A1. A series of PEA precursors was obtained by a three-step polycondensation reaction between
ethylene glycol and adipic acid. The obtained precursors were characterized by NMR (1H and
13C), SEC, Maldi-Tof, DSC and ATG. For the next activity, a PEA with a Mn of 2480 g/mol
was selected.
A2: A series of PCL-PEA-PCL block copolymers were prepared starting from the previously
obtained PEA precursors. Also in this activity was optimized the preparation procedure of
nanoparticles (NPs) based on PCL by an emulsion/evaporation method. A series of parameters
were studied, such as: organic/aqueous ratio; surfactant concentration; sonication time; addition
rate of the organic phase; molecular characteristics of PCL and PVA. After the optimization of
this procedure, drug-loaded NPs were obtained at different drug/PCL ratios (wt/wt). The
obtained NPs were analyzed by DLS and FTIR. Finally, electrospun fibers were also obtained
and the encapsulation of a drug model was investigated. The diameters of the fibers were
determined by SEM in the absence and in the presence of drug.
A3. During this stage, the effect of drug administration systems on blood components was
determined. Considering the fact that nanoparticles (NPs) obtained starting from PCL-PEA-PCL
copolymers will be administered by intravenous injection, it was necessary to determine the
degree of hemolysis of these systems. These tests were performed, depending on time and
concentration, both for empty NPs and for those loaded with different proportions of Nystatin.
Also, their hemolytic effect was compared with that of NPs obtained using a commercial PCL
homopolymer. It is found that regardless of the sample, the degree of hemolysis is lower than the
maximum allowed value of 5%, which means that these systems do not cause blood coagulation.
A4. Cell viability was analyzed at 24 and 48 hours both for the systems, based on PCL-PEA-
PCL copolymers, free as well as those loaded with different amounts of Nystatin. It was found
that the systems based on PCL-PEA-PCL have cell viability higher than 80% for concentrations
equal to or lower than 200 µg/ml both at 24 and at 48 hours. The testing of the antifungal effect
of the systems loaded with Nystatin was carried out by determining the diameter of the inhibition
zone by the diffusimetric method, using as reference strains Candida albicans, Candida glabrata,
Candida parapsilopsis, Candida tropicalis. The antifungal activity of nanoparticles loaded with
nystatin increases with the increase of the amount of nystatin encapsulated in the NPs. At the
highest concentration in nystatin, an antifungal activity comparable to that of free nystatin or
even higher is observed for Candida albicans, Candida glabrata and Candida tropicalis.