Purpose: Following anterior cruciate ligament (ACL) injury and reconstruction (ACL-R), quadriceps muscle weakness and low hamstring-to-quadriceps muscle strength ratios have been associated with post-traumatic osteoarthritis (PTOA) as well as primary OA. However, this relationship has not been examined using tibiofemoral joint space width difference (JSW-D) as a measurement of PTOA onset. JSW-D is a measurement technique that is sensitive to PTOA changes prior to the clinical manifestation of the disease. It is important to determine risk factors associated with the onset and progression of PTOA at the earliest time point in the disease process when intervention can alter the progression of the disease. Consequently, the focus of this study was to examine the relationship between isokinetic knee strength and JSW-D at one and four years post-ACL-R compared to a group of healthy, non-injured subjects of similar sex, age, BMI, and activity level. Methods: A secondary analysis of data collected during a longitudinal study of biomarkers of PTOA following ACL-R was utilized for this exploratory study. Entry criteria for injured subjects included: age at time of ACL-R = 14-55; BMI = 18.5-30; Tegner activity score > 4; < 2/3 meniscectomy; < grade IIIa articular cartilage lesions; no history of joint surgeries, knee injections, or other knee pathologies; no abnormal knee laxity or evidence of radiographic OA at baseline. Additional criteria for controls included: no knee pain/dysfunction, and normal clinical evaluation and baseline MRI. At 1- and 4-years post ACL-R, standing, bilateral metatarsal phalangeal (MTP) view knee x-rays were obtained for JSW assessment. Subjects were considered to have significant narrowing if their injured minus normal (contralateral) knee JSW difference (JSW-D) fell below the 95% confidence interval of controls. At the same time point, isokinetic knee strength was assessed at 60, 180, and 300 deg/sec, and normalized to body weight. Relationships between strength and JSW-D were evaluated between ACL subgroups (normal and narrowed) and controls using analysis of covariance adjusted for age, sex, BMI, and time since surgery. Post-hoc group comparisons were made using Fisher's LSD. Results: Of the 34 ACL-R patients included, 6 had significant JSW-D narrowing at 1-year follow-up, and 8 had narrowing at 4-year follow-up. The mean strength values for all ACL-R subjects (“normal” and “narrowed JSW-D” groups combined) were found to be significantly less than the control group at both time points. However, ACL-R subjects with narrowed JSW-D did not differ from normal JSW-D subjects (all p > 0.20) with one exception: extension at 60 degrees/second at four-year follow-up (p = 0.005). The pattern of knee extension strength at 60 deg/sec at 1-year follow-up was similar to changes observed at 4-year follow-up. Extension to flexion strength ratios were not found to have a significant association with JSW-D at any speed. Conclusions: Strength deficits present as early as one year following ACL-R and persist at 4-years post. In general, ACL-R group subjects were not found to have significantly different strength values within JSW subgroups. In contrast to previous studies, the current investigation did not reveal an association between PTOA and low extension to flexion strength ratios. It should be noted that no subjects in this study reported appreciable pain/dysfunction or reduced physical activity, which is indicative of the clinical manifestation of OA. However, subjects with narrowed JSW-D had significantly lower extension strength at 60 degrees/second than controls and injured subjects with normal JSW-D. It may be that individuals that loose a significant amount of knee extension strength are at greater risk for tibiofemoral JSW narrowing following ACL trauma.