Prolonged sitting is a potential risk factor for musculoskeletal disorders in office workers. This study aims to evaluate the effect of active breaks on reducing muscle overload in subjects who sit for long periods using infrared thermography (IRT). A sample of 57 office workers participated in this study and were divided into two groups: active breaks (ABs) and no active breaks (NABs). The NAB group sat continuously for 90 min without standing up, while the AB group performed stretching and mobility exercises every 30 min. IRT measurements were taken every 30 min before the active breaks. The results highlight that the skin temperature of the back increased significantly in both groups after 30 min of sitting; however, in the subsequent measurements, the AB group showed a decrease in temperature, while the NAB group maintained a high temperature. Exercise and time point of measurement all reported -values < 0.001; there were no statistically significant differences between the Δ of the NAB and AB groups, while the Δ and Δ of the NAB and AB groups showed statistically significant differences for all back regions. The clinical relevance of this study confirms the negative effects of prolonged sitting on the health of the back, demonstrating that active breaks can reduce back strain, emphasizing the need for workplace interventions. In addition, IRT represents a non-invasive method to assess back muscle overload and monitor the effectiveness of interventions in all categories of workers who maintain a prolonged sitting position. The main limitation of this study is the absence of a questionnaire for the assessment of back pain, which does not allow a direct correlation between temperature changes and back pain outcomes.