Small linear microgrooves have been observed on root surfaces associated with osseous defects when videoscope-assisted minimally invasive surgery (V-MIS) is performed. The aim of this study was to evaluate the frequency of occurrence of these microgrooves associated with osseous defects. Videos of dry skulls from an anatomy department collection and videos made during consecutive V-MIS procedures on patients were evaluated to determine the presence or absence of microgrooves associated with osseous defects. In the dry skulls, 69 areas of apparent periodontal osseous defects were evaluated. In the V-MIS videos, 63 osseous defects were evaluated. In the dry skulls, 34 sites were positive for microgrooves on the root surfaces (49%), while in the V-MIS procedures 50 sites were positive for microgrooves (79%). Microgrooves of unknown origin were detected in a high percentage of osseous defects. Microgrooves may represent risk factors for site-specific progression of periodontal destruction.