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Dextran grafting on PTFE surface for cardiovascular applications. 聚四氟乙烯表面葡聚糖接枝心血管应用。
Pub Date : 2014-01-01 DOI: 10.4161/biom.28805
Eléonore C Michel, Vanessa Montaño-Machado, Pascale Chevallier, Amélie Labbé-Barrère, Didier Letourneur, Diego Mantovani

The modification of biomaterial surfaces with biomolecules influences the biological response. In this work, caboxymethyldextrans (CMD) with different degrees of substitution have been grafted to surfaces by introduction of amino moieties directly onto the substrate surface. Polytetrafluoroethylene was selected as a model substrate for biomaterial as it is already largely used for cardiovascular clinical applications. Firstly, CMD polymers were characterized by FTIR, (1)H-NMR, and conductimetric titration. Then, the coatings have been analyzed by XPS to confirm the grafting and determine the composition. Once characterized, biological performances of CMD coatings were investigated. The hemocompatibility was ascertained using the free hemoglobin method. The effects on endothelial and smooth muscle cell adhesion were also studied. Results indicated that CMD at a 0.2 substitution degree, significantly influenced the biological property of PTFE by exhibiting non-thrombogenic properties as well as enhancing endothelial cell adhesion along with limiting smooth muscle cell adhesion. This work suggested the creation of versatile pro-active biomaterials suitable for different biomedical applications.

生物分子修饰生物材料表面影响生物反应。在这项工作中,不同取代度的羧甲基右旋糖酐(CMD)通过直接在底物表面引入氨基片段而接枝到表面。选择聚四氟乙烯作为生物材料的模型底物,因为它已经广泛用于心血管临床应用。首先,用FTIR、(1)H-NMR和电导滴定对CMD聚合物进行了表征。然后,用XPS对涂层进行了分析,确定了接枝反应和组成。在对其进行表征后,对其生物性能进行了研究。用游离血红蛋白法测定血液相容性。并对其对内皮细胞和平滑肌细胞粘附的影响进行了研究。结果表明,0.2取代度的CMD显著影响聚四氟乙烯的生物学性能,表现出非血栓性,增强内皮细胞粘附,限制平滑肌细胞粘附。这项工作建议创造适合不同生物医学应用的多功能活性生物材料。
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引用次数: 16
Interfacing polymeric scaffolds with primary pancreatic ductal adenocarcinoma cells to develop 3D cancer models. 将聚合物支架与原发性胰腺导管腺癌细胞结合,建立三维肿瘤模型。
Pub Date : 2014-01-01 DOI: 10.4161/21592527.2014.955386
Claudio Ricci, Carlos Mota, Stefania Moscato, Delfo D'Alessandro, Stefano Ugel, Silvia Sartoris, Vincenzo Bronte, Ugo Boggi, Daniela Campani, Niccola Funel, Lorenzo Moroni, Serena Danti

We analyzed the interactions between human primary cells from pancreatic ductal adenocarcinoma (PDAC) and polymeric scaffolds to develop 3D cancer models useful for mimicking the biology of this tumor. Three scaffold types based on two biocompatible polymeric formulations, such as poly(vinyl alcohol)/gelatin (PVA/G) mixture and poly(ethylene oxide terephthalate)/poly(butylene terephthalate) (PEOT/PBT) copolymer, were obtained via different techniques, namely, emulsion and freeze-drying, compression molding followed by salt leaching, and electrospinning. In this way, primary PDAC cells interfaced with different pore topographies, such as sponge-like pores of different shape and size or nanofiber interspaces. The aim of this study was to investigate the influence played by the scaffold architecture over cancerous cell growth and function. In all scaffolds, primary PDAC cells showed good viability and synthesized tumor-specific metalloproteinases (MMPs) such as MMP-2, and MMP-9. However, only sponge-like pores, obtained via emulsion-based and salt leaching-based techniques allowed for an organized cellular aggregation very similar to the native PDAC morphological structure. Differently, these cell clusters were not observed on PEOT/PBT electrospun scaffolds. MMP-2 and MMP-9, as active enzymes, resulted to be increased in PVA/G and PEOT/PBT sponges, respectively. These findings suggested that spongy scaffolds supported the generation of pancreatic tumor models with enhanced aggressiveness. In conclusion, primary PDAC cells showed diverse behaviors while interacting with different scaffold types that can be potentially exploited to create stage-specific pancreatic cancer models likely to provide new knowledge on the modulation and drug susceptibility of MMPs.

我们分析了来自胰腺导管腺癌(PDAC)的人类原代细胞与聚合物支架之间的相互作用,以开发可用于模拟该肿瘤生物学的3D癌症模型。基于两种生物相容性聚合物配方,如聚乙烯醇/明胶(PVA/G)混合物和聚(环氧对苯二甲酸乙酯)/聚(对苯二甲酸丁二酯)(PEOT/PBT)共聚物,通过不同的技术,即乳化和冷冻干燥、压缩成型后盐浸和静电纺丝,获得了三种支架类型。通过这种方式,原代PDAC细胞与不同的孔隙形貌相连接,如不同形状和大小的海绵状孔隙或纳米纤维间隙。本研究的目的是研究支架结构对癌细胞生长和功能的影响。在所有支架中,原代PDAC细胞表现出良好的活力,并合成肿瘤特异性金属蛋白酶(MMPs),如MMP-2和MMP-9。然而,只有通过基于乳化和基于盐浸的技术获得的海绵状孔隙才允许有组织的细胞聚集,这与天然PDAC的形态结构非常相似。不同的是,在PEOT/PBT电纺丝支架上没有观察到这些细胞团。MMP-2和MMP-9作为活性酶,分别导致PVA/G和PEOT/PBT海绵的升高。这些发现表明海绵支架支持胰腺肿瘤模型的生成,具有增强的侵袭性。总之,原代PDAC细胞在与不同支架类型相互作用时表现出不同的行为,这可能被用于创建特定阶段的胰腺癌模型,可能为MMPs的调节和药物敏感性提供新的知识。
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引用次数: 46
Magnetic force microscopy: quantitative issues in biomaterials. 磁力显微镜:生物材料的定量问题。
Pub Date : 2014-01-01 Epub Date: 2014-07-22 DOI: 10.4161/biom.29507
Daniele Passeri, Chunhua Dong, Melania Reggente, Livia Angeloni, Mario Barteri, Francesca A Scaramuzzo, Francesca De Angelis, Fiorenzo Marinelli, Flavia Antonelli, Federica Rinaldi, Carlotta Marianecci, Maria Carafa, Angela Sorbo, Daniela Sordi, Isabel Wce Arends, Marco Rossi

Magnetic force microscopy (MFM) is an atomic force microscopy (AFM) based technique in which an AFM tip with a magnetic coating is used to probe local magnetic fields with the typical AFM spatial resolution, thus allowing one to acquire images reflecting the local magnetic properties of the samples at the nanoscale. Being a well established tool for the characterization of magnetic recording media, superconductors and magnetic nanomaterials, MFM is finding constantly increasing application in the study of magnetic properties of materials and systems of biological and biomedical interest. After reviewing these latter applications, three case studies are presented in which MFM is used to characterize: (i) magnetoferritin synthesized using apoferritin as molecular reactor; (ii) magnetic nanoparticles loaded niosomes to be used as nanocarriers for drug delivery; (iii) leukemic cells labeled using folic acid-coated core-shell superparamagnetic nanoparticles in order to exploit the presence of folate receptors on the cell membrane surface. In these examples, MFM data are quantitatively analyzed evidencing the limits of the simple analytical models currently used. Provided that suitable models are used to simulate the MFM response, MFM can be used to evaluate the magnetic momentum of the core of magnetoferritin, the iron entrapment efficiency in single vesicles, or the uptake of magnetic nanoparticles into cells.

磁力显微镜(MFM)是一种基于原子力显微镜(AFM)的技术,该技术使用带有磁性涂层的原子力显微镜尖端,以典型的原子力显微镜空间分辨率探测局部磁场,从而获得反映样品在纳米尺度上的局部磁性的图像。作为磁性记录介质、超导体和磁性纳米材料表征的成熟工具,MFM在生物和生物医学领域的材料和系统的磁性研究中得到了越来越多的应用。在回顾了后一种应用之后,提出了三个案例研究,其中MFM用于表征:(i)利用载铁蛋白作为分子反应器合成磁铁蛋白;(ii)装载纳米粒的磁性纳米颗粒,用作药物递送的纳米载体;(iii)利用叶酸包被的核壳超顺磁性纳米颗粒标记白血病细胞,以利用细胞膜表面叶酸受体的存在。在这些例子中,对MFM数据进行了定量分析,证明了目前使用的简单分析模型的局限性。如果使用合适的模型来模拟MFM响应,则MFM可以用于评估磁性铁蛋白核心的磁动量,单个囊泡中的铁捕获效率或磁性纳米颗粒进入细胞的摄取。
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引用次数: 46
Nanocarbon surfaces for biomedicine. 用于生物医学的纳米碳表面。
Pub Date : 2014-01-01 Epub Date: 2014-03-19 DOI: 10.4161/biom.28537
Giacomo Reina, Emanuela Tamburri, Silvia Orlanducci, Stefano Gay, Roberto Matassa, Valeria Guglielmotti, Teresa Lavecchia, Maria Letizia Terranova, Marco Rossi

The distinctive physicochemical, mechanical and electrical properties of carbon nanostructures are currently gaining the interest of researchers working in bioengineering and biomedical fields. Carbon nanotubes, carbon dendrimers, graphenic platelets and nanodiamonds are deeply studied aiming at their application in several areas of biology and medicine.   Here we provide a summary of the carbon nanomaterials prepared in our labs and of the fabrication techniques used to produce several biomedical utilities, from scaffolds for tissue growth to cargos for drug delivery and to biosensors.

碳纳米结构独特的物理化学、机械和电学特性,目前正引起生物工程和生物医学领域研究人员的兴趣。碳纳米管、碳树状大分子、石墨血小板和纳米金刚石正在深入研究,旨在将其应用于生物学和医学的几个领域。在这里,我们总结了在我们的实验室中制备的碳纳米材料,以及用于生产几种生物医学用途的制造技术,从组织生长的支架到药物输送的货物和生物传感器。
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引用次数: 20
Influence of ECAP process on mechanical and corrosion properties of pure Mg and ZK60 magnesium alloy for biodegradable stent applications. ECAP工艺对纯Mg和ZK60镁合金生物可降解支架机械性能和腐蚀性能的影响
Pub Date : 2014-01-01 DOI: 10.4161/biom.28283
Ehsan Mostaed, Maurizio Vedani, Mazdak Hashempour, Massimiliano Bestetti

Equal channel angular pressing (ECAP) was performed on ZK60 alloy and pure Mg in the temperature range 150-250 °C. A significant grain refinement was detected after ECAP, leading to an ultrafine grain size (UFG) and enhanced formability during extrusion process. Comparing to conventional coarse grained samples, fracture elongation of pure Mg and ZK60 alloy were significantly improved by 130% and 100%, respectively, while the tensile strength remained at high level. Extrusion was performed on ECAP processed billets to produce small tubes (with outer/inner diameter of 4/2.5 mm) as precursors for biodegradable stents. Studies on extruded tubes revealed that even after extrusion the microstructure and microhardness of the UFG ZK60 alloy were almost stable. Furthermore, pure Mg tubes showed an additional improvement in terms of grain refining and mechanical properties after extrusion. Electrochemical analyses and microstructural assessments after corrosion tests demonstrated two major influential factors in corrosion behavior of the investigated materials. The presence of Zn and Zr as alloying elements simultaneously increases the nobility by formation of a protective film and increase the local corrosion damage by amplifying the pitting development. ECAP treatment decreases the size of the second phase particles thus improving microstructure homogeneity, thereby decreasing the localized corrosion effects.

在150 ~ 250℃的温度范围内,对ZK60合金和纯Mg进行了等通道角挤压(ECAP)。在ECAP后,发现了明显的晶粒细化,导致超细晶粒尺寸(UFG)和挤压过程中的成形性增强。与常规粗晶试样相比,纯Mg和ZK60合金的断裂伸长率分别提高了130%和100%,而抗拉强度仍保持在较高水平。在ECAP加工的坯料上进行挤压,生产出小管(外径/内径为4/2.5 mm)作为可生物降解支架的前体。对挤压管的研究表明,即使挤压后,UFG ZK60合金的显微组织和显微硬度也基本稳定。此外,纯Mg管在挤压后晶粒细化和力学性能方面都有进一步的改善。腐蚀试验后的电化学分析和显微组织评价表明了影响材料腐蚀行为的两个主要因素。Zn和Zr作为合金元素的存在通过形成保护膜增加了合金的高贵性,同时通过扩大点蚀的发展增加了合金的局部腐蚀损伤。ECAP处理减小了第二相颗粒的尺寸,从而改善了微观组织的均匀性,从而降低了局部腐蚀效应。
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引用次数: 83
Properties of titanium-alloyed DLC layers for medical applications. 医用钛合金DLC层的性能。
Pub Date : 2014-01-01 Epub Date: 2014-08-05 DOI: 10.4161/biom.29505
Ludek Joska, Jaroslav Fojt, Ladislav Cvrcek, Vitezslav Brezina

DLC-type layers offer a good potential for application in medicine, due to their excellent tribological properties, chemical resistance, and bio-inert character. The presented study has verified the possibility of alloying DLC layers with titanium, with coatings containing three levels of titanium concentration prepared. Titanium was present on the surface mainly in the form of oxides. Its increasing concentration led to increased presence of titanium carbide as well. The behavior of the studied systems was stable during exposure in a physiological saline solution. Electrochemical impedance spectra practically did not change with time. Alloying, however, changed the electrochemical behavior of coated systems in a significant way: from inert surface mediating only exchange reactions of the environment in the case of unalloyed DLC layers to a response corresponding rather to a passive surface in the case of alloyed specimens. The effect of DLC layers alloying with titanium was tested by the interaction with a simulated body fluid, during which precipitation of a compound containing calcium and phosphorus--basic components of the bone apatite--occurred on all doped specimens, in contrast to pure DLC. The results of the specimens' surface colonization with cells test proved the positive effect of titanium in the case of specimens with a medium and highest content of this element.

由于其优异的摩擦学性能、耐化学性和生物惰性特性,dlc型层在医学上具有良好的应用潜力。本研究验证了钛合金DLC层的可能性,制备了含三种钛浓度的涂层。钛主要以氧化物的形式存在于表面。其浓度的增加也导致碳化钛的存在增加。在生理盐水溶液中,所研究的系统的行为是稳定的。电化学阻抗谱几乎不随时间变化。然而,合金化在很大程度上改变了涂层系统的电化学行为:在非合金化DLC层的情况下,惰性表面只介导环境的交换反应,而在合金样品的情况下,响应相对于被动表面。DLC层与钛合金化的影响是通过与模拟体液的相互作用来测试的,在此过程中,与纯DLC相比,所有掺杂样品上都发生了含有钙和磷的化合物(骨磷灰石的基本成分)的沉淀。标本表面细胞定植试验结果证明,在钛元素含量中等和最高的标本中,钛具有积极作用。
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引用次数: 15
The effects of a SiO2 coating on the corrosion parameters cpTi and Ti-6Al-7Nb alloy. 研究了SiO2涂层对cpTi和Ti-6Al-7Nb合金腐蚀参数的影响。
Pub Date : 2014-01-01 DOI: 10.4161/biom.28535
Marcin Basiaga, Witold Walke, Zbigniew Paszenda, Paweł Karasiński, Janusz Szewczenko

The aim of this paper was to evaluate the usefulness of the sol-gel method application, to modificate the surface of the Ti6Al7Nb alloy and the cpTi titanium (Grade 4) with SiO2 oxide, applied on the vascular implants to improve their hemocompatibility. Mechanical treatment was followed by film deposition on surface of the titanium samples. An appropriate selection of the process parameters was verified in the studies of corrosion, using potentiodynamic and impedance method. A test was conducted in the solution simulating blood vessels environment, in simulated body fluid at t = 37.0 ± 1 °C and pH = 7.0 ± 0.2. Results showed varied electrochemical properties of the SiO2 film, depending on its deposition parameters. Correlations between corrosion resistance and layer adhesion to the substrate were observed, depending on annealing temperature.

本文的目的是评估溶胶-凝胶法应用的有效性,用二氧化硅氧化物修饰Ti6Al7Nb合金和cpTi钛(4级)表面,应用于血管植入物,以改善其血液相容性。机械处理后,在钛样品表面沉积薄膜。在腐蚀研究中,采用电位动力学和阻抗法验证了工艺参数的合理选择。在模拟血管环境的溶液中,在t = 37.0±1℃,pH = 7.0±0.2的模拟体液中进行试验。结果表明,SiO2薄膜的电化学性能随其沉积参数的不同而变化。根据退火温度的不同,观察到耐腐蚀性和层与衬底的附着力之间的相关性。
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引用次数: 12
Alternative technique for calcium phosphate coating on titanium alloy implants. 钛合金植入物上磷酸钙涂层的替代技术。
Pub Date : 2014-01-01 Epub Date: 2014-03-19 DOI: 10.4161/biom.28534
Van Quang Le, Geneviève Pourroy, Andrea Cochis, Lia Rimondini, Wafa I Abdel-Fattah, Hadeer I Mohammed, Adele Carradò

As an alternative technique for calcium phosphate coating on titanium alloys, we propose to functionalize the metal surface with anionic bath containing chlorides of palladium or silver as activators. This new deposition route has several advantages such as controlled conditions, applicability to complex shapes, no adverse effect of heating, and cost effectiveness. A mixture of hydroxyapatite and calcium phosphate hydrate is deposited on the surface of Ti-6Al-4V. Calcium phosphate coating is built faster compared with the one by Simulated Body Fluid. Cell morphology and density are comparable to the control one; and the results prove no toxic compound is released into the medium during the previous seven days of immersion. Moreover, the cell viability is comparable with cells cultivated with the virgin medium. These experimental treatments allowed producing cytocompatible materials potentially applicable to manufacture implantable devices for orthopedic and oral surgeries.

作为在钛合金表面涂覆磷酸钙的替代技术,我们提出用含钯或银的氯化物作为活化剂的阴离子浴对金属表面进行功能化。这种新的沉积路线有几个优点,如可控的条件,适用于复杂的形状,没有不利的影响加热,和成本效益。羟基磷灰石和磷酸钙水合物的混合物沉积在Ti-6Al-4V表面。与模拟体液法相比,磷酸钙涂层的构建速度更快。细胞形态和密度与对照相当;结果证明,在前七天的浸泡过程中,没有有毒化合物释放到介质中。此外,细胞活力与用原始培养基培养的细胞相当。这些实验性治疗方法允许生产细胞相容性材料,可能适用于制造骨科和口腔手术的植入式装置。
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引用次数: 17
Poly(vinyl alcohol) hydrogel coatings with tunable surface exposure of hydroxyapatite. 羟基磷灰石表面暴露可调的聚乙烯醇水凝胶涂层。
Pub Date : 2014-01-01 DOI: 10.4161/biom.28764
David Moreau, Arthur Villain, David N Ku, Laurent Corté

Insufficient bone anchoring is a major limitation of artificial substitutes for connective osteoarticular tissues. The use of coatings containing osseoconductive ceramic particles is one of the actively explored strategies to improve osseointegration and strengthen the bone-implant interface for general tissue engineering. Our hypothesis is that hydroxyapatite (HA) particles can be coated robustly on specific assemblies of PVA hydrogel fibers for the potential anchoring of ligament replacements. A simple dip-coating method is described to produce composite coatings made of microscopic hydroxyapatite (HA) particles dispersed in a poly(vinyl alcohol) (PVA) matrix. The materials are compatible with the requirements for implant Good Manufacturing Practices. They are applied to coat bundles of PVA hydrogel fibers used for the development of ligament implants. By means of optical and electronic microscopy, we show that the coating thickness and surface state can be adjusted by varying the composition of the dipping solution. Quantitative analysis based on backscattered electron microscopy show that the exposure of HA at the coating surface can be tuned from 0 to over 55% by decreasing the weight ratio of PVA over HA from 0.4 to 0.1. Abrasion experiments simulating bone-implant contact illustrate how the coating cohesion and wear resistance increase by increasing the content of PVA relative to HA. Using pullout experiments, we find that these coatings adhere well to the fiber bundles and detach by propagation of a crack inside the coating. These results provide a guide to select coated implants for anchoring artificial ligaments.

骨锚定不足是结缔性骨关节组织人工替代品的主要限制。使用含骨导电陶瓷颗粒的涂层是通用组织工程中积极探索的改善骨整合和增强骨-种植体界面的策略之一。我们的假设是羟基磷灰石(HA)颗粒可以被牢固地包裹在PVA水凝胶纤维的特定组件上,用于韧带置换的潜在锚定。本文描述了一种简单的浸涂方法,用于制备分散在聚乙烯醇(PVA)基体中的微观羟基磷灰石(HA)颗粒复合涂层。材料符合植入物良好生产规范的要求。它们被应用于包裹用于韧带植入物发育的PVA水凝胶纤维束。通过光学显微镜和电子显微镜,我们发现涂层厚度和表面状态可以通过改变浸渍液的组成来调节。基于后向散射电子显微镜的定量分析表明,通过将PVA与HA的重量比从0.4降低到0.1,可以将HA在涂层表面的暴露量从0调整到55%以上。模拟骨与种植体接触的磨损实验表明,相对于HA,增加PVA的含量可以提高涂层的粘聚性和耐磨性。通过拉拔实验,我们发现这些涂层可以很好地粘附在纤维束上,并通过涂层内部裂纹的扩展而分离。这些结果为人工韧带锚定涂层植入物的选择提供了指导。
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引用次数: 14
Design of 2D chitosan scaffolds via electrochemical structuring. 基于电化学结构的二维壳聚糖支架设计。
Pub Date : 2014-01-01 Epub Date: 2014-06-18 DOI: 10.4161/biom.29506
Lina Altomare, Elena Guglielmo, Elena Maria Varoni, Serena Bertoldi, Andrea Cochis, Lia Rimondini, Luigi De Nardo

Chitosan (CS) is a versatile biopolymer whose morphological and chemico-physical properties can be designed for a variety of biomedical applications. Taking advantage of its electrolytic nature, cathodic polarization allows CS deposition on electrically conductive substrates, resulting in thin porous structures with tunable morphology. Here we propose an easy method to obtain CS membranes with highly oriented micro-channels for tissue engineering applications, relying on simple control of process parameters and cathodic substrate geometry. Cathodic deposition was performed on two different aluminum grids in galvanostatic conditions at 6.25 mA cm(-2) from CS solution [1g L(-1)] in acetic acid (pH 3.5). Self-standing thin scaffolds were cross linked either with genipin or epichlorohydrin, weighted, and observed by optical and electron microscopy. Swelling properties at pH 5 and pH 7.4 have been also investigated and tensile tests performed on swollen samples at room temperature. Finally, direct and indirect assays have been performed to evaluate the cytotoxicity at 24 and 72 h. Thin scaffolds with two different oriented porosities (1000 µm and 500 µm) have been successfully fabricated by electrochemical techniques. Both cross-linking agents did not affected the mechanical properties and cytocompatibility of the resulting structures. Depending on the pH, these structures show interesting swelling properties that can be exploited for drug delivery systems. Moreover, thanks to the possibility of controlling the porosity and the micro-channel orientation, they should be used for the regeneration of tissues requiring a preferential cells orientation, e.g., cardiac patches or ligament regeneration.

壳聚糖(CS)是一种多用途的生物聚合物,其形态和化学物理性质可用于多种生物医学用途。利用其电解性质,阴极极化允许CS沉积在导电衬底上,从而产生具有可调谐形貌的薄多孔结构。在这里,我们提出了一种简单的方法来获得具有高度定向微通道的CS膜,用于组织工程应用,依赖于简单的工艺参数控制和阴极衬底几何形状。在恒电流条件下,从CS溶液[1g L(-1)]到乙酸(pH为3.5)中,在6.25 mA cm(-2)下对两个不同的铝栅格进行阴极沉积。独立的薄支架与吉尼平或环氧氯丙烷交联,称重,并通过光学和电子显微镜观察。还研究了pH 5和pH 7.4下的膨胀特性,并在室温下对膨胀样品进行了拉伸试验。最后,进行了直接和间接测定,以评估24和72 h时的细胞毒性。通过电化学技术成功制备了两种不同取向孔隙度(1000µm和500µm)的薄支架。两种交联剂均不影响所得结构的力学性能和细胞相容性。根据pH值的不同,这些结构显示出有趣的膨胀特性,可以用于药物输送系统。此外,由于可以控制孔隙度和微通道取向,它们可以用于需要优先细胞取向的组织的再生,例如心脏补片或韧带再生。
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引用次数: 11
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