This study explores the bioactive potential of banana leaf extracts and their innovative integration into knitted hemp fabrics. To obtain the extracts, distinct extraction methodologies were employed, namely conventional extraction, ultrasound-assisted extraction, and pressurized-liquid extraction. Aqueous and hydroethanolic solvents, namely 20% (v/v) and 50% (v/v), were employed during the extraction process. Furthermore, the cationization and functionalization of knitted hemp fabrics with the banana leaf extracts was achieved through padding. The extracts’ phenolic content and antioxidant activity were evaluated using the Folin–Ciocalteu (FC) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays, respectively. The results indicated that both ultrasound-assisted extraction and pressurized-assisted extraction substantially enhanced the yield of phenolic compounds in comparison to conventional extraction, while employing 50% EtOH as a solvent also improved extraction yields for all extraction methodologies. The functionalized knits were further characterized concerning their antioxidant activity by DPPH, assessing their antimicrobial properties through ATCC TM-100 standard against three microorganisms (Staphylococcus aureus, Candida Krusei, and Candida albicans), and UV protection according to the standard AS/NZS 4399:2017. Antioxidant activity was highest in knits functionalized with extracts obtained via ultrasound-assisted extraction, while antimicrobial properties were most pronounced in knits treated with hydroalcoholic extracts, particularly those derived from assisted methods. The UV protection was enhanced in extracts with higher ethanol concentrations obtained through ultrasound-assisted extraction, with these knits exhibiting the highest Ultraviolet Protection Factor (UPF). This research not only highlights the efficacy of the alternative extraction technologies but also offers valuable insights for the development of innovative, biocompatible materials with enhanced bioactive properties for diverse applications in the textile and healthcare sectors, paving the way for sustainable applications.