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Biomedical subjects

A Bantjes

Publications and source records attributed to A Bantjes.

At least 37 records · Page 2Linked to original sources

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↗

Large-scale purification of factor VIII by affinity chromatography: optimization of process parameters.

The optimization of a new process for the extraction of human coagulation factor VIII (FVIII) from plasma with the tailor-made affinity matrix dimethylamino-propylcarbamylpentyl-Sepharose CL-4B (C3-C5 matrix) is described. First, plasma is applied to DEAE-Sephadex A-50 anion exchanger in order to separate a number of proteins, including coagulation factors II, IX and X (prothrombin complex), from FVIII. Subsequently, the unbound fraction of the ion exchanger, containing FVIII, is contacted with the C3-C5 affinity matrix. Optimization of the FVIII affinity chromatographic procedure is accomplished in terms of the ligand density of the matrix, adsorption mode (batch-wise versus column-wise adsorption and matrix to plasma ratio), and conditions of pH and conductivity to be applied on washing and desorption. In scale-up experiments, by processing 20 l of plasma, the recovery (340 U VIII:C/kg plasma) and the specific activity (s.a.) (1.2 U VIII:C/mg protein) are better than those obtained by cryoprecipitation (recovery 300 U VIII:C/kg plasma, s.a. 0.3 U VIII:C/mg protein). The newly developed process using the specially designed C3-C5 affinity matrix has potential application in the process-scale purification of FVIII.

Buffers↗

Detection of surface-adsorbed (lipo)proteins by means of a two-step enzyme-immunoassay: a study on the Vroman effect.

In view of reports on the involvement of high-molecular-weight (HMW) kininogen and high-density lipoprotein (HDL) in the Vroman effect, we studied the adsorption of fibrinogen, HMW kininogen, HDL and several other proteins from pooled human plasma and congenitally HMW kininogen-deficient plasma onto glass and low-density polyethylene, both as a function of the plasma concentration and the contact time. Mixtures of purified (lipo)proteins were also included in the study. Protein adsorption was determined by means of a two-step enzyme-immunoassay. Our results support the hypothesis that HMW kininogen is involved in the displacement of fibrinogen, which is almost instantly adsorbed from normal plasma onto glass. On hydrophobic polymers like polyethylene, the low amounts of adsorbed fibrinogen and HMW kininogen from plasma and concentrated plasma solutions may be due to a preferential adsorption of HDL.

Adsorption↗

Platelet deposition studies on copolyether urethanes modified with poly(ethylene oxide).

Pellethane 2363 80A films and tubings were chemically modified and the effect of these modifications on platelet deposition was studied. Grafting of high molecular weight poly(ethylene oxide) and graft polymerization of methoxy poly(ethylene glycol) 400 methacrylate resulted in surfaces with a good water wettability. The increased hydrophilicity of these modified surfaces could be demonstrated by contact angle measurements. The platelet deposition was investigated with tubings in a capillary flow system, using different types of perfusates. Platelet deposition from a buffer-containing perfusate on surfaces modified with either high molecular weight poly(ethylene oxide) or methoxy poly(ethylene glycol) 400 methacrylate was almost absent and less than on Pellethane 2363 80A. Using a citrated plasma-containing perfusate the amount of deposited platelets on Pellethane 2363 80A modified with high molecular weight poly(ethylene oxide) was low and about the same as on unmodified surfaces. However, a marked reduced platelet deposition compared to unmodified Pellethane 2363 80A was found when the platelets were activated by Ca2+ ionophore. The improved blood compatibility of the modified Pellethane 2363 80A tubings obviously indicates the favourable effect of the presence of grafted PEO on the surface.

Biocompatible Materials↗

The effect of vascular cell seeding on platelet deposition in an in vitro capillary perfusion model.

In view of the reported beneficial effects of cell seeding on the performance of synthetic vascular grafts, we determined the deposition of 111Indium-labelled human platelets in polyethylene capillary tubes covered with human endothelial cells (HEC) or smooth muscle cells (SMC) using an in vitro perfusion model. Platelet deposition decreased with increasing vascular cell coverage and was virtually absent when the number of adherent vascular cells of both types exceeded 50,000/cm2. Platelet deposition increased with increasing shear rate (300-900 s-1) only when surface coverage was less than 50,000 vascular cells/cm2. Deposition of Ca2+-ionophore A 23187-activated platelets in capillary tubes completely covered with SMC was significantly higher compared to capillaries covered with a similar number of HEC. When HEC-lined capillaries were treated with aspirin, only the deposition of activated platelets increased slightly, but significantly. This platelet reactivity was more evident when the endothelial lining was not confluent. These results demonstrate that, although seeding of HEC and SMC may both prevent the deposition of non-activated platelets to surfaces, non-aspirin-treated confluent HEC linings offer the best protection against deposition of activated platelets.

Aspirin↗

Surface modification of copolyether-urethane catheters with poly(ethylene oxide).

Pellethane 2363 80A catheters were modified with poly(ethylene oxide) in order to improve their blood compatibility. Contact angle measurements showed that Pellethane 2363 80A surfaces had increased wettability after this modification. The results of in vitro blood compatibility tests showed that surface modification with poly(ethylene oxide) resulted in a five-fold reduction of platelet deposition. Activation of coagulation was not affected.

Biocompatible Materials↗

Deposition of endothelial fibronectin on polymeric surfaces.

Cellular fibronectin is deposited on tissue culture polystyrene during the adhesion and spreading of cultured human endothelial cells (HEC). Following the seeding of HEC upon this polymer, larger amounts of fibronectin are deposited as both cell density and incubation time increase. Our results indicate that the ability to deposit cellular fibronectin onto a polymeric surface is a condition for the spreading and proliferation of HEC.

Adsorption↗

Platelet deposition in a capillary perfusion model: quantitative and morphological aspects.

The capillary perfusion model according to Cazenave and co-workers was characterized by investigating the effects of protein precoating, perfusion time and shear rate on platelet deposition using 111Indium labelling of human platelets and scanning electron microscopy (SEM). Compared with uncoated polyethylene, platelet deposition was increased after precoating with purified human von Willebrand factor, fibrinogen or fibronectin, and decreased by preadsorbed immunoglobulin G, albumin or whole plasma. Platelet aggregates were observed on immunoglobulin G-coated polyethylene, whereas all other surfaces showed single adherent platelets. Complete platelet spreading was only observed after precoating with fibronectin. The quantitative data concerning platelet deposition were evaluated by using the convective-diffusion theory. Our results indicate the applicability of this perfusion model for the in vitro testing of biomaterials.

Adsorption↗

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↗

Removal of low density lipoprotein from blood plasma using cross-linked, sulfated polyvinylalcohol.

The properties of a new synthetic LDL binding material, consisting of fragments of loosely crosslinked hydrogel, based on sulfated polyvinylalcohol are described. When incubated with 20 times its volume of plasma, this material binds up to 95% of the LDL, even from plasma with severely elevated LDL cholesterol levels (up to 20 mM). In addition a cholesterol-rich subfraction of VLDL is bound but HDL is not bound. After about 10 min binder/plasma contact the LDL removal is complete and no other additives are required. LDL binding capacity is dependent on the average binder particle size, indicating a restricted penetration of LDL particles into the binder matrix.

Adult↗

Small diameter blood vessel prostheses from blends of polyethylene oxide and polypropylene oxide.

Studies on the relationship between blood platelet adhesion and type and amount of polyether segments in copolyetherurethanes report a reduced platelet adhesion with increasing polyether content. We therefore assumed that combinations of polyethylene oxide (PEO) and polypropylene oxide (PPOX) might give materials with a good blood compatibility. Water-soluble PEO was attached to PPOX by u.v.-initiated crosslinking. Films were tested for hydrophilicity, mechanical properties, protein adsorption and blood compatibility. The hydrophilicity was determined by swelling experiments. A compromise between hydrophilicity (PEO) and mechanical strength (PPOX) was met at a swelling of 0.5 (PPOX/PEO ratio: 90/10). In protein adsorption studies only small amounts of adsorbed proteins were found. Three blood material interaction in vitro tests gave good results: a low platelet adhesion and kallikrein generation and a high APTT value. Porous tubings (inner diameter 1.3 mm) were fabricated, by spinning from solution, for implantation in the abdominal aorta of rats. Stress-strain diagrams were comparable to those reported for natural blood vessels.

Adsorption↗

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↗

Crosslinked agarose encapsulated sorbents resistant to steam sterilization. Preparation and mechanical properties.

The application of agarose in hemoperfusion is hampered by the lack of a suitable sterilization method. A technique has been developed for the crosslinking of agarose encapsulated sorbents by the reaction with 1,3-dichloro-2-propanol (DCP) under strong alkaline conditions. A twofold molar excess of DCP with respect to agarose and an equimolar amount of sodium hydroxide at a concentration of 0.3 mol/L with a reaction time of 1-4 h at 50 degrees C are found to be the optimal conditions. The compressive strength of crosslinked beads is increased by a factor of 4. Agarose capsules are found to degrade by the influence of gamma radiation, but are resistant to steam sterilization at 134 degrees C during at least 30 min when crosslinked.

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↗

Preferential adsorption of high density lipoprotein from blood plasma onto biomaterial surfaces.

In the present study a two step enzyme immuno assay (EIA) was used for the investigation of the adsorption of proteins and lipoproteins from solutions and from blood plasma onto polymer surfaces. It was found that only a small adsorption of the major blood proteins occurred from plasma. Evidence is presented that the reason for this adsorption behaviour is a preferential adsorption of high density lipoprotein (HDL).

Adsorption↗

Grafting of a synthetic heparinoid polyelectrolyte onto silicone rubber.

Using a [3H]-labelled polyelectrolyte as a tracer, the 60Co grafting of a synthetic heparinoid polyelectrolyte onto silicone rubber was studied. The polyelectrolyte was labelled by the coupling of a radioactive amine to carboxylate groups of the polyelectrolyte. The amount of grafted polyelectrolyte was found to be dependent on the radiation dose.

Adsorption↗