Cell Adhesion and Cytotoxicity Assessment of Collagen-Coated Electrospun PCL Membranes Functionalized with Silver Nanoparticles

Chronic and complex wounds require biomaterials that are both cytocompatible and antimicrobial. Herein, electrospun polycaprolactone (PCL) nanofiber membranes were coated with Type I collagen and functionalized with silver nanoparticles (AgNPs). The main objective was to assess fibroblast adhesion, proliferation, and cytotoxicity. Membrane morphology and surface characteristics were analyzed in a previous work by SEM, AFM, and wettability measurements, confirming the transformation from hydrophobic PCL to fully wettable collagen-coated surfaces. In this study, Murine 3T3 fibroblasts were cultured on PCL, PCL-Collagen, PCL-Collagen-Citrate, and PCL-Collagen-AgNPs membranes. Cellular activity was quantified using Alamar Blue assays at 24, 48, and 72 h, while cytotoxicity was determined by LDH release. Cellular viability and adhesion were studied using confocal microscopy. All membrane types supported fibroblast growth, with collagen-coated samples exhibiting the highest metabolic activity. AgNPs-functionalized membranes sustained overall cell viability above 90%, with cytotoxicity values of approximately 10% at 24 h and 20% at 48 h. Antimicrobial evaluations demonstrated complete inhibition of <i>Pseudomonas aeruginosa</i> and <i>vancomycin-resistant Enterococcus</i>, and partial inhibition of <i>Staphylococcus aureus</i>. These results indicate that collagen-coated, AgNPs-functionalized electrospun PCL membranes exhibit both high cytocompatibility and significant antimicrobial activity, supporting their potential as advanced wound-dressing materials.