In this study, computational fluid dynamics (CFD) analysis of a rotating-wall perfused-vessel (RWPV) bioreactor is performed to characterize the complex hydrodynamic environment for the simulation of cartilage development in RWPV bioreactor in the presence of tissue-engineered cartilage constructs, i.e., cell-chitosan scaffolds. Shear stress exerted on chitosan scaffolds in bioreactor was calculated for different rotational velocities in the range of 33–38rpm. According to the calculations, the lateral and lower surfaces were exposed to 0.07926–0.11069dyne/cm² and 0.05974–0.08345dyne/cm², respectively, while upper surfaces of constructs were exposed to 0.09196–0.12847dyne/cm². Results validate adequate hydrodynamic environment for scaffolds in RWPV bioreactor for cartilage tissue development which concludes the suitability of operational conditions of RWPV bioreactor.