利用胶原-肝素自组装多层膜改善纯钛表面血液相容性的研究

为了改善钛表面的血液相容?用层层自组装技术将抗凝药物肝素和细胞外基质成分之一的胶原层层交替吸附到钛表面,形成多层薄膜,使具有良好的抗凝血功能.钛通过氢氧化钠处理表面带负电,在聚赖氨酸溶液中会吸附一层带正电的聚赖氨酸,然后将其交替浸泡于带负电的肝素和带正电的胶原中形成自组装薄膜.通过傅立叶红外漫反射(FTIR)检测材料表面基团的变化,通过测水接触角的方法跟踪组装过程,通过扫描电子显微镜(SEM)观察材料表面形貌的变化.血小板黏附实验显示钛改性后表面血小板黏附量明显减少,且反映血小板激活程度的P-selectin的表达量结果也显示,钛改性后表面血小板激活程度显著降低,活化部分凝血活酶时间(APTT)和外源性凝血系统的凝血酶原时间(PT)的实验显示钛改性后表面凝血时间延长,血液相容性得到改善.研究说明这种方法对于心血管材料钛的改性有重要价值.     

钛表面层层组装胶原与肝素改善血液相容性的研究

为了改进钛表面的血液相容性，本实验通过电荷作用层层组装带正电的细胞外基质胶原和带负电的肝素，在钛表面形成多层仿生膜，以改善钛表面的血液相容性。钛通过氢氧化钠处理，表面形成多孔结构，并产生碱性羟基，之后将其浸泡于多聚赖氨酸溶液中，然后将材料交替浸泡于肝素和胶原中，最后一层为肝素，通过电荷吸引形成多层膜．通过傅立叶红外漫反射（FTTR）检测各步反应后表面基团的变化，通过扫描电子显微镜（SEM）观察表面形貌的变化，初步说明肝素已经结合在材料表面；血小板黏附实验直观反应组装膜有良好的血液相容性。通过本实验说明这种改性方法对于材料血液相容性的改善有一定价值。     

钛材料表面固定多聚赖氨酸-肝素纳米颗粒以改善血液相容性的研究

通过将多聚赖氨酸(PLL)-肝素纳米颗粒固定在多巴胺涂覆的钛表面,以改善其血液相容性。利用zeta电位仪及甲苯胺蓝法检测纳米颗粒的粒径及成分,通过傅立叶变换红外光谱(FT-IR)、X射线光电子能谱(XPS)及水接触角等对颗粒固定前后表面理化性质的变化进行表征。通过体外血小板粘附实验、肝素释放及活化部分凝血活酶时间(APTT)检测对改性样品的血液相容性进行评价。结果表明,PLL-肝素纳米颗粒成功固定在多巴胺沉积的钛表面,纳米颗粒的固定有效降低钛材料表面血小板的粘附行为,大大提高了血液相容性。     

退火温度对P离子注入TiO2薄膜材料物理与生物学性能的影响

利用离子注入技术在TiO2薄膜中注入P元素,然后分别在不同温度进行退火处理,得到一系列钛氧化物掺杂材料.随着退火温度的上升,P注入的TiO2薄膜材料电阻降低,与水的接触角在70度左右波动.通过LDH及血小板粘附实验,发现高温退火样品的血液相容性得到改善.     

磷酰胆碱类似物在有机硅材料表面的常压等离子体引发接枝改性研究

采用He常压辉光放电等离子体(APGDP)处理有机硅薄膜材料表面并引发2-甲基丙烯酰氧乙基磷酰胆碱(MPC)在其表面接枝聚合.通过ATR-FTIR对接枝前后膜表面的结构进行表征分析,确定MPC共价接枝到材料表面.改性后膜表面的接触角由101°下降到54°,在室温下保存15天后仍维持在58°左右,表明接枝MPC后有机硅材料获得高亲水性的表面,并能使这一性质较好地保持.接枝前后膜的力学性质变化不大.体外血小板粘附实验表明,接枝MPC后的材料表面能够显著抑制血小板的粘附和聚集,具有优良的血液相容性,可以作为一种新型医用生物弹性体.     

涤纶材料表面固定水蛭素及其血液相容性研究

利用紫外辐照仪在聚对苯二甲酸乙二醇酯(涤纶,PET)膜表面接枝聚丙烯酸(PET-AA)长链分子,再通过化学方法进一步固定水蛭素,获得的改性表面的X光电子能谱分析表明水蛭素分子被有效地固定在PET表面.改性表面的水接触角降低,亲水性得到改善.体外血小板粘附实验的结果显示,固定了水蛭素的PET薄膜的血液相容性比未改性的PET膜有明显的改善.     

肝素和Ⅳ胶原在Ti表面组装及不同组装层数对内皮祖细胞的影响

利用层层自组装技术将带有相反电荷的肝素和Ⅳ型胶原交替组装到材料表面,研究了组装前后及不同组装层数对内皮祖细胞的影响。Ti通过碱活化处理后,表面带上羟基,与多聚赖氨酸中的羧基结合,使材料形成带正电荷的氨基表面,接着将带有异种电荷的肝素和Ⅳ型胶原交替滴加到材料表面形成多层自组装层。通过傅立叶红外掠射检测各步处理后材料表面基团的变化,通过甲苯胺蓝定量表征材料表面肝素组装量,通过原子力显微镜观察组装前后材料表面形貌变化,通过水接触角的测定跟踪组装过程中材料表面亲疏水性能变化。结果显示肝素和Ⅳ型胶原成功地组装到材料表面,修饰后的表面较Ti表面内皮祖细胞的粘附数量多,随着组装层数的增加,细胞在材料表面粘附数量也逐渐增加,且其生长与增殖逐步加快。     

不同晶体结构氧化钛薄膜性能研究

用非平衡磁控溅射技术分别制备了锐钛矿、金红石、锐钛矿和金红石共存结构的氧化钛薄膜.用X射线衍射(XRD)分析薄膜的晶体结构,血小板粘附和凝血因子实验研究薄膜的血液相容性,分光光度计和接触角测量法测试薄膜表面物化性质.研究表明,氧化钛薄膜具有宽禁带的半导体特性,血液相容性优于热解碳.金红石结构氧化钛薄膜由于其与血浆白蛋白、血浆纤维蛋白原、血液及水之间有较小的界面张力,自身较低的表面能色散分量与极性分量的比值,加之其宽禁带宽度的n型半导体特性使其具有最优的血液相容性.     

医用钛表面生物分子自组装薄膜的制备

采用分子自组装(SAM)方法在医用钛表面制备了硫酸软骨素薄膜.通过激光共聚焦显微镜,傅立叶红外光谱,X射线光电子能谱以及接触角测定仪等分析测试手段,比较研究了表面预处理方法的活化作用以及成膜反应时间对自组装薄膜的影响规律.实验结果表明,用等体积比的浓硫酸和双氧水混合溶液在75C对钛表面处理1h后的钛片能获得最高的表面自由能,而自组装膜的反应在12h内即可完成.     

仿贻贝磷脂聚合物的合成及其仿生涂层的构建

为改善纯钛表面的抗凝血性能,合成了一系列磷酸胆碱仿生聚合物,并在其表面构建了稳定的仿生涂层,通过一系列实验、表征和评价优化出更有利于钛材表面改性的磷酸胆碱聚合物。首先采用简单的自由基聚合法和羧基与氨基的反应,分两步合成了5种不同比例的2-甲基丙烯酰氧乙基磷酰胆碱(MPC)、甲基丙烯酸(MA)和多巴胺(DA)的无规共聚(PMMDA3-Q、PMMDA3-T、PMMDA5-Q、PMMDA5-T、PMMDA7),并通过FTIR、1 H NMR、GPC以及UV-Vis等完成了定性、定量表征和分析。通过自组装的方式,将5种不同聚合物分别组装到钛表面,并采用QCM-D、AFM及血小板粘附实验考察了5种聚合物在钛表面的组装量、表面粗糙度以及改性后的钛表面抗血小板粘附及抑制血小板激活的能力。结果表明,聚合物PMMDA7更适合作为钛材表面改性聚合物,为构建磷酸胆碱仿生多功能涂层提供了实验基础和方法。     

Study on protein conformation and adsorption behaviors in nanodiamond particle-protein complexes

In the present research, the conformation of bovine serum albumin (BSA) in the nanodiamond particle (ND)-BSA complex was studied by Fourier transform infrared spectroscopy, fluorescence spectroscopy, UV-vis spectroscopy, and circular dichroism spectroscopy. The spectroscopic study revealed that most BSA structural features could be preserved in the complex though the BSA underwent conformational changes in the complex due to ND-BSA interaction. In addition, BSA adsorption isotherms and zeta-potential measurements were employed to investigate the pH dependence of the ND-BSA interaction. The changes in surface charge of the ND-BSA complex with pH variations indicated that the binding of BSA to ND might lead to not only the adsorption of BSA onto the ND surface but also the partial breakup of ND aggregates into relatively small ND-BSA aggregates because of the strong binding force between ND and BSA. The results show that ND is an excellent platform for protein immobilization with high affinity and holds great potential to be used for biosensor applications.     

Electrical characterization of protein molecules by a solid-state nanopore

The authors measured ionic current blockages caused by protein translocation through voltage-biased silicon nitride nanopores in ionic solution. By calculating the mean amplitude, time duration, and the integral of current blockages, they estimated the relative charge and size of protein molecules at a single molecule level. The authors measured the change in protein charge of bovine serum albumin (BSA) protein induced by pH variation. They also confirmed that BSA molecules indeed traverse nanopores using an improved chemiluminescent analysis. They demonstrated that a larger protein fibrinogen could be distinguished from BSA by a solid-state nanopore measurement.     

Sonochemical synthesis of (3-aminopropyl)triethoxysilane-modified monodispersed silica nanoparticles for protein immobilization

3-Aminopropyltriethoxysilane modified monodispersed silica nanoparticles were synthesized by a rapid sonochemical co-condensation synthesis procedure. The chemical nature of surface organic modifier on the obtained modified silica nanoparticle was characterized by ¹³C and ²⁹Si MAS Nuclear Magnetic Resonance (NMR) spectroscopies, Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA)- differential scanning calorimetry (DSC). Due to the strengthened positive surface charge of the silica nanoparticles by the modification with aminopropyl groups, the capability for bovine serum albumin (BSA) adsorption was significantly increased as compared with bare silica nanoparticles. 80 mg/g BSA was adsorbed on modified silica nanoparticles, whereas only 20 mg/g BSA could be loaded on pure silica nanoparticles. The enhanced positive surface charge repelled proteins with net positive charge and the modified silica nanoparticles exhibited negligible adsorption of lysozyme, thus a selective adsorption of proteins could be achieved.     

Carbon nanotubes induce secondary structure changes of bovine albumin in aqueous phase

Interaction of nanomaterials to protein molecules is one of the most important issues to deeply understand the influences of the nanomaterials upon physiological processes and protein functions. So far most of investigations focused on the protein molecules adsorbed on the nanomaterials surface, less is known about those in the aqueous phase (not absorbed). In this work, luminescent spectroscopy analysis, circular dichroism measurement, atomic force microscopy, matrix-assisted laser desorption/ionization time of flight mass spectrometry, isoelectric focusing and sulfate polyacrylamide gel electrophoresis were used to investigate the influence of oxidized water-soluble multiwalled carbon nanotubes (CNT) dispersing in aqueous solution upon the structures of bovine serum albumin (BSA) through co-incubation. We focused on BSA molecules that stayed in the aqueous phase instead of those adsorbed by CNT. Experimental results show that the fractions of beta-sheet decreased from 33.3% to 29.8% and beta-turn increased from 2% to 5% in reference with native BSA. There was a slight increase of alpha-helix and a slight reduction of random coil. BSA molecules that had been encountered with CNT and were left in the solution formed a loose and flatten morphology on graphite substrates instead of their native tight and round morphology observed by AFM. The value of isoelectric point for BSA after exposed to CNT moved towards to a higher pH position compared with native BSA. All together, it was concluded that the oxidized water-soluble multiwalled carbon nanotubes not only adsorb bovine serum albumin molecules to their surface, but also induces albumin molecules in the aqueous solution undergo secondary structure changes, which lead to a conformation change.     

Competitive adsorption of bovine serum albumin and lysozyme on characterized calcium phosphates by polyacrylamide gel electrophoresis method

Characterizations of hydroxyapatite (HA), biphasic calcium phosphate (BCP) and beta tricalcium phosphate (β-TCP) ceramic particles were carried out using X-ray diffusion (XRD), Scanning electron micrograph (SEM), Particle Sizer and Zeta potential analyzer. Competitive adsorption of bovine serum albumin (BSA) and lysozyme (LSZ) on the three calcium phosphates were investigated by polyacrylamide gel electrophoresis (PAGE) method. The results showed that HA, BCP and β-TCP ceramic particles with irregular shapes and similar size distributions all had negative surface net charges in pH7.4 phosphate buffered saline (PBS) solution and exhibited alike behaviors of BSA and LSZ adsorption. LSZ had higher affinity for calcium phosphate ceramics than BSA and its adsorption on them didn’t be almost influenced by the increasing of BSA concentration in the solution. Electrostatic interaction played an important role on the competitive adsorption of BSA and LSZ on the surface of calcium phosphate ceramic particles.     

Preliminary study of adsorption behavior of bovine serum albumin(BSA) protein and its effect on antibacterial and corrosion property of Ti-3Cu alloy

The adsorption behavior,antibacterial,and corrosion properties of a Ti-3Cu alloy were studied in a phosphate-buffered saline solution containing 0,1,3,and 6 g L-1 bovine serum albumin protein at 37 ℃and pH =7.4 (±0.2).The protein adsorption behavior was examined via cyclic voltammetry,secondary ions mass spectroscopy (SIMS),and angle-resolved X-ray photoelectron spectroscopy (ARXPS).The cor-rosion property was analyzed by the open circuit potential (OCP),potentiodynamic polarization (PD),and electrochemical impedance spectroscopy (EIS) examinations.The antibacterial test was conducted according to the GB/T 21510 China Standard.It was observed that the surface charge density (Q～s) was directly proportional to the amount of the adsorbed BSA protein,signifying that the protein adsorption was accompanied by the charge transfer,pointing to chemisorptions phenomena.BSA amino groups and other organic species were observed in the surface analysis examinations.It was shown that the formation of barrier complexes between the TiO2 oxide-layer and PBS solution resulted in decreasing the release of Cu-ions,which consequently reduced the antibacterial activity.On the other hand,these barrier complexes improved the corrosion resistance by increasing the charge transfer resistance and double-layer capacitance of the Ti-3Cu alloy.     

BSA和FN在纳米化钛表面的蛋白吸附及释放行为

通过阳极氧化技术, 在钛表面制备一层管径为100nm左右的氧化钛纳米管. 选取小牛血清白蛋白(BSA)和纤维连接蛋白(FN)两种蛋白质进行蛋白吸附实验, 并在仿生条件下进行蛋白质的体外释放. 对吸附了蛋白的纳米管试样表面进行红外和荧光定性分析, 同时采用考马斯亮蓝法对纳米管表面的蛋白吸附进行定量检测, 实验发现纳米管试样更有利于蛋白质的吸附, 且FN在试样表面吸附时的吸附率大于BSA. 氧化钛纳米管的蛋白释放分为突释和缓释两个阶段, 其释放机制符合Fickian扩散.     

Incorporation of bovine serum albumin into biomimetic coatings on titanium with high loading efficacy and its release behavior

Bovine serum albumin (BSA) was employed as a model protein to study its loading efficiency into a calcium phosphate (CaP) coating on titanium substrates. It is found that the protein loading efficiency can be adjusted by varying the specific configurations of the coating system such as simulated body fluid (SBF) volume, solution height and container selection for the SBF. A BSA loading efficiency as high as 90% was achieved when the ratio of the substrate surface area to modified SBF (m-SBF) volume was as high as 0.072. The release of BSA from the biomimetic coatings was also investigated in vitro. A sustained release was achieved although a large quantity of BSA was still trapped in the coating after 15 days of immersion in a phosphate buffer solution. A much faster release rate would be expected when the coating is implanted in vivo due to the active involvement of osteoclast cells and enzymes.     

Dynamic Adsorption of Albumin on Nanostructured TiO(2)Thin Films

Spectroscopic ellipsometry was used to characterize the optical properties of thin (<5 nm) films of nanostructured titanium dioxide (TiO(2)). These films were then used to investigate the dynamic adsorption of bovine serum albumin (BSA, a model protein), as a function of protein concentration, pH, and ionic strength. Experimental results were analyzed by an optical model and revealed that hydrophobic interactions were the main driving force behind the adsorption process, resulting in up to 3.5 mg/m(2) of albumin adsorbed to nanostructured TiO(2). The measured thickness of the adsorbed BSA layer (less than 4 nm) supports the possibility that spreading of the protein molecules on the material surface occurred. Conformational changes of adsorbed proteins are important because they may subsequently lead to either accessibility or inaccessibility of bioactive sites which are ligands for cell interaction and function relevant to physiology and pathology.     

携载蛋白质的氧化钛纳米管表面成骨细胞行为

通过阳极氧化技术,在光滑钛表面制备一层管径约100nm的纳米管,并选用小牛血清白蛋白（BSA）和纤维连接蛋白（FN）以及两种蛋白的混合溶液（BF）进行蛋白吸附试验。将吸附蛋白后的试样进行细胞培养,所用细胞为MC-3T3E1成骨细胞株,结果表明,较光滑钛吸附蛋白前后的纳米管试样显示出更好的成骨活性,同时,吸附单组分纤维连接蛋白的纳米管试样能更好地促进成骨细胞的黏附和生长,具有更高的生物活性。