The aim of the study was to assess results from a large cohort of patients undergoing otosclerosis surgery with respect to the impact of intra-operative variables on post-operative hearing function and complications. We enrolled 384 patients affected by otosclerosis who were subjected to stapes surgery between 2004 and 2013 at a single institution. Surgery was performed in all cases under local anaesthesia, using a manual perforator and/or microdrill. Teflon-piston prosthesis was used in all patients. Audiological data obtained preoperatively and at last follow up examination (minimum 12 months) were compared. Statistical analysis was performed using the multiple regression model. Peripheral rim otosclerosis and diffuse otosclerosis were associated with better functional results compared to the obliterative pattern (p < 0.05). Mean postoperative Air-Bone Gap was significantly higher in the 0.4 mm, compared to 0.6 mm piston group at 0.5 kHz (p < 0.001) and 1 kHz (p < 0.02); in the stapedotomy group a statistically significant difference was found between 0.4 and 0.6 mm piston groups, in favour of the latter p < 0.05). No differences were encountered in terms of average hearing threshold and complications. Intra-operative variables cannot be fully predictable and our data could help in stratification of the results and as a landmark for the surgeon’s decisions.