Reconstruction of bone tissue defects is a problematic area of the modern world. Temporary “platforms” of various materials for improving cell adhesion and proliferation have been extensively researched in recent decades. β-tricalcium phosphate (β-TCP) is a suitable biocompatible, biodegradable material used for bone regeneration. The creation of scaffolds with specifically designed surface structures will enable bone engineering applications that require navigated cell proliferation on a substrate with pre-set geometric limits. In this study, an innovative laser-based technique for surface modification was applied to improve the morphological properties of the surface of β-TCP pellets for proper cell surface environment creation. The obtained topographies with diverse processing parameters were compared. Homogenous microgroove structures, less than 100 µm, without the onset of melting and crack formation, were produced. The contribution from the accumulation effect of a diverse number of laser pulses (N = 1–100) on the final structure dimensions was examined. The microstructured scaffolds were investigated by confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) analyses. We studied the effect of the patterned surface of the material on the mouse calvaria osteoblast (MC3T3) cells’ viability and cytotoxicity from 1 to 7 days. The results indicated that cell behavior was affected by microscale dimensions of the surface.