This study develops and evaluates electrospun polycaprolactone (PCL) nanofiber membranes coated with collagen and functionalized with silver nanoparticles (AgNPs) as advanced dressings for complex wounds. Membranes were produced by electrospinning (12% w/v PCL), plasma-activated to increase hydrophilicity, then coated with Type I collagen; AgNPs were synthesized by citrate reduction (Lee–Meisel) and applied as the antimicrobial layer. Morphological and surface characterization (SEM, AFM, confocal) showed homogeneous fibrous networks (≈350–600 nm) and uniform coatings without nanoarchitectural changes. Physicomechanical testing indicated a compliant, non-rigid material suitable for cutaneous motion; fluid management was appropriate (WVTR within the desirable range). In vitro, 3T3 fibroblasts exhibited improved adhesion and higher metabolic signal on PCL+collagen versus bare PCL (MTT, on-disc Alamar Blue), while AgNPs-bearing membranes maintained short-term cytocompatibility. Antimicrobial activity was quantified by broth microdilution (MIC) against Staphylococcus aureus ATCC 29213, Pseudomonas aeruginosa ATCC 27853, and VRE Enterococcus faecium Ef-5, and by on-membrane biofilm inhibition assays, demonstrating reduced planktonic growth and adherent biomass. Cytotoxicity was assessed using suspension Alamar Blue (contact exposure) and LDH release at 24–48 h, yielding high viability with low–moderate LDH, clearly below the positive control, consistent with a safe operating window at the applied AgNPs loading. Overall, the integrated PCL/collagen/AgNPs system provides compliant mechanical support, pro-regenerative bioactive cues, and a measurable antimicrobial advantage, establishing the basis for ex vivo/in vivo studies toward clinical wound-care applications.
Questo studio sviluppa e valuta membrane nanofibrose di policaprolattone (PCL) rivestite con collagene e funzionalizzate con nanoparticelle d’argento (AgNPs) come medicazioni avanzate per ferite complesse. Le membrane sono state ottenute per elettrofilatura (PCL 12% p/v), attivate al plasma per aumentare l’idrofilia e successivamente rivestite tramite deposito controllato di collagene di Tipo I; le AgNP sono state sintetizzate con riduzione al citrato (Lee–Meisel) e applicate come strato antimicrobico. La caratterizzazione morfologica e superficiale (SEM, AFM, confocale) ha mostrato reti fibrose omogenee (≈350–600 nm) e coating uniformi senza alterazioni della nano‐architettura. Le proprietà fisico-meccaniche hanno indicato materiale conforme e non rigido, idoneo al movimento cutaneo; la gestione dei fluidi è risultata adeguata (WVTR nell’intervallo desiderabile). In vitro, i fibroblasti 3T3 hanno mostrato migliore adesione e incremento di segnale metabolico su PCL+collagene rispetto al PCL nudo (MTT, Alamar Blue on-disc), mentre le membrane con AgNPs hanno mantenuto citocompatibilità a breve termine. L’attività antimicrobica è stata quantificata mediante microdiluizione in brodo (MIC) contro Staphylococcus aureus ATCC 29213, Pseudomonas aeruginosa ATCC 27853 ed Enterococcus faecium VRE (Ef-5), e mediante saggi di inibizione del biofilm su membrana, evidenziando riduzione della crescita planktonica e della biomassa aderente. La citotossicità è stata indagata con Alamar Blue in sospensione (contatto) e rilascio di LDH a 24–48 h, con valori elevati di vitalità e LDH basso-moderato, inferiori al controllo positivo, suggerendo una finestra operativa sicura ai carichi di AgNPs impiegati. Nel complesso, l’integrazione PCL/collagene/AgNPs fornisce un supporto meccanico conforme, segnali bioattivi pro-rigenerativi e un vantaggio antimicrobico misurabile, ponendo le basi per ulteriori studi ex vivo/in vivo verso applicazioni cliniche in wound care avanzato.
UTILIZZO DI NANOFIBRE DI POLICAPROLATTONE RIVESTITE DI COLLAGENE PER MIGLIORARE LA GUARIGIONE DI FERITE DIFFICILI
IURILLI, MARTIN
2026
Abstract
This study develops and evaluates electrospun polycaprolactone (PCL) nanofiber membranes coated with collagen and functionalized with silver nanoparticles (AgNPs) as advanced dressings for complex wounds. Membranes were produced by electrospinning (12% w/v PCL), plasma-activated to increase hydrophilicity, then coated with Type I collagen; AgNPs were synthesized by citrate reduction (Lee–Meisel) and applied as the antimicrobial layer. Morphological and surface characterization (SEM, AFM, confocal) showed homogeneous fibrous networks (≈350–600 nm) and uniform coatings without nanoarchitectural changes. Physicomechanical testing indicated a compliant, non-rigid material suitable for cutaneous motion; fluid management was appropriate (WVTR within the desirable range). In vitro, 3T3 fibroblasts exhibited improved adhesion and higher metabolic signal on PCL+collagen versus bare PCL (MTT, on-disc Alamar Blue), while AgNPs-bearing membranes maintained short-term cytocompatibility. Antimicrobial activity was quantified by broth microdilution (MIC) against Staphylococcus aureus ATCC 29213, Pseudomonas aeruginosa ATCC 27853, and VRE Enterococcus faecium Ef-5, and by on-membrane biofilm inhibition assays, demonstrating reduced planktonic growth and adherent biomass. Cytotoxicity was assessed using suspension Alamar Blue (contact exposure) and LDH release at 24–48 h, yielding high viability with low–moderate LDH, clearly below the positive control, consistent with a safe operating window at the applied AgNPs loading. Overall, the integrated PCL/collagen/AgNPs system provides compliant mechanical support, pro-regenerative bioactive cues, and a measurable antimicrobial advantage, establishing the basis for ex vivo/in vivo studies toward clinical wound-care applications.| File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.14242/357306
URN:NBN:IT:UNITS-357306