The purpose of this study was to evaluate the influence of different longitudinal bending stiffnesses (LBS) on running economy and lower extremity joint biomechanics in male and female runners. Thirty participants (15 F;15M) performed a treadmill protocol of five-minute randomised aerobic runs with four different footwear conditions varying in LBS: 16.1 N/mm; 32.7 N/mm; 46.1 N/mm; 90.1 N/mm. Biomechanical data was collected at the metatarsophalangeal (MTP) and ankle joints. Running economy was quantified via oxygen consumption data. Oxygen consumption observed no significant differences between footwear conditions (p = 0.960) or significant interaction between footwear and sex (p = 0.126). Stance time observed a significant difference between footwear conditions (p < 0.001), and a significant interaction of footwear and sex (p = 0.008), increasing in females as stiffness increased. At the MTP joint, a shoe effect was present for peak bending angle (p < 0.001), peak extension (p = 0.040) and flexion (p < 0.001) angular velocity, and energy generated at the joint (p = 0.010). There was also an interaction effect of peak MTP extension angular velocity (p = 0.044), with females slightly decreasing as stiffness increased, with no reaction from males. At the ankle joint, a shoe effect was present for peak dorsiflexion (p < 0.001) and plantarflexion (p < 0.001) angular velocity, peak negative power (p < 0.001) and energy absorbed at the joint (p < 0.001). Comparing the interaction between shoe condition and male/female runners, male and female runners reacted similarly to the increased LBS with no effect on running economy and subtle changes in the stance time and MTP angular velocity for female runners. While further research is needed in this area investigating aspects related to sex-specific optimal stiffness element location and geometry, it does not appear that the stiffness magnitude has a sex dependence.