PubMed HealthSearch

Biomedical subjects

J Feijen

Publications and source records attributed to J Feijen.

At least 19 recordsLinked to original sources

Relations between in vitro cytotoxicity and crosslinked dermal sheep collagens.

Collagen-based biomaterials have found various applications in the biomedical field. However, collagen-based biomaterials may induce cytotoxic effects. This study evaluated possible cytotoxic effects of (crosslinked) dermal sheep collagen (DSC) using a 7-d-methylcellulose cell culture with human skin fibroblasts. Non-crosslinked DSC (NDSC), hexamethylene-diisocyanate-crosslinked DSC (HDSC), and glutaraldehyde-crosslinked DSC (GDSC), their extracts (1 x 10 d to 4 x 10 d extracts), or the corresponding extracted DSC samples were tested. Cell growth was evaluated by cell counting, while cell morphology was assessed by light microscopy and transmission-electron microscopy. Both GDSC and, to a lesser extent, HDSC, induced cytotoxicity, observed as inhibited cell growth and deviant cell morphology. The deviant morphology consisted of extensive accumulations of lipid, reduction in the amount and dilatation of rough endoplasmatic reticulum, increased inclusions of cell remnants, and relatively rounded cell membranes. With HDSC, both primary cytotoxicity, due to extractable products from the material, and secondary cytotoxicity, possibly due to a release of cytotoxic products resulting from enzymatic cell-biomaterial interactions, could be discriminated. With GDSC, however, no clear distinction between primary and secondary cytotoxicity could be made. With NDSC, only primary cytotoxicity, measured as low inhibition of cell proliferation, but without deviant morphology, was observed. These remarkable differences in cytotoxicity are discussed in relation to residual agents and specific crosslinks present in DSCs as a consequence of processing and the crosslinking agents used. The residual agents and the specific crosslinks give rise to differences in direct release of products and in sensitivity to hydrolysis and enzymatic breakdown.

Animals

Physicochemical characterization of Escherichia coli. A comparison with gram-positive bacteria.

Eight Escherichia coli strains were characterized by determining their adhesion to xylene, surface free energy, zeta potential, relative surface charge, and their chemical composition. The latter was done by applying X-ray photoelectron spectroscopy (XPS) and infrared spectroscopy (IR). No relationship between the adhesion to xylene and the water contact angles of these strains was found. Three strains had significantly lower surface free energies than the other strains. Surface free energies were either obtained from polar and dispersion parts or from Lifshitz-van der Waals and acid/base parts of the surface free energy. A correlation (r = 0.97) between the polar parts and the electron-donor contributions to the acid/base part of the surface free energy was found. The zeta potentials of all strains, measured as a function of pH (2-11), were negative. Depending on the zeta potential as a function of pH, three groups were recognized among the strains tested. A relationship (r = 0.84) was found between the acid/base component of the surface free energy and the zeta potential measured at pH = 7.4. There was no correlation between results of XPS and IR studies. Data from the literature of XPS and IR studies of the gram-positive staphylococci and streptococci were compared with data from the gram-negative E. coli used in this study. It appeared that in these three groups of bacteria, the polysaccharide content detected by IR corresponded well with the oxygen-to-carbon ratio detected by XPS.

Anion Exchange Resins

In vivo fragmentation of microporous polyurethane- and copolyesterether elastomer-based vascular prostheses.

A previous study showed that microporous, compliant and (bio)degradable vascular prostheses prepared from a polyurethane/poly(L-lactic acid) mixture can function as a temporary scaffold for the regeneration of small-calibre arteries. In this study the mechanism of fragmentation of vascular prostheses made of polyurethane, copolyesterether and blends of either polyurethane or copolyesterether with polymers differing in biodegradability, crystallinity and glass transition temperature is investigated. Animal studies revealed that after 6 wk of implantation only the prostheses made of blends containing a second polymer which was non-elastic at 37 degrees C were fragmented extensively, whether the second polymer was (bio)degradable or not. It is concluded that fragmentation of the prostheses is mainly caused by alternating stresses induced by the arterial pulsations and that (bio)degradation plays a minor role.

Animals

In vitro leucocyte adhesion to modified polyurethane surfaces. I. Effect of ionizable functional groups.

To study the effect of ionizable functional groups on the adhesion of leucocytes to surfaces, both poly(ethyleneimine) and poly(acrylic acid) were immobilized on polyurethane films, resulting in the introduction of amine and carboxylic acid groups, respectively. This was confirmed by contact angle measurements and XPS analysis. In vitro adhesion of granulocytes and lymphocytes on untreated and modified surfaces was compared. The number of adherent cells on modified surfaces as a function of time was significantly higher than on untreated surfaces. This effect was most pronounced for the adhesion of lymphocytes to surfaces modified with amine groups. In this case, the number of adherent cells after 1 h of exposure was three times higher than on untreated surfaces. A moderate enhancement of leucocyte adhesion was observed in the case of surfaces modified with carboxylic acid groups. There is evidence that these groups were not ionized under the experimental conditions used. The modification procedures described may be used to improve polyurethane filters for the removal of leucocytes from blood.

Cell Adhesion

Secondary cytotoxicity of cross-linked dermal sheep collagens during repeated exposure to human fibroblasts.

We investigated commercially available dermal sheep collagen either cross-linked with hexamethylenediisocyanate, or cross-linked with glutaraldehyde. In previous in vitro studies we could discriminate primary, i.e. extractable, and secondary cytotoxicity, due to cell-biomaterial interactions, i.e. enzymatic actions. To develop dermal sheep collagen for clinical applications, we focused in this study on the release, e.g. elimination, of secondary cytotoxicity over time. We used the universal 7 d methylcellulose cell culture with human skin fibroblasts as a test system. Hexamethylenediisocyanate-cross-linked dermal sheep collagen and glutaraldehyde-cross-linked dermal sheep collagen were tested, with intervals of 6 d, over a culture period of 42 d. With hexamethylenediisocyanate-cross-linked dermal sheep collagen, cytotoxicity, i.e. cell growth inhibition and deviant cell morphology, was eliminated after 18 d of exposure. When testing glutaraldehyde-cross-linked dermal sheep collagen, the bulk of cytotoxic products was released after 6 d, but a continuous low secondary cytotoxicity was measured up to 42 d. As a control, non-cross-linked dermal-sheep collagen was tested over a period of 36 d, but no secondary cytotoxic effects were observed. The differences in release of secondary cytotoxicity between hexamethylenediisocyanate-cross-linked dermal sheep collagen, glutaraldehyde-cross-linked dermal sheep collagen and non-cross-linked dermal sheep collagen are explained from differences in cross-linking agents and cross-links obtained. We hypothesize that secondary cytotoxicity results from enzymatic release of pendant molecules from hexamethylene-diisocyanate-cross-linked dermal sheep collagen, e.g. formed after reaction of hydrolysis products of hexamethylenediisocyanate with dermal sheep collagen.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Bacterial migration along solid surfaces.

An in vitro system was developed to study the migration of uropathogenic Escherichia coli strains. In this system an aqueous agar gel is placed against a solid surface, allowing the bacteria to migrate along the gel/solid surface interface. Bacterial strains as well as solid surfaces were characterized by means of water contact angle and zeta potential measurements. When glass was used as the solid surface, significantly different migration times for the strains investigated were observed. Relationships among the observed migration times of six strains, their contact angles, and their zeta potentials were found. Relatively hydrophobic strains exhibited migration times shorter than those of hydrophilic strains. For highly negatively charged strains shorter migration times were found than were found for less negatively charged strains. When the fastest-migrating strain with respect to glass was allowed to migrate along solid surfaces differing in hydrophobicity and charge, no differences in migration times were found. Our findings indicate that strategies to prevent catheter-associated bacteriuria should be based on inhibition of bacterial growth rather than on modifying the physicochemical character of the catheter surface.

Bacterial Adhesion

Growth of uropathogenic Escherichia coli strains at solid surfaces.

The adhesion and growth of two catheter-associated (O2K2 and O83K?) and two non catheter-associated (O111K58 and O157K-) uropathogenic Escherichia coli strains on glass, poly(methyl methacrylate) (PMMA), a negatively charged copolymer of MMA and methacrylic acid (MAA) and a positively charged copolymer of MMA and trimethylaminoethyl methacrylate chloride (TMAEMA-Cl) were studied. The solid surfaces were placed in a parallel plate perfusion system. After preadhesion of the bacteria onto the surfaces, growth was initiated by perfusing the system with MacConkey broth. Growth was measured by counting adherent bacteria as a function of time. Bacterial strains were characterized by means of water contact angle, microbial adhesion to hydrocarbon (MATH), anion exchange resin retention (ARR) and zeta potential measurements. Solid surfaces were characterized by means of water contact angle and zeta potential measurements. The catheter-associated strains had significantly higher water contact angles, zeta potentials and ARR values than the non catheter-associated strains. Non catheter-associated strains did not grow at the surfaces used. Catheter-associated strains did not grow at the positively charged surface but exhibited growth at the other surfaces. Strains grew more rapidly at surfaces with a relatively high negative zeta potential and a low water contact angle than at surfaces with a relatively low negative zeta potential and a high water contact angle. The growth of strain O2K2 on glass was significantly reduced when urine instead of MacConkey broth was used as perfusion medium.

Anion Exchange Resins

Improved adhesion and proliferation of human endothelial cells on polyethylene precoated with monoclonal antibodies directed against cell membrane antigens and extracellular matrix proteins.

Endothelial cell seeding may improve the patency of synthetic vascular grafts provided that platelet reactivity of nonendothelialized sites is not increased. We have investigated if surface-adsorbed monoclonal antibodies directed against endothelial cell membrane proteins and against extracellular matrix proteins promote the adhesion and proliferation of cultured human endothelial cells, without causing platelet deposition at non-endothelialized sites. Adhesion of endothelial cells onto polyethylene coated with monoclonal antibodies directed against endothelial cell-specific membrane antigens, integrin receptors and glycoprotein CD31 was equal to or higher than adhesion onto fibronectin-coated polyethylene. Endothelial cells did not proliferate on these surface-adsorbed antibodies. However, pre-coating of polyethylene with mixtures of endothelial cell-specific monoclonal antibodies and monoclonal antibodies directed against fibronectin or von Willebrand factor, resulted in relatively high adhesion and optimal proliferation. Platelet reactivity of the polyethylene surface was found to significantly increase after adsorption of fibronectin, endothelial cell-specific monoclonal antibody or its Fc fragments. In contrast, adsorption of F(ab')2 fragments of endothelial cell-specific monoclonal antibody did not promote platelet deposition. Therefore, it is concluded that coating of vascular graft materials with mixtures of F(ab')2 fragments of monoclonal antibodies specifically directed against endothelial cells and against extracellular matrix proteins may be an effective way to both promote the growth of seeded endothelial cells and limit platelet-graft interaction.

Adsorption

Asymmetric membrane filters for the removal of leukocytes from blood.

As part of a study on the mechanisms of leukocyte filtration, the influence of pore size distribution on filter efficiency was investigated. Conventional leukocyte filters are not suitable for model studies, as these filters are composed of tightly packed synthetic fibers, with a poorly defined porous structure. Therefore, open cellular polyurethane membranes with pore size distributions varying from approximately 15 to 65 microns were prepared. Filtration experiments with stacked packages of these membranes showed that leukocytes are best removed (greater than 99%) by filters with a pore size distribution of 11-19 microns. These pore sizes approach the size of leukocytes (6-12 microns). However, due to fast clogging, blood flow through these filters is rapidly reduced, which results in a low filter capacity. With an asymmetric membrane filter, in which the pore size decreases from about 65 to 15 microns in the direction of blood flow, both moderate removal of leukocytes (greater than 80%) and maintenance of flow (approximately 0.2 mL/s) are obtained. This results in efficient leukocyte removal. From cell analysis of both filtrate and filter, it is concluded that adhesion rather than sieving is the major filtration mechanism. Thus, further optimization of the filter may be achieved by surface modification.

Cell Separation

Adhesion of Escherichia coli on to a series of poly(methacrylates) differing in charge and hydrophobicity.

The adhesion of three Escherichia coli strains on to six poly(methacrylates) differing in hydrophobicity and surface charge was measured as a function of time under laminar flow conditions. Polymers used were poly(methyl methacrylate) (PMMA), poly(hydroxyethyl methacrylate) (PHEMA) and copolymers of MMA or HEMA with either 15% methacrylic acid (MAA) or 15% trimethylaminoethyl methacrylate-HCl salt (TMAEMA-Cl). Bacterial and polymer surfaces were characterized by means of water contact angles and zeta potentials. Both the sessile drop contact angles and the zeta potentials of the bacterial surfaces were significantly different. No significant differences in the sessile drop contact angles of the polymer surfaces were observed. Using the Wilhelmy plate technique large contact angle hysteresis was observed for the different polymer surfaces. Surfaces of copolymers with MAA had more negative zeta potentials than those of the corresponding homopolymers. Surfaces of copolymers with TMAEMA-Cl had positive zeta potentials. The highest numbers of adherent bacteria were found on materials with positive zeta potentials, irrespective of the bacterial strain used. Bacterial adhesion on to copolymers with MAA was less than on to the corresponding homopolymers. Bacterial equilibrium adhesion values correlate with the zeta potentials of the polymer surfaces (r greater than 0.85). On substrates with less negative zeta potentials high numbers of adhered bacteria were observed. Additionally, the equilibrium bacterial adhesion values could be related with receding contact angles of polymer surfaces with negative zeta potentials (r greater than 0.86). High equilibrium adhesion values were obtained for polymers with high contact angles. No correlation between the zeta potentials and contact angles of the bacteria with the adhesion values was found.

Bacterial Adhesion

Adhesion of endothelial cells and adsorption of serum proteins on gas plasma-treated polytetrafluoroethylene.

From in vitro experiments it is known that human endothelial cells show poor adhesion to hydrophobic polymers. The hydrophobicity of vascular prostheses manufactured from Teflon or Dacron may be the reason why endothelialization of these grafts does not occur after implantation in humans. We modified films of polytetrafluoroethylene (Teflon) by nitrogen plasma and oxygen plasma treatments to make the surfaces more hydrophilic. Depending on the plasma exposure time, modified polytetrafluoroethylene surfaces showed water-contact angles of 15-58 degrees, versus 96 degrees for unmodified polytetrafluoroethylene. ESCA measurements revealed incorporation of both nitrogen- and oxygen-containing groups into the polytetrafluoroethylene surfaces, dependent on the plasma composition and exposure time. The thickness of the modified surface layer was approximately 1 nm. The adhesion of cultured human endothelial cells from 20% human serum-containing culture medium to modified polytetrafluoroethylene surfaces with contact angles of 20-45 degrees led to the formation of a monolayer of cells, which was similar to the one formed on tissue culture polystyrene, the reference surface. This was not the case when endothelial cells were seeded upon unmodified polytetrafluoroethylene. Surface-modified expanded polytetrafluoroethylene prosthesis material (GORE TEX soft tissue) also showed adhesion of endothelial cells comparable to cell adhesion to the reference surface. The amounts of serum proteins, including fibronectin, adsorbed from serum-containing medium to modified polytetrafluoroethylene surfaces were larger than those adsorbed to unmodified polytetrafluoroethylene. Moreover, the modified surfaces probably allow the exchange of adsorbed serum proteins with cellular fibronectin.

Adsorption

In vivo degradation of processed dermal sheep collagen evaluated with transmission electron microscopy.

The in vivo degradation of hexamethylenediisocyanate-tanned dermal sheep collagen was studied with transmission electron microscopy. Discs of hexamethylenediisocyanate-tanned dermal sheep collagen were subcutaneously implanted in rats. Both an intra- and an extracellular route of degradation could be distinguished. In addition to normal components of a typical foreign body reaction, remarkable phenomena, such as locally deviant neutrophil morphology, infiltration of basophil-like cells, indications of foreign body multinucleate giant cells formed from different cell types, aluminium silicate accumulations and calcium phosphate depositions, were observed. Foreign body multinucleate giant cells intracellularly degraded hexamethylenediisocyanate-tanned dermal sheep collagen after internalization. Both internalized and cellularly enveloped hexamethylenediisocyanate-tanned dermal sheep collagen degraded by the detachment of fibrils. Another extracellular route of degradation was characterized by calcium phosphate depositions in large bundles of hexamethylenediisocyanate-tanned dermal sheep collagen. From 6 wk, the hexamethylenediisocyanate-tanned dermal sheep collagen implant was replaced by rat connective tissue, which was subsequently also degraded. After 15 wk, the presence of basophil-like foreign body multinucleated giant cells containing aluminium/silicon-crystalline accumulations still persisted. These phenomena were related to the specific nature of the material used and suggest cytotoxicity. They emphasize the need for detailed evaluation at the ultrastructural level of newly developed biomaterials before they can be used for medical applications.

Animals

Synthesis of poly(hydroxypropylglutamine-prazosin carbamate) and release studies.

Prazosin, an antihypertensive drug with postsynaptic alpha 1-adrenergic blocking activity, has been coupled to poly-N5-(3-hydroxypropyl-L-glutamine) (PHPG) via a carbamate linkage. PHPG was activated by p-nitrophenyl chloroformate and then reacted with prazosin to form p(HPG-prazosin carbamate) conjugate. Drug loading was 23.9% (w/w). Activated polymer and conjugates were characterized by infrared spectroscopy and differential scanning calorimetry. In vitro studies proceeded in pH 7.4 isotonic phosphate-buffered saline solution. Prazosin was released at a rate of 0.92 mg/day/100 mg conjugate from p(HPG-prazosin carbamate) particles. In vivo studies were performed with New Zealand White rabbits. P(HPG-prazosin carbamate) conjugate particles (100 mg) were suspended in 2 ml saline and injected subcutaneously into both flanks of rabbits. P(HPG-prazosin carbamate) conjugates, following an initial burst, demonstrated a nearly constant plasma prazosin concentration profile above 2 ng/ml, which was maintained for 10 days.

Animals

Secretory IgA adsorption and oral streptococcal adhesion to human enamel and artificial solid substrata with various surface free energies.

In this paper, secretory IgA adsorption from a single component sIgA solution and from human whole saliva onto human enamel and artificial solid substrata with various surface free energies was studied as a function of time. ELISA indicated that screening or displacement of adsorbed sIgA by other salivary proteins occurred only on low surface free energy substrata, not on high surface free energy substrata such as enamel. In addition, the adhesion of three oral streptococcal strains (Streptococcus mitis BMS, S. sanguis 12, and S. mutans NS), also having widely different surface free energies, to sIgA-coated surfaces was studied. The adhesion of all three streptococcal strains was significantly reduced in the presence of a sIgA coating. However, ranking the adhesion data with respect to the various substrata revealed a similar order to that in the case of uncoated substrata, indicating that substratum properties were at least partly transferred by the adsorbed protein film to the interface with adhering micro-organisms. For S. sanguis 12 and S. mitis BMS, adhesion decreased proportionally with the amounts of sIgA detected by ELISA, but for S. mutans NS such relations with the amounts of sIgA detected on the protein-coated substrata were not found. Thus, for S. mutans NS a specific antibody effect seems to exist in addition to a non-specific protein effect like that observed for S. sanguis 12 and S. mitis BMS.

Adsorption

Protein adsorption from plasma onto poly(n-alkyl methacrylate) surfaces.

Protein adsorption of human serum albumin (HSA), human fibrinogen (Fg), human immunoglobulin G (IgG), high density lipoprotein (HDL) and high molecular weight kininogen (HMWK) from plasma onto poly (n-alkyl methacrylate) (PAMA) surfaces was measured using a semi-quantitative enzyme-immunoassay. Adsorption was investigated for PAMA(n = 1) (n is the number of C-atoms in the n-alkyl side chain), PAMA(n = 8) and PAMA(n = 18). PAMA(n = 1) has a relatively hydrophilic surface as compared to the more hydrophobic PAMA(n = 8) surface. Both polymers have surface chains which do not reorient after contact with water. The PAMA(n = 18) surface is relatively hydrophobic but in this case polymer surface chains and segments are able to reorient after contact with water. Protein adsorption was measured both as a function of time and as a function of the plasma dilution. If adsorption from plasma was measured as a function of time no exchange of proteins could be observed. The amount of adsorbed protein was always larger in the case of the hydrophobic PAMA(n = 8) as compared to PAMAS(n = 1 and 18), probably due to hydrophobic interactions between the proteins and the PAMA(n = 8) surface. At high plasma concentration relatively large amounts of HDL adsorb onto PAMA(n = 8), indicating that this lipoprotein preferentially adsorbs onto this surface.

Adsorption

Adherence of coagulase-negative staphylococci onto polyethylene catheters in vitro and in vivo: a study on the influence of various plasma proteins.

Bacterial adherence on PE catheters inserted into the carotid artery of rabbits was assessed at 5, 30, and 240 min after injection with bacteria of five coagulase-negative staphylococci (CN Staph). These studies revealed that CN Staph adhere onto PE catheters 5 min after injection with bacteria. At 240 min after injection with bacteria almost all catheters were sterile, indicating that initially adherent bacteria on PE catheters disappear with time. EM studies revealed high numbers of platelets and leukocytes and many fibrin deposits on the surface of the PE catheter. In addition, the adherence of the five test strains on PE catheters was determined in vitro. In these in vitro studies the bacteria and catheters were pretreated with either fibrinogen, fibronectin, albumin or citrated whole plasma or the liquid adherence medium was supplemented with these proteins or the adherence assay was done in citrated whole plasma. The presence of proteins in the adherence medium or pretreatment of the catheter or bacteria with either fibronectin, albumin or whole citrated plasma markedly inhibited bacterial adherence. In contrast, pretreatment of bacteria or both bacteria and catheters with fibrinogen enhanced bacterial adherence suggesting the presence of ligands for fibrinogen on the cell surface of CN Staph.

Animals