Background: Osteoarthritis is the most prevalent joint disease worldwide, affecting over 600 million people. Cartilage-penetrating systems, such as avidin, are emerging as a therapeutic approach for the delivery of disease-modifying osteoarthritis drugs, improving drug uptake and retention in the cartilage, thereby helping to alleviate symptoms and slow disease progression. Methods: This study aims to investigate whether an avidin-based nanocarrier delivering kartogenin (Av-bKGN) can promote cartilage matrix regeneration in anterior cruciate ligament transection with hemi-meniscectomy (ACLT-hMnX) rat models. We compared outcomes from two ACLT-hMnX rat models of OA which differ in the timing of disease induction, representing the early and late stages of OA. Results: The results demonstrate that Av-bKGN leads to greater cartilage regeneration compared to the free drug, particularly in the early-stage OA model, whereas the effect is less pronounced in late-stage disease model. Conlusion: Taken together, these in vivo findings suggest that cartilage-penetrating delivery systems for disease-modifying osteoarthritis drugs are more effective as a preventive strategy in early disease stages rather than as a curative approach in advanced osteoarthritis.