The clinical implication of cefepime HCl (CEF) is compromised owing to restricted oral bioavailability and harmful adverse effects without any authorized oral formulation available. The present investigation provides an innovative sustained-release oral drug delivery strategy that tackles the challenges of limited oral bioavailability and prolongs the half-life of CEF. Accordingly, CEF was loaded into a bilosome, a liposome or noisome-based vesicle employing bile salt as a permeation enhancer. Despite its hydrophilic nature, the drug was effectively loaded into bilosomes. Nine various formulas were fabricated by a reverse phase evaporation method. The resulting vesicles increased the encapsulation efficiency (EE %) from 39.31 ± 0.03 % to 63.09 ± 0.01 %, drug loading capacity (DLC %) from 6.99 ± 0.25 to 42.91 ± 0.11 %, the particle size (PS) from 264 ± 13.52 nm to 405.40 ± 8.91 nm, and the polydispersity index (PDI) values ranged from 0.243 ± 0.040 to 0.430 ± 0.050. The zeta potential (ZP) changed from 􀀀 35.67 ± 3.73 mV to 􀀀 62.21 ± 2.21 mV. Further, the release profile exhibited dual release pattern within 24 h, with the percentage of release (CR %) expanding from 42 ± 0.13 % to 69.16 ± 0.09 %. The selected formula was found to be B3 with EE % = 56.61 ± 0.02 %, PS = 264 ± 13.52 nm, ZP = 􀀀 62.21 ± 2.21 mV, PDI = 0.430 ± 0.050, CR % = 52.94 ± 0.06 %, and IC50 of 3.4 ± 0.40 μg/ml against MCF-7 cells with scattered spherical non-agglomerated vesicles. Additionally, it exhibited higher anti-MRSA biofilm, relative bioavailability (5.1 fold), and antimicrobial capacity against P. aeruginosa, E. coli, B. subtilis, and S. aureus compared to pure CEF. Our data demonstrate that bilosome is a powerful nanocarrier for oral delivery of cefepime, boosting its biological impacts and pharmacokinetic profile.