Biological and Biophysical Characterization of Hybrid PLCL Nanofibers Incorporating Stem Cell-Derived Secretome

The design of multifunctional biomaterials that offer both structural support and biochemical cues is essential for enhancing tissue regeneration. In this study, hybrid nanofibrous scaffolds composed of poly(L-lactide-co-ε-caprolactone) (PLCL) and bioactive factors secreted by Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) were fabricated via co-electrospinning. Nanofibers were produced in aligned and random configurations following an optimized protocol developed at the Institute for Bioengineering of Catalonia (IBEC). Their morphology and topography were characterized by light microscopy, scanning electron microscopy (SEM), and atomic force microscopy (AFM), and fiber orientation was quantified via Fast Fourier Transform (FFT) analysis. The scaffolds showed fiber diameters of 542.9 ± 62.3 nm, with aligned fibers predominantly oriented within 20° of the principal axis. Human AD-MSCs were used to assess biocompatibility and cell-material interactions. Aligned and random nanofiber architectures elicited distinct cellular responses. AD-MSCs on aligned fibers exhibited smaller spreading areas (~320 μm²) vs. on random nanofibers (~500 μm²) and substantially higher proliferation, resulting in a shorter cell-doubling time (~25 h) than those on random nanofibers (~130 h) or control substrates (~70 h). In addition, aligned nanofibers promoted markedly faster migration, reaching rates of ~5000 μm²/h surface coverage, compared with random nanofibers (~770 μm²/h) and controls (~1800 μm²/h). Together, the results show that nanofiber alignment and biochemical functionalization jointly influence MSC behavior and improve regeneration, highlighting the potential of these PLCL-based hybrid secretome/PLCL nanofibers for advanced wound healing.