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Biomedical subjects

R Barbucci

Publications and source records attributed to R Barbucci.

46 records · Page 3Linked to original sources

A material (PUPA) presenting both the properties of polyurethanes and the capacity of adsorbing a high quantity of heparin.

Polyurethanes are widely used for biomedical applications, but there is still a constant search for improved blood-compatible materials. We studied a material (PUPA) obtained by the interconnection between a poly(amido-amine), N2LL, capable of forming stable complexes with heparin and a commercial polyurethane, Pellethane 2363-80AE, using hexamethylenediisocyanate as the crosslinking agent. The amount of absorbed heparin (evaluated by biological tests) was generally much higher than that found on the poly(amido-amine) surface-grafted polyurethane.

Adsorption↗

Heparinizable materials (III). Heparin retention power of a poly(amido-amine) either as crosslinked resin, or surface-grafted on PVC.

The retentive power of a poly(amido-amine), in the form of a highly hydrophilic crosslinked resin, has been evaluated at different pH's. The resin releases heparin quantitatively in a very narrow pH range (10.8-11.4). This poly(amido-amine) has also been grafted on PVC tubes, and the heparin-adsorbing capacity of the materials so obtained has been tested biologically. In this case heparin is only released at pH greater than 10 so confirming the strong interaction between our polymer and heparin.

Acrylic Resins↗

Heparin adsorbing capacities at physiological pH of three poly(amido-amine) resins, and of poly(amido-amine)-surface-grafted glass microspheres.

Three poly(amido-amine)s of similar structure in the form of highly hydrophilic crosslinked resins, have been prepared, and tested for their heparin-adsorbing capacity at physiological pH. They showed different capacities, and their capacities were related to their basicities. One of the same polymers was grafted on the surface of glass microspheres. After treatment, it was shown that the microspheres could adsorb significant amounts of heparin. In all cases most of the adsorbed heparin was hardly eluted with saline, plasma, or blood, but could be recovered by eluting with 0.1 M NaOH. The resins were found to have some haemolytic properties, but no haemolysis was observed with the grafted microspheres.

Adsorption↗

Preparation and ESCA characterization of poly(vinyl chloride) surface-grafted with heparin-complexing poly(amido amine) chains.

By a simple process poly(amido-amine) chains have been grafted on the surface of poly(vinyl chloride). Grafted poly(vinyl chloride) is able to adsorb heparin, thus providing potentially non-thrombogenic surfaces. The grafting of poly(amido-amine), and the heparin adsorption have been studied by ESCA. It has been found that the total amount of grafted poly(amido-amine) depends on the molecular weight of the poly(amido-amine) used in the grafting reaction, but the amount of heparin adsorbed on the grafted material is relatively independent of the length of the poly(amino-amine) grafted chains.

Heparin↗

The influence of molecular weight on the biological activity of heparin like sulphated hyaluronic acids.

Sulphated hyaluronic acids having a sulphation degree of 3.5 per disaccharide unit, HyalS3.5, were prepared with different molecular weights corresponding to 21 x 10(3), 320 x 10(3) and 3500 x 10(3). The thrombin inhibition in plasma and in the presence of purified molecules, i.e. fibrinogen, antithrombin III (AT III) and heparin cofactor II (HC II), were studied for the three different MW compounds at different concentrations. The thrombin time in plasma depended on the length of the chain, and the two lower MW HyalS3.5 inhibited thrombin both by direct aspecific interaction and via HC II, whereas the activity of the highest MW compound was mainly related to the electrostatic interaction with HC II. The inactivation of FXa serine protease was only attributed to HyalS3.5-AT III complex.

Antithrombin III↗

Photoimmobilization of sulfated hyaluronic acid for antithrombogenicity.

Anticoagulant polymer sulfated hyaluronic acid was patterned immobilized on a poly(ethylene terephthalate) (PET) film in a specific pattern by photolithography. Hyaluronic acid was sulfated by a sulfur trioxide-pyridine complex. The polymer was coupled with azidoaniline. The derivatized polymer was cast on a PET film from aqueous solution. After drying, the film was photoirradiated in the presence or absence of a photomask. The micropatterning was confirmed by staining with a dye, brilliant green. Since the anticoagulant polymer has negative charges, the cationic dye was adsorbed on the regions where the anticoagulant polymer was immobilized. Platelet adhesion was reduced on the sulfated hyaluronic acid-immobilized areas. The immobilized sulfated hyaluronic acid significantly reduced thrombus formation.

Blood Coagulation↗

Fibroblast cell behavior on bound and adsorbed fibronectin onto hyaluronan and sulfated hyaluronan substrates.

The effect of fibronectin protein (Fn) coating onto polysaccharide layers of hyaluronic acid (Hyal) and its sulfated derivative (HyalS) on fibroblast cell adhesion was analyzed. The Hyal or HyalS were coated and grafted on the glass substrate by a photolithographic method. The Fn coating was achieved by two different routes: the immobilization of Fn by covalent bond to the polysaccharide layers and the simple adsorption of Fn onto Hyal and HyalS surfaces. AFM, SEM, and ATR-FTIR techniques were used for the chemical and topographical characterization of the surfaces. According to AFM and SEM data, the surface topography was dependent on the method used to cover the polysaccharide layers with the protein. ATR-FTIR analysis supplied information about the rearrangement of Fn after the interaction (adsorption or binding) with the Hyal and the HyalS. The conformational changes of the Fn were minimal when it was simply adsorbed on HyalS surfaces and larger once bound, whereas on the Hyal layer the protein underwent a bigger conformational change once adsorbed and covalently grafted. Then, the biological characterization was carried out by analyzing the human diploid skin fibroblasts adhesion on these surfaces. The morphology of fibroblasts was evaluated by SEM, whereas the dynamics of fibroblasts movement were recorded by a time-lapse system. Cell variations in area, perimeter, and length were analyzed at 2, 4, and 6 h. It was found that the addition of Fn (covalently bound or merely adsorbed) was fundamental in the promotion of fibroblasts adhesion and spreading. The greatest adhesion occurred onto HyalS layers covered by the adsorbed Fn.

Adsorption↗

Blood tubing and cytokine production: effect of sterilization.

Blood tubings commonly represent an integral component of hemodialysis circuits. Different factors may influence their biocompatibility, such as the type of material, the sterilization mode and the geometry. In vivo the final biocompatibility may be further complicated by the individual host response, the flow parameters, and the impact of mechanical trauma on blood's cellular components (i.e. erythrocytes). In this in vitro study we evaluated some commercially available blood tubings sterilized by different methods as to their interaction with normal leukocyte population and tested the response of these cells in terms of cytokines (IL-1beta, IL-1Ra, TNF-alpha). As a positive control, leukocytes were incubated with 0.5 ng/mL of bacterial lipopolysaccharide (LPS) or with Cuprophan of comparable surface. The results showed that cytokine production was markedly reduced, particularly in the case of gamma-ray-sterilized tubings. Of interest, it was not always related to the adherence. However in some cases, particularly of gamma-ray sterilization, adherence was none, despite the cytokine production.

Apoptosis↗