Resonance frequency analysis (RFA) has been used as a diagnostic method to measure implant stability at all stages of healing. In addition to evaluating the status of the peri-implant marginal bone, it can also indicate the most appropriate time to load the implant. This in vitro study aimed to evaluate the efficacy of RFA as a diagnostic method for the detection of peri-implant marginal bone loss (MBL). Forty bone-level Klockner Vega implants were placed in a polyurethane block with elastic properties similar to those of the maxillary bone. The insertion torque and primary implant stability at the time of placement were measured using an RFA device. A circumferential peri-implant defect was created by removing the cortical bone portion in each implant using a trephine. The stability values were measured again using RFA. The stability values measured using RFA were lower after the creation of the circumferential peri-implant defect, indicating a statistically significant decrease in implant stability. The results of the study tend to show a relationship between peri-implant marginal bone loss and modifications in implant stability measured by RFA.