The structure (surface topography, pore size, and porosity) of 3D scaffolds plays an important role in tissue regeneration. Although surface topography, the porosity and pore size of 3D porous scaffolds have been frequently studied, these parameters of 3D printed SF-based scaffolds for tracheal epithelium growth have not been investigated. In this study, we investigated the effects of the structure properties of 3D printed silk fibroin/Hydroxypropyl methyl cellulose(SF/HPMC) scaffolds on the adhesive and proliferative behaviors of tracheal epithelium in vitro. This work fabricated six types of 3D printed SF/HPMC scaffolds with different surface properties, pore size and porosity. The surface topography, pore size, and porosity of the different scaffolds were examined. Normal human bronchial epithelial cell lines (BEAS-2B cells) were cultured on the different scaffolds for 7 days. The proliferation of cells on the different scaffolds was tested by CCK-8 assay. The morphology of BEAS-2B cells on different scaffolds was observed by scanning electron microscopy(SEM). The porosity of 20 wt% SF/HPMC scaffolds with rough surface and smooth surface, and 30 wt% SF/HPMC scaffolds with rough surface and smooth surface were 70.5 ±2.0%、65.5 ±6.1%、63.9 ±2.1%、59.6 ±2.1%, respectively; The pore size of 20 wt% SF/HPMC scaffolds was 443.9±104.1μm and 681.1±115.1μm, respectively. Results showed that BEAS-2B cells proliferated better on the rough surface than that on the smooth surface; BEAS-2B cells proliferated better on the scaffold with higher porosity (65.5 ±6.1%) than that with lower porosity (59.6 ±2.1%); In addition, the cells proliferated well on the SF/HPMC scaffold with small pore size (443.9±104.1μm). SEM showed that cells grew in a sheet on the rough surface and tended to grow in clusters on the smooth surface; BEAS-2B cells tended to grow in clusters on the scaffold with large pore size, while cells could spread into a sheet and form connections between pores on the scaffold with small pore size. In summary, SF/HPMC scaffolds with rough surface, high porosity, and small pore size facilitated cell growth. This provides a preliminary experimental basis for selecting the suitable structure of 3D printed SF/HPMC scaffolds for repairing tracheal defects.