Aims Bi-Unicondylar Arthroplasty (Bi-UKA) is a bone and anterior cruciate ligament (ACL) preserving alternative to Total Knee Arthroplasty (TKA) when the patellofemoral joint is preserved. The aim of this study is to investigate the clinical outcomes and biomechanics of Bi-UKA. Methods Bi-UKA subjects (N = 22) were measured on an instrumented treadmill, using standard gait metrics, at top walking speeds. Age, sex and BMI-matched healthy (N = 24) and primary TKA (N = 22) subjects formed control groups. TKA subjects with pre-operative patellofemoral or tricompartmental arthritis or ACL dysfunction were excluded. The Oxford Knee Score (OKS) and EuroQol-5D (EQ-5D) were compared. Bi-UKA then TKA were performed on eight fresh frozen cadaveric knees, to investigate knee extensor efficiency under controlled laboratory conditions, using a repeated measures study design. Results Bi-UKA walked 20% faster than TKA (Bi-UKA 6.70.9km/h, TKA 5.60.7km/h p<0.001), exhibiting nearer-normal vertical Ground Reaction Forces in maximum weight-acceptance and mid-stance, with longer step and stride lengths compared to TKA (p<0.05). Bi-UKA subjects reported higher OKS (p=0.004) and EQ-5D (p<0.001). In vitro, Bi-UKA generated the same extensor moment as native knees at low flexion angles, whilst reduced extensor moment was measured following TKA (p<0.003). Conversely, at higher flexion angles, the extensor moment of TKA was normal. Over the full range, the extensor mechanism was more efficient following Bi-UKA than TKA (p<0.05). Conclusion Bi-UKA had more normal gait characteristics and improved patient reported outcomes, compared to matched TKA subjects. This can, in part, be explained by differences in extensor efficiency.