Bicomponent electrospun nanofibers based on the combination of synthetic (i.e., aliphatic polyesters such as polycaprolactone (PCL)) and natural proteins (i.e., gelatin) have been extensively investigated as temporary platforms to instruct cells by the release of molecular/pharmaceutical signals for the regeneration of several tissues. Here, water soluble proteins (i.e., gelatin), strictly embedded to PCL, act as carriers of bioactive molecules, thus improving bioavailability and supporting cell activities during in vitro regeneration. However, these proteins are rapidly digested by enzymes, locally produced by many different cell types, both in vitro and in vivo, with significant drawbacks in the control of molecular release. Hence, we have investigated three post-processing strategies based on the use of different crosslinking agents—(1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) (EDC), glyceraldehyde (GC), and 1,4-butanediol diglycidyl ether (BDDGE)—to delay the dissolution time of gelatin macromolecules from bicomponent fibers. All of the qualitative (i.e., SEM, TGA) and quantitative (i.e., Trinitrobenzene sulfonate (TNBS) and bicinchoninic acid (BCA) assays) morphological/chemical analyses as well as biocompatibility assays indicate that EDC crosslinking improves the chemical stability of bicomponent fibers at 37 °C and provides a more efficient encapsulation and controlled sustained release of drug, thus resulting in the best post-treatment to design bio-inspired fibrous platforms for the extended in vitro release of drugs.
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