Ti6Al4V scaffolds with pore sizes between 300 and 600 μm are deemed suitable for bone tissue engineering. However, a significant proportion of pores in human bones are smaller than 300 μm, playing a crucial role in cell proliferation, differentiation, and bone regeneration. Ti6Al4V scaffolds with these small‐sized pores have not been successfully fabricated, and their cytocompatibility remains unknown. The paper presents a novel ink formula specifically tailored for the fabrication of Ti6Al4V scaffolds featuring precise and unobstructed sub‐300 μm structural pores, achieved by investigating the rheological properties and printability of five inks containing 60–77.5 vol.% Ti6Al4V powders and PS/DCM/NMP binders. Ti6Al4V scaffolds with 50 μm to 600 μm pores are fabricated via direct ink writing and subsequently subjected to in vitro MC3T3‐E1 and BMSCs experiments. The live/dead assay, FITC‐ phalloidin/DAPI staining, and CCK‐8 assay indicate that the 100 μm pore‐sized scaffolds exhibit the highest cell adhesion and proliferation capacity. The alizarin red staining, RT‐qPCR assay, and immunocytochemical staining demonstrate the superior osteogenic differentiation potential of 100 μm and 200 μm pore‐sized scaffolds. The study highlights the importance of sub‐300 μm structural pores and redefines the optimal pore size for Ti6Al4V scaffolds, advancing the development of bone tissue engineering and clinical medicine.This article is protected by copyright. All rights reserved