Radiation grafting of hydrophilic monomers onto poly[bis(trifluoroethoxy)phosphazene].

The hydrophilic monomers dimethylacrylamide (DMAA) and acrylamide (AAm) were radiation grafted onto poly[bis-(trifluoroethoxy)phosphazene] (PTFP) for biocompatibility enhancement through direct irradiation of the polymer films immersed in monomer-solvent mixtures, and the progress of grafting was followed up to a large monomer depletion. The water absorption of the samples was found to increase as the amount of the grafted chains increased. The grafted chains, mainly confined in the film surface at high dose rates, progressively extend to the interior of the sample as the dose rate decreases. In the grafted samples the ultimate mechanical properties epsilon r and sigma r were essentially preserved. The in vivo tests indicated an enhancement of surface biocompatibility in the PTFP-g-AAm samples and not in the PTFP-g-DMAA samples.

Biocompatible polyphosphazenes by radiation-induced graft copolymerization and heparinization.

Investigations were carried out on the radiation-induced graft copolymerization by direct irradiation of dimethylaminoethyl methacrylate on to poly(bis(trifluoroethoxy)phosphazene) and on to poly (bis(phenoxy)phosphazene). Kinetics of grafting were followed with the polyphosphazenes immersed in monomer - methanol mixtures of various composition. The grafted film samples were quaternized with methyl iodide and, to the produced ammonium group, heparin was ionically bonded with high yield. On the grafted and heparinized-grafted film samples an evaluation of hydrophilicity, mechanical properties, biocompatibility and anticoagulating properties was carried out.

Controlled release of biomolecules from temperature-sensitive hydrogels prepared by radiation polymerization.

Poly(acryloyl-L-proline methyl ester)-based hydrogels containing 1 and 5% of a crosslinking agent were studied as drug delivery systems. The drug loading properties were investigated by matrix incubation into solutions containing biomolecules with molecular weight ranging between 300 and 65,000 Da. The loading yield was found to depend on both the crosslinking degree and the molecular weight of the drug. In vitro release studies were carried out with both swollen and dry matrices loaded with gentamicin, isoniazid and insulin. Gentamicin and isoniazid were released by a bimodal Fickian diffusion with a remarkable burst that was found to depend on both matrix crosslinking degree and physical state. In vivo, the subcutaneous implantation into mice of the isoniazid loaded matrices allowed for an efficient drug release for 800 h. In vitro insulin was released from the swollen matrices for 1500 h by diffusional Fickian mechanism while the dry ones displayed a lag time followed by Fickian diffusion release. The subcutaneous implantation of the insulin-loaded matrices into diabetic mice induced a remarkable decrease in the glucose concentrations in blood. In particular, the dry 1% matrices were found to maintain a low glucose level for 700 h.

Radiation-induced polymerization for the immobilization of penicillin acylase.

The immobilization of Escherichia coli penicillin acylase (EC 3.5.1.11) was investigated by radiation-induced polymerization of 2-hydroxyethyl methacrylate at low temperature. A leak-proof composite that does not swell in water was obtained by adding the cross-linking agent trimethylolpropane trimethacrylate to the monomer-aqueous enzyme mixture. Penicillin acylase, which was immobilized with greater than 70% yield, possessed a higher Km value toward the substrate 6-nitro-3-phenylacetamidobenzoic acid than the free enzyme form (Km = 1.7 X 10(-5) and 1 X 10(-5) M, respectively). The structural stability of immobilized penicillin acylase, as assessed by heat, guanidinium chloride, and pH denaturation profiles, was very similar to that of the free-enzyme form, thus suggesting that penicillin acylase was entrapped in its native state into aqueous free spaces of the polymer matrix.

Controlled release of proteins and peptides from hydrogels synthesized by gamma ray-induced polymerization.

Hydrogels prepared by radiation-induced polymerization at a low temperature have been used as carriers for the controlled release of peptides and proteins. It was found that polymerization of 2-hydroxyethyl methacrylate in the presence of poly (ethylene glycol) methyl ether (MPEG) enabled the more porous and swellable matrics to be obtained, the higher the molecular weight of MPEG. As a consequence, protein release took place at an increasing extent and, provided that MPEG molecular weight was high enough, high molecular weight proteins could also be released. Such a state of affairs was not met in the case of hydrogels based on poly (2-hydroxyethyl acrylate). SEM analysis revealed that even high molecular weight MPEG did not give rise to any porosity, even though the degree of swelling was very high. As a result, no protein release was observed. It was therefore concluded that control of hydrogel porosity for the controlled release of large proteins is of overwhelming importance.

In vitro and in vivo behaviour of narciclasine released from matrices based on poly (2-hydroxyethyl methacrylate).

Narciclasine (1,2,3,7-tetrahydroxy-8,9-methylendioxy-1,2,3,4-tetrahydrophena ntridone) is a natural substance with strong antimitotic effects on cells and potential antitumor activity. Its release form a hydrogel matrix was studied with the purpose of avoiding the concentration spikes of the parenteral administration. The matrix prepared by gamma ray polymerization of a mixture of 2-hydroxyethyl methacrylate (85%) and trimethylolpropane trimethacrylate (15%) was found to release narciclasine for several days, according to a diffusion controlled mechanism. In agreement with its antimitotic effect, narciclasine inhibited the growth rate of healthy mice, when the drug-loaded matrix was introduced subcutaneously. Antitumor effect was observed in an experimental model of Erlich ascitic tumor when low amounts of tumor cells were inoculated. No effect was observed at high concentrations of inoculum or towards solid tumors (Sarcoma 180). This behaviour was related to the rapid clearance of narciclasine from the body which prevented the reaching of sufficient therapeutical concentrations. A pharmacokinetic investigation carried out by an original method of assay demonstrated that narciclasine was accumulated in significant amounts in the kidney only and eliminated in urine with a half time of less than 20 min.