Electrospun pectin/poly(vinyl alcohol) (PVA) nanofibers incorporating turmeric and bioactive keratin hydrolysate (BKH) were developed as multifunctional systems combining antioxidant and antimicrobial properties. The fibers exhibited homogeneous morphology with diameters ranging from 405 to 518 nm. Thermal characterization indicated a three-stage degradation profile, with maximum mass loss occurring between 200 and 400 °C, corresponding to disruption of the fibrous network and cleavage of covalent bonds within the matrix. Mechanical evaluation revealed that turmeric incorporation reduced tensile strength from 12.23 MPa to 2.18-3.04 MPa, while elongation at break decreased from 57% to 15-34%, depending on BKH content, evidencing compositional modulation of structural integrity. Antimicrobial assays showed the inhibition of Staphylococcus aureus and Escherichia coli in both agar plates and milk. The ABTS radical scavenging ranged from 77% (1700 μM TEAC) to 90% (1993 μM), while DPPH activity reached 30% (512 μM TEAC) in formulations containing only BKH and 92% (1461 μM) in nanofibers incorporating both turmeric and BKH, indicating enhanced antioxidant capacity in combined systems. Cytotoxicity assessment using L929 cells demonstrated preserved cell viability for NF1, whereas NF2 exhibited reduced biocompatibility in the MTT assay. Hemolytic activity remained below 2% across all formulations, confirming minimal erythrocyte membrane disruption. The combined results demonstrate that variations in BKH content influence mechanical properties, antioxidant capacity, antimicrobial performance, and cellular viability in a composition-dependent manner. These findings indicate the developed nanofibers show potential as multifunctional materials for active food packaging applications.
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