The effect of heat- or ultra violet ozone-treatment of titanium on complement deposition from human blood plasma.

Titanium (Ti) is a well known metallic biomaterial extensively used in dental, orthopaedic-, and occasionally also in blood contacting applications. It integrates well to bone and soft tissues, and is shown upon blood plasma contact to activate the intrinsic pathway of coagulation and bind complement factor 3b. The material properties depend largely on those of the nm-thick dense layer of TiO(2) that becomes rapidly formed upon contact with air and water. The spontaneously formed amorphous Ti-oxide has a pzc approximately 5-6 and its water solubility is at the order of 1-2 micromolar. It is often subjected to chemical- and heat treatments in order to increase the anatase- and rutile crystallinity, to modify the surface topography and to decrease the water solubility. In this work, we prepared sol-gel derived titanium and smooth PVD titanium surfaces, and analysed their oxide and protein deposition properties in human blood plasma before and after annealing at 100-500 degrees C or upon UVO-treatment for up to 96 hours. The blood plasma results show that complement deposition vanished irreversibly after heat treatment at 250-300 degrees C for 30 minutes or after UVO exposure for 24 hours or longer. XPS and infrared spectroscopy indicated change of surface water/hydroxyl binding upon the heat- and UVO treatments, and increased Ti oxidation. XRD analysis confirmed an increased crystallinity and both control (untreated) and annealed smooth titanium displayed low XRD-signals indicating some nanocrystallinity, with predominantly anatase phase. The current results show that the behaviour of titanium dioxide in blood contact can be controlled through relatively simple means, such as mild heating and illumination in UV-light, which both likely irreversibly change the stoichiometry and structure of the outmost layers of titanium dioxide and its OH/H(2)O binding characteristics.

Sol-gel derived titania coating with immobilized bisphosphonate enhances screw fixation in rat tibia.

A variety of surface modifications have been tested for the enhancement of screw fixation in bone, and locally delivered anti-osteoporosis drugs such as bisphosphonates (BP) are then of interest. In this in vivo study, the impact of surface immobilized BP was compared with systemic BP delivery and screws with no BP. After due in vitro characterization, differently treated stainless steel (SS) screws were divided into four groups with 10 rats each. Three of the groups received screws coated with sol-gel derived TiO(2) and calcium phosphate (SS+TiO(2)+CaP). One of these had no further treatment, one had alendronate (BP) adsorbed to calcium phosphate mineral, and one received systemic BP treatment. The fourth group received uncoated SS screws and no BP (control). The screw pullout force was measured after 4 weeks of implantation in rat tibiae. The immobilized amount and release rate of alendronate could be controlled by different immersion times. The SS+TiO(2)+CaP coating did not increase the pullout force compared to SS alone. Surface delivered alendronate enhanced the pullout force by 93% [p = 0.000; 95% Confidence Interval (CI): 67-118%] compared to SS, and by 39% (p = 0.044; 95% CI: 7-71%) compared to systemic alendronate delivery. Both surface immobilized and systemically delivered alendronate improved implant fixation. Also, locally delivered, that is, surface immobilized alendronate showed a better fixation than systemically delivered. Using sol-gel derived TiO(2) as a platform, it is possible to administer controllable amounts of a variety of BPs.

Cell viability in a wet silica gel.

A modified two-step sol-gel route using silicon ethoxide (TEOS) has been used to synthesize amorphous sol-gel-derived silica, which has been successfully used as a cell encapsulation matrix for 3T3 mouse fibroblasts and CRL-2595 epithelial cells due to its non-toxicity. The sol-gel procedure comprised a first, low pH hydrolysis step, followed by a neutral condensation-gelation step. A high water-to-TEOS ratio and the addition of d-glucose as a porogen and source of nutrients were chosen to minimize silica dissolution and improve the biocompatibility of the process. Indeed, the cell integrity in the encapsulation process was preserved by alcohol removal from the starting solution. Cells were then added in a buffered medium, causing rapid gelation and entrapment of the cells within a randomly structured siloxane matrix in the shape of a monolith, which was maintained in the wet state. MTT and alamarBlue assays were used to check the cytotoxicity of the silica gels and the viability of entrapped cells at initial times in contact with silica. To improve cell attachment, cell clumping experiments - where groups of cells were formed - were designed, rendering improved viability. The obtained materials are therefore excellent candidates for designing tissue-culture scaffolds and implantable bioreactors for biomedical applications.

Comparison of biomaterials and extracellular matrices as a culture platform for multiple, independently derived human embryonic stem cell lines.

Long-term in vitro culture of undifferentiated human embryonic stem cells (hESCs) traditionally requires a fibroblast feeder cell layer. Using feeder cells in hESC cultures is highly laborious and limits large-scale hESC production for potential application in regenerative medicine. Replacing feeder cells with defined human extracellular matrix (ECM) components or synthetic biomaterials would be ideal for large-scale production of clinical-grade hESCs. We tested and compared different feeder cell-free hESC culture methods based on different human ECM proteins, human and animal sera matrices, and a Matrigel matrix. Also selected biomaterials were tested for feeder cell-free propagation of undifferentiated hESCs. The matrices were tested together with conventional and modified hESC culture media, human foreskin fibroblast-conditioned culture medium, chemically defined medium, TeSR1, and modified TeSR1 media. The results showed the undefined, xenogeneic Matrigel to be a superior matrix for hESC culture compared with the purified human ECM proteins, serum matrices, and the biomaterials tested. A long-term, feeder cell-free culture system was successful on Matrigel in combination with mTeSR1 culture medium, but a xeno-free, fully defined, and reproducible feeder cell-free hESC culture method still remains to be developed.

Sol-gel-derived TiO(2)-SiO (2) implant coatings for direct tissue attachment. Part I: design, preparation and characterization.

A series of sol-gel derived TiO(2)-SiO(2) mixed oxide coatings were prepared by carefully controlling the process parameters to obtain silica-releasing coatings consisting of nanoparticles. These features are of paramount importance for enhanced cell adhesion and activation. To achieve both these goals the Ti-alkoxide and Si-alkoxide were first separately hydrolysed and the titania-silica mixed sol was further reacted before the dipping process to obtain the desired particle sizes resulting to the biologically favourable topographical features. Silica release was observed from all the prepared coatings and it was dependent on SiO(2) amount added to the sols, i.e., the higher the added amount the higher the release. In addition, calcium phosphate was able to nucleate on the coatings. From the obtained SiO(2) dissolution data, together with the detailed XPS peak analysis, the mixed oxide coatings are concluded to be chemically heterogeneous, consisting of TiO(2) and SiO(2) species most likely linked together by Ti-O-Si bonds. TiO(2) is chemically stable making long-term implant coating possible and the desired nanoscale dimensions were well preserved although the composition was changed as a consequence of SiO(2) dissolution under in vitro conditions.

Sol-Gel-derived TiO2-SiO2 implant coatings for direct tissue attachment. Part II: Evaluation of cell response.

Silica-releasing sol-gel derived TiO2-SiO2 coatings with tailored nanostructure were evaluated in fibroblast and osteoblast cell cultures. The adhesion of both fibroblasts and osteoblasts proceeded within two hours. The highest fibroblast proliferation activities were observed on the TiO2-SiO2 (70:30) and (30:70) coatings. However, the cell layer on TiO2-SiO2 (30:70) coating was disordered. Prolonged osteoblast activity was observed on the coatings as a function of increased amount of released silica. At day 21 the surfaces were fully covered by the calcified nodules and extracellular matrix except for the coatings TiO2-SiO2 (10:90) i.e. having the highest SiO2 amount. The results suggested that TiO2-SiO2 (70:30) was the best for fibroblasts and TiO2-SiO2 (30:70) for osteoblasts. The applicability of the sol-gel derived TiO2 and TiO2-SiO2 coatings as an alternative for the calcium phosphate based implant coatings are discussed.

Wet powder processing of sol-gel derived mesoporous silica-hydroxyapatite hybrid powders.

This paper describes a method by which a porous silica coating layer can be obtained on different apatite particles through a simple sol-gel synthesis route. Sol-gel derived powders of hydroxyapatite (HAP) and beta tricalciumphosphate (beta-TCP) were coated with a mesoporous silica using C16TAB (hexadecyltrimethylammonium bromide) as a template in order to induce mesophase formation. Further calcination of the material removes the template from the mesophase and leaves a highly ordered hexagonal arranged mesoporous silica structure with a core of HAP/beta-TCP. The phase purity of the SiO2/apatite composites has been thoroughly investigated by the means of FT-IR, XRD, and solid state 31P MAS NMR. The phase purity of these materials is shown to be dependent on the solubility properties of the used apatites. The hybrid materials are suitable as a multifunctional biomaterial where osteoconductive properties can be combined with drug delivery.

Topography and surface energy dependent calcium phosphate formation on Sol-Gel derived TiO2 coatings.

Heterogeneous nucleation and growth of calcium phosphate (CaP) on sol-gel derived TiO(2) coatings was investigated in terms of surface topography and surface energy. The topography of the coatings was derived from AFM measurements, while the surface energy was determined with contact angle measurements. The degree of precipitation was examined with scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The precipitation of CaP was found to be dependent on both topography and surface energy. A high roughness value when combining the RMS roughness parameter S(q) with the number of local maxima per unit area parameter S(ds) enhances CaP formation. The hydrophilicity of the coating was also found to be of importance for CaP formation. We suggest that the water contact angle, which is a direct measure of the hydrophilicity of the surface, may be used to evaluate the surface energy dependent precipitation kinetics rather than using the often applied Lewis base parameter.

Structure of self-assembled multilayers prepared from water-soluble polythiophenes.

We have studied the structure and morphology of self-assembled polyelectrolyte multilayers prepared using poly(styrenesulfonate) (PSS) and four different cationic poly(alkoxythiophene) derivatives bearing methylimidazolium-terminated ionic side chain at the 3-position of the thiophene ring: poly(1-methyl-3-[3-[3-thienyloxy]-propyl]-1H-imidazolium) (P3TOPIM), poly(1-methyl-3-[6-[3-thienyloxy]-hexyl]-1H-imidazolium) (P3TOHIM), poly(1-methyl-3-[2-[(4-methyl-3-thienyl)oxy]-ethyl]-1H-imidazolium ) (P4Me-3TOEIM), and poly(1-methyl-3-[6-[(4-methyl-3-thienyl)oxy]-hexyl]-1H-imidazolium ) (P4Me-3TOHIM). All the multilayers exhibited regular growth. The thickness of the multilayers was measured with ellipsometry, their layer-by-layer growth was followed by polarization modulation infrared reflection-absorption spectroscopy (PM-IRRAS) and ellipsometry, and the morphology of the films was studied by atomic force microscopy (AFM). The length of the methylimidazolium-terminated side chain (C(n), n = 2, 3, 6) and the substituent (H or Me) at the 4-position of the thiophene ring were varied. All multilayers were inhomogeneous in the sub-micrometer scale and contained aggregates of two kinds. The large ones with a low and constant surface number density were attributed to PSS, whereas the small aggregates were polythiophene-based. The surface density of these organic semiconducting nanoparticles greatly depended on the structure of polythiophene, being favored by polymer regioregularity and the length of the side chain. The side chains remained disordered in all the multilayers, but with polythiophenes having hexyl chains both the imidazolium and thiophene rings tended to orient themselves more perpendicular to the surface than in films containing shorter chains (C2 or C3). The relative water content of the multilayers (at 7.1% relative humidity) did not depend on the film thickness and was the lowest for P4Me-3TOHIM. As the number of bilayers increased the methylimidazolium-sulfonate ion pairs gradually weakened and became more individually hydrated.

Destruction of Deinococcus geothermalis biofilm by photocatalytic ALD and sol-gel TiO2 surfaces.

The aim of the present work was to explore possibilities of photocatalytic TiO2 coating for reducing biofilms on non-living surfaces. The model organism, Deinococcus geothermalis, known to initiate growth of durable, colored biofilms on machine surfaces in the paper industry, was allowed to form biofilms on stainless steel, glass and TiO2 film coated glass or titanium. Field emission electron microscopy revealed that the cells in the biofilm formed at 45 degrees C under vigorous shaking were connected to the surface by means of numerous adhesion threads of 0.1-0.3 microm in length. Adjacent cells were connected to one another by threads of 0.5-1 microm in length. An ultrastructural analysis gave no indication for the involvement of amorphous extracellular materials (e.g., slime) in the biofilm. When biofilms on photocatalytic TiO2 surfaces, submerged in water, were exposed to 20 W h m(-2) of 360 nm light, both kinds of adhesion threads were completely destroyed and the D. geothermalis cells were extensively removed (from >10(7) down to below 10(6) cells cm(-2)). TiO2 films prepared by the sol-gel technique were slightly more effective than those prepared by the ALD technique. Doping of the TiO2 with sulfur did not enhance its biofilm-destroying capacity. The results show that photocatalytic TiO2 surfaces have potential as a self-cleaning technology for warm water using industries.

Layer-by-layer electrostatic self-assembly of single-wall carbon nanotube polyelectrolytes.

We have used anionic and cationic single-wall carbon nanotube polyelectrolytes (SWNT-PEs), prepared by the noncovalent adsorption of ionic naphthalene or pyrene derivatives on nanotube sidewalls, for the layer-by-layer self-assembly to prepare multilayers from carbon nanotubes with polycations, such as poly(diallyldimethylammonium) or poly(allylamine hydrochloride) (PDADMA or PAH, respectively), and polyanions (poly(styrenesulfonate), PSS). This is a general and powerful technique for the fabrication of thin carbon nanotube films of arbitrary composition and architecture and allows also an easy preparation of all-SWNT (SWNT/SWNT) multilayers. The multilayers were characterized with vis-near-IR spectroscopy, X-ray photoelectron spectroscopy (XPS), surface plasmon resonance (SPR) measurements, atomic force microscopy (AFM), and imaging ellipsometry. The charge compensation in multilayers is mainly intrinsic, which shows the electrostatic nature of the self-assembly process. The multilayer growth is linear after the initial layers, and in SWNT/polyelectrolyte films it can be greatly accelerated by increasing the ionic strength in the SWNT solution. However, SWNT/SWNT multilayers are much more inert to the effect of added electrolyte. In SWNT/SWNT multilayers, the adsorption results in the deposition of 1-3 theoretical nanotube monolayers per adsorbed layer, whereas the nominal SWNT layer thickness is 2-3 times higher in SWNT/polyelectrolyte films prepared with added electrolyte. AFM images show that the multilayers contain a random network of nanotube bundles lying on the surface. Flexible polyelectrolytes (e.g., PDADMA, PSS) probably surround the nanotubes and bind them together. On macroscopic scale, the surface roughness of the multilayers depends on the components and increases with the film thickness.

Sol-gel synthesis of a multifunctional, hierarchically porous silica/apatite composite.

In this study, a degradable, hierarchically porous silica/apatite composite material is developed from a simple low-temperature synthesis. Mesoporosity is induced in the silica portion by the use of supramolecular templating. The template is further removed by calcination. Firstly, hydroxyapatite is synthesized through a sol-gel method at near room temperature conditions. After the mineralization process, the crystal surface is coated with a mesoporous silica matrix using the templates already present in the bulk solution. The material is characterized by XRD, N(2)-sorption, FT-IR, SEM/EDS, and TEM. The coating layer is distributed fairly homogeneously over the apatite surface and the coating thickness is easily adjustable and dependent on the amount of added silica precursor. The hybrid material is shown to efficiently induce calcium phosphate formation under in vitro conditions and simultaneously work as a carrier system for drugs.

TF-XRD examination of surface-reactive TiO2 coatings produced by heat treatment and CO2 laser treatment.

When surface-reactive (bioactive) coatings are applied to medical implants by means of CO2 laser processing, the bioactivity of the surface of the implant can be locally modified to match the properties of the surrounding tissues to provide a firm fixation of the implant. The aim of this study was to compare the heat treated TiO2 coatings with the laser-treated TiO2 coatings in terms of amorphous-crystalline-phase development. The coatings were characterized with thin-film X-ray diffraction (TF-XRD), atomic force microscopy (AFM) and scanning electron microscopy (SEM). The TiO2 coatings heat treated at 500 degrees C known to be bioactive in SBF (simulated body fluid) consisted mainly of anatase with some rutile-phase, suggesting a predominant effect of anatase on reactivity of coatings. However, the coatings preheat-treated at 500 degrees C with further laser treatment exhibited enhanced bioactivity while consisting mainly of rutile. These findings indicated a key role of both rutile and anatase for the reactivity of the coatings. Without preheat treatment, by laser treatment alone, the amorphous titania coatings developed into mixed anatase/rutile containing coatings. This structural organization and the increase in crystal size are thus considered to be the reasons for their bioactivity. The SBF results indicate the possibility to control bioactivity by altering laser power used through the anatase/rutile crystallinity enhancement.

Topographical parameters for specifying a three-dimensional surface.

The importance of different surface geometries and thereby the need for versatile surface identification by describing a number of different surface features is emphasized. A set of topographical parameters for the description of the amplitude and spatial and hybrid properties of surfaces was utilized for a versatile three-dimensional surface characterization of sol-gel samples with different topographies. The image data were measured by atomic force microscopy. The results demonstrate the power of the roughness parameters to identify surfaces according to their specific characteristics. An example is also given about how certain surface topographical properties may control the material reactivity.

Use of sol-gel-derived titania coating for direct soft tissue attachment.

A firm bond between an implant and the surrounding soft tissue is important for the performance of many medical devices (e.g., stents, canyls, and dental implants). In this study, the performance of nonresorbable and reactive sol-gel-derived nano-porous titania (TiO(2)) coatings in a soft tissue environment was investigated. A direct attachment between the soft tissue and the sol-gel-derived titania coatings was found in vivo after 2 days of implantation, whereas the titanium control implants showed no evidence of soft tissue attachment. The coated implants were in immediate contact with the connective tissue, whereas the titanium controls formed a gap and a fibrous capsule on the implant-tissue interface. The good soft tissue attachment of titania coatings may result from their ability to initiate calcium phosphate nucleation and growth on their surfaces (although the formation of poorly crystalline bonelike apatite does not occur). Thus, the formation of a bonelike CaP layer is not crucial for their integration in soft tissue. The formation of bonelike apatite was hindered by the adsorption of proteins onto the initially formed amorphous calcium phosphate growth centers, thus preventing the dissolution/reprecipitation processes required for the formation of poorly crystalline bonelike apatite. These findings might open novel application areas for sol-gel-derived titania-based coatings.

Mineralization of dentin induced by treatment with bioactive glass S53P4 in vitro.

Dentin hypersensitivity can be managed to occlude dentin tubules, but none of the agents used are components of natural dentin. Using a calcium phosphate precipitation (CPP) method, dentin tubules can be occluded with a calcium phosphate (CaP) layer similar to the major inorganic component of dentin. The CPP method utilizes acidic pH conditions, such as etching of dentin, over the course of several dental treatments. A gentler method can be used to produce a CaP layer on the surface of dentin. By treating with bioactive glass S53P4 (BAG), or regular commercial glass (CG), mineralization occurs in physiologically neutral solutions such as simulated body fluid (SBF) and remineralization solution (RMS). After a short period of immersion, silica is dissolved from both types of glass, but the amount of silica released is much greater from BAG than from CG. The dissolved silica is adsorbed on the surface of dentin during the pretreatment procedure and enhances the mineralization of dentin in SBF. After 14 days' mineralization the dentin is fully covered by the CaP layer, but after 14 days' immersion in RMS decalcification of the dentin occurs. Pretreatment with BAG decreases the degree of decalcification of dentin during the mineralization process. These findings suggest that bioactive glass S53P4 can be used as a therapeutic material for mineralization of dentin and its tubules in a physiological environment.

The Hofmeister anion effect and the growth of polyelectrolyte multilayers.

The influence of a variety of counteranions on the properties of polyelectrolyte multilayers deposited by layer-by-layer technique is studied by using ellipsometry and AFM. We found out that in thin dry multilayers (20-90 nm) ofpoly(4-styrenesulfonate) (PSS) and poly(diallyldimethylammonium) (PDADMA), the thickness follows reasonably well the position of the counteranion in the Hofmeister series. The polyelectrolyte-counteranion interaction is studied by means of viscosity measurements of semidilute solutions of PDADMA in the presence of different anions. The dynamic viscosities follow the Hofmeister series of anions and correlate with the thickness of multilayers. Two parameters describing the interaction of ions with water, the Jones-Dole viscosity B coefficient and the hydration entropy, are used to explain the anion effect on the developing multilayer thickness. Reasonably smooth and monotonic functional dependence is observed between the layer thickness and these two parameters.