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1.
Layer-by-layer deposition of polyelectrolyte multilayer (PEM) thin films has recently been applied to biomaterial applications. This simple and versatile technique provides a wide variety of potential utilization by insertion of biomolecules such as cell adhesion peptides. In this work dual peptides containing RGD (a cell-binding domain) and LHRRVKI (a heparin-binding domain) were immobilized onto polystyrene by the PEM technique and the effects on osteoblast cell culture were investigated. These peptides were conjugated to the amino groups of poly(allylamine hydrochloride) and then adsorbed onto the top of a 10 layer poly(allylamine hydrochloride)/poly(acrylic acid) film assembled at either pH 2.0 or pH 6.5. Osteoblasts, isolated from neonatal rat calvariae, were then seeded and cultured on the peptide-conjugated surfaces. We found that the cells adhered and grew better on the RGD-conjugated PEM films. The osteoblasts exhibited a better differentiated phenotype on the pH 2.0 films than the pH 6.5 films with respect to calcium deposition. The incorporation of LHRRVKI did not support cell adhesion, growth and matrix mineral deposition. Our results showed that the efficacy of RGD conjugation on osteoblast behavior was affected by the base PEM film.  相似文献   

2.
This paper reports a precise, live cell-patterning method by means of patterning a silicon or glass substrate with alternating cytophilic and cytophobic self-assembled monolayers (SAMs) deposited via molecular vapor deposition. Specifically, a stack of hydrophobic heptadecafluoro-1,1,2,2-tetrahydrodecyltrichlorosilane SAMs and a silicon oxide adhesion layer were patterned on the substrate surface, and a hydrophilic SAM derived from 3-trimethoxysilyl propyldiethylenetriamine was coated on the remaining non-treated areas on the substrate surface to promote cell growth. The primary characteristics of the reported method include: (i) single-cell resolution; (ii) easy alignment of the patterns with the pre-existing patterns on the substrate; (iii) easy formation of nanoscale patterns (depending on the exposure equipment); (iv) long shelf life of the substrate pattern prior to cell culturing; (v) compatibility with conventional, inverted, optical microscopes for simple visualization of patterns formed on a glass wafer; and (vi) the ability to support patterned cell (osteoblast) networks for at least 2 weeks. Here, we describe the deposition technique and the characterization of the deposited layers, as well as the application of this method in the fabrication of multielectrode arrays supporting patterned neuronal networks.  相似文献   

3.
Many engineers and surgeons trace implant failure to poor osseointegration (or the bonding of an orthopedic implant to juxtaposed bone) and/or bacteria infection. By using novel nanotopographies, researchers have shown that nanostructured ceramics, carbon fibers, polymers, metals, and composites enhance osteoblast adhesion and calcium/phosphate mineral deposition. However, the function of bacteria on materials with nanostructured surfaces remains largely uninvestigated. This is despite the fact that during normal surgical insertion of an orthopedic implant, bacteria from the patient's own skin and/or mucosa enters the wound site. These bacteria (namely, Staphylococcus epidermidis) irreversibly adhere to an implant surface while various physiological stresses induce alterations in the bacterial growth rate leading to biofilm formation. Because of their integral role in determining the success of orthopedic implants, the objective of this in vitro study was to examine the functions of (i) S. epidermidis and (ii) osteoblasts (or bone-forming cells) on ZnO and titania (TiO(2)), which possess nanostructured compared to microstructured surface features. ZnO is a well-known antimicrobial agent and TiO(2) readily forms on titanium once implanted. Results of this study provided the first evidence of decreased S. epidermidis adhesion on ZnO and TiO(2) with nanostructured when compared with microstructured surface features. Moreover, compared with microphase formulations, results of this study showed increased osteoblast adhesion, alkaline phosphatase activity, and calcium mineral deposition on nanophase ZnO and TiO(2). In this manner, this study suggests that nanophase ZnO and TiO(2) may reduce S. epidermidis adhesion and increase osteoblast functions necessary to promote the efficacy of orthopedic implants.  相似文献   

4.
Shim J  Hagerman E  Wu B  Gupta V 《Acta biomaterialia》2008,4(6):1657-1668
A previously developed laser spallation technique to determine the tensile strength of thin film interfaces was successfully adopted to determine the tensile strength of interfaces between three different live mammalian cells (osteoblast, chondrocyte and fibroblast) and polystyrene (untreated and fibronectin coated) and titanium surfaces. No noticeable differences in the interfacial tensile strength values were found across the three cell types on the same substrate although osteoblasts showed slightly lower adhesion strength when cultured on untreated polystyrene surfaces. Significant differences were, however, measured for cells treated on different surfaces. Use of fibronectin increased the interfacial tensile strength for all cell types, and cells bonded much better to titanium than to untreated polystyrene surfaces. Cell interfacial strength was higher when cultured with serum than in a serum-free environment. The results demonstrate the remarkable sensitivity of the laser spallation experiment in determining the effects of local interfacial microstructure and chemistry on cell adhesion.  相似文献   

5.
Ti being bioinert shows poor bone cell adhesion with an intervening fibrous capsule. Ti could be made bioactive by several methods including growing in situ TiO2 layer on Ti-surface. TiO2 nanotubes were grown on Ti surface via anodization process and the bone cell-material interactions were evaluated. Human osteoblast cell attachment and growth behavior were studied using an osteoprecursor cell line for 3, 7, and 11 days. An abundant amount of extracellular matrix (ECM) between the neighboring cells was noticed on anodized nanotube surface with filopodia extensions coming out from cells to grasp the nanoporous surface of the nanotube for anchorage. To better understand and compare cell-materials interactions, anodized nanoporous sample surfaces were etched with different patterns. Preferential cell attachment was noticed on nanotube surface compare to almost no cells in etched Ti surface. Cell adhesion with vinculin adhesive protein showed higher intensity, positive contacts on nanoporous surface and thin focal contacts on the Ti-control. Immunochemistry study with alkaline phosphatase showed enhanced osteoblastic phenotype expressions in nanoporous surface. Osteoblast proliferation was significantly higher on anodized nanotube surface. Surface properties changed with the emergence of nanoscale morphology. Higher nanometer scale roughness, low contact angle and high surface energy in nanoporous surface enhanced the osteoblast-material interactions. Mineralization study was done under simulated body fluid (SBF) with ion concentration nearly equal to human blood plasma to understand biomimetic apatite deposition behavior. Although apatite layer formation was noticed on nanotube surface, but it was nonuniform even after 21 days in SBF.  相似文献   

6.
目的 通过对涂层的表面结构及其细胞相容性的研究,对新型的钛基溶胶凝胶HA涂层技术进行评价.方法 在纯钛材料表面制备新型的溶胶凝胶HA涂层,采用SEM对涂层表面特征进行测试.体外成骨细胞(MC-3T3)培养测试涂层的细胞相容性,并将钛基HA溶胶凝胶涂层的细胞相容性与传统的钛基等离子喷涂HA涂层做比较.结果 经过水热处理的钛基涂层表面为均一的晶体颗粒表面,细胞学实验发现与等离子HA涂层相比较,水热处理溶胶凝胶表面增强了细胞粘附作用,具有较好的成骨细胞粘附(P<0.05).结论 本实验结果提示这种新型的水热处理钛基HA溶胶凝胶表面具有良好的成骨细胞粘附特性,因而有待于作进一步研究.  相似文献   

7.
Extracellular matrix (ECM)-like coating was developed on biodegradable biomaterials based on the electrostatic self-assembly (ESA) technique to promote osteoblast growth. Poly(ethylenimine) (PEI) was first employed to obtain a stable positively charged surface on poly (DL-lactide) (PDL-LA) substrate. Gelatin was selected as ECM-like biomacromolecule to deposit on the activated PDL-LA substrate using the ESA technique. zeta-Potential results showed alternating charge of polyelectrolytes (PEI/gelatin) layering on PDL-LA microspheres. Quartz crystal microbalance (QCM) measurement further verified the gradual deposition of PEI/gelatin on PDL-LA thin film. Osteoblast cells (MC3T3) were chosen to test the cell behavior on modified PDL-LA substrates. The osteoblast test about cell activity, intracellular total DNA content, total protein content and cell morphology by SEM investigation on ECM-like multilayer-modified PDL-LA substrate showed to promote osteoblast growth. Comparing conventional coating methods, polyelectrolyte multilayers are easy and stable to prepare. It may be a good choice for the surface modification of complex biomedical devices. These very flexible systems allow broad medical applications for drug delivery and tissue engineering.  相似文献   

8.
热解碳人工心脏瓣膜材料表面改性研究   总被引:2,自引:0,他引:2  
对氮离子注入热解碳及在热解碳表面离子束增强沉积钛氧化物薄膜进行了研究,采用X光电子能谱仪,俄歇电子能谱仪测定了表面改性层的成价和价态,用四探针方法测定了表面电阻率,用接触角方法测定了材料表面能,用动态凝血时间及血小板粘附评价了表面改性层的血液相容性。研究表明,热解碳经表面N离子注入后有碳氧化合物形成,其血液相容性有所改善,而采用离子束增强沉积合成TiO2-x薄膜则使热解碳备注相 容性获得了显著改善  相似文献   

9.
Surface engineering approaches that alter the topological chemistry of a substrate could be used as an effective tool for directing cell interactions and their subsequent function. It is well known that the physical environment of nanotopography has positive effects on cell behavior, yet direct comparisons of nanotopographic surface chemistry have not been fully explored. Here we compare TiO(2) nanotubes with carbon-coated TiO(2) nanotubes, probing osteogenic cell behavior, including osteoblast (bone cells) and mesenchymal stem cell (MSC) (osteo-progenitor cells) interactions with the different surface chemistries (TiO(2) vs. carbon). The roles played by the material surface chemistry of the nanotubes did not have an effect on the adhesion, growth or morphology, but had a major influence on the alkaline phosphatase (ALP) activity of osteoblast cells, with the original TiO(2) chemistry having higher ALP levels. In addition, the different chemistries caused different levels of osteogenic differentiation in MSCs; however, it was the carbon-coated TiO(2) nanotubes that had the greater advantage, with higher levels of osteo-differentiation. It was observed in this study that: (a) chemistry plays a role in cell functionality, such as ALP activity and osteogenic protein gene expression (PCR); (b) different cell types may have different chemical preferences for optimal function. The ability to optimize cell behavior using surface chemistry factors has a profound effect on both orthopedic and tissue engineering in general. This study aims to highlight the importance of the chemistry of the carrier material in osteogenic tissue engineering schemes.  相似文献   

10.
The corrosion behavior and cell adhesion property of nanostructured TiO2 films deposited electrolytically on Ti6Al4V were examined in the present in vitro study. The nanostructured TiO2 film deposition on Ti6Al4V was achieved via peroxoprecursors. SEM micrographs exhibit the formation of amorphous and crystallite TiO2 nanoparticles on Ti6Al4V before and after being annealed at 500 degrees C. Corrosion behavior of TiO2-deposited and uncoated Ti6Al4V was evaluated in freely aerated Hank's solution at 37 degrees C by the measurement and analysis of open-circuit potential variation with time, Tafel plots, and electrochemical impedance spectroscopy. The electrochemical results indicated that nano-TiO2 coated Ti6Al4V showed a better corrosion resistance in simulated biofluid than uncoated Ti6Al4V. Rat bone cells and human aortic smooth muscle cells were grown on these substrates to study the cellular responses in vitro. The SEM images revealed enhanced cell adhesion, cell spreading, and proliferation on nano-TiO2 coated Ti6Al4V compared to those grown on uncoated substrates for both cell lines. These results suggested that nanotopography produced by deposition of nanostructured TiO2 onto Ti alloy surfaces might enhance corrosion resistance, biocompatibility, and cell integration for implants made of Ti alloys.  相似文献   

11.
Extracellular matrix (ECM)-like coating was developed on biodegradable biomaterials based on the electrostatic self-assembly (ESA) technique to promote osteoblast growth. Poly(ethylenimine) (PEI) was first employed to obtain a stable positively charged surface on poly(DL-lactide) (PDL-LA) substrate. Gelatin was selected as ECM-like biomacromolecule to deposit on the activated PDL-LA substrate using the ESA technique. ζ-Potential results showed alternating charge of polyelectrolytes (PEI/gelatin) layering on PDL-LA microspheres. Quartz crystal microbalance (QCM) measurement further verified the gradual deposition of PEI/gelatin on PDL-LA thin film. Osteoblast cells (MC3T3) were chosen to test the cell behavior on modified PDL-LA substrates. The osteoblast test about cell activity, intracellular total DNA content, total protein content and cell morphology by SEM investigation on ECM-like multilayer-modified PDL-LA substrate showed to promote osteoblast growth. Comparing conventional coating methods, polyelectrolyte multilayers are easy and stable to prepare. It may be a good choice for the surface modification of complex biomedical devices. These very flexible systems allow broad medical applications for drug delivery and tissue engineering.  相似文献   

12.
Hydroxyapatite coatings have been proven to improve the osteointegration of metal implants through a tight binding to the bone mineral phase as well as through favorable osteoblast adhesion and proliferation onto the implant surface. However, hydroxyapatite coatings are not stable and they tend to delaminate from the metal surface when challenged by the mechanical stresses experienced by the implant. Recently, a new multiphase anodic spark deposition (ASD) method has been optimized where the formation of a thick oxide film is followed by the deposition of a calcium phosphate mineral phase and its etching by alkali. The data in this paper demonstrate that this novel type of coating, BioSpark, improves the material osteointegration potential when compared to conventional ASD while offering more mechanical stability. A faster mineralization was obtained by incubation in simulated body fluids and osteoblasts showed better adhesion, proliferation, differentiation, and collagen production. These performances were related to the surface morphology, to the film calcium/phosphate ratio and its surface oxygen content, as well as to a preferential binding of structural proteins such as fibronectin.  相似文献   

13.
Sargeant TD  Rao MS  Koh CY  Stupp SI 《Biomaterials》2008,29(8):1085-1098
Surface modification enables the creation of bioactive implants using traditional material substrates without altering the mechanical properties of the bulk material. For applications such as bone plates and stents, it is desirable to modify the surface of metal alloy substrates to facilitate cellular attachment, proliferation, and possibly differentiation. In this work we present a general strategy for altering the surface chemistry of nickel-titanium (NiTi) shape memory alloy in order to covalently attach self-assembled peptide amphiphile (PA) nanofibers with bioactive functions. Bioactivity in the systems studied here includes biological adhesion and proliferation of osteoblast and endothelial cell types. The optimized surface treatment creates a uniform TiO(2) layer with low levels of Ni on the NiTi surface, which is subsequently covered with an aminopropylsilane coating using a novel, lower temperature vapor deposition method. This method produces an aminated surface suitable for covalent attachment of PA molecules containing terminal carboxylic acid groups. The functionalized NiTi surfaces have been characterized by X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectroscopy (ToF-SIMS), and atomic force microscopy (AFM). These techniques offer evidence that the treated metal surfaces consist primarily of TiO(2) with very little Ni, and also confirm the presence of the aminopropylsilane overlayer. Self-assembled PA nanofibers presenting the biological peptide adhesion sequence Arg-Gly-Asp-Ser are capable of covalently anchoring to the treated substrate, as demonstrated by spectrofluorimetry and AFM techniques. Cell culture and scanning electron microscopy (SEM) demonstrate cellular adhesion, spreading, and proliferation on these functionalized metal surfaces. Furthermore, these experiments demonstrate that covalent attachment is crucial for creating robust PA nanofiber coatings, leading to confluent cell monolayers.  相似文献   

14.
Surface modifications and cell-materials interactions with anodized Ti   总被引:2,自引:0,他引:2  
The objective of this study was to investigate in vitro cell-materials interactions using human osteoblast cells on anodized titanium. Titanium is a bioinert material and therefore becomes encapsulated after implantation into the living body by a fibrous tissue that isolates it from the surrounding tissues. In this work, a bioactive TiO(2) layer was grown on commercially pure titanium substrate by an anodization process using different electrolyte solutions, namely H(3)PO(4), HF and H(2)SO(4). These electrolytes produced bioactive TiO(2) films with a nonporous structure showing three distinctive surface morphologies. Human osteoblast cell growth behavior was studied with as-received and anodized surfaces using an osteoprecursor cell line (OPC 1) for 3, 5 and 11days. When anodized surfaces were compared for cell-materials interaction, it was noticed that each of the surfaces has different surface properties, which led to variations in cell-materials interactions. Colonization of the cells was noticed with a distinctive cell-to-cell attachment in the HF anodized surface. Good cellular adherence with extracellular matrix extensions in between the cells was noticed for samples anodized with H(3)PO(4) electrolyte. The TiO(2) layer grown in H(2)SO(4) electrolyte did not show significant cell growth on the surface, and some cell death was also noticed. Cell adhesions and differentiation were more pronounced with vinculin protein and alkaline phosphatase, respectively, on anodized surfaces. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium assays also showed an increase in living cell density and proliferation with anodized surfaces. It was clear that rough surface morphology, high surface energy and low values of contact angles were important factors for better cell materials interaction. A mineralization study was done in simulated body fluid with ion concentrations nearly identical to those of human blood plasma to further understand biomimetic apatite deposition behavior. Similar to cell-materials interaction, variations in mineral deposition behavior were also noticed for films grown with different electrolytes.  相似文献   

15.
Polyetheretherketone (PEEK) is used as an alternative to titanium in medical devices. Previous in vitro studies examining PEEK have differed in their choice of polymer variant [PEEK or carbon-fiber reinforced PEEK (CFR-PEEK)], source of polymer (some of which are no longer available or for implantation) and cell type. While all studies demonstrated favorable cytocompatibility of the PEEK material, no studies are available which reflect the current state of the art of the material. Here, we use different forms of the only implantable grade PEEK available. These are compared with commercially pure titanium (cpTi) Grade 1 using a human primary osteoblast model. Sample materials were presented as industrially relevant surfaces. Machined or injection molded PEEK and CFR-PEEK were evaluated along with polished (Ra=0.200microm) and rough (Ra=0.554microm) cpTi. Osteoblast adhesion at 4h on injection molded variants of PEEK (Ra=0.095microm) and CFR-PEEK (Ra=0.350microm) material was comparable to titanium. Machined variants of PEEK (Ra=0.902microm) and CFR-PEEK (Ra=1.106microm) materials were significantly less. Proliferation at 48h determined by [(3)H]-thymidine incorporation was the greatest on the smoothest of all materials, the injection molded unfilled PEEK, which was significantly higher than the rough titanium control. The machined unfilled PEEK had the lowest DNA synthesis. RT-PCR for alkaline phosphatase, Type I collagen and osteocalcin normalized to glyceraldehyde-3-phosphate dehydrogenase revealed different patterns of mRNA levels. High mRNA levels for Type I collagen showed that CFR-PEEK stimulated osteoblast differentiation, whilst injection molded unfilled PEEK was less differentiated. Machined unfilled PEEK had comparable message levels of bone matrix proteins as rough titanium. All material variants permitted a degree of mineralization. Scanning electron microscopy at 3 days and 2 weeks in differentiation medium showed that human osteoblasts were well spread on all the different substrates. The varied response reported here at different time points during the study suggests that material formulation (unfilled PEEK or CFR-PEEK), subjection to industrial processing, surface roughness and topography may all influence the cellular response of osteoblasts to PEEK. Thus, differences in human osteoblast responses were found to the various samples of PEEK, but implantable grade PEEK, in general, was comparable in vitro to the bone forming capacity of rough titanium.  相似文献   

16.
17.
An adhesion test procedure applied to plasma-sprayed hydroxyapatite (HA) coatings to measure the "LASAT threshold" (LAser Shock Adhesion test) is described. The good repeatability and minimal discrepancy of the laser-driven adhesion test data were ascertained for conventional plasma sprayed HA coatings. As a further demonstration, the procedure was applied to HA coatings with diverse characteristics on the ceramic/metal interface. Different preheating and grit blasting conditions and the presence of a thick plasma-sprayed Ti sublayer or a thin TiO(2) layer prepared by oxidation were investigated through LASAT. It was assessed that a rough surface can significantly improve the coating's bond strength. However, it was also demonstrated that a thin TiO(2) layer on a smooth Ti-6Al-4V substrate can have a major influence on adhesion as well. Preheating up to 270°C just prior to the first HA spraying pass had no effect on the adhesion strength. Further development of the procedure was done to achieve an in situ LASAT with in vitro conditions applied on HA coatings. To that end, different crystalline HA contents were soaked in simulated body fluid (SBF). Beyond the demonstration of the capability of this laser-driven adhesion test devoted to HA coatings in dry or liquid environment, the present study provided empirical information on pertinent processing characteristics that could strengthen or weaken the HA/Ti-6Al-4V bond.  相似文献   

18.
Abstract

A strategy developed for obtaining positive cellular responses remains to be focused in the filed of functional biomimetics. In this study, a hydrogel covered simvastatin-loaded polyetheretherketone (PEEK) bio-composites was constructed with the purpose of bone tissue regeneration therapy. Briefly, a three-dimensional (3D) porous structure was fabricated on PEEK surface; then the substrate was functionalized with the poly(L-lactic acid)/simvastatin porous film and hyaluronic acid hydrogel subsequently. In vitro cell attachment, proliferation, and cytoskeletal observation experiments reveal that our scaffolds show better bio-affinity due to the layer of hyaluronic acid hydrogel compared with control. Furthermore, the alkaline phosphatase activity, calcium mineral deposition evaluation, and gene expression for osteogenic potential all exhibit that the superior osteogenic differentiation of MC3T3-E1 pre-osteoblasts on our scaffolds. Therefore, our PEEK samples loaded with simvastatin and covered with hyaluronic acid hydrogel hold great potential in clinical applications for bone repair.  相似文献   

19.
Surface topography is one of the most important factors influencing the attachment and spreading of cells. In the present study, layer-by-layer assembled titanium dioxide (TiO2) nanoparticle thin films were chosen for attachment, proliferation and spreading studies on mouse mesenchymal stem cells (MSC). Increasing surface roughness was observed with increasing number of layer-by-layer assembled TiO2 thin films. Four layer TiO2 thin film showed higher number of attached cells than a one layer thin film and control surfaces. MSCs experienced no cytotoxic effects after culture on the TiO2 coated substrates as observed from the cytotoxicity tests. Cell spreading, visualized with scanning electron microscopy, showed a faster rate of spreading on a rougher surface. Cells on a four-layer substrate, at 12 h showed complete spreading, where as most of the cells on a control surface and a one-layer surface, at 24 h, retained a rounded morphology. In conclusion, TiO2 nanoparticle thin films were successfully assembled in alternation with polyelectrolytes and in-vitro studies with MSC showed an increase in the attachment and faster spreading of cells on rougher surfaces.  相似文献   

20.
Mn(2+)-doped carbonated hydroxyapatite (Mn-CHA) thin films were obtained by pulsed laser deposition on Ti substrates. The results of the performed complementary diagnostic techniques, X-ray diffraction, infrared spectroscopy, X-ray photoelectron spectroscopy, and energy dispersive X-ray spectroscopy investigations indicate that the films are crystalline with a Ca/P ratio of about 1.64-1.66. The optimum conditions, when nearly stoichiometric crystalline thin films were deposited, were found to be 10 Pa oxygen pressure, 400 degrees C substrate temperature, and postdeposition heat treatment in water vapors at the same substrate temperature. The films were seeded with L929 fibroblast and hFOB1.19 osteoblast cells and subjected to in vitro tests. Both fibroblast and osteoblast cells have a good adherence on the Mn-CHA film and on the Ti or polystyrene references. Proliferation and viability tests showed that osteoblast cells growth on Mn-CHA-coated Ti was enhanced as compared to uncoated pure Ti surfaces. Caspase-1 activity was not affected significantly by the material, showing that Mn-CHA does not induce apoptosis of cultured cells. These results demonstrate that Mn-CHA films on Ti should provoke a faster osteointegration of the coated implants as compared to pure Ti. (c) 2004 Wiley Periodicals, Inc. J Biomed Mater Res 71A: 353-358, 2004.  相似文献   

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