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Biomedical subjects

N Barbani

Publications and source records attributed to N Barbani.

13 recordsLinked to original sources

Transglutaminase reactivity with gelatine: perspective applications in tissue engineering.

Gelatine was crosslinked by means of an enzymatic treatment using tissue transglutaminase (tTGase) (Sigma) and microbial transglutaminase (mTGase) (Ajinomoto) which catalyses the formation of isopeptide bonds between the gamma-carbonyl group of a glutamine residue and the epsilon-amino group of a lysine residue. The reaction is an interesting alternative to the traditional glutaraldehyde crosslinking, which has several drawbacks (e.g., in medical application) due to the toxicity of the chemical reagent. To further investigate the possibility to utilize the modified protein for tissue engineering application, TGase crosslinked gelatine was incorporated in a gellan matrix, a polysaccharide, to enhance the stability in aqueous media. Films obtained by casting were characterized by thermal analysis, chemical imaging, swelling behaviour and cell adhesion.

Animals↗

Composites between hydroxyapatite and poly(epsilon-caprolactone) synthesized in open system at room temperature.

The hydroxyls present on the surface of hydroxyapatite (HA) granules, annealed at 700 composite function, 900 composite function and 1,100 composite function C, are able to initiate the polymerization of epsilon-caprolactone (CL), not only at 185 composite function C under vacuum, but also at room temperature in open system. A polymer layer ionically linked to the substrate is formed on HA surface, enhancing the compatibility between the organic phase and the inorganic one in composite biomaterials. We studied the characteristics of the polymer, produced by the reaction carried out at room temperature in open system, as well as the percentages of the poly(epsilon-caprolactone) (PCL) ionically bonded to the HA structure and of the "free" one. Both percentages appear very dependent on the annealing temperature; in particular, HA annealed for 1 h at 1,100 composite function C is the most efficient initiator of the reaction leading to ionically bonded PCL. The percentages of "free" polymer are much higher than at 185 composite function C under vacuum. Its formation is attributed to the role of water in opening the CL rings, and to the presence of CO(3) (2-) and HPO(4) (2-) ions in the HA annealed at lower temperatures. The presence of water appears to be the limiting factor for the production of PCL not bonded to the HA structure.

Caproates↗

The relevance of the transfer of molecular information between natural and synthetic materials in the realisation of biomedical devices with enhanced properties.

Past and recent attempts to introduce in synthetic polymers molecular information from natural substances through simple blending, template polymerization and molecular imprinting are reviewed. The most promising approaches that can open the way to the realisation of new materials with improved biocompatibility, antibody- or enzyme-like performances are analysed more deeply. The realisation of bioartificial blends from natural and synthetic polymers, molecularly imprinted nanospheres or membranes that can act as recognition element in (bio)sensing devices, as synthetic enzymes or as key constituents of body fluids purification tools is presented in order to make the reader aware of the fascinating possibilities that these techniques make available to the biomedical science and engineering in the close future. The last part of the paper describes recent attempts to insert recognition elements for large molecules as proteins, DNA segments, viruses or whole cells in synthetic polymer systems, in order to develop new systems in the treatments of diseases and for tissue-engineering applications.

Animals↗

Molecularly imprinted bioartificial membranes for the selective recognition of biological molecules. Part 2: release of components and thermal analysis.

Molecularly imprinted membranes imprinted for a large-molecular-weight protein were realised using a blend of natural and synthetic polymers. Bioartificial membranes of synthetic (poly(ethylene-co-vinyl alcohol)-EVAL, Clarene) and biological (Dextran) polymers, molecularly imprinted with alpha-amylase as the template, were prepared and investigated. Dimethyl sulfoxide (DMSO) solutions of the alpha-amylase template, Clarene and Dextran were mixed under stirring in the desired proportions and dipped in DMSO (solvent)/water (non solvent) mixture, to obtain the phase separation. The release of Clarene, Dextran and alpha-amylase in the inversion baths was quantified by spectrophotometric methods and final composition of membranes was established. To study the interactions between the polymer components and between polymeric materials and the template, differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) were carried out. Results indicated that stable and continuous bioartificial membranes of Clarene and Dextran can be obtained, whereby calorimetric analysis suggested the presence of high interaction between alpha-amylase and the Clarene component.

Biocompatible Materials↗

Acrylic polymeric nanospheres for the release and recognition of molecules of clinical interest.

Cross-linked poly(methylmethacrylate-co-methacrylic acid) nanospheres were imprinted with theophylline through template radical polymerisation in diluted acetonitrile solution. This study will focus on the effect of functional monomer nature used (methylmethacrylate and/or methacrylic acid) in the recognition and in the release of template in order to develop a material with combined properties of drug delivery and rebinding for clinical applications. After template extraction the nanospheres showed satisfactory recognition properties (up to 1mg template/g of polymer). Moreover polymers prepared selectively removed theophylline with a theophylline rebinding of 5.1 times higher than that of caffeine, a compound of similar structure. Drug release properties were also satisfactory (up to 95% of loaded theophylline in 7 days).

Acrylic Resins↗

Molecularly imprinted bioartificial membranes for the selective recognition of biological molecules.

Membranes of a synthetic (poly(ethylene-co-vinyl alcohol), Clarene) and a biological (dextran) polymer, imprinted with alpha-amylase, of different compositions were prepared by the phase-inversion process. Membrane morphologies were shown to be dependent on the synthetic-biological components composition. The removal of the template from the membranes was performed by extraction with water, while an aqueous solution of alpha-amylase was permeated across the membranes under pressure to obtain the rebinding of the template. The selectivity of alpha-amylase-imprinted membranes was investigated by the same uptake experiment using pepsin, albumin and amyloglucosidase, and the rebinding of these proteins was compared with that of the print molecule. Before and after template extraction and after the rebinding experiment, kinetic measurements of the imprinting molecule were conducted to estimate the activity of the enzyme immobilised in the polymer matrix. Results obtained revealed that the immobilised enzyme maintains a good functionality while in the membrane compared to the free enzyme and the imprinted 'bioartificial' dextran and Clarene membranes, obtained by the phase-inversion method, can establish efficient interaction with alpha-amylase as template molecule, as confirmed by the fair selectivity in rebinding tests.

Carbohydrate Sequence↗

Bioartificial polymeric materials based on polysaccharides.

Bioartificial polymeric materials, based on blends of polysaccharides with synthetic polymers such as poly(vinyl alcohol) (PVA) and poly(acrylic acid) (PAA), were prepared as films or hydrogels. The physico-chemical, mechanical, and biological properties of these materials were investigated by different techniques such as differential scanning calorimetry, dynamic mechanical thermal analysis, scanning electron microscopy, and in vitro release tests, with the aim of evaluating the miscibility of the polymer blends and to establish their potential applications. The results indicate that while dextran is perfectly miscible with PAA, dextran/PVA, chitosan/PVA, starch/PVA, and gellan/PVA blends behave mainly as two-phase systems, although interactions can occur between the components. Cross-linked starch/PVA films could be employed as dialysis membranes: they showed transport properties comparable to, and in some cases better than, those of currently used commercial membranes. Hydrogels based on dextran/PVA and chitosan/PVA blends could find applications as delivery systems. They appeared able to release physiological amounts of human growth hormone, offering the possibility to modulate the release of the drug by varying the content of the biological component.

Biocompatible Materials↗

Effect of chitosan and dextran on the properties of poly(vinyl alcohol) hydrogels.

Hydrogels are three-dimensional polymeric networks very similar to biological tissues and potentially useful as drug delivery systems. Poly(vinyl alcohol)-based hydrogels containing different amounts of dextran or chitosan were prepared using the freezing-thawing method. Repeated freezing-thawing cycles of a poly(vinyl alcohol) (PVA) aqueous solution lead to the formation of crystallites which act as cross-linking sites, and a hydrogel with a high capacity to swell is obtained. The effects of the two different polysaccharides on the properties of the obtained materials were investigated by differential scanning calorimetry, dynamic mechanical analysis and scanning electron microscopy. In addition the release with time of poly(vinyl alcohol) in aqueous medium, was monitored and evaluated. On the basis of the obtained results it seems that the presence of dextran favors the crystallization process of PVA, allowing the formation of a more ordered and homogeneous structure. Instead, chitosan seems to perturb the formation of PVA crystallites leading to a material with a less regular structure.

Journal Article↗

Bioartificial materials based on collagen: 1. Collagen cross-linking with gaseous glutaraldehyde.

The effect of exposure time of thin films of soluble collagen to glutaraldehyde (GTA) vapour was studied at 37 degrees C, and was evaluated by examining the thermal and biological stability and the swelling ratio. It was found that the collagen films treated with GTA vapour for 18 h showed the highest denaturation temperature, the lowest swelling ratio, and an enhanced proteolytic stability. This study shows that soluble collagen can be effectively cross-linked with GTA vapour and that the degree of cross-linking can be controlled by varying the exposure time.

Animals↗

Bioartificial materials based on collagen: 2. Mixtures of soluble collagen and poly(vinylalcohol) cross-linked with gaseous glutaraldehyde.

Thin films of both pure soluble collagen (CLG) and poly(vinylalcohol) (PVA) and mixtures of the two, ranging from 20-80% PVA composition were studied to test the effects of PVA content and of glutaraldehyde vapour cross-linking. Both the thermal and mechanical behavior and, in addition, proteolytic stability were clearly influenced by the ratio of CLG/PVA. The experimental results indicate that no thermodynamic compatibility occurs between the two homopolymers. However, there is evidence that strong interactions, probably due to hydrogen bond formation, occur between the biological and synthetic polymers. The interactions appear stronger in those films with a lower PVA content and which were not cross-linked. Both the thermal and biological stability are increased and there is an improvement of the mechanical properties. The mutual intermolecular influence appears to allow the attainment of a good mechanical compatibility between CLG and PVA.

Animals↗

The activation of human plasma prekallikrein as a hemocompatibility test for biomaterials. II. Contact activation by EVAL and EVAL-SMA copolymers.

The activation of human plasma prekallikrein (PKK) to kallikrein (KK), induced by the contact of blood with foreign materials, is a useful in vitro hemocompatibility test. Kallikrein is easily detected by its reaction with the chromogenic substrate H-D-Pro-Phe-Arg-pNA, which releases p-nitroaniline, revealed by its absorption at 405 nm. This test, which was already carried out by evaluating PKK activation by the 'end-point' method, has been carried out in this work by the more accurate 'initial velocity' method, i.e. by evaluating the activation from the initial rates of the KK-substrate reaction. The tests were carried out on the following materials: borosilicate glass (as a high-activation reference material), silicone (as a low-activation reference material), the commercial biomaterial Cardiothane 51, three graft copolymers synthesized in our laboratory by reacting ethylene-vinyl alcohol copolymer (EVAL) with styrene-maleic anhydride copolymer (SMA), and EVAL itself. A mathematical treatment based on a simple kinetic model has been used for a first-approximation evaluation of the PKK-activating power of the materials tested. The quite low activating power of the EVAL-SMA copolymers, which are easily processable into water-permeable hollow fibers, suggests the possibility of their use in blood dialyzers.

Amino Acid Sequence↗

Transport properties of EVAl-starch-alpha amylase membranes.

We investigated the influence of various physicochemical parameters on the morphology and time-porosity formation of membranes composed of ethylene-vinyl alcohol, starch, and alpha-amylase. In particular, we determined that (1) it is possible to obtain a membrane with desired porosity by phase inversion in an appropriate water-ethanol mixture and (2) the enzymatic bioerosion is controlled by the amount of alpha-amylase present in the blend. Although no experiments involving drugs were carried out, the delivery properties of the film were determined by measuring the Darcy permeability, the effective diffusivity, and the mean reaction rate of the membranes, relating them to the modality of membrane preparation, the amount of enzyme present within the membrane, and the incubation time of the samples in a buffer solution. Simple theoretical models of the delivery properties of the membranes were developed, leading to predictions that were in good agreement with the experimental results.

Biological Transport↗

Fibers by bioresorbable poly(ester-ether-ester)s as potential suture threads: a preliminary investigation.

Fibers made by a bioresorbable poly(epsilon-caprolactone)-block-poly(oxyethylene)-block-poly(epsilon-caprolactone) copolymer, having a number average molecular mass of about 200,000 Da and an average molar composition of 66% oxycaproyl units and 34% oxyethylene units, were melt-spun, with the aim at using them as suture threads. Their properties were investigated by the stress-strain test and by differential scanning calorimetry (DSC). The results obtained show that the properties of this material depend very strongly on the alignment of its macromolecules. In particular, the only partial alignment, obtainable by a relatively moderate drawing just after the extrusion, leads to values of elongation at break too high for use of the fibers as suture threads. The DSC analysis reveals interesting properties of the material, but also confirms their strong dependence on the extrusion procedure and on the mechanical treatment. In conclusion, the results of this preliminary study show that the spinning technique must be improved, and that further investigations are necessary to ascertain the possibility of using these poly(ester-ether-ester)s for the fabrication of suture threads.

Journal Article↗