One-step electrospun scaffold of dual-sized gelatin/poly-3-hydroxybutyrate nano/microfibers for skin regeneration in diabetic wound.

This work aimed to implement an electrospinning protocol that allows simultaneous production of micro- and nanofibers in a single scaffold to mimic the extracellular matrix (ECM) combining biodegradable polymers and proteins, and to evaluate its capability to manage diabetic wounds. Poly-3-hydroxybutyrate (PHB) and gelatin (Ge) were chosen to prepare micro- and nanofibers, respectively. Electrospinning conditions were optimized testing various polymer concentrations, voltages, and flow rates. One-step dual-size fibers were obtained from 8%w/v PHB in chloroform (microfibers, 1.25 ± 0.17 μm) and 30%w/v gelatin in acetic acid (75%w/v) (nanofibers, 0.20 ± 0.04 μm), at 0.5 mL/h and 25 kV. A chemical characterization, swelling, hydrophilicity of scaffolds made of PHB-microfibers, Ge-nanofibers and their combination (Ge-PHB) were evaluated before and after crosslinking with genipin. All scaffolds showed excellent fibroblasts viability and attachment after incubation for 1, 3, and 7 days, and low levels of hemolysis. In vivo wound healing was evaluated in diabetic rats for 21 days. Ge-containing scaffolds promoted faster healing. The wounds treated with the Ge-PHB scaffolds proved to be in a late proliferative stage showing higher content of hair follicles and sweat glands and lower content in fibroblast compared with the control wounds.