Individuals with various lower-limb neuromuscular and musculoskeletal impairments are often prescribed passive-dynamic ankle-foot orthoses (PD-AFOs) to compensate for impaired ankle muscle weakness. Several studies have demonstrated the beneficial effects of PD-AFOs on pathological gait, but few studies have examined the influence of the AFO stiffness characteristics on gait performance. One challenge to performing such studies is the difficulty of manufacturing custom AFOs with a wide range of controlled stiffness levels. However, selective laser sintering (SLS) is a wellsuited additive manufacturing technique for generating subject-specific PD-AFOs of varied stiffness. Therefore, the overall goal of this study was to use SLS manufactured PD-AFOs to identify the relationships between AFO stiffness and gait performance in patients with various lower-limb neuromuscular and musculoskeletal impairments. Six subjects with unilateral impairments were enrolled in this study. For each subject, one subject-specific PD-AFO equivalent to the subject’s clinically prescribed carbon fiber PD-AFO (nominal), one 20% more compliant and one 20% more stiff were manufactured using SLS. Three-dimensional kinematic and kinetic data were collected from each subject while ambulating with each PD-AFO at two different speeds to allow a comprehensive biomechanical analysis to assess the influence of PD-AFO stiffness on gait performance. The results showed that in the compliant AFO condition, the AFO limb vertical ground reaction force (GRF) impulse during loading and the non-AFO limb medial GRF impulse during push-off decreased. In addition, the AFO limb braking GRF impulse during loading and the non-AFO limb braking GRF impulse in early single-limb stance decreased. Furthermore, in the compliant AFO condition, negative knee work during early single-limb stance increased while positive hip work in early swing decreased in the AFO limb. Overall, as AFO stiffness decreased, the AFO limb contributed less to body support and braking. In addition, a decreased medial GRF impulse coupled with an increased vertical GRF impulse during non-AFO single-limb stance suggests that walking stability may be compromised as AFO stiffness decreases. Thus, a tradeoff may exist between preserving stability and increasing net propulsion, which should be considered when assessing the mobility needs of individuals prescribed PD-AFOs as a result of various neuromuscular and musculoskeletal impairments.