Surface modification of titanium substrates with silver nanoparticles embedded sulfhydrylated chitosan/gelatin polyelectrolyte multilayer films for antibacterial application

To develop Ti implants with potent antibacterial activity, a novel “sandwich-type” structure of sulfhydrylated chitosan (Chi-SH)/gelatin (Gel) polyelectrolyte multilayer films embedding silver (Ag) nanoparticles was coated onto titanium substrate using a spin-assisted layer-by-layer assembly technique. Ag ions would be enriched in the polyelectrolyte multilayer films via the specific interactions between Ag ions and –HS groups in Chi-HS, thus leading to the formation of Ag nanoparticles in situ by photo-catalytic reaction (ultraviolet irradiation). Contact angle measurement and field emission scanning electron microscopy equipped with energy dispersive X-ray spectroscopy were employed to monitor the construction of Ag-containing multilayer on titanium surface, respectively. The functional multilayered films on titanium substrate [Ti/PEI/(Gel/Chi-SH/Ag) _n /Gel] could efficiently inhibit the growth and activity of Bacillus subtitles and Escherichia coli onto titanium surface. Moreover, studies in vitro confirmed that Ti substrates coating with functional multilayer films remained the biological functions of osteoblasts, which was reflected by cell morphology, cell viability and ALP activity measurements. This study provides a simple, versatile and generalized methodology to design functional titanium implants with good cyto-compatibility and antibacterial activity for potential clinical applications.     

Fabrication and in vitro evaluation of stable collagen/hyaluronic acid biomimetic multilayer on titanium coatings

Layer-by-layer (LBL) self-assembly technique has been proved to be a highly effective method to immobilize the main components of the extracellular matrix such as collagen and hyaluronic acid on titanium-based implants and form a polyelectrolyte multilayer (PEM) film by electrostatic interaction. However, the formed PEM film is unstable in the physiological environment and affects the long-time effectiveness of PEM film. In this study, a modified LBL technology has been developed to fabricate a stable collagen/hyaluronic acid (Col/HA) PEM film on titanium coating (TC) by introducing covalent immobilization. Scanning electron microscopy, diffuse reflectance Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy were used to characterize the PEM film. Results of Sirius red staining demonstrated that the chemical stability of PEM film was greatly improved by covalent cross-linking. Cell culture assays further illustrated that the functions of human mesenchymal stem cells, such as attachment, spreading, proliferation and differentiation, were obviously enhanced by the covalently immobilized Col/HA PEM on TCs compared with the absorbed Col/HA PEM. The improved stability and biological properties of the Col/HA PEM covalently immobilized TC may be beneficial to the early osseointegration of the implants.     

Effects of immobilization of hyaluronic acid and carboxymethyl chitosan onto a -NH2 functionalized titanium surfaces on MG 63 cell proliferations

The purpose of this study is the development of bioactive functionalized titanium surface by immobilizing hyaluronic acid (HA) and carboxymethyl chitosan (CMCH) onto -NH2 functionalized titanium surfaces to improve biological and chemical properties of titanium. The in vitro biological evaluation showed that introducing the CMCH and HA to the Ti/NH2 enhanced initial cell proliferation compared to untreated Ti surface.     

Probing the stability of biocompatible sodium hyaluronate/chitosan nanocoatings against changes in salinity and pH

The stability of polyelectrolyte multilayer assemblies was investigated with emphasis on the effects of solution ionic strength, pH, and polymer molecular weight on the film thickness and surface topography. The multilayers consisting of two polysaccharides, the polyanion sodium hyaluronate (HA) and the polycation chitosan (CH) were studied using surface plasmon resonance (SPR) spectroscopy, impedance quartz crystal microbalance (QCM), and atomic force microscopy (AFM). SPR/QCM experiments show that coatings consisting of four HA/CH bilayers assembled at pH 4.5 in the presence of 0.15 M NaCl are stable in NaCl solutions of concentration less than 0.8 M. These multilayers are stable when placed in contact with aqueous solutions ranging in pH from 3.5 to 9. The molecular weight of the polysaccharides has only a marginal effect on the stability of the films in the range explored here (HA: Mn = 360,000 or 31,000 g/mol; CH: Mn = 160,000 or 30,000 g/mol). AFM imaging reveals that different mechanisms may account for the multilayers stability versus salt and pH treatments. While increasing the ionic strength induces reorganization of the surface topography from isolated spherical islets to elongated worm-like features, changes in pH have no appreciable effects on the coating topography prior to complete disintegration.     

Cytotoxicity and antibacterial property of titanium alloy coated with silver nanoparticle-containing polyelectrolyte multilayer

Silver nanoparticle (AgNP) was incorporated into dopamine-modified alginate/chitosan (DAL/CHI) polyelectrolyte multilayer to modify the surface of titanium alloy and improve its antibacterial property. Scanning electron microscopy showed that AgNP with the size of 50 nm embedded in DAL/CHI multilayers homogeneously. X-ray photoelectron spectroscopy analysis indicated that the nanoparticles were silver (0) with peaks at 368.4 and 374.4 eV, respectively. The formation of silver (0) without the addition of reductants was due to the self-polymerization of dopamine, which can reduce the silver cation into neutral metal. The polyelectrolyte multilayer coating enhanced the wettability of titanium alloy and promoted the fibroblast proliferation significantly, which could be attributed to the excellent biocompatibility of DAL/CHI. Despite the slight fall of L929 cell activity after AgNP incorporation, AgNP-DAL/CHI multilayer inhibited the growth of both Escherichia coli and Staphylococcus aureus. The above results demonstrate that dopamine decoration is a simple and effective way to induce the in-situ formation of AgNP within polyelectrolyte multilayer. Furthermore, the AgNP-containing multilayer considerably enhances the antibacterial activity of titanium alloy. The fabrication of AgNP-DAL/CHI multilayer on the surface of titanium implant might have great potential in orthopedic use.     

Preparation of HA/chitosan composite coatings on alkali treated titanium surfaces through sol-gel techniques

In this paper we demonstrated for the first time the feasibility to generate well crystallized hydroxyapatite/chitosan (HA/CS) composite coatings on alkali treated titanium surfaces through combining the in situ hydrothermal precipitation and sol-gel dip coating technique without introducing any other coupling agent. Alkali treatment converting hydrophobic Ti surfaces into super-hydrophilic surfaces was the pre-requirement of uniform coatings. Preliminary cell culturing results revealed that the coatings were biocompatible and supported osteoblasts attachment.     

Novel heat spreader coatings for high power electronic devices

A new set of heat spreader coatings consisting of multilayers of diamond/AlN/diamond were deposited on high heat capacity substrates of molybdenum and silicon nitride. Bonding of the heat spreaders to the device wafers using gold-tin eutectic solder was carried out after metallization layers of titanium, gold and copper were deposited on diamond. Prior to bonding, backside of the silicon wafers was also metallized with titanium, gold and copper and the gallium arsenide wafers with titanium, copper-germanium alloy and gold. Characterization of the multilayer diamond films was carried out by Raman spectroscopy, X-ray diffraction and scanning electron microscopy. The bonded wafers were tested for adhesion strength, resistance against peeling due to thermal cycling and failure under stress. Further, the bonded regions were characterized by scanning electron microscopy, energy dispersive spectroscopy and X-ray mapping of different elements. The heat spreader characteristics of the single layer diamond and the multilayer diamond substrates were tested by infrared imaging. The results illustrate that the multilayer diamond heat spreader coatings provide better heat dissipation and also possess better adhesion strength and resistance against peeling under thermal cycling. These novel multilayer diamond/AlN/diamond heat spreaders are expected to considerably improve the life of high frequency power devices.     

Corrosion behavior of nanolayered TiN/NbN multilayer coatings prepared by reactive direct current magnetron sputtering process

TiN, NbN and TiN/NbN multilayer coatings were deposited on tool steel substrates using a reactive DC magnetron sputtering process. The coatings were characterized using X-ray diffraction, nanoindentation, atomic force microscopy, scanning electron microscopy (SEM) and energy-dispersive X-ray analysis. The corrosion behavior of TiN/NbN multilayer coatings was studied in 0.5 M HCl and 0.5 M NaCl solutions using potentiodynamic polarization and compared with single layered TiN and NbN coatings. Approximately 1.5 μm thick coatings of TiN, NbN and TiN/NbN multilayers showed good corrosion protection of the tool steel substrate and multilayer coatings performed better than single layered coatings. The corrosion behavior of the multilayers improved with total number of interfaces in the coatings. In order to conclusively demonstrate the positive effect of layering, corrosion behavior of 40-layer TiN/NbN multilayers was studied at lower coating thicknesses (32–200 nm) and compared with single layer TiN coatings of similar thicknesses. The polarization data and SEM studies of these coatings indicated that the corrosion behavior improved with coating thickness and multilayers showed better corrosion resistance as compared to the single layer coatings. Other studies such as intrinsic corrosion, effects of Ti interlayer and post-deposition annealing on the corrosion behavior of the multilayer coatings are also presented in this paper. The results of this study demonstrate that nanolayered multilayers can effectively improve the corrosion behavior of transition metal nitride hard coatings.     

Investigation of conductive and transparent Al-doped ZnO/Ag/Al-doped ZnO multilayer coatings by electron beam evaporation

Multilayer coatings consisting of thin silver layer sandwiched between layers of Al-doped ZnO (AZO) were prepared by electron beam evaporation. The optical and electrical performances of AZO/Ag/AZO multilayers were investigated. Optimization of the multilayer coatings resulted with low sheet resistance of 7.7 Ω/sq and transmittance of 85%. The influence of thickness of each layer on the optic and electrical performance was analyzed. The sheet resistance of the multilayer was reduced to 5.34 Ω/sq. and the average transmittance was improved to 90% by the thermal treatment. The coatings had satisfactory properties of low resistance, high transmittance and thermal stability.     

Bioactive TiOB‐Coating on Titanium Alloy Implants Enhances Osseointegration in a Rat Model

The surface properties of titanium alloy implants for improved osseointegration in orthopaedic and dental surgery have been modified by many technologies. Hydroxyapatite coatings with a facultative integration of growth factors deposited by plasma spraying showed improved osseointegration. Our approach in order to enhance osseointegration was carried out by a surface modification method of titanium alloy implants called plasma chemical oxidation (PCO). PCO is an electrochemical procedure that converts the nm‐thin natural occurring titanium‐oxide layer on an implant to a 5 µm thick ceramic coating (TiOB‐surface). Bioactive TiOB‐surfaces have a porous microstructure and were loaded with calcium and phosphorous, while bioinert TiOB‐surfaces with less calcium and phosphorous loadings are smooth. A rat tibial model with bilateral placement of titanium alloy implants was employed to analyze the bone response to TiOB‐surfaces in vivo. 64 rats were randomly assigned to four groups of implants: (i) pure titanium alloy (control), ii) titanium alloy, type III anodization, (iii) bioinert TiOB‐surface, and (iv) bioactive TiOB‐surface. Mechanical fixation was evaluated by pull out tests at 3 and 8 weeks. The bioactive TiOB‐surface showed significantly increased shear strength at 8 weeks compared to all other groups.     

Mechanical and tribological properties of Ti/TiAlN duplex coatings on high and low alloy tool steels

In this research work, different steels were hardened using plasma nitriding process in an Arc-PVD device. Afterwards, on both non-nitrided and plasma nitrided substrates, different Ti/TiAlN multilayer coatings were deposited by a magnetron sputtering device. Scratch tests were performed on the duplex systems in order to characterize their adhesive properties. The failure modes of individual coating systems under various normal loads were described using light optical microscope. Furthermore, to study the effect of plasma nitriding on the friction coefficient and wear rate of the systems, ball-on-disc tests were carried out. It was shown that high and low alloy tool steels obtained different levels of hardness after plasma nitriding process. The results of scratch tests showed that both Ti/TiAlN multilayer coatings have a higher adhesion to plasma nitrided steels compared to non-nitrided steels. In addition, plasma nitriding of the substrates resulted in an increase in the wear rates of multilayer coatings. It was also revealed that the adhesion and the tribology of multilayers depend strongly on the amount of titanium interlayers. An increase of the thickness of titanium interlayers enhanced the adhesion of multilayers. However, increasing the thickness of titanium interlayers decreased the hardness and wear resistance of multilayers.     

钛表面制备羟基磷灰石/壳聚糖复合涂层研究

通过原位水热合成和溶胶-凝胶浸提涂敷法在碱处理的钛表面制备了HA/CS复合涂层. 接触角检测表明碱处理使钛表面具有超亲水性.X射线衍射分析表明复合涂层成分为HA和CS, 各组分含量由热重分析确定. 用扫描电镜对复合涂层的形貌进行观察,发现不同HA含量的复合涂层具有不同的形貌. 通过培养成骨细胞考察了复合涂层的细胞相容性.Alamar Blue检测表明HA/CS复合涂层表面细胞粘附及增殖能力较好. ALP检测表明HA/CS复合涂层表面的细胞分化能力较好. 综合研究结果表明, 复合涂层有较好的细胞相容性.     

Chemically tunable electrochemical dissolution of noncontinuous polyelectrolyte assemblies: an in situ study using ecAFM

The electrochemically triggered dissolution of noncontinuous polyelectrolyte assemblies presenting distinct nanomorphologies and its tuning by chemical cross-linking were monitored locally, in situ, by electrochemical atomic force microscopy. Poly-l-lysine and hyaluronic acid deposited layer-by-layer on indium tin oxide electrodes at specific experimental conditions formed well-defined nanostructures whose morphologies could be easily and precisely followed along the dissolution process. In addition to shrinkage of polyelectrolyte nanodroplets, ecAFM images revealed the faster dissolution of coalesced structures compared to droplet-like complexes, and the readsorption of dissolved polyelectrolytes onto slower dissolving neighboring structures. Covalently cross-linked PLL/HA assemblies dissolved only partially, and exhibited slower dissolution rates compared to native multilayers, with a clear dependence on the cross-link density. Tuning the electrochemical dissolution of polyelectrolyte multilayers through chemical cross-linking opens new prospects for future biomedical applications, such as the development of advanced drug or gene delivery platforms allowing for tightly controlled releases of different compounds at specific rates.     

Preparation and characterization of novel sinomenine microcapsules for oral controlled drug delivery

Background: Developing a sustained release drug to cure arthritis is needed. Sinomenine (SIN) is abstracted from sinomenium acutum and widely used in the treatment of various rheumatism and arrhythmia with few side effects. The primary aim of this study is to develop SIN microcapsules with polyelectrolyte multilayers for controlled drug release. Method: SIN microcrystals were encapsulated with chitosan, gelatin, and alginate by layer-by-layer technique, such as (gelatin/alginate)₄ and (chitosan/alginate)₆. The size distribution, zeta-potential, stability, and morphology of the microcapsules were characterized by a particle size analyzer, zetasizer, ultraviolet spectroscopy, and transmission electron microscope, respectively. The in vitro controlled release pattern of SIN was studied using a diffusion cell assembly at physiological pH of 6.8 or 1.4. Results: Light stability of these microcapsules was improved after microencapsulation. Compared with release rate of the SIN microcapsules coated by the poly(dimethyldiallyl ammonium chloride)/alginate and gelatin/alginate multilayers, release rate of the SIN microcapsules coated with chitosan/alginate multilayers was fast. Release rate progressively decreased with the increase of chitosan/alginate bilayer number and the decrease of pH value of release medium. Conclusion: These novel SIN microcapsules may be developed into oral controlled drug delivery for rheumatism and arthritis.     

Surface analysis and osteoblasts response of a titanium oxi-carbide film deposited on titanium by ion plating plasma assisted (IPPA)

Titanium is the most widely used material in orthopaedic and dental implantoprosthesis due to its superior physical properties and enhanced biocompatibility due to the spontaneous formation of a passivating layer of titanium oxides which, however, does not form good chemical bonds with bone and tends to brake exposing bulk titanium to harsh body fluids releasing titanium particles which may prime an inflammation response and a fibrotic tissue production. In order to avoid these possible problems and to enhance the biocompatibility of titanium implants, modifications of titanium surfaces by many different materials as hydroxyapatite, titanium nitride, titanium oxide and titanium carbide have been proposed. The latter is shown to be an efficient protection for the titanium implant in the harsh conditions of biological tissues and, compared to untreated titanium, acting like an osteoblast stimulation factor increasing in vitro production of proteins involved in osteogenesis. These results were confirmed by in vivo experiments in rabbits: implants covered by the titanium carbide (TiC) layer were faster and better osseointegrated than untreated titanium implants. The TiC layer was deposited by a Pulsed Laser Deposition (PLD) device which allowed only one deposition per cycle, shown to be unsuitable for industrial applications. Therefore the main objective of the present work was to replace PLD process with an Ion Plating Plasma Assisted (IPPA) deposition process, which is suitable for industrial upgrading. By this technique, nanostructured TiOx-TiCy-C has been deposited on titanium after sandblasting with 120 micron zirconia spheres. XPS analyses revealed the presence of about 33% carbon (50% of which is present as free carbon), 39% oxygen and 28% titanium (37% of which is bound to carbon to form TiC and 63% is bound to oxygen to form non stoichiometric oxides). Surface mechanical response of as-deposited coatings has been performed by nanoindentation techniques. Focused Ion Beam micrographs showed bigger differences on the obtained nanostructure compared to the PLD coating structure; in vitro tests confirm for IPPA produced coatings an improvement in stimulating osteoblasts to produce mRNA's of proteins involved in the ossification process, this latter case they resulted to be faster and more efficient. The proposed treatement is expected to improve the good results obtained by PLD, in vivo as well.     

Quantification of carbon nanotube induced adhesion of osteoblast on hydroxyapatite using nano-scratch technique

This paper explores the nano-scratch technique for measuring the adhesion strength of a single osteoblast cell on a hydroxyapatite (HA) surface reinforced with carbon nanotubes (CNTs). This technique efficiently separates out the contribution of the environment (culture medium and substrate) from the measured adhesion force of the cell, which is a major limitation of the existing techniques. Nano-scratches were performed on plasma sprayed hydroxyapatite (HA) and HA-CNT coatings to quantify the adhesion of the osteoblast. The presence of CNTs in HA coating promotes an increase in the adhesion of osteoblasts. The adhesion force and energy of an osteoblast on a HA-CNT surface are 17 ± 2 μN/cell and 78 ± 14 pJ/cell respectively, as compared to 11 ± 2 μN/cell and 45 ± 10 pJ/cell on a HA surface after 1 day of incubation. The adhesion force and energy of the osteoblasts increase on both the surfaces with culture periods of up to 5 days. This increase is more pronounced for osteoblasts cultured on HA-CNT. Staining of actin filaments revealed a higher spreading and attachment of osteoblasts on a surface containing CNTs. The affinity of CNTs to conjugate with integrin and other proteins is responsible for the enhanced attachment of osteoblasts. Our results suggest that the addition of CNTs to surfaces used in medical applications may be beneficial when stronger adhesion of osteoblasts is desired.     

Electrospray deposition of nanohydroxyapatite coatings: A strategy to mimic bone apatite mineral

The biological performance of orthopaedic and oral metallic implants can be enhanced significantly by the application of bioactive coatings. In this work, a cost-efficient alternative to the traditional technique to produce a hydroxyapatite (HA) coating with a nanostructured feature onto a metallic implant surface at room temperature via electrospray deposition, is presented. To evaluate the bioactive capacity of these nanoHA (nHA) coatings in vitro, an acellular simulated body fluid soaking experiment and a human osteoblast (HOB) cell culture work were conducted. Under these physiological conditions, the accelerated apatite precipitation process occurred on the nHA-coated titanium surfaces as compared to the uncoated titanium surfaces. HOB cells developed mature cytoskeletons with distinct evidence of actin stress fibres and vinculin adhesion plaques, on these nHA coatings. Hence, this deposition technique holds great potential in producing high quality nHA coatings for biomedical applications.     

羟基磷灰石/钛合金骨替代材料体外培养成骨细胞的实验研究

实验选用小鼠头盖骨成骨细胞,采用体外细胞培养技术对具有羟基磷灰石涂层的钛合金(HA/Ti)与未经过表面改性的钛合金两种骨替代材料进行细胞相容性评价,动态观察两种骨替代材料对成骨细胞生长、附着的影响。结果表明两种骨替代材料对成骨细胞生长无抑制作用,未发生细胞毒性反应,细胞在两种材料表面均能正常粘附、生长、增殖,均具有良好的细胞附着形态和细胞增殖率,而HA/钛合金材料具有更好的成骨性,是一种骨细胞相容性良好的骨替代材料。     

In Situ ATR‐FTIR Spectroscopy on the Deposition and Protein Interaction of Polycation/Alginate Multilayers

We report in situ ATR‐FTIR spectroscopy studies of the deposition of biocompatible polyelectrolyte multilayers (PEMs) consisting of the polycation chitosan (CHT) or poly(ethyleneimine) (PEI) and the polyanion sodium alginate (ALG) and their interactions with model‐protein human serum albumin (HSA). HSA‐adsorption data for PEI/ALG and CHT/ALG PEMs show the importance of the outermost polyelectrolyte (PEL) layer for protein interactions: CHT‐ or PEI‐terminated PEMs are highly attractive to HSA, while ALG‐terminated PEMs are repulsive, relevant for the generation of protein‐active or protein‐inert biomedical surfaces. The bound HSA is found to be located at the surface rather than the inner region of the PEM.     

钛表面仿生矿化及其对成骨细胞行为的影响

采用仿生矿化法对商用纯钛进行表面处理,经过处理的样品表面形成一层薄的钙磷涂层,SEM和EDX分析表明,涂层晶体构型均一;XRD和FTIR分析晶体组成主要为碳酸羟基磷灰石.采用成骨细胞体外培养方法,探讨仿生矿化涂层对于细胞初期附着、增殖的影响.认为钙磷涂层可以提高成骨细胞的初期附着率,而对于细胞的增殖行为影响不大.