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Selection of two reliable parameters to evaluate the impact of the mobile-phase composition on capillary electrochromatography performance with monolithic and particle-packed capillary columns.

Different models have been described in the literature to evaluate the total porosity of CEC columns: gravimetric, flow as well as conductivity-based methods. In this study, these models have been compared for two kinds of CEC columns: two mixed-mode silica particle stationary phases and different monolithic columns (acrylate or polystyrene divinylbenzene-based). The total porosities measured from the conductivity-based methods were lower than the total column porosities obtained by gravimetric or flow methods for all the investigated columns while the wide distribution of observed values shows that conductivity-based methods discriminate columns more efficiently with very different properties. We propose a conductivity-based method taking into account the actual length proposed by Horvath, to evaluate what we call an "actual electrokinetic" porosity (AEP). This parameter, based on electrokinetic theory only, affords the most consistent evaluation of porosity under experimental CEC conditions for the packed- and acrylate-based monolithic columns. To illustrate the potential of AEP and actual EOF for the estimation of the performances of a CEC system (stationary and mobile phases) we studied the influence of the mobile-phase composition on these parameters for CEC separations with an ammonium embedded packed stationary phase. The AEP and the actual electroosmotic mobility should allow a better understanding of the perfusive EOF and stationary-phase wettability. For neutral compounds (substituted phenols), AEP evaluation allowed us to predict the mobile-phase conditions able to enhance the efficiency while both AEP and actual EOF had to be considered in the case of peptide analysis.

Acrylates↗

Fractionation of proteins in reconstituted skim-milk.

Properties of the dried milk were improved when calcium chloride was added to the raw milk prior to the drying process. Solubility, wettability, undenatured whey protein and protein retention indices of the dried milk were increased and the reconstitutability was improved. The elution profile of the protein fractions on a Sephadex G-100 column differed according to the forward preheating of the raw milk and the addition of calcium chloride.

Animals↗

Plasma surface modification of synthetic absorbable sutures.

The aim of the study was to examine the feasibility of using plasma surface modification technology to alter the hydrolytic degradation rate of commercial synthetic absorbable sutures. Size 2-0 Dexon, Vicryl, PDSII, and Maxon sutures were tested. They were treated by two different surface modification techniques: parylene deposition and plasma gases (Methane, trimethylsilane, and tetrafluoroethene). The thickness of surface treatment ranged from 200 to 1000 A. The treated sutures were subject to in vitro hydrolytic degradation in phosphate buffer of pH = 7.4 at 37 degrees C for up to 120 days. The tensile breaking strength, weight loss, surface wettability, bending stiffness, and surface morphology were evaluated. The results indicated that the concept of plasma surface treatment for altering the hydrolytic degradation of synthetic absorbable sutures was feasible, and the level of improvement depended on the type of sutures, the treatment conditions, and the duration of hydrolysis. Vicryl and PDSII sutures showed overall the best improvement in tensile strength retention among the four commercial sutures. Dexon and Maxon sutures, however, exhibited only marginal improvement. The observed improvement in tensile strength retention appeared to be related to the increasing hydrophobicity of the sutures. The surface treatments did not adversely affect the bending stiffness of the sutures and no visible surface morphological changes were observed. Refinements and optimization of the surface treatment conditions are needed for achieving the maximum advantage of the proposed concept, particularly shielding the harmful effect of uv during plasma treatment.

Biodegradation, Environmental↗

Visible-light-induced surface graft polymerization via camphorquinone impregnation technique.

A surface modification method that is particularly applicable to complexly shaped fabricated devices has long been awaited. In this article, we describe the visible-light-induced surface photograft polymerization technique by which an inner surface of the device is modified by visible-light irradiation through the external surface. Comphorquione, as a photoradical initiator, was impregnated on a segmented polyurethane surface by solvent soaking, followed by visible-light irradiation in the presence of monomers such as acrylamide and poly(ethylene glycol) methacrylate. The resultant surfaces were highly wettable with water, and surface chemical compositional analysis by X-ray photoelectron spectroscopy revealed that the surface was graft-polymerized with these monomers. The simple and widespread applicability of this surface modification technique to biomedical devices is discussed.

Biocompatible Materials↗

Ultrathin coating by plasma polymerization applied to corneal contact lens.

Plasma coating (deposition of polymer under the influence of plasma) is utilized to modify the surface properties of corneal contact lens. An ultrathin layer (thickness of roughly 200 A) of plasma polymer of acetylene/H20/N2 is applied to poly(methyl methacrylate) (PMMA) corneal contact lenses. The surface becomes highly wettable with water compared to uncoated lens. When coated and uncoated contact lenses are placed on rabbit eyes, a remarkable difference in accumulation of mucous matter is observed. With the control lenses, the accumulation of mucous matter in a week is sufficient to affect the optical clarity of the lenses, whereas the coated lenses show no change after three months continuous wearing. The comparative degree of adhesion of the corneal epithelium cells onto glass, modified glass, PMMA, and coated PMMA surfaces is studied using tissue cultures and phase contrast microscopy. The coated PMMA surface exhibits a degree of tissue adhesion lower than that of control PMMA and higher than that of glass surface, and no sign of toxicity of the coated surface is observed by the tissue cultures.

Acetylene↗

Effects of polymer surface molecular structure and force-field characteristics on blood interfacial phenomena.

To quantify the effects of major surface structural factors influencing interfacial reactions induced by polymers in native blood, model surfaces of solvent-cast films of two analogous poly(ether urethanes) and three homologous polyamides (nylon 4, 6/6, and 12) were exposed ex vivo to canine blood under the well-defined hemodynamic conditions of the Stagnation Point Flow Experiment. The selected surfaces allow for incremental changes in properties and were characterized by their "Composite Surface Free ENergy Function," gamma'S, which describes the surface force field as the sum of the mean dispersion (gammaSd) and polar (gammaSp) contributions and is computed from wettability spectra obtained with ultrapure diagnostic liquids. Blood interfacial effects were measured by the shear-limited diameter of the white cell circle formed around the stagnation point, the flow parameter at which symmetric aggregation occurred, and the surface-number density of platelets, [P s], remaining adherent under fixed conditions. At identical flows, within each group of polymers, both the WBC-circle diameter and [P s] scale with gamma Sp/gamma'S, implying that 1) only the magnitude but not the interaction mechanism varies as a function of incremental structural and surface changes, 2) the primary determinant of surface-induced effects is the polar force contribution, and 3) the magnitude of gamma'S is secondary if gammaSd/gamma'S is sufficiently great.

Adsorption↗

New prospects for a prolonged functional life-span of artificial hip joints by using the material combination polyethylene/aluminium oxide ceramin/metal.

Investigations over the years have shown that the mirror-finished Al2O3 ceramic is a much more suitable frictional counterpart to ultrahigh molecular weight (UHMW) polyethylene than metal. Despite the extremely gread hardness difference between polyethylene and Al2O3 ceramic, a considerable lower wear rate is obtained for the polyethylene socked with this new low-friction material combination. The unexpectedly favorable tribological behavior of this ceramic material in contact with polyethylene may be attributed to the following factors: better values for corrosion resistance characteristics, wettability with liquids, surfact gloss, hardness, and scratch resistance of the ceramic material in comparison with those of the hitherto used metallic implant materials (AISI-316L steel or cast Co-Cr-Mo alloy). It appears that, by using this new combination of materials for the socket and the ball, it will be possible to prolong the service life of artificial hip joints considerably without having effecy any fundamental changes in the present design and implantation principle retaining the hitherto used anchorage shaft made of wrought Co-Ni-Cr-Mo-Ti alloy Protasul-10 of extremely high corrosion fatigue strength.

Alloys↗

Adsorption characteristics of plasma fibronectin in relationship to biological activity.

We studied the adsorption properties of plasma fibronectin (pFN) on wettable tissue culture (TC) dishes and nonwettable bacteriological (BAC-T) dishes in relationship to its biological activity of promoting fibroblast spreading. The binding of pFN to the dish surfaces was found to be very tight and partially resistant to treatments with 1M NaOH, 2% SDS, 8M urea, or 6M guanidine HCl. Only the combination of trypsin (1 mg/ml) followed by 1M NaOH resulted in complete recovery of surface bound material. The amount of surface bound pFN did not directly correlate with its activity when comparing TC and BAC-T dishes. At low concentrations, more pFN adsorbed onto the surfaces of BAC-T dishes than TC dishes, but the adsorbed material was biologically less active on BAC-T dishes. At high concentrations, pFN adsorbed similarly on both dish surfaces reaching a maximum level of 320 ng/cm2 or 4.4 x 10(11) molecules/cm2. The possibility was considered that pFN unfolded into an inactive conformation when adsorbed on BAC-T dishes at a low concentration but that at high concentrations, molecular packing requirements prevented unfolding. Evidence in favor of this hypothesis was the observation that addition of 50 micrograms/ml of serum albumin simultaneously with low pFN concentrations resulted in adsorption of pFN on BAC-T dishes in an active conformation even though the amount of adsorbed pFN decreased. Competition experiments between pFN and other proteins were carried out, and it was found that pFN has a much higher affinity for BAC-T or TC dish surfaces than albumin or fibrinogen. It was also found that mild heat denaturation of albumin increased its affinity for the surface by an order of magnitude.

Adsorption↗

Ex vivo shunt testing of hydrogel-silicone rubber composite materials.

Hydrogel polymers consisting of HEMA (2-hydroxyethyl methacrylate) alone or co-polymerized with NVP N-(vinyl pyrrolidone) were incorporated into silicone rubber tubing to produce hydrophilic composite materials. Polymerization methods of the interpenetrating network (IPN) and radiation-induced type were used to impregnate the silicone rubber substrates. These methods produced materials with varied wettability (contact angle with water ranged from 57 degrees to 95 degrees) but similar texture. The surface morphology as observed in the scanning electron microscope was the same for all the materials. Four types of composite materials and two sizes of silicone rubber control tubing in a total of 19 dogs were used as ex vivo femoral artery-to-vein (A-V) shunts and were sampled at 5, 15, 30, 45, and 60 min. Data obtained by using scanning electron microscopy (SEM) were used to determine the type and amount of adhering blood cells at the time intervals described. Large platelet aggregates and/or a variable initial response were found to be predictors of shunt failure (occlusion). Most samples showed only varied levels of platelet adhesion, but other cell types were seen in some samples. The adhesion was found to peak in all cases by 30 min and to reach lower levels by 60 min, suggesting that an initial phase of reaction was complete. The course of platelet adhesion for a 60-min period is presented.

Animals↗

Adhesion of coagulase-negative staphylococci to methacrylate polymers and copolymers.

Adhesion of coagulase-negative staphylococci (CNS) was studied onto a homologous series of methacrylate polymers and copolymers. The materials varied in wettability (contact angles) and were either positively or negatively charged (zeta-potential). Bacterial adhesion experiments performed in a parallel-plate perfusion system showed that positively charged TMAEMA-Cl copolymers significantly promoted the adhesion of CNS as compared with all other methacrylate (co)polymers tested. The bacterial adhesion rates onto the positively charged surfaces are diffusion-controlled, whereas those onto the surfaces with a negative zeta-potential are more surface-reaction-controlled due to the presence of a potential energy barrier. The bacterial adhesion rates onto various poly (alkyl methacrylates) were similar. The number of adhering bacteria onto the negatively charged MMA/MAA copolymer did not differ from that onto pMMA, indicating that sufficient sites on the copolymer surface with the same potential energy barrier as that on pMMA, were available for adhesion. Decreasing rates of adhesion of CNS were observed onto MMA/HEMA copolymers with increasing HEMA content coinciding with increasing hydrophilicity. Low plateau values for the bacterial adhesion were observed on 50MMA/50HEMA, pHEMA, and 85HEMA/15MAA, indicating that the adhesion onto these materials was reversible. Four CNS strains with different surface characteristics all showed higher numbers of adhering bacteria onto 85MMA/15TMAEMA-Cl than onto 85MMA/15MAA and pMMA.

Binding Sites↗

The influence of protein adsorption on interactions of cultured human endothelial cells with polymers.

A systematic study of the effects of polymer surface properties on the interaction with human endothelial cells (HEC) may lead to the development of small-diameter vascular grafts. HEC, suspended in culture medium containing 20% serum adhered and spread onto moderately wettable polymers such as TCPS (tissue culture polystyrene). Reduced or no adhesion of HEC was observed upon the hydrophobic polymers PETP (polyethyleneterephthalate, Dacron) and FEP (fluoroethylenepropylene copolymer, Teflon). Polymers precoated with the proteins albumin (Alb), high density lipoprotein (HDL), and immunoglobulin G (IgG) inhibited the adhesion of HEC, whereas fibronectin (Fn) coatings promoted cell adhesion. Endothelialization of PETP and FEP only occurred after precoating of these materials with Fn. The adsorption of Fn, Alb, HDL, and IgG from solutions of different serum concentrations onto TCPS, PETP, and FEP was related to the adhesion of HEC. Serum Fn only adsorbed onto TCPS, with the maximum at 0.1% serum concentration. Maximal cell adhesion onto TCPS was also observed after pretreatment with a solution containing 0.1% serum. The cell adhesion inhibiting proteins Alb and HDL preferentially adsorbed at higher serum concentrations. Desorption of these proteins and exchange for, e.g., cellular Fn may result in cell spreading and proliferation of HEC upon TCPS.

Adsorption↗

Adsorption of fibronectin derived from serum and from human endothelial cells onto tissue culture polystyrene.

Human endothelial cells (HEC) suspended in a culture medium containing 20% human serum (CMS) adhere and spread on(to) moderately wettable polymers, such as tissue culture polystyrene (TCPS). We have previously shown that serum derived-fibronectin, which is a cell adhesion promoting protein, has a high affinity for TCPS, but that the amount of fibronectin which adsorbed from CMS was relatively small. In this study we investigated whether fibronectin derived from HEC contributes to the adhesion and spreading of the cells on(to) TCPS. Therefore, HEC were seeded in the presence of fibronectin-depleted CMS. The amount of fibronectin detected on TCPS increased with both cell seeding density and incubation time. Although initial HEC adhesion is delayed on TCPS which has been precoated with albumin (Alb), high density lipoprotein (HDL) or immunoglobulin G (IgG), maximal numbers of adhering and spreading HEC were found on these surfaces 6 h after seeding of HEC. Fibronectin was detected on these surfaces, but an exchange of preadsorbed Alb, HDL, or IgG for fibronectin could not be demonstrated. We conclude that HEC deposit fibronectin onto TCPS, irrespective of the presence of a preadsorbed layer of proteins which delay cell adhesion.

Adsorption↗

Serotonin and beta-thromboglobulin release reaction from platelet as triggered by interaction with polypeptide derivatives.

The effect of wettability of 14 polypeptide derivatives upon adhesion and activation of platelets was investigated with reference to release reactions from adhered platelets, using radioisotope labeling and radioimmunoassay method. The serotonin release was more significant from platelets adhered to polymer materials to which a large number of platelets are adhered. However, no clear relationship was found between adhesion of platelets and beta-thromboglobulin release from adhered platelets. Therefore, stimuli inducing serotonin release and beta-thromboglobulin release were considered to be from different origins. The trend in beta-thromboglobulin release was well correlated with the extent of morphological change of adhered platelets as observed by scanning electron microscope. It was suggested that the determination of released beta-thromboglobulin in association with the measurement of platelet adhesion could be useful for evaluation of blood compatibility of materials.

Blood Platelets↗

Attachment and growth of fibroblast cells on polypeptide derivatives.

Attachment and growth of mouse fibroblast cells on polypeptide derivatives with different wettabilities were studied in the presence or absence of serum proteins. In the presence of serum, a peak level of cell attachment was observed for substrates with a contact angle of around 70 degrees. However, no relationship was found between cell attachment and water contact angle of the substrate in the absence of serum. Ca2+ -dependent cell attachment was observed on hydrophobic polypeptides in the absence of serum proteins. This suggests that Ca2+ -dependent membrane proteins function as mediators for cell attachment to hydrophobic polypeptides. In the presence of serum proteins, it was found that cell attachment is affected by the metabolism, Ca2+, and the cytoskeleton of the cell. Cell growth rate on hydrophilic polypeptide substrates was higher than that on intermediate or hydrophobic polypeptide substrates. This demonstrates that the strong interaction between cells and substrates will be unfavorable for a dewebbing process during mitosis.

Animals↗

Human erythrocyte adhesion and spreading on protein-coated polymer surfaces.

Protein adsorption is the first event which occurs when polymer surfaces are exposed to blood. The adsorption of proteins modifies the surface properties of the substrates and therefore influences subsequent cell-surface interactions. In an attempt to elucidate the fundamental mechanisms governing cell-proteinated-surface interactions, the extent of fresh human erythrocyte adhesion and spreading on protein-coated surfaces was examined. Five human serum proteins (albumin, fibrinogen, immunoglobulin G, fibronectin, and transferrin) were used at bulk concentrations ranging from 0.01 mg/mL to 50 mg/mL. Polymer substrates covering a wide range of wettability were employed. Protein adsorption significantly reduces erythrocyte adhesion and spreading on all test surfaces with minimum adhesion observed on fibrinogen: IgG greater than albumin greater than fibronectin greater than transferrin greater than fibrinogen. The extent of these effects is dependent on the nature of the adsorbed protein, the protein bulk concentration, and the surface properties of the underlying polymer substrates.

Biopolymers↗

Dental implant materials. I. Some effects of preparative procedures on surface topography.

The effect of different treatments for preparing implant materials was examined by scanning electron microscopy and by contact angle measurements. The materials examined were Ti6A14V alloy, Co-Cr-Mo alloy, A12O3, and synthetic hydroxyapatite. Samples were prepared with solid or porous surfaces of these materials. These were detergent-cleaned and then either autoclaved (steam sterilization), radiation-sterilized, nitric acid-etched, or plasma-cleaned. The results of wettability studies indicated marked changes in surface energy corresponding to the different preparation methods, and differences in surface morphology were also observed. These differences could have significant consequences on in vivo implant behaviour as mediated by tissue-implant interactions.

Alloys↗

Protein adsorption to poly(ethylene oxide) surfaces.

Surfaces containing poly(ethylene oxide) (PEO) are interesting biomaterials because they exhibit low degrees of protein adsorption and cell adhesion. In this study different molecular weight PEO molecules were covalently attached to poly(ethylene terephthalate) (PET) films using cyanuric chloride chemistry. Prior to the PEO immobilization, amino groups were introduced onto the PET films by exposing them to an allylamine plasma glow discharge. The amino groups on the PET film were next activated with cyanuric chloride and then reacted with bis-amino PEO. The samples were characterized by scanning electron microscopy, water contact angle measurements, gravimetric analysis, and electron spectroscopy for chemical analysis (ESCA). The adsorption of 125I-labeled baboon fibrinogen and bovine serum albumin was studied from buffer solutions. Gravimetric analysis indicated that the films grafted with the low-molecular-weight PEO contained many more PEO molecules than the surfaces grafted with higher-molecular-weight PEO. The high-molecular-weight PEO surfaces, however, exhibited greater wettability (lower water contact angles) and less protein adsorption than the low-molecular-weight PEO surfaces. Adsorption of albumin and fibrinogen to the PEO surfaces decreased with increasing PEO molecular weight up to 3500. A further increase in molecular weight resulted in only slight decreases in protein adsorption. Protein adsorption studies as a function of buffer ionic strength suggest that there may be an ionic interaction between the protein and the allylamine surface. The trends in protein adsorption together with the water contact angle results and the gravimetric analysis suggest that a kind of "cooperative" water structuring around the larger PEO molecules may create an "excluded volume" of the hydrated polymer coils. This may be an important factor contributing to the observed low protein adsorption behavior.

Adsorption↗

Surface energy characterization of unalloyed titanium implants.

Osteointegration is dependent on a variety of biomechanical and biochemical factors. One factor is the wettability of an implant surface that is directly influenced by its surface energy. This investigation used the Zisman plot to determine critical surface tension as one representative measurement of surface energy. The effects of surface treatment, bulk grain size, and surface roughness on the critical surface tension of unalloyed titanium (Ti) were examined. Radio frequency glow discharge-treated Ti had the highest critical surface tension, followed by the passivated and heat-sterilized conditions. Titanium with no surface treatment had the lowest critical surface tension. The surface energy of Ti with an average grain size of 23 microns was not significantly different from that with a grain size of 70 microns. Surface roughness was shown to cause significant difference in measurements and definitely should be considered in studies of this kind.

Prostheses and Implants↗