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Electrochemical polymerization of aniline monomers infiltrated into well-ordered truncated eggshell structures of polyelectrolyte multilayers.

The use of nanosphere lithography to construct two-dimensional arrays of polystyrene (PS) particles coated with multilayered polyelectrolyte (PE) shells and truncated eggshell structures composed of PE thin layers is reported. The truncated eggshell PE structures were produced by extraction of the PS particle cores with toluene. The core-extraction process ruptures the apex of the PE coating and causes a slight expansion of the PE thin layers. Aniline hydrochloride was infiltrated into the PE shells and subsequently electropolymerized to yield an array of a composite containing polyaniline (PAni) and PE thin shells. Voltammetric, quartz crystal microbalance, and reflectance Fourier transform infrared spectroscopic measurements indicate that aniline monomers were confined within the thin PE shells and the electropolymerization occurred in the interior of the PE shell. The PE thickness governs the amount of infiltrated monomer and the ultimate loading of the PAni in the truncated eggshell structure. Surface-structure imaging by atomic force microscopy and scanning electron microscopy, carried out after each step of the fabrication process, shows the influence of the PE thickness on the organization and dimensions of the arrays. Thus, the PE thin shells composed of different layers can function as nanometer-sized vessels for the entrapment of charged species for further construction of composite materials and surface modifications. This approach affords a new avenue for the synthesis of new materials that combine the unique properties of conductive polymers and the controllability of template-directed surface reactions.

Aniline Compounds↗

In vivo evaluation of bone tissue behavior on ion implanted surfaces.

The ion implantation process offers several unique advantages over other surfaces modifications techniques, in regard to ion release and material mechanical characteristics. The aim of this study was to evaluate the in vivo bone tissue response to ion implanted surfaces. Untreated and nitrogen-ion-implanted stainless steel implants were inserted in the tibia diaphysis (cortical bone) and proximal tibia epiphysis (trabecular bone) of 12 New Zealand White rabbits. The animals were divided into three groups of four animals each, which were maintained for 4, 12 and 24 weeks according to internationally accepted and standardized procedures. At sacrifice, the implants were retrieved with surrounding bone and fixed in 4% neutral buffered formaldehyde and embedded in polymethylmethacrylate (PMMA). The samples were reduced in slices and stained with hematoxylin-eosin, light-green, fuchsin acid and giemsa solution for histological evaluation; fluorescent markers were also used to assess bone apposition. Histomorphometric evaluation was used to determine the extent of bone-material contact. Results from histological and morphometrical analyses revealed active remodeling of bone around both types of implants (control and ion implanted). However, faster bone deposition was observed around the treated material (12 weeks). Both materials reached similar endpoints, as no significant differences between them were evident at 24 weeks. The results demonstrate that ion implanted stainless steel has similar, or slightly enhanced, biological compatibility in contact with bone compared to untreated material; thus it may be a useful material in biomedical applications where reduced ion release or enhanced mechanical properties (as provided by ion implantation) are required.

Journal Article↗

Disappearance of an epithelial cell surface-specific glycoprotein (Epith-1) associated with epithelial-mesenchymal conversion in sea urchin embryogenesis.

Cell surface modification during mesenchyme ingression was examined using a monoclonal antibody (mAb), anti-Epith-1 mAb, raised against a protein (Epith-1) that was confined to the lateral surface of the epithelial cells in embryo of the sea urchin, Temnopleurus hardwicki. The mAb epitope was N-glycosylated oligosaccharides of 160 kDa monomeric Epith-1 protein. The glycoprotein was negatively charged, and its isoelectric point (IP) was 4.98. The mAb, however, is not immunologically cross-reactive with other sea urchin embryos including Hemicentrotus pulcherrimus, Strongylocentrotus nudus, and Scaphechinus mirabilis. Epith-1 is present initially in the cytoplasm of unfertilized eggs. Cytoplasmic Epith-1 shifted to the cell surface to be integrated in plasma membrane during the first cleavage, and remained there during early embryogenesis by retaining the same relative molecular mass (Mr). During primary and secondary mesenchyme ingression periods, however, Epith-1 disappears from the presumptive mesenchyme cell surface that was associated with internalization of the protein. In plutei, an additional anti-Epith-1 mAb-positive protein appears at the 142 kDa region, which was not associated with any visible alteration of the histologic localization of the protein in larvae. Anti-Epith-1 mAb IgG did not inhibit the reaggregation of epithelial cells in vitro, which suggests that either the protein is not involved in cell-cell adhesion or that the mAb is not recognizing the active site of the protein.

Amidohydrolases↗

Overview of surface variability of metallic endosseous dental implants: textured and porous surface-structured designs.

A variety of successful endosseous dental implants with different surface forms are currently available for clinical use. These all achieve implant-to-bone fixation primarily (if not totally) through mechanical interlock of bone with implant surface features introduced by design or chance during implant fabrication. Equally important to establishment of rigid fixation is the rate at which it is achieved, because faster rates allow earlier implant loading and less chance of inadvertent early loading that might prevent implant "osseointegration." Investigations of surface modification to favorably affect osteoconductivity and bone bonding represent an active area of research in the field of dental implant development. This article presents a review of available surface designs and future research directions for improved devices.

Coated Materials, Biocompatible↗

Scanning electron microscopic investigations of surface treated large-bore catheters used for extracorporeal detoxification methods.

Typical complications caused by surface properties of synthetic catheter implants are infection, thrombosis, and stenosis. New methods for surface modification with the aim of reducing such complications are ion beam-based technologies. In our study 109 large-bore catheters without (n = 42) and with treated surfaces with silver (n = 39) or silicone (n = 28) were inserted into the interna jugular and the subclavian veins and were used for extracorporeal detoxification methods. After removal, the catheters were investigated with scanning electron microscopy (SEM) and for bacterial colonization. In 42 large-bore catheters without surface treatment deposits of fibrin, protein and blood cells were seen on the inner and outer surface. Bacterial colonization was observed in 38.1%. In contrast, the catheters with treated outer surfaces showed a very low thrombogenicity and a low contamination rate of 8.9%. The ion beam-based technologies reduce the thrombogenicity and infection rates of the catheter surfaces. In comparison to catheters without treated surfaces, catheters with surface treatment are good alternatives in blood contacting applications ranging from hemodialysis to oncology.

Adsorption↗

A novel soft and cotton-like chitosan-sugar nanoscaffold.

A novel type of chitosan nanoscaffold with a soft and cotton-like appearance is proposed. The key to success is based on two points: (i) the change in morphology of chitin whisker to chitosan nanoscaffold and (ii) the surface modification of the nanoscaffold chitosan with a sugar unit. Simple deacetylation of chitin whisker gives a colloidal solution of chitosan, of which the chitosan is in a nanoscaled scaffold. Surface functionalization of the chitosan nanoscaffold with lactose or maltose via a heterogeneous system in water at room temperature results in a soft and cotton-like chitosan containing mesopores. As all steps are organic solvent free, this chitosan-sugar nanoscaffold might be a promising material for biopolymer-supported tissue engineering.

Acetylation↗

Surface functional group dependent apatite formation on bacterial cellulose microfibrils network in a simulated body fluid.

The apatite forming ability of biopolymer bacterial cellulose (BC) has been investigated by soaking different BC specimens in a simulated body fluid (1.5 SBF) under physiological conditions, at 37 degrees C and pH 7.4, mimicking the natural process of apatite formation. From ATR-FTIR spectra and ICP-AES analysis, the crystalline phase nucleated on the BC microfibrils surface was calcium deficient carbonated apatite through initial formation of octacalcium phosphate (OCP) or OCP like calcium phosphate phase regardless of the substrates. Morphology of the deposits from SEM, FE-SEM, and TEM observations revealed the fine structure of thin film plates uniting together to form apatite globules of various size (from <1 mum to 3 mum) with respect to the substrates. Surface modification by TEMPO (2,2,6,6-tetramethylpyperidine-1-oxyl)-mediated oxidation, which can readily form active carboxyl functional groups upon selective oxidation of primary hydroxyl groups on the surface of BC microfibrils, enhanced the rate of apatite nucleation. Ion exchanged treatment with calcium chloride solution after TEMPO-mediated oxidation was found to be remarkably different from other BC substrates with the highest deposit weight and the smallest apatite globules size. The role of BC substrates to induce mineralization rate differs according to the nature of the BC substrates, which strongly influences the growth behavior of the apatite crystals.

Acetobacter↗

Post-testicular change in the reptile sperm surface with particular reference to the snake, Natrix fasciata.

Sperm surface changes occurring in the reptile Wolffian duct have been explored with particular references to the snake, Natrix fasciata. In the snake Wolffian duct there are several proteins not present in serum, the pattern of which changes in concert with the seasonal testicular cycle. Whereas testicular spermatozoa did not bind antibody to duct secretions, all Wolffian duct spermatozoa did so over both head and tail, according to immunofluorescence patterns. Thus, on entering the Wolffian duct, the entire surface of N. fasciata spermatozoa acquires one of more of the duct's secretory components. As indicated by immunofluorescence, immunoelectrophoresis, and immunodiffusion, epitopes on at least some molecules that bind to spermatozoa or that remain free in the duct fluid are shared with those in other Natrix species, but not in more distant reptiles (turtle, anole lizard), nor chicken, rat, or rabbit. In regard to glycoproteins, one prominent con A-reactive band was present in polyacrylamide gel electrophoresis (PAGE) of snake fluid and more were evident in fluid collected from the turtle duct. However, such lectin-reactive elements did not bind to spermatozoa as judged by an absence of any change in snake, turtle and lizard sperm lectin-binding patterns in passing from the testis into and through the Wolffian duct. In all, evidence from these and other species studied begins to suggest that the nature of the post-testicular sperm surface modification displayed in most vertebrates that fertilize internally may differ in sub-therian and therian groups, respectively. There appears to be a relative emphasis on glycosyl-rich surface elements in the latter. The possible significance of these changes for sperm function in the different groups is discussed briefly in terms of sperm survival/storage, as well as capacitation and sperm binding to the zona.

Animals↗

Stealth PEG-PHDCA niosomes: effects of chain length of PEG and particle size on niosomes surface properties, in vitro drug release, phagocytic uptake, in vivo pharmacokinetics and antitumor activity.

A series of novel niosomes with the amphiphilic copolymer of poly (methoxy-polyethyleneglycol cyanoacrylate-co-n-hexadecyl cyanoacrylate) (PEG-PHDCA) acted as surface modification materials were prepared and Hydroxycamptothecin (HCPT) was used as a model drug. This work concentrated on the effects of PEG chain length and particle sizes on the niosomes surface properties, in vitro drug release, phagocytic uptake, in vivo pharmacokinetics and antitumor activity. Within the range of PEG Mw from 2000 to 10000, the increasing zeta potential (from -16.08 to -5.25 mv) and thicker fixed aqueous layer (3.82 to 5.78 nm) would facilitate the niosomes' stealth effects, while the reduced PEG chain density (from 0.53 to 0.17 PEG/nm2) and the quickened speed of drug release would diminish the effects. As a result, the PEG5000-PHDCA niosomes had the least phagocytic uptake, the longest half-life of 11.46 h and the best tumor inhibition rate of 97.1%. In the groups different in particle size (PEG5000-PHDCA niosomes from 92.5 to 204.6 nm), the bigger particles could be uptaken by macrophages more quickly, regardless of the changes of other physicochemical parameters. Correspondingly, PEG5000-PHDCA niosomes with particle sizes of 92.5, 144.2, 204.6 nm could extend the half-life of HCPT to 11.46, 6.33, 4.46 h, respectively. At last, the tumor inhibition rate of PEG5000-PHDCA niosomes (92.5 nm) at a dose of 2 mg/kg was five times that of HCPT injection at 4 mg/kg. The stealth effects of the PEG-PHDCA niosomes and the enhanced stability of lactone form of HCPT were accountable for the powerful antitumor effects of niosomes.

Algorithms↗

Thyroid stimulating hormone assays based on the detection of gold conjugates by scanning force microscopy.

Low bulk concentrations of thyroid stimulating hormone (TSH) were detected by scanning force microscopy (SFM) using gold-labeled conjugates. Anti-TSH antibodies were covalently bound onto amino-modified silicon oxide wafers. Surface modification was examined by contact-angle measurements, ellipsometry, X-ray photoelectron spectroscopy, and SFM. Antibodies were found to form a monolayer of prone molecules with an average surface density of 5000 IgG/mum2. TSH molecules were then allowed to bind to immobilized antibodies. The immunological reaction was quantified by SFM using gold-labeled species. Two scanned force microscopic immunoassays (SFMIA) were compared: first, a competitive test which used gold-labeled TSH molecules mixed with free TSH antigens was performed . Afterward, a sandwich assay was carried out, using gold-labeled anti-TSH antibodies. This latter method was found to be far more sensitive than competitive SFMIA. Gold conjugates were also found to be of great use to quantify antigens in large volumes by a sandwich test: a sensitivity threshold as low as 0.015 ng of TSH/ml (0.075 UI/ml or 6 x 10(-13) M) was estimated.

Binding, Competitive↗

Antiinfective and encrustation-inhibiting materials--myth and facts.

Catheters, urethral and ureteral stents and other urological implants are frequently affected by encrustration and infection due to their permanent contact with urine. Indwelling urinary catheters provide a haven for microorganisms and thus require extensive monitoring. Several surface modification techniques have been proposed to improve the performance of devices including the immobilization of biomolecules, the incorporation of hydrophilic grafts to reduce protein adsorption, the creation of hydrophobic surfaces, the creation of microdomains to regulate cellular and protein adhesion, new polymers and antimicrobial coatings. Physico-chemical explanation to elucidate the mechanism of such encrustation or infection inhibiting materials is still not available. Our series of experiments showed a marked decrease of silver-activity in biological fluids which corresponds with the controversial clinical results obtained with silver coated urinary catheters. Rifampicin/minocycline coated catheters had very low activity against Gram-negative rods, enterococci and Candida spp., the main causing organisms of urinary catheter infection. Surface engineered materials and antimicrobial drug delivery systems will be the next generation of sophisticated urinary catheters and stents, if both efficacy as well as efficiency has been proved clinically.

Anti-Bacterial Agents↗

Immobilization of amine-modified oligonucleotides on aldehyde-terminated alkanethiol monolayers on gold.

Chemistry is described for the fabrication of DNA arrays on gold surfaces. Alkanethiols modified with terminal aldehyde groups are used to prepare a self-assembled monolayer (SAM). The aldehyde groups of the monolayer may be reacted with amine-modified oligonucleotides or other amine-bearing biomolecules to form a Schiff base, which may then be reduced to a stable secondary amine by treatment with sodium cyanoborohydride. The surface modifications and reactions are characterized by polarization modulation Fourier transform infrared reflection absorption spectroscopy (PM-FTIRRAS), and the accessibility, binding specificity, and stability of the DNA-modified surfaces are demonstrated in hybridization experiments.

Aldehydes↗

Two-level antibacterial coating with both release-killing and contact-killing capabilities.

Using a combination of an aqueous layer-by-layer deposition technique, nanoparticle surface modification chemistry, and nanoreactor chemistry, we constructed thin film coatings with two distinct layered functional regions: a reservoir for the loading and release of bactericidal chemicals and a nanoparticle surface cap with immobilized bactericides. This results in dual-functional bactericidal coatings bearing both chemical-releasing bacteria-killing capacity and contact bacteria-killing capacity. These dual-functional coatings showed very high initial bacteria-killing efficiency due to the release of Ag ions and retained significant antibacterial activity after the depletion of embedded Ag because of the immobilized quaternary ammonium salts.

Anti-Bacterial Agents↗

Coating and selective deposition of nanofilm on silicone rubber for cell adhesion and growth.

A recently developed method for surface modification, layer-by-layer (LbL) assembly, has been applied to silicone, and its ability to encourage endothelial cell growth and control cell growth patterns has been examined. The surfaces studied consisted of a precursor, with alternating cationic polyethyleneimine (PEI) and anionic sodium polystyrene sulfonate (PSS) layers followed by alternating gelatin and poly-D-lysine (PDL) layers. Film growth increased linearly with the number of layers. Each PSS/PEI bilayer was 3 nm thick, and each gelatin/PDL bilayer was 5 nm thick. All layers were more hydrophilic than the unmodified silicone rubber surface, as determined from contact angle measurements. The contact angle was primarily dictated by the outermost layer. Of the coatings studied, gelatin was the most hydrophilic. A film of (PSS/PEI)4/(gelatin/PDL)4/ gelatin was highly favorable for cell adhesion and growth, in contrast to films of (PSS/PEI)8 or (PSS/PEI)8/PSS. Cell growth patterns were successfully controlled by selective deposition of microspheres on silicone rubber, using microcontact printing with a silicone stamp. Cell adhesion was confined to the region of microsphere deposition. These results demonstrate that the LbL self-assembly technique provides a general approach to coat and selectively deposit films with nanometer thickness on silicone rubber. Furthermore, they show that this method is a viable technique for controlling cellular adhesion and growth.

Animals↗

Quantitative bacterial analysis of porous, fabric, and smooth non-blood contacting implant surfaces and their tissue interfaces in a 169 day pneumatic total artificial heart animal recipient.

All long-term total artificial heart (TAH) survivals are subject to sepsis. Survival can be prolonged, but the source of the infection cannot be eliminated with any known course of antibiotics or treatment regimen. Sambo, a U-100 pTAH calf, survived 169 days. At week 6, he became septic, growing a Pseudomonas species (Ps). Weekly blood cultures were intermittently positive until week 13 when they became continuously positive until his demise, from a ruptured left ventricular pumping diaphragm. Spatially specific porous silicone rubber (SSP) was used for surface modifications on the drive lines and as cuffs around the Dacron TAH graft to large vessel anastomoses. This gave an excellent opportunity to examine two types of porous implants surfaces (Dacron grafts and SSP) to the smooth Biomer ventricular surfaces with their respective adjoining tissue interfaces for bacterial colonization. Nine tissue samples and 13 implant surfaces were processed with Costerton's quantitative bacterial techniques. The largest numbers of bacteria (> 10(6)/cm2 Ps.) were grown from the smooth ventricular surface and in the cul-de-sac where the SSP delaminated from the driveline (two smooth implant surfaces in contact but without tissue apposition). The Dacron grafts were intermediate in bacterial concentrations and SSP surface modified drivelines and tissues were sterile. In this model, the more intimate biointegration found in the porous implants showed improved bacterial resistance in a chronically infected pTAH. The more completely biointegrated and neo-vascularized porosity SSP was the only implant surface and opposing implant tissue interface sampled to remain sterile.

Animals↗

Modification of the complement binding properties of polystyrene: effects of end-point heparin attachment.

In recent years, conjugation of heparin to biomaterials has been shown to improve its biocompatibility. The purpose of the present work was to compare complement activation and binding of C3 to unmodified and heparin-treated polystyrene surfaces of microtitre plates. When polystyrene was incubated with human serum, C3 was deposited on the surface by both adsorption and binding dependent on activation of the classical (CPW) and alternative (APW) pathways. After end-point attachment of heparin, significant C3 deposition, although at reduced levels, occurred only by CPW-mediated mechanisms, while adsorption and APW-mediated binding were strongly reduced. Generally, the modified surface bound lower amounts of protein, e.g. serum albumin and IgG, than the unmodified. By contrast, it had increased affinity for C1q which leads to binding of C1 and activation of complement via the CPW. Nevertheless, the net effect of the surface modification on the complement reaction was an overall reduction of C3 binding due to obliteration of APW. This can be related to an enhanced factor H/I-dependent down-regulation of C3b and to the lowered protein-adsorbing property of the surface, both of which have inhibitory effects on APW and on the C3 shunt-dependent activation of the complement system.

Complement Activation↗

Glow discharge plasma treatment for surface cleaning and modification of metallic biomaterials.

Glow discharge plasma treatment is a frequently used method for cleaning, preparation, and modification of biomaterial and implant surfaces. The merits of such treatments are, however, strongly dependent on the process parameters. In the present work the possibilities, limitations, and risks of plasma treatment for surface preparation of metallic materials are investigated experimentally using titanium as a model system, and also discussed in more general terms. Samples were treated by different low-pressure direct current plasmas and analyzed using Auger electron spectroscopy (AES), x-ray photoelectron spectroscopy (XPS), atomic force microscopy, scanning electron microscopy, and light microscopy. The plasma system is a home-built, ultra-high vacuum-compatible system that allows sample introduction via a load-lock, and precise control of pressure, gas composition and flow rate, etc. This system allows uniform treatment of cylindrical and screw-shaped samples. With appropriate plasma parameters, argon plasma remove all chemical traces from former treatments (adsorbed contaminants and other impurities, and native oxide layers), in effect producing cleaner and more well-controlled surfaces than with conventional preparation methods. Removal (sputtering) rates up to 30 nm/min are possible. However, when inappropriate plasma parameters are used, the result may be increased contamination and formation of unintentional or undesired surface layers (e.g., carbides and nitrides). Plasma-cleaned surfaces provide a clean and reproducible starting condition for further plasma treatments to form well-controlled surface layers. Oxidation in pure O2 (thermally or in oxygen plasmas) results in uniform and stoichiometric TiO2 surface oxide layers of reproducible composition and thicknesses in the range 0.5-150 nm, as revealed by AES and XPS analyses. Titanium nitride layers were prepared by using N2 plasmas. While mild plasma treatments leave the surface microstructure unaffected, heavy plasma treatment can give rise to dramatic morphologic changes. Comparison of these results with corresponding analyses of commercial implants and electropolished and/or anodically oxidized samples shows that the plasma treatment offers superior control of the surface status. However, it is also shown that improper control of the plasma process can produce unwanted and irreproducible results.

Argon↗

Cell mimetic monolayer supported chitosan-haemocompatibility studies.

Chitosan is a natural polymer, widely explored for biomedical and tissue engineering applications. However the thrombogenic nature limits their application in blood contacting devices and implants. Here, we have attempted to understand the haemocompatibility of chitosan by immobilizing a monolayer of cell mimetic lipid compositions. The phosphatidylcholine/cholesterol/galactocerebroside lipid composition (PC/Chol/GalC, 1:0.35:0.125) was deposited onto the chitosan films. Characterization of the modified surface was done by sessile drop contact angle measurement. The contact angle of the chitosan film reduced from 80.65 +/- 1.4 to 23.5 +/- 1.9 after the surface modification. Swelling nature of chitosan seemed to influence the orientation and packing of the lipid monolayer. In vitro calcification studies with metastable salt solution indicated increased calcification on the modified surface. This may be due to formation of nuclei for calcification on the expanding monolayer. The preliminary haemocompatibility studies with washed platelets, leukocytes and erythrocytes showed overall reduction in blood cell adhesion to the modified surfaces. Scanning electron microscopy was used for morphological characterization of platelet adhesion and activation on the surfaces. On the bare chitosan surface, fully spread platelets with extending pseudopodia indicated platelet activation. The smooth surface of the modified film did not activate platelets. These studies showed that, though the lipid monolayer on chitosan film is able to reduce the over all blood cell adhesion and platelet activation it is prone to calcification.

Biocompatible Materials↗