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J Feijen

Publications and source records attributed to J Feijen.

At least 145 records · Page 8Linked to original sources

An in vitro study of the adhesion of blood platelets onto vascular catheters. Part I.

The adhesion of human blood platelets onto vascular catheters was studied using a specially designed perfusion chamber. Polyurethane catheters were exposed to citrated human blood for different periods (up to 20 min) and at different wall shear rates (190, 260, 330 sec-1). The rate of platelet adhesion was determined using 111In-labeled platelets, while the morphology of adhering platelets was investigated using scanning electron microscopy. A linear increase in platelet adhesion was found within the first 10 min of perfusion, after which a plateau value was reached. The number of adhering platelets did not vary significantly with the shear rates applied, which may indicate that within the range of shear rates studied, the adhesion of platelets onto the catheter surface is mainly determined by the rate of the reaction between the platelets and the material surface. Catheters coated with a conjugate of heparin and albumin showed a four- to five-fold reduction in platelet adhesion as compared to uncoated catheters. This reduction in platelet adhesion was not only due to the presence of albumin moieties at the surface but also to the presence of heparin residues in the adsorbed albumin-heparin conjugate.

Albumins↗

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↗

Adhesion of cultured human endothelial cells onto methacrylate polymers with varying surface wettability and charge.

The adhesion of human endothelial cells (HEC) onto a series of well-characterized methacrylate polymer surfaces with varying wettabilities and surface charges was studied either in serum-containing (CMS) or in serum-free (CM) culture medium. HEC adhesion in CMS onto (co)polymers of hydroxyethyl methacrylate (HEMA) and methyl methacrylate (MMA) was found to be optimal on the moderately wettable copolymer (mol ratio 25 HEMA/75 MMA). Positively-charged copolymers of HEMA or MMA with trimethylaminoethyl methacrylate-HCl salt (TMAEMA-Cl), both with mol ratios of 85/15 and a negatively-charged copolymer of MMA with methacrylic acid (MAA), mol ratio 85/15, showed high numbers of adhering HEC. In CM, HEC adhered onto the three charged copolymers mentioned above, but neither onto the copolymer of HEMA and MAA (mol ratio 85/15) nor onto the HEMA/MMA co- and homopolymers. Complete cell spreading in CM was only observed on the positively-charged copolymers.

Acrylates↗

In vivo quantification of cell-polymer interactions.

An in vivo rat model was developed to determine cell-polymer interactions under physiological conditions. Microporous tubular grafts, made of polytetrafluoroethylene, a polyetherurethane, a polyesterurethane and also a modified polyetherurethane were implanted intraperitoneally in rats. The grafts were filled with cultured rat smooth muscle cells prior to implantation. At t = 0, 2 and 48 h, the grafts were evaluated macroscopically and also prepared for light microscopy and for cell count of their contents. At t = 0 no cellular attachment was observed on the lumenal side of the capsules. At t = 2 h a monolayer of smooth muscle cells could be observed on all materials except PTFE, on which only small patches of cells were observed. At t = 48 h a multilayer of cells was seen on all materials except PTFE. Cell counts at 48 h demonstrated no multiplication in the PTFE graft but a 1.4, 2.3 and 2.0-fold multiplication in the polyetherurethane, polyesterurethane and the modified polyurethane grafts respectively. These in vivo results show a clear linear relationship with our in vitro results in which it has been proved that cell spreading increased with increasing substratum surface free energy. This rat model allows the study of cell-polymer interactions in vivo, in a standardized way, under controlled physiological conditions.

Animals↗

Coupling of naltrexone to biodegradable poly(alpha-amino acids).

The narcotic antagonist naltrexone (I) was modified at the 3 and 14 hydroxyl positions and covalently coupled to a biodegradable poly(alpha-amino acid) backbone through a labile bond. Selective acetylation of I with acetic anhydride gave naltrexone-3-acetate (II), which was subsequently succinoylated to naltrexone-3-acetate-14-hemisuccinate (III) with succinic anhydride. The polymeric backbone chosen for initial coupling experiments was poly-N5-(3-hydroxypropyl)-L-glutamine (PHPG). The side-chain hydroxyl functionality permitted covalent bonding of III through an ester linkage. Hydrolysis of covalently bound drug to give naltrexone or its derivatives (II and III) should be much slower than diffusion of drug through the polymer matrix. While hydrolysis of naltrexone from the polymer side chain is first order, release of drug from the matrix can be zero order due to the geometry of the device and the physical and chemical interactions between naltrexone and the polymer matrix. In vitro studies of PHPG-naltrexone conjugate in disk form did not show constant release because of the hydrophilic nature of the polymer backbone and the changing local chemical environment upon hydrolysis of drug-polymer linkages. The conjugated system was made more hydrophobic by coupling drug to copolymers of hydroxypropyl-L-glutamine (HPG) and L-leucine. Conjugates of III coupled with copoly(HPG-70/Leu-30) demonstrated a nearly constant, but slightly declining release rate of naltrexone and its derivatives for 28 days in vitro.

Amino Acids↗

Competitive adsorption of plasma proteins at solid-liquid interfaces.

The competitive adsorption of human serum albumin (HSA), human immuno-gamma-globulin (HIgG) and human fibrinogen (HFb) onto polystyrene (PS) at 20 degrees C and a pH of 7.35 (phosphate-buffered saline) was studied. Protein adsorption was studied using enzyme immunoassay. The results obtained with the immunoassay were compared with those obtained using radiolabelled proteins. Recent studies revealed that the adsorption behaviour of radiolabelled proteins onto surfaces differs from that of the non-labelled proteins, which may lead to misinterpretation of adsorption data. Differences in the adsorption behaviour of the labelled proteins as compared to non-labelled proteins can possibly be explained by the formation of modified proteins during the labelling procedure as shown by ion-exchange high-performance liquid chromatography (HPLC). The competitive adsorption of HSA, HIgG and HFb onto a PS latex was studied by measuring the depletion of proteins in solution. The decrease in protein concentration in solution was determined by HPLC techniques. A strong preferential adsorption of HFb was observed with maximum adsorption values of 0.6 micrograms/cm2.

Adsorption↗

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↗

Cell surface characteristics of coagulase-negative staphylococci and their adherence to fluorinated poly(ethylenepropylene).

The ability of 21 nonencapsulated and 15 encapsulated coagulase-negative staphylococci (CNS) to adhere to xylene in xylene-water emulsions and to fluorinated poly(ethylenepropylene) (FEP) films revealed remarkable differences. Nonencapsulated CNS strains adhered well to FEP, whereas their adherence to xylene ranged widely. Encapsulated strains with low adherence to xylene showed slight adherence to FEP. Encapsulated strains which adhered well to xylene ranged widely in their adherence to FEP. It was concluded that results obtained from the xylene adherence test were not predictive of the adherence of CNS to the hydrophobic FEP surface. The number of nonwashed, slime-producing CNS strains adhering to FEP was similar to that of washed bacteria of the same strains. Bacterial adherence to FEP was decreased when FEP films were exposed to a solution containing extracellular products (EP) obtained from a slime-producing CNS strain. Bacterial adherence to xylene also decreased when the bacterial suspensions contained EP. Apparently, initial adherence of CNS to FEP and xylene is hampered by EP. Nonencapsulated and encapsulated CNS pretreated with proteolytic enzymes failed to adhere to xylene and FEP, indicating that intact surface proteins or constituents associated with surface proteins mediated their adherence to xylene and FEP. Freeze-etch replicas of a CNS strain adhering to FEP showed a smooth, flattened area on the bacterial surface at the contact site of the bacteria with the FEP, indicating that an external layer was present at the bacterial surface.

Adhesiveness↗

Interaction of cultured human endothelial cells with polymeric surfaces of different wettabilities.

The in vitro interaction of human endothelial cells (HEC) and polymers with different wettabilities in culture medium containing serum was investigated. Optimal adhesion of HEC generally occurred onto moderately wettable polymers. Within a series of cellulose type of polymers the cell adhesion increased with increasing contact angle of the polymer surfaces. Proliferation of HEC occurred when adhesion was followed by progressive flattening of the cells. Our results suggest that moderately wettable polymers exhibit a serum and/or cellular protein adsorption pattern that is favourable for growth of HEC.

Cell Adhesion↗

Adhesion of Staphylococcus epidermidis and Staphylococcus saprophyticus to a hydrophobic biomaterial.

The relative surface charge and hydrophobicity of 16 strains of Staphylococcus epidermidis showed large variations. For this species no relationship between the two surface parameters was found. A highly negative surface charge was observed in all seven encapsulated strains (one S. epidermidis and six Staphylococcus saprophyticus strains). The adhesion of the staphylococci to fluorinated polyethylene-propylene films was not related to the relative surface charge and the hydrophobicity of the bacteria. On films pre-exposed to human plasma, the bacterial adhesion was substantially reduced. Mechanisms involved in the adhesion of coagulase-negative staphylococci to this biomaterial are discussed.

Adhesiveness↗

Complement inhibitory and anticoagulant activities of fractionated heparins.

Almost monodisperse heparin fractions (Mw/Mn less than 1.1) were obtained by gel filtration of a commercial heparin. These fractions were assayed for anticoagulant activity (thrombin times and APTT), chromogenic anti-factor Xa activity, inhibitory activity for the human classical complement pathway, carboxyl group content and total sulfate content. Linear relationships were observed between the molecular weight of the heparin fractions and the anti-coagulant activities as determined by thrombin time- and APTT-assay and the classical complement pathway inhibitory activity. On the other hand a hyperbolic-like relationship was observed between the molecular weight of the heparin fractions and the chromogenic anti-factor Xa activity. The heparin fractions did not show significant differences with respect to the carboxyl group and total sulfate content. Low- and high affinity heparin fractions were obtained by affinity chromatography using immobilized AT III. High- and low-affinity fractions greatly differed not only with respect to their APTT activity, but also where their complement-inhibitory activities were concerned. The latter in contrast to literature data available. These differences could not be explained by the observed differences in molecular weight of high and low affinity heparin respectively.

Animals↗

Inhibition of surface induced coagulation by preadsorption of albumin-heparin conjugates.

Surface coatings of the albumin-heparin conjugates were developed to improve the blood compatibility of polymeric materials. Glass, PVC, Biomer and cellulose acetate were coated with albumin-heparin conjugate and its adsorption and desorption behavior on glass in particular was studied using 3H and 51Cr radiolabeled conjugates. Precoated materials showed a significant prolongation of the Lee-White clotting time as compared with noncoated ones. It was demonstrated that the prolonged clotting time for pretreated glass was due to surface bound conjugate. Prolonged recalcification times of plasma exposed to glass, Biomer, and PVC were obtained using albumin-heparin conjugate precoated surfaces. Albumin-heparin conjugates with high affinity for antithrombin III gave more prolonged clotting times as low affinity conjugates when used as coatings for glass. This indicates that the behaviour of heparin in preadsorbed conjugates resembles that of heparin in solution.

Adsorption↗

The adsorption behaviour of two commercial IgG-preparations onto a polystyrene latex surface.

The adsorption behaviour of two commercial preparations of human IgG onto a polystyrene latex surface was studied. The adsorption isotherms obtained differed markedly; one preparation showed a plateau value of 0.4 microgram cm-2 which was reached at 0.1 g l-1, whereas the other preparation showed no plateau value within the concentration range studied (0.1-7.0 g l-1). Characterization by means of iso-electric focusing and HPLC also showed differences between the two preparations. No differences were observed when immuno-electrophoresis was carried out. These results stress the necessity for proper characterization of proteins used in adsorption studies.

Adsorption↗

Interaction of antithrombin III with preadsorbed albumin-heparin conjugates.

The adsorption of antithrombin III (AT III) onto polystyrene surfaces preadsorbed with albumin or albumin-heparin conjugates was studied using a two step enzyme immuno assay. When AT III-buffer solutions were used, the highest adsorption values were measured on high affinity albumin-heparin conjugate pretreated surfaces. Less AT III adsorption was found on nonfractionated albumin-heparin conjugate preadsorbed surfaces. AT III adsorption could also be detected on low affinity conjugate and albumin coated surfaces. When AT III was adsorbed from plasma or plasma dilutions with buffer, only AT III on surfaces preadsorbed with high affinity or nonfractionated albumin-heparin conjugate was found. These results demonstrate that the heparin moiety of the conjugate is directed to the solution phase whereas the albumin moiety contacts the polystyrene surface.

Adsorption↗

New perspectives in myringoplasty.

Until 1950 the treatment of a perforated eardrum consisted of covering the drum permanently with artificial material. Since then a surgical technique to establish a functional reconstruction of the eardrum was developed (myringoplasty). A survey of the biological grafting materials used in this technique is given. Biodegradable and non-degradable synthetic materials may prove to be a valuable supplement of the existing biological grafting materials. Artificial eardrums made from several biodegradable poly(alpha-hydroxy acids) and poly(alpha-amino acids) and made from a number of microporous poly(tetrafluorethylene) membranes and from a microporous bisphenol-A poly(carbonate) membrane were implanted into the ears of rats and dogs and as a reference subcutaneously. The implants were histologically examined for periods up to one year. From the biodegradable polymers studied poly(beta-benzyl-L-aspartate-co-L-leucine) 50/50 evoked the least tissue reaction and the newly formed eardrums were the best in terms of thickness and overall integrity. The formation of a reinforced eardrum may be accomplished by the support of an inert, very thin, highly porous poly(tetrafluoroethylene membrane) preferably implanted as a composite graft with a biodegradable polymer.

Animals↗

Covalently bound conjugates of albumin and heparin: synthesis, fractionation and characterization.

Covalently bound conjugates of human serum albumin and heparin were prepared as compounds which could improve the blood-compatibility of polymer surfaces either by preadsorption or by covalent coupling of the conjugates onto blood contacting surfaces. The conjugates (10-16 weight % of heparin) were obtained by a condensation reaction between albumin and heparin using 1-ethyl-3-(dimethylaminopropyl)-carbodiimide. Unreacted albumin and heparin were removed by diethyl-aminoethyl (DEAE)-cellulose and Cibacron Blue Sepharose chromatography respectively. The activity of the heparin component incorporated in the albumin-heparin conjugates (Ac) was compared with that of the heparin used for the synthesis of the conjugates (Anat) by thrombin time, inhibition of Factor Xa and the activated partial thromboplastin time (APTT) assays. The Ac/Anat ratio for the above assays was as follows: Thrombin time 1.25, Factor Xa inhibition 0.5. and APTT 0.5. Gel filtration chromatography showed broad-molecular weight distributions. The conjugates were fractionated using immobilized antithrombin III (ATIII). High ATIII and low ATIII affinity conjugate fractions showed the same behavior as ATIII fractionated heparin with respect to thrombin times and Factor Xa inhibition.

Animals↗