Immobilization of NaIO4-treated heparin on PEG-modified 316L SS surface for high anti-thrombin-III binding

Poor compatibility between blood and metallic coronary artery stents is one reason for arterial restenosis; however, the immobilization of anticoagulant agents on the surface of the stent is a feasible method of improving stent compatibility. Heparin, a well-known anticoagulant, has been frequently used to coat the surfaces of certain biomaterials to attain blood compatibility. The compound 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide has often been utilized for the immobilization of heparin, but the critical carboxyl groups of heparin (with regards to heparin's anticoagulant activity) will be reduced by this method. This study examined possible methods of heparin immobilization without consuming these carboxyl groups. The 316L stainless steel surface was first activated with hexamethylene diisocyanate and then coupled with bis-amine-terminated poly (ethylene glycol) (BA-PEG) so as to create active amine groups. Sodium periodate (NaIO(4); SP) was then used to oxidize heparin to form aldehyde groups. The treated heparin could then be grafted onto the activated surface of the test material without losing its carboxyl groups. Effective surface modification of the hexamethylene diisocyanate-activated and BA-PEG-grafted 316L SS surface was confirmed using Fourier Transform Infrared Spectroscopy, electron spectroscopy for chemical analysis and a water contact angle test. After the heparin was immobilized on the BA-PEG-grafted 316L SS surface by SP, the surface showed an improvement in antithrombrin III (AT III) binding ability, its anticoagulant property, and hemocompatibility in comparison with heparin grafted by 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide.     

316L不锈钢表面电镀Rh膜的腐蚀行为及血液相容性研究

通过电镀Rh对316L不锈钢进行表面改性,以提高不锈钢的抗腐蚀性和血液相容性,用XPS对铑镀层的成分进行了表征,运用电化学方法研究了镀有Rh和未进行表面处理的316L不锈钢在Tyrode‘s生理溶液中的腐蚀行为,并通过溶血率测试对表面镀有Rh的316L不锈钢初步进行了血液相容性评价,结果表明：表面镀Rh的不锈钢的抗体液腐蚀性提高,。且溶血率下降,说明镀Rh可以提高316L不锈钢的血液相容性。     

316L-Cu抗菌不锈钢种植体中Cu离子的抗菌活性

背景：316L-Cu抗菌不锈钢是通过在不锈钢中添加一定量的铜元素,并采取特殊的热处理使不锈钢基体中均匀弥散分布富铜析出相,从而赋予抗菌不锈钢优异的抗菌性能。 目的：评价新型316L-Cu抗菌不锈钢种植体对牙龈卟啉单胞菌的抗菌性能,并检测316L-Cu抗菌不锈钢的Cu离子溶出行为,为其进一步临床应用提供生物医学证据。 方法：以医用316L不锈钢为成分基础,添加3.5%-4.0%的Cu元素制成316L-Cu抗菌不锈钢。将其按       0.1 cm2/L表面积对容量比置于模拟体液中,在37 ℃震荡水浴中浸泡1-10 d。采用石墨炉原子吸光光度计每天测定浸提液中的铜离子含量。通过覆膜法抗菌实验及扫描电镜形态学观察评价316L-Cu抗菌不锈钢的杀菌性能。 结果与结论：316L-Cu抗菌不锈钢在模拟体液中浸泡1-10 d后的Cu离子溶出量远高于普通医用316 L不锈钢。随着作用时间的延长,316L-Cu抗菌不锈钢的杀菌率逐步提高,10 h时杀菌率已达100%。316L-Cu抗菌不锈钢作用3 h后牙龈卟啉单胞菌的菌体形态即出现明显的变化,6 h后菌体出现明显的裂解,9 h后材料表面绝大多数细菌即崩解为碎片。说明316L-Cu抗菌不锈钢能够有效地杀灭种植体周围炎的主要致病菌,同时能够缓慢释放Cu离子,改变种植体周围微环境,说明其为一种极具潜力的生物材料,具有良好的临床应用价值。 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

白蛋白固定等离子体聚烯丙胺薄膜及体外血小板粘附行为

采用脉冲射频等离子体聚合技术,在医用不锈钢表面制备了等离子体聚烯丙胺薄膜,并进一步在聚合薄膜表面固定白蛋白(BSA)分子。采用傅立叶红外光谱(FTIR)、X光电子能谱(XPS)、台阶仪、原子力显微镜和淌滴法分别表征结构成分、厚度、表面形貌和水接触角。结果表明:随着放电功率增加,聚烯丙胺薄膜中氮含量及伯胺基浓度减小,薄膜厚度和表面粗糙度增加,接触角增大。XPS分析证明白蛋白分子被固定于聚烯丙胺薄膜表面。体外血小板粘附评价表明,不锈钢表面固定白蛋白的聚烯丙胺薄膜能显著抑制血小板粘附及激活。     

Immobilization of the direct thrombin inhibitor-bivalirudin on 316L stainless steel via polydopamine and the resulting effects on hemocompatibility in vitro

Bivalirudin (BV), a peptidic direct thrombin inhibitor, derived from hirudin, has gained increasing interest in clinical anticoagulant therapy in the recent years. In this work, a hemocompatible surface was prepared by immobilization of BV on 316L stainless steel (SS) using a bonding layer of polydopamine (DA). X-ray photoelectron spectroscopy (XPS) was used to determine the chemical composition of the surfaces to characterize polydopamine intermediate layer and the immobilized BV. The quantity of bound BV was measured by quartz crystal microbalance (QCM). The hemocompatibility in vitro was evaluated by coagulating time of activated partial thromboplastin time (aPTT) and prothrombin time (PT) assay, platelet adhesion and activation, fibrinogen adsorption, and activation and whole blood test. The effect of sterilizing method on the bioactivity of immobilized BV was also evaluated. The results showed that BVs were successfully immobilized on SS surface with the DA interlayer at a density of 98 ng/cm(2) . BV coating surface prolonged aPTT and PT, inhibited the activation of platelet and fibrinogen significantly. Sterilization by ultraviolet radiation was possible with only marginal loss of activity. Thus, the approach described here may provide a basis for the preparation of 316L SS surface modification for use in cardiovascular implants.     

Preparation and characteristics of the sulfonated chitosan derivatives electrodeposited onto 316l stainless steel surface

In order to ameliorate the properties of corrosion resistance and achieve applications in anti-biofouling of 316L stainless steel (SS), a sulfated derivative of chitosan was deposited onto stainless steel surface by an electrochemical method. In detail, chitosan-catechol (CS-CT) was synthesised in the hydrochloric acid solution by the Mannich reaction and then electrodeposited on the surface of the polished 316L stainless steel. The chitosan-catechol deposited SS sample was further modified with maleic anhydride and sulfite. The grafting progress was monitored by FTIR, UV spectrophotometer and X-ray photoelectron spectroscopy. Hydrophilicity and corrosion resistance of modified SS were characterized by water contact angle measurements, Tafel curves and electrochemical impedance spectroscopy. The morphology of the SS surface before and after the modification was investigated by atomic force microscopy and scanning electron microscope. Further, the anti-biofouling performance in terms of the anti-adsorption protein and anti-bacteria effects of all modified SS samples were estimated, and the modified 316L exhibits the capability of lower protein adsorption and improved antibacterial effect.     

Properties of super stainless steels for orthodontic applications

Orthodontic stainless-steel appliances are considered to be corrosion resistant, but localized corrosion can occur in the oral cavity. This study was undertaken to evaluate the properties of super stainless steels in orthodontic applications. Accordingly, the metallurgical properties, mechanical properties, corrosion resistance, amount of the released nickel, cytotoxicity, and characteristics of the passive film were investigated. Corrosion resistances of the specimens were high and in the following order: super austenitic stainless steel (SR-50A) > super ferritic stainless steel (SFSS) = super duplex stainless steel (SR-6DX) > 316L SS > super martensitic stainless steel (SR-3Mo) in artificial saliva, 37 degrees C. At 500 mV (SCE), current densities of SR-50A, SFSS, SR-6DX, 316L SS, and SR-3Mo were 5.96 microA/cm(2), 20.3 microA/cm(2), 31.9 microA/cm(2), 805 microA/cm(2), and 5.36 mA/cm(2), respectively. Open circuit potentials of SR-50A, 316L SS, SR-6DX, SR-3Mo, and SFSS were - 0.2, - 0.22, - 0.24, - 0.43, and - 0.46 V (SCE), respectively. SR-50A, SFSS, and SR-6DX released below 3 ng/ml nickel for 8 weeks, and increased a little with immersion time, and 316L SS released about 3.5 ng/ml nickel, but SR-3Mo released a large amount of nickel, which increased with immersion time. The study demonstrated that SR-50A, SR-6DX, and SFSS have high corrosion resistance and mild or no cytotoxicity, due to the passive film enhanced by synergistic effect of Mo + N or by high addition effect of Cr + W. All super stainless steels showed very low cytotoxicity regardless of their nickel contents, although SR-3Mo was found to be relatively cytotoxic. From these studies, these steels are considered suitable for orthodontic applications.     

Enhanced human osteoblast cell adhesion and proliferation on 316 LS stainless steel by means of CO2 laser surface treatment

An effective and novel technique for improving the biocompatibility of a biograde 316 LS stainless steel through the application of CO(2) laser treatment to modify the surface properties of the material is described herein. Different surface properties, such as surface roughness, surface oxygen content, and surface energy for CO(2) laser-treated 316 LS stainless steel, untreated, and mechanically roughened samples were analyzed, and their effects on the wettability characteristics of the material were studied. It was found that modification of the wettability characteristics of the 316 LS stainless steel following CO(2) laser treatment was achieved. This improvement was identified as being mainly due to the change in the polar component of the surface energy. One-day cell adhesion tests showed that cells not only adhered and spread better, but also grew faster on the CO(2) laser-treated sample than on either the untreated or mechanically roughened sample. Further, compared with the untreated sample, MTT cell proliferation analysis revealed that the mechanically roughed surface resulted in a slight enhancement, and CO(2) laser treatment brought about a significant increase in cell proliferation. An increase in the wettability of the 316 LS stainless steel was observed to positively correlate with the cell proliferation.     

医用不锈钢表面生物活性纳米多层膜的制备及血液相容性

为改善人体内金属植入物的生物相容性,采用静电自组装的方法在医用316L不锈钢表面制备出聚乙烯亚胺(PEI)与透明质酸(HA)复合的生物活性纳米多层膜.小角X射线衍射结果显示多层膜呈现出层层规则排列,运用原子力显微镜对涂层的表面形貌以及粗糙度分析证明随着层数的增加表面形貌得到改善,由粗糙逐渐变得平滑.对涂层进行血小板黏附实验表明该纳米多层膜血小板黏附明显降低、血液相容性明显得到改善.     

Fibrinogen adsorption onto 316L stainless steel under polarized conditions

Adsorption of the plasma protein fibrinogen onto electrically polarized 316L stainless steel was observed and quantified using both in situ and ex situ atomic force microscopy (AFM) techniques. Significant differences in fibrinogen adsorption were observed across voltages. Ex situ studies showed significantly lower area coverage (theta) and height of adsorbed Fb on cathodically polarized surfaces when compared to anodically polarized surfaces. Conformational differences in the protein may explain the distinctions in Fb surface area coverage (theta) and height between the anodic and cathodic cases. In situ studies showed significantly slower kinetics of Fb adsorption onto surfaces below -100 mV (vs. Ag/AgCl) compared to surfaces polarized above -100 mV. Electrochemical current density data showed large charge transfer processes (approximately 1 x 10(-5) to 1 x 10(-4) A/cm(2)) taking place on the 316L SS surfaces at voltages below -100 mV (vs. Ag/AgCl). These relatively large current densities point to flux of ionic species away from the surface as a major source of the reduction in adsorption kinetics rather than just hydrophilic or electrostatic effects.     

Construction and hemocompatibility study of highly bioactive heparin-functionalized surface

A simple method is developed to construct anticoagulant surfaces via passive adsorption of heparin onto the protonated plasma-polymerized allylamine (PPAam) films from phosphate-buffered saline (PBS). These protonated PPAam surfaces are found to have high affinity to heparin. Importantly, the heparin-functionalized PPAam (Hep-PPAam) surfaces show good retention of heparin after long-term immersion in PBS. The Hep-PPAam surface prolongs the activated partial thromboplastin time for about 20 s as compared to 316L stainless steel even though the adsorption amount of heparin is only about 300 ng/cm(2) . This indicates that the heparin bound to the protonated PPAam surfaces in this way maintains a high bioactivity. Blood platelet adhesion and activation on this surface is remarkably reduced and adsorption and activation of fibrinogen is inhibited. Thus, Hep-PPAam surface modification leads to a significant improve of the hemocompatibility. ? 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 100A:3124-3133, 2012.     

新型医用不锈钢研究

现有医用植入不锈钢由于其优良的综合性能广泛应用于医疗领域，但其中含有的镍元素由于腐蚀溶出，除对人体产生过敏反应外，还存在致畸、致癌的危害性。本文研究了新型医用无镍不锈钢（BIOSSN4）的力学性能、耐蚀性能和血液相容性等。与传统使用的医用316L不锈钢相比，BIOSSN4具有更好的强韧性配合，优良的耐蚀性和血液相容性，这种优势将会为其提供广阔的应用前景。     

Adhesion enhancement of steel fibers to acrylic bone cement through a silane coupling agent

The use of a silane coupling agent (methacryloxypropyl-trichlorosilane) to improve the mechanical properties of steel fiber-reinforced acrylic bone cements was assessed. Changes to the tensile and fracture properties of bone cements reinforced with silane-coated or uncoated 316L stainless steel fibers of different aspect ratios were studied. Contact-angle measurements indicated that the coupling agent coats the metal surface through room temperature treatments in a short time (within 2 h). Push-out tests indicated that the interfacial shear strength of silane-coated 316L stainless steel rods is 141% higher than the uncoated rods. The elastic moduli, ultimate stresses, and fracture toughness of the silane-coated, steel fiber-reinforced bone cements are significantly higher than the bone cements reinforced with uncoated steel fibers. There were no differences in the tensile mechanical properties of the silane-coated or uncoated, steel fiber-reinforced cements after aging in a physiological saline solution, indicating that the bonding effectiveness is decreased by the intrusion of water at the metal-polymer interface. Because of possible biocompatibility issues with leaching of the silane coupling agent and no long-term mechanical benefit in simulated aging experiments, the use of these agents is not recommended for in vivo use.     

A study on grafting and characterization of HMDI-modified calcium hydrogenphosphate

It is known that the organic molecules can provide an effective means to manipulate the surface properties of the biodegradable ceramic. There are two ways to modify the surface of the biodegradable ceramic by organic molecules. The first one is through surface adsorption but organic molecules will easily be washed out in the physiological environment. The second approach is to graft organic molecules through covalent bond to the hydroxyl groups that are available on the surface of the ceramics. Isocyanate group has been reported as a coupling agent for hydroxyapatite and organic molecule. The studies showed that the isocyanate could react with hydroxyl groups of hydroxyapatite and form a covalent bond between isocyanate and hydroxyapatite. In the study, hexamethylene diisocyanate (HMDI) was used as coupling agent and calcium hydrogenphosphate (CaHPO4, CHP) was the candidate ceramic. CHP will react with HMDI at the temperature of 20 degrees C, 30 degrees C, 40 degrees C, 50 degrees C, 60 degrees C, and 70 degrees C for 4h. Dibutyltin dilaurate and hydroquinone were used as catalyst and inhibitor, respectively. The effect of reaction temperature on the grafted yield will be described. The linkage between CHP and HMDI will be characterized by DTA, TGA, FTIR, XRD, and 31P, 13C liquid state NMR. From the results, we successfully modified the surface of CHP with coupling agent of HMDI. The grafted yield of HMDI on CHP was increasing with the reaction temperature. The best temperature for CHP modified by HMDI is around 50 degrees C. The linkage between HMDI and the surface of CHP is a urethane linkage as CHP-O-CO-NH-(CH2)6-N=C=O. After further treatment, the terminal group of CHP treated with HMDI (MCHP) will be converted into a primary amine group as the formula of CHP-O-CO-NH-(CH2)6-NH2. If reaction temperature is 60 degrees C, long extension chain will occur with a urea linkage between the isocyanate groups as the formula of CHP-O-CO-NH-(CH2)6-(NH-CO-NH-(CH2)6)n-NH2. At reaction temperature higher than 60 degrees C, the HMDI will become prepolymerized forms in solution. The prepolymerized forms such as allophanate, biuret, uretidione and urea linkage will turn the solution into gel type mixture, which will lead to low grafted yield of HMDI on CHP. When MCHP prepared at the temperature 20 degrees C, there is no evidence of long extension but the grafted yield is the lowest only 0.9 wt% around.     

Preparation and characterization of sol-gel hydroxyapatite and its electrochemical evaluation for biomedical applications

Metallic materials having good mechanical properties, high corrosion resistance, and good compatibility with biological materials are used as orthopedic devices. Type 316L stainless steel is the most widely used material for implant fabrication in India for orthopedic applications owing to their good corrosion resistance, hot and cold workability, excellent mechanical properties, and availability at low cost. However, it faces corrosion-related problems in physiological environment and thus releases toxic metal ions into the tissues surrounding the implants. Hence, hydroxyapatite (HAP) coatings over the metal implant have been developed as an alternative method to improve the surface conditions of the base metal. In the present investigation, the development of a sol-gel-derived hydroxyapatite coating on 316L SS is being explored. The corrosion resistance behavior of the coating was assessed through electrochemical studies involving cyclic polarization experiments and impedance analysis in Ringer's solution. The results have indicated that the sol-gel-derived HAP coatings exhibited excellent resistance to localized attack on pristine 316L SS.     

等离子体预处理对316L不锈钢-EVAL界面结合强度的影响

研究丁Ar、N2及Ar＋N2等离子体预处理对316L不锈钢与其表面EVAL涂层之间结合强度的影响。采用全反射丌IR、AFM测试研究了等离子体预处理前后316L不锈钢表面物化状态的变化；利用X-光电子能谱研究了316L不锈钢与EVAL涂层之间的界面反应。研究结果表明：Ar、N2及Ar＋N2等离子体预处理后，316L不锈钢与EVAL涂层之间的结合强度分别高达0．92MPa、1．2MPa、1．5MPa（未经处理的试样为0．15MPa）。全反射FTIR和AFM分析结果表明Ar、N2等离子体均对基体表面有较好的清洗作用，Ar等离子体对基体表面具有较强的轰击、刻蚀作用。XPS测试表明EVAL在经Ar等离子体处理的基体表面仅以物理吸附状态存在，推测与N2等离子体处理的基体之间形成了metal-N-O形式的氢键结合。     

金属粉末注射成型制备316L不锈钢植入材料

研究金属粉末注射成型(MIM)法制备的316L不锈钢植入材料的生物相容性。以目前临床应用的钛合金植入材料为对照,将MIM法制备的植入材料浸提液与L929细胞混合培养后用流式细胞仪测定其DNA期的细胞百分比,将二种材料分别植入动物体内通过病理组织学观察的方法来评价其生物相容性。统计学分析两组数据无明显差异,结果表明,MIM法制备的316L不锈钢植入材料具有良好的生物相容性。     

The influences of electrical potential and surface finish on the fatigue life of surgical implant materials.

The influence of both imposed anodic potential in Ringer's solution and surface finish on the fatigue lives of annealed 316 type stainless steel and annealed pure titanium were measured and statistically compared to fatigue data run in air. The applied potentials in simulated extracellular fluid approximated conditions existing within the body while also producing the types of surface defects actually found on removed long time implants within the time interval of the accelerated R. R. Moor fatigue tests. Differentiating tests were run at single levels of applied cyclic stress well above the endurance limits. In Ringer's solution, the fatigue life of the 316 stainless steel decreased with increasing applied potential, and at +500 mV was significantly shorter than when run in air. At each condition, the 316 stainless steel was independent of initial surface finish. In contrast, the fatigue life of titanium improved rapidly with increasingly fine surface finishes. Furthermore, compared to air, the application of +500 mV in Ringer's solution improved the life of the rough surface finished material and markedly increased to number of cycles to failure for the electopolished specimens.     

Model surfaces engineered with nanoscale roughness and RGD tripeptides promote osteoblast activity

Cell adhesion to biomaterials is a prerequisite for tissue integration with the implant surface. Herein, we show that we can generate a model silica surface that contains a minimal-length arginine-glycine-aspartic acid (RGD) peptide that maintains its biological activity. In the first part of this study, attachment of MC3T3-E1 osteoblast-like cells was investigated on silicon oxide, amine terminated substrates [i.e., 3-aminopropyl triethoxysilane (APTS)], grafted RGD, and physisorbed RGD control. The APTS layer exhibited nanoscale roughness and presented amine functional groups for grafting a minimal RGD tripeptide devoid of any flanking groups or spacers. Contact angle measurements indicated that the hydrophobicity of the APTS surface was significantly lower than that of the surface with grafted RGD (RGD-APTS). Atomic force microscopy showed that surfaces covered with RGD-APTS were smoother (Ra = 0.71 nm) than those covered with APTS alone (Ra = 1.59 nm). Focusing mainly on cell morphology, experiments showed that the RGD-APTS hybrid provided an optimum surface for cell adhesion, spreading, and cytoskeletal organization. Discrete focal adhesion plaques were also observed consistent with successful cell signaling events. In a second set of experiments, smooth, monolayers of APTS (Ra = 0.1 nm) were used to prepare arginine-glycine-aspartic acid-serine (RGDS)-APTS and arginine-glycine-glutamic acid-serine (RGES)-APTS (control) substrates. Focusing mainly on cell function, integrin and gene expression were all enhanced for rate osteosarcoma cells on surfaces containing grafted RGDS. Both sets of studies demonstrated that grafted molecules of RGD(S) enhance both osteoblast-like cell adhesion and function.     

Interactions between stainless steel, shear stress, and monocytes

Angioplasty with stent placement is commonly used to treat coronary atherosclerosis. However, 20-40% of stainless steel stents restenose within 6 months via a prolonged inflammatory response mediated by monocytic infiltration and cytokine secretion. In the current study, we tested a hypothesis that blood flow and monocytes interact to alter stent corrosion. We assessed the effects of THP1 monocytes on the corrosion rate of 316L stainless steel (316LSS) under shear stress (0.5-50 dyn/cm(2)). In addition, THP1 cytokine secretion was determined using cytokine arrays and ELISA analyses. Data were compared using ANOVA and Tukey post hoc analysis (alpha = 0.05). Monocytes significantly lowered 316LSS corrosion rates without limiting current density. However, shear stress alone did not alter the corrosion rate of 316LSS. THP1 cells adhered to the 316LSS surface at all flow rates. Exposure to the 316LSS/corrosion test under high fluid flow rates increased (>twofold) the secretion of 7 of the 42 cytokines tested (angeogenin, GRO, I309, interleukin 8, interleukin 6, interleukin 1beta, and macrophage chemoattractant protein-1). Each of these cytokines play a role in wound healing, macrophage differentiation, and cell proliferation, all hallmarks of in-stent restenosis. Furthermore, only IL8 levels were significantly higher than any of the system controls during the 316LSS/corrosion test conditions. The IL8 levels from the 316LSS/corrosion tests were not significantly different from the +LPS control. Together, these data suggest that monocytic cells maybe activated by exposure to 316LSS stents and could contribute to in-stent restenosis and altered corrosion of the stent.