The present study delineates the formulation of niosomes from biocompatible surfactant Tyloxapol and their potential as drug delivery system for anti-tuberculosis drugs. Drug loaded niosomes have a size of 150 nm with a loading efficiency of 97.95±0.2, 98.89±0.2 and 99.50±0.2% for rifampicin (RIF), isoniazid (INH), pyrazinamide (PZA), respectively. Fourier transform infrared spectroscopic studies infer that the drugs are in harmony with the fabricated niosomes since no visible interactions between the drug and niosomes have been detected. The prepared formulations are quite stable as assessed using absorption spectroscopy. TEM images and photoluminescence results reveal that RIF and INH are located in the film bilayer whereas PZA is adsorbed mainly on the surface head groups. In vitro dissolution studies at physiological conditions have been undertaken to compare the release behavior of drugs from the prepared niosomes. Sustained release has been achieved for hydrophilic drugs and an acceptable release in case of RIF. Comparison of regression coefficients of different kinetic models reveal that INH release follows Fickian diffusion mechanism whereas RIF and PZA, a non-Fickian release mechanism. Such a versatile system is expected to reduce dose-related drug toxicity and reach the atelectatic areas.