Cartilage regeneration is still a highly challenging field due to its low self-healing ability. This study used a core-shell electrospinning technique to enhance cartilage tissue engineering by incorporating cartilage extracellular matrix (ECM). The core of fibers included poly(3-hydroxybutyrate)-Chitosan (PHB-Cs) and Halloysite nanotubes. The shell of fibers consisted of Cs and ECM (0, 1, 3, 5 wt%). Subsequently, the scaffolds were named 0E, 1E, 3E, and 5E. The study aimed to assess the impact of ECM on cellular behavior and chondrogenesis. Our findings indicate that ECM reduced fiber diameter from 775 nm for the 0E scaffold to 454 nm for the 1E scaffold. Water contact angle measurements revealed an increasing trend by ECM addition, from 42° for 0E to 67° for 1E. According to mechanical analysis, the 1E scaffold represented the highest strength (5.81 MPa) and strain (3.17%). Based on these analyses, the 1E was considered the optimum scaffold. MTT analysis showed cell viability of over 80% for the 0E and 1E. Also, the gene expression level was assessed for Collagen II, Aggrecan, SOX 9, and Collagen X. The results represented that in the 1E scaffold Collagen II, Aggrecan, and SOX 9 were more upregulated at the end of the 21st day. However, in the 1E scaffold collagen X, as a hypertrophy marker, was downregulated at the end of the experiment. Overall, these results confirmed the potential of the 1E scaffold to be introduced as a promising cartilage tissue engineering scaffold for further studies.