No abstract

Recent efforts in scientific research in the field of peripheral nerve regeneration have been directed towards the development of artificial nerve guides. We have studied various materials with the aim of obtaining a biocompatible and biodegradable two layer guide for nerve repair. The candidate materials for use as an external layer for the nerve guides were poly(caprolactone) (PCL), a biosynthetic blend between PCL and chitosan (CS) and a synthesised poly(ester-urethane) (PU). Blending PCL, which is a biocompatible synthetic polymer, with a natural polymer enhanced the system biocompatibility and biomimetics, fastened the degradation rates and reduced the production costs. Various novel block poly(ester-urethane)s are being synthesised by our group with tailored properties for specific tissue engineering applications. One of these poly(ester-urethane)s, based on a low molecular weight poly(caprolactone) as the macrodiol, cycloesandimethanol as the chain extender and hexamethylene diisocyanate as the chain linker, was investigated for the production of melt extruded nerve guides. We studied natural polymers such as gelatin (G), poly(L-lysine) (PL) and blends between chitosan and gelatin (CS/G) as internal coatings for nerve guides. In vitro and in vivo tests were performed on PCL guides internally coated either with G or PL to determine the differences in the quality of nerve regeneration associated with the type of adhesion protein. CS/G natural blends combined the good cell adhesion properties of the protein phase with the ability to promote nerve regeneration of the polysaccharide phase. Natural blends were crosslinked both by physical and chemical crosslinking methods. In vitro neuroblast adhesion tests were performed on CS/G film samples, PCL/CS and PU guides internally coated with G to evaluate the ability of such materials towards nerve repair.

Bioartificial blends of poly-(epsilon-caprolactone) (PCL) with a polysaccharide (starch, S; dextran, D; or gellan, G) (PCL/S, PCL/D, PCL/G 90.9/9.1 wt ratio) were prepared by a solution-precipitation technique and widely characterized by differential scanning calorimetry analysis (DSC), Fourier transform infrared-attenuated total reflectance spectroscopy (FTIR-ATR), optical microscopy (OM), wide-angle X-ray diffraction analysis (WAXD), and thermogravimetry (TGA). DSC showed that the polysaccharide reduced the crystallinity of PCL and had a nucleation effect, which was also confirmed by OM analysis. Hoffman-Weeks analysis was performed on PCL and blend samples allowing calculation of their equilibrium melting temperatures (). WAXD showed that the crystalline unit cell type was the same for PCL and blends. FTIR-ATR did not evidence interactions between blend components. Thermal stability was affected by the type of polysaccharide. Microparticles (<125 microm) were produced from blends by cryogenical milling and characterized by scanning electron microscopy analysis (SEM). Selective laser sintering (SLS), a new rapid prototyping technology for scaffold fabrication, was applied to sinter blend microparticles according to a PC-designed two-dimensional geometry (strips and 2 x 2 mm(2) square-meshed grids). The optimal experimental conditions for sintering were established and laser beam parameters (beam speed, BS, and power, P) were found for each blend composition. Morphology of sintered objects was analyzed by SEM and found to be dependent on the morphology of the sintered powders. Sintered samples were analyzed by chemical imaging (CI), FTIR-ATR, DSC, and contact angle analysis. No evidence of the occurrence of degradation phenomena was found by FTIR-ATR for sintered samples, whereas DSC parameters of PCL and blends showed changes which could be attributed to some molecular weight decrease of PCL during sintering. CI of sintered samples showed that the polysaccharide phase was homogeneously dispersed within the PCL matrix, with the only exception being the PCL/D blend. The contact angle analysis showed that all samples were hydrophilic. Fibroblasts were then seeded on scaffolds to evaluate the rate and the extent of cell adhesion and the effect of the polysaccharides (S, D, G) on the bioactivity of the PCL-based blends.