Objective: To evaluate the etiologic heterogeneity between medial and lateral tibiofemoral radiographic osteoarthritis (ROA). Methods: Knees without medial or lateral tibiofemoral ROA at baseline were followed for 60-month in Multicenter Osteoarthritis Study (MOST) and for 48-month in Osteoarthritis Initiative (OAI). We examined the relation of previously reported risk factors to incident medial and lateral tibiofemoral ROA separately and determined the etiology heterogeneity with a ratio of rate ratios (RRs) (i.e., the RR for medial tibiofemoral ROA divided by the RR for lateral tibiofemoral ROA) using a duplication method for Cox proportional hazard regression. Results: Of 2,016 participants in MOST, 436 and 162 knees developed medial or lateral tibiofemoral ROA, respectively. Obesity and varus malalignment were 95% and 466% more strongly associated with incident medial tibiofemoral ROA than with lateral tibiofemoral ROA, respectively (ratios of RRs, 1.95 [95% confidence interval (CI):1.05–3.62] and 5.66 [95% CI:3.20–10.0]). In contrast, the associations of female sex and valgus malalignment with incident medial tibiofemoral ROA were weaker or in an opposite direction compared with lateral tibiofemoral Osteoarthritis (OA) (ratios of RRs, 0.40 [95% CI:0.26–0.63] and 0.20 [95% CI:0.12–0.34], respectively). Older age tended to show a weaker association with incident medial tibiofemoral ROA than with incident lateral tibiofemoral ROA. No heterogeneity was observed for the relation of race, knee injury, or contralateral knee ROA. These findings were closely replicated in OAI. Conclusion: Risk factor profiles for medial and lateral tibiofemoral ROA are different. These results can provide a framework for the development of targeted prevention and potential treatment strategies for specific knee OA subtypes.