This study explores the impact of different bicycle saddle designs on saddle pressure distribution and comfort in female cyclists suffering from medical conditions affecting the groin area, including Lichen Sclerosus, Lichen Planus, and Vulvar Carcinoma. These conditions can cause discomfort and pain during cycling, leading to a decrease in physical activity and quality of life. The research involved a comparison of four distinct saddle designs, each aimed at reducing pressure in specific areas through partial cutouts or surface depressions. Participants were divided into a case group, comprising 23 women with the aforementioned medical conditions, and a control group of 50 women with no such conditions. Pressure measurements were collected using a stationary bicycle and a pressure mapping saddle cover containing 64 sensors. Additionally, participants completed questionnaires to evaluate perceived comfort and symptom severity. The quality of life (QoL) score was significantly lower in the case group (0.83 ± 0.09) compared to the control group (0.96 ± 0.09, p < 0.001). No significant differences were found between the case and control group in terms of cycling habits. Complaints during cycling were reported by 100% of the participants in the case group and only 8% of the participants in the control group. Pain and numbness were the most reported symptoms for cycling at home. The results indicated significant differences in loaded weight per saddle between the case and control groups (p = 0.001, p = 0.010, p = 0.003 & p = 0.013). In the case group, participants experienced higher pressure. Saddle 2 was favored by 44% of the case group participants for home testing. These findings underscore the need for specialized saddle designs to improve cycling comfort and potentially alleviate pain for women with medical conditions in the groin area.