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Progression of primary knee osteoarthritis (OA) is often quantified by measuring structural alterations of the joint such as those in tibiofemoral joint space width (JSW) over time. Limited information is available regarding changes that occur during the onset and progression of posttraumatic OA (PTOA) that are often associated with anterior cruciate ligament (ACL) injury. Furthermore, there is a paucity of information regarding JSW changes in healthy patients, making JSW interpretation challenging during early PTOA progression.To evaluate tibiofemoral JSW after ACL injury, ACL reconstruction, and rehabilitation compared with healthy, matched controls.Cohort study; Level of evidence, 2.A total of 39 ACL-injured patients and 32 matched controls were evaluated. Injured patients were assessed at presurgical baseline and after ACL reconstruction (mean follow-up, 46 months), as were controls. Bilateral, standing, fluoroscopy-assisted, and posterior-anterior metatarsal-phalangeal view knee radiographs were obtained at each visit and JSW was measured.The JSW differences between knees in control patients were not significantly different and did not change over time. Baseline JSW differences in the ACL group were significantly different than in controls. Three patients (7.9% of total) had an increased JSW difference, and 1 patient (2.6%) had a decreased medial JSW difference. In the lateral compartment, 6 patients (15.8%) had a decreased JSW difference. At follow-up, 2 patients in the ACL group (5%) had a decreased medial JSW difference, and 3 (7.9%) had a significantly increased difference in relation to controls. Lateral compartment analyses revealed 7 (18.4%) patients with a significantly decreased JSW difference and no patients with an increased difference compared with controls.One third of ACL-injured knees underwent significant JSW change soon after injury; consequently, evaluation of within-knee JSW changes over time in ACL-injured patients may not be appropriate with a study based on case-control analysis. The JSWs in the healthy knee of ACL-injured patients do not change over time, allowing this knee to be used as a control for the injured knee. This is important when evaluating the earliest stages of PTOA after ACL injury, when patients are asymptomatic and intervention may be most beneficial.