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The goal of this study was to characterize the protein composition of the conditioning film deposited onto the surface of ureteral stents during in vivo implantation and to relate its presence to the precipitation of calcium crystals. The protein pattern of the conditioning film of implanted nonencrusted and encrusted urological stents was assessed by SDS-PAGE and Western blot of the desorbed species. The results obtained highlighted different electrophoresis profiles between nonencrusted and encrusted stents. Western blot showed the ubiquitous presence of albumin, while Tamm-Horsfall Protein and alpha1-microglobulin adsorption was limited to nonencrusted devices. By an in vitro dynamic model in which artificial urine was flowed through the lumen of control and retrieved nonencrusted stents, we demonstrated that the organic layer remarkably enhanced crystal precipitation and aggregation events on the surface.

A recent study in our laboratory has identified the potential role of urine-derived alpha-1-microglobulin (alpha-1-m) in mediating Pseudomonas aeruginosa adhesion to polystyrene, while other workers have suggested a possible role of the protein in the immunological response. Due to the ubiquitous presence of alpha-1-m in body fluids, the adsorption of the protein from serum, cerebrospinal fluid, urine and used continuous ambulatory peritoneal dialysis fluid onto polystyrene was investigated. The treated surfaces were sequentially immersed in water and increasingly concentrated isopropanol-water solutions in order to selectively desorb bound proteins on the basis of their binding strength. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of the wash supernatants showed different protein desorption profiles for each biological fluid, despite the qualitative similarity between the protein composition of the fluids, and highlighted the uptake of alpha-1-m from each fluid to the surface. In the case of urine, the analysis was extended to commercial polyurethane and silicone stents. The ease of desorption of urine-derived alpha-1-m could be correlated with surface hydrophobicity of the stent biomaterial.

Ideally an active bone biomaterial should increase the mineralisation rate at the bone healing sites, keeping at the same time the inflammation process to levels required for tissue regeneration. Our studies suggest that in addition to improving the nucleation process for new bone formation, coating titanium with phospholipids may reduce the inflammatory response, which was shown to vary depending on the formulation employed. As phosphatidylserine reduced the inflammatory response to the greatest extent, in the second part of this study we examined its effect on osteoblast mineralisation. These studies demonstrated that phosphatidylserine improves the nucleation process for bone formation, by promoting the formation of bone-like tissue, so the high mineralisation potential of phosphatidylserine-coated titanium, together with the lower level of inflammatory response, supports the further development of this technology for coating osteointegrative devices.

Poly(2-hydroxyethylmethacrylate) [poly(HEMA)] is a widely used biomaterial which does not allow cell adhesion and growth on its surface, limiting its use in biomedical applications in which cell cohesion is detrimental. We have prepared a poly(HEMA)-gelatin composite hydrogel using a sequential interpenetrating polymer network technique. The properties of this material were compared with poly(HEMA) freeze-dried sponges in terms of morphology, mechanical properties and biocompatibility. Moreover, in vivo biocompatibility experiments highlighted the occurrence of cellular interactions on the surface of the poly(HEMA)-gelatin interpenetrating polymer network, which are usually absent when unmodified poly(HEMA) hydrogels are implanted in the same host organism. These tests also showed a progressive gelatin degradation from the surface to the bulk of the poly(HEMA)-gelatin specimens during short-term (7 d) implantation. Finally, in vitro tests confirmed an improved ability of this composite to scaffold for the cells.

According to the modern definition of biocompatibility, a biocompatible material need not be inert but be bioactive. A benign reactivity implies that the reactivity has to be appropriate for the intended use. Chitosan, a non-acetylated or partially deacetylated chitin (a linear homopolymer of beta (1-4)-linked N-acetylglucosamine) has been proposed as a biomaterial because of its apparent satisfactory biocompatibility. The present investigation demonstrates that chitosan has an in vitro stimulatory effect on both macrophage nitric oxide (NO) production and chemotaxis. The macrophage NO secretion is attributed to the N-acetylglucosamine unit of the chitosan molecule rather than to the glucosamine residue (28 and 15 microM NO respectively). Moreover, the immune stimulatory effect of chitosan was very specific since other glycosaminoglycans, such as N-acetyl-D-mannosamine and N-acetyl-D-galactosamine, had no effects on NO production (5 and 8 respectively). In vivo experiments strengthen this hypothesis. Transmission electron microscopy analysis identifies the presence of many leucocytes in the specimens after 14 d post-implantation, showing poor healing processes (i.e. fibroblast proliferation and collagen deposition) that characterize the tissue repair at this time in our animal model.