骨髓间充质干细胞在真空和大气喷涂镁黄长石涂层表面粘附和铺展良好, 在两种涂层表面的增殖速度均明显高于HA涂层。本研究表明真空等离子喷涂的镁黄长石陶瓷涂层因其显著提高的生物活性及化学稳定性, 可能更适合用作人工关节涂层材料。     

Development of nano-hydroxyapatite coating by electrohydrodynamic atomization spraying

Electrohydrodynamic atomisation (EHDA) spraying of a hydroxyapatite (HA) suspension consisting of nano-particles (nHA) has been used to produce a HA coating comprising of nanostructured surface topography. In EHDA the suspension is jetted from a needle under an electric field. Obtaining the stable cone-jet mode of EHDA is critical to improve the quality and optimise the morphology of HA coatings, therefore a systematic investigation of the effects of several key processing parameters, such as suspension flow rate, applied voltage and distance between the needle and substrate, and needle size was carried out in this work. The HA coatings processed under different spraying parameters were compared and then scored according to uniformity and microstructural integrity. It was found that all of these parameters had a very significant influence on the morphology of nHA coating prepared. Under an optimised processing condition, where a needle orifice diameter of 300 μm, kept at a distance of 20 mm from the substrate, a flow rate of 20 μL/min, and the applied voltage kept within 4.3 kV and 5.2 kV, a uniform nHA coating was obtained. This is a crucial step forward in obtaining advanced nano-hydroxyapatite coatings of high quality for biomedical applications by using EHDA spraying.     

Electrophoretic deposition of HA/MWNTs composite coating for biomaterial applications

A composite coating of hydroxyapatite (HA)/multi-walled carbon nanotubes (MWNTs) has been fabricated by electrophoretic deposition (EPD). The nano powders of HA and MWNTs were dispersed in ethanol with total concentration of 0.005 g/mL and MWNTs 20% and 30% contents (wt). And the pH value of suspension was adjusted in a range from 4 to 5. After stabilization the mixture was ultrasonically treated for 3 h to form a stable suspension. Prior to the electrophoretic deposition, the titanium substrate was hydrothermally treated at 140 in NaOH (10 mol/L) solution for 6 h. A titanium sheet and circinal net of stainless steel were used as a cathode and an anode respectively, and a constant deposition voltage of 30 V was applied for 50-60 s in the EPD process. The thickness of the coatings was controlled from 10 mum to 20 mum. The samples of composite coating were then sintered in a resistance tube furnace in flowing argon at 700 for 2 h. The structure of the as prepared coating was characterized by SEM and XRD, and the bonding force of the coating/substrate was measured by an interfacial shear strength test. It is shown that the bonding strengths between the coating and the titanium substrate is as high as 35 MPa. The cell culture experiments indicate that the prepared composite coating of HA/MWNTs possesses good biocompatibility.     

Preparation of an electrodeposited hydroxyapatite coating on titanium substrate suitable for in-vivo applications

In this paper the porous hydroxyapatite coating on Ti implant materials was prepared by the process of electrodeposition, hydrothermal and sinter. The surface morphology, bond strength and thickness of HA coatings were investigated by SEM, AFM, and its biocompatibility was evaluated by cytotoxicity experiments and implant experiments, respectively. Results showed that (1) The HA coatings was 50 μm thickness and adhered on the Ti substrate strongly, which bond strength reached 38MPa. AFM analysis showed that the HA coating was porous structure, in which the mean pore size was 236.5 μm, (2) Cytotoxicity experiments and implant experiments showed that HA-coated Ti implant materials has little cytotoxicity in vitro and little inflammatory reaction in vivo, and there were no statistically disparity between HA-coated Ti implant and titanium implant materials of clinical application (p > 0.05), which demonstrated that HA-coated Ti has a good biocompatibility.     

Investigation of metallic nanoparticles adsorbed on the QCM sensor by SEM and AFM techniques

Quartz crystal microbalance (QCM) is known as a very sensitive device used for determination of mass quantity adsorbed on sensor surface. Its detection limits are in the range of ng cm\(^{-2}\). The adsorption mechanism of metallic nanoparticles on QCM sensor was investigated by scanning electron microscopy (SEM) and atomic force microscopy (AFM). This study aims to highlight the importance of QCM applications in nanoparticles deposition field. The layers formed through adsorption process, induced by the oscillations of the QCM sensor, were investigated by AFM for surface topography and for particle mean size values. The morphology of layers and nanoparticles dimensions were determined by SEM. For a more complex investigation of the nanoparticles adsorption mechanism, the chemical composition of layers was achieved using SEM coupled with energy dispersive X-ray spectrometer (SEM-EDS). This preliminary research involved a new approach in characterization of metallic nanoparticles layers to achieve functional assembled monolayers.     

Quartz crystal microbalance with integrated surface plasmon grating coupler

We have integrated a surface plasmon grating coupler into a quartz crystal microbalance (QCM) for studying surface association/dissociation reactions. In the integrated system only QCM measurement is needed to record both the optical and the acoustic signals in the same association/dissociation reaction. This integration considerably simplifies a conventional combination instrument of a grating-coupled surface plasmon resonance (SPR) spectrometer and a quartz crystal microbalance by eliminating a number of SPR components. Moreover, in the integrated system detection of the light reflections is not needed by which one bypasses the interference problem caused by two coherent light reflections off the glass window used to seal the fluid sample and off the sensor surface. The utility of the integrated system is demonstrated using a layer-by-layer polyelectrolyte multilayer deposition protocol, in which the complete features of a conventional grating-coupled SPR/QCM combination instrument are retained, including detection of optical and acoustic changes, as well as monitoring of adsorption kinetics.     

生物活性钛涂层

真空等离子喷涂的钛涂层经 ５．０ｍｏｌ／Ｌ ＮａＯＨ溶液处理后,将其浸泡在含 Ｃａ^２＋、ＨＰＯ_４^２－的模拟生理体液（ＦＣＳ和ＳＢＦ）中,考察涂层诱导羟基磷灰石生长过程,并评价其生物活性．用ＳＥＭ观察碱处理前后和在模拟生理体液中浸泡后钛涂层的形貌,用ＡＥＳ分析了碱处理前后钛涂层的表面成分;用ＸＲＤ、ＦＴ－ＩＲ和ＥＤＳ表征浸泡后涂层表面生长物的结构和成分;并测量了处理后钛涂层在浸泡过程中溶液中离子浓度和ｐＨ值的变化．结果表明,经处理的钛涂层在模拟生理体液中能诱导羟基磷灰石在其表面生长;在ＳＢＦ和ＦＣＳ分别形成碳酸羟基磷灰石层和含氧磷灰石的羟基磷灰石层．钛涂层的活性是由于碱处理后表面形成了网状和纤维状结构的Ｎａ－Ｔｉ－Ｏ化合物．这种化合物在模拟生理溶液中释放Ｎａ^＋,吸收Ｈ^＋;形成水化钛酸盐,诱导羟基磷灰石成核生长．     

低温熔盐电沉积羟基磷灰石复合涂层的研究

为了提高羟基磷灰石涂层的结合强度,在AlCl3-NaCl-TiCl3低温熔盐体系中加入HA微粒,复合电沉积制备Al-Ti/HA复合涂层,并对涂层的表面形貌、结构和结合强度进行了研究.结果表明: HA微粒均匀分散在Al-Ti合金镀层中,其共沉积量随电流密度的减小和熔盐中HA浓度的增大而增强;涂层的结合强度随HA共沉积量的增大而增强,当HA的共沉积量为40.1%(质量分数)时,Al-Ti/HA复合涂层的结合强度达到28.1 MPa.