Silk has been used commercially as biomedical sutures for decades. Recently silk fibroin, especially from Bombyx mori silkworm, has been explored for many tissue engineering applications such as bone and cartilage due to its impressive biological compatibility and mechanical properties. In Thailand, Thai native silkworms have been long cultivated. Distinct characteristics of cocoon Thai silk are its yellow color and coarse filament. There is more sericin in Thai silk than in other Bombyx mori silks. These characteristics provide Thai silk a unique texture for textile industry. It is therefore the aim of this study to develop three-dimensional silk fibroin-based scaffolds from Thai yellow cocoon “Nangnoi-Srisaket” of Bombyx mori silkworms using salt-leaching method. To enhance the biological properties, type A gelatin, the denature form of collagen having good biocompactibility, was used to conjugate with silk fibroin scaffolds. The pore size of salt-leached silk fibroin scaffold structure represented the size of salt crystals used (600-710µm). After gelatin conjugation, gelatin was partly formed fibers inside the pores of silk fibroin scaffolds resulting in fiber-like structure with highly interconnection. Gelatin conjugation enhanced the compressive modulus of silk fibroin scaffolds by 93%. The results on in vitro culture using mouse osteoblast-like cells (MC3T3-E1) showed that gelatin conjugation could promote the cell proliferation in silk fibroin scaffolds. Moreover, the observed morphology of cells proliferated inside the scaffold after 14 days of culture showed the larger spreading area of cells on conjugated gelatin/silk fibroin scaffolds, compared to round-shaped cells on silk fibroin scaffolds. The results implied that Thai silk fibroin looked promising to be applied in tissue engineering and gelatin conjugation on Thai silk fibroin scaffolds could enhance the biological properties of scaffolds.