Small-sized stainless steel hand files are conventionally employed in root canal treatment procedures for canal scouting and for glide path establishment, owing to their superior flexibility and proficiency in navigating confined spaces. Given the diversity of brands available in the market, there exists potential variability in their physical characteristics, thereby influencing clinical performance. Consequently, this study aims to conduct a comparative analysis of the design, metallurgy, and mechanical characteristics among seven stainless steel hand file brands across ISO sizes 06, 08, and 10. A total of 315 new 25 mm length stainless steel hand files with apical sizes of 0.06, 0.08, and 0.10 from seven distinct brands were included in the study. A meticulous inspection of all instruments was undertaken to identify any structural deformations that might render them ineligible for the study. The design inspection involved the random selection of instruments from each group, which were examined under various microscopes, including a dental operating microscope, optical microscope, and scanning electron microscope. Furthermore, two instruments from each group underwent energy-dispersive X-ray spectroscopy analysis for elemental composition documentation. Mechanical tests were conducted to evaluate the instruments' resistance to lateral deformation (buckling) and their microhardness. Statistical analysis was executed using the nonparametric Mood's median test, with a predetermined significance level of 0.05. Regarding the instruments design, all files exhibited an active blade length ranging from 16 to 17 mm. However, variations were observed in the number of spirals, tip designs, and sizes, with the API K-File notably larger in sizes 0.06 and 0.08 compared to the other instruments. Despite uniform elements composition, differences in geometric features and mechanical properties were evident. Concerning buckling strength, the API K-File demonstrated superior performance across all tested sizes, while the Dentsply ReadySteel, SybronEndo, and Mani K-Files exhibited lower results (p < 0.05). In microhardness assessments, both the API and Oro K-Files displayed the lowest outcomes, with medians of 531 HVN and 532 HVN, respectively, whereas the SybronEndo K-File exhibited the highest microhardness (657 HVN). Despite similar metallurgical composition, the observed distinctions in geometric features and mechanical properties underscore the impact of the manufacturing process on the characteristics of glide path stainless steel endodontic files. These disparities may ultimately influence their clinical performance.