Tissue engineering (TE) seeks to employ regeneration techniques on damaged tissues by developing biocompatible scaffolds, enabling tissue growth and repair. For that, specific cell culture conditions and stimuli are required. These requirements can be ensured through a perfusion bioreactor that simulates the biological and mechanical environment of native tissue. This work presents the methodology behind the development of an innovative bioreactor concept, which integrates two independent chambers for cell culturing of double-layered scaffolds to regenerate osteochondral tissues. This concept presents a geometry integrating two chambers separated by a superhydrophobic membrane, assuring medium separation. Scaffolds are placed on the membrane and submitted to mechanical stimulation through orthogonally placed electrically driven pistons. The generated compressive stress intends to simulate in vivo loading. The chambers profile was designed to avoid turbulence flow of the medium for some speed range. To avoid any alteration of the prescribed parameters as ideal for the tissue growth, temperature, pH, mechanical compressive forces, and static pressure values are monitored and recorded during the whole experiment and can be adjusted at any time whenever necessary. The innovative bioreactor concept presented here is expected to contribute to overcoming engineering constraints associated with biotissue production, namely osteochondral tissues.