Two series of well-defined lignin fractions derived from birch and spruce alkaline lignin (AL) by sequential solvent fractionation (i-PrOH-EtOH-MeOH) were engaged in a structure–property-application relationship study. The bacterial-derived alkaliphilic laccase (MetZyme) extensively catalyzed the oxidation and polymerization of AL fractions in an aqueous alkaline solution (pH 10). Lignin fractions with low molar mass reached a higher polymerization degree due to more phenolic-OH groups serving as reactive sites of oxidation and better lignin-laccase accessibility arose from a lower lignin condensation degree than the high molar mass ones. In comparison, AL fractions from spruce were found to be less reactive toward the laccase-catalyzed polymerization than those from birch, which was attributed to the much pronounced aryl-vinyl moieties’ oxidation. Furthermore, in situ polymerization of birch AL fractions using microfibrillated cellulose as a structural template was conducted in an aqueous medium and a dispersion of nanocellulose with its fiber network evenly coated by aligned lignin nanoparticles was obtained. The present study not only provides fundamental insights on the laccase-assisted oxidation and polymerization of lignin but also presents a new perspective for valorizing lignin in biobased fiber products through green processing of solvent fractionation and enzymatic treatment.

Tissue engineering is currently one the fastest growing engineering fields, requiring fabrication of advanced and multifunctional materials to be used as scaffolds or dressing for tissue regeneration. In this work, a bilayer matrix was fabricated by electrospinning of a hybrid cellulose acetate nanofibers (CA) containing bioactive latex or Ciprofloxacin over highly interconnected collagen (CSPG) 3D matrix previously obtained by a freeze-drying process. The bilayer matrix was fabricated with a nanofibrous part as the primary (top) layer and a spongy porous part as the secondary (bottom) layer by combining electrospinning and freeze-drying techniques to enhance the synergistic effect of both materials corresponding to physical and biological properties. The final material was physicochemically characterized using Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The bilayer matrix exhibited nanofibrous and 3D porous structure with properties such as high porosity, swelling, and stability required for soft-tissue-engineering applications. Furthermore, the in vitro biological and fluorescence properties of the matrix were tested against NIH 3T3 fibroblast and human keratinocyte (HaCaT) cell lines and showed good cell adhesion and proliferation over the bilayer matrix. Thus, the synergistic combination of nanofibrous material deposition onto to the collagenous porous material has proved efficient in the fabrication of a bilayer matrix for skin-tissue-engineering applications.