Considering the role of glucosamine sulfate (GS) in the biosynthetic pathways of chondrocytes, an attempt was made to design an electrospun poly-3-hydroxybutyrate (PHB) scaffold loaded with GS to develop cartilage tissue engineering. The study was initiated using the optimal electrospun scaffold conditions for the synthesis of PHB/GS. The resulting scaffolds have shown excellent pore architectures and mechanical behavior compared to pure PHB. UV spectrophotometric analysis, for the evaluation of the GS release behavior, showed zero-order kinetics release. In vitro results indicated excellent cell viability, cell adhesion, and cell penetration of PHB/GS scaffolds compared to pure PHB. [GRAPHICS] .