To optimize the structural, mechanical, and bioactivity properties, several bioactive-glass/chitosan/zeolite bionanocomposites were prepared using different methods and various compositions. A series of bionanocomposites containing 25, 50, and 70 wt% of bioactive-glass, 25, 45, and 70 wt% of chitosan and 5 wt% of natural zeolite were fabricated through three methods, including liquid phase, ultrasonic and microwave. Structural and morphological properties of the bio-nanocomposites were studied using several techniques, X-Ray Diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), Dynamic light scattering (DLS), Field Emission Scanning Electron Microscopy (FE-SEM) and energy dispersive X-ray spectroscopy (EDX). Mechanical tests of the bio-nanocomposites revealed that the samples with 50% bioactive-glass had the highest strength. The bio-nanocomposite composed of 50 wt% bio-glass, 45 wt% chitosan and 5 wt% zeolite prepared by liquid-phase route exhibited the highest fractural and compressive strength. Moreover, to study the bioactivity behavior of the bio-nanocomposites the growth of hydroxyapatite was investigated in the Simulated body fluid (SBF) solution. SEM and XRD findings revealed that composites prepared through the ultrasound-assisted route had the highest bio-activity potential.