Twenty‐five microliters of a 2% saline solution of levofloxacin (LVFX) or ciprofloxacin (CPFX) was injected every other day for 2 wk into the knee joint space of CD rats (weighing 62.7–86.7 g) from the age of 3 wk. Early in the course of injection, histologic examination revealed chondrocyte necrosis without marked matrix change in the articular cartilage of the femoral condyles adjacent to the intercondylar groove. After 7 injections, the surface and intermediate zones of the articular cartilage showed extensive necrosis, sometimes with cavity formation in the center of the same portion. Papain completely depleted matrix basophilia in all zones throughout the condyle and caused cartilage necrosis with cavity formation. One injection of iodoacetic acid caused necrosis of almost all chondrocytes over the entire condyle, but chondrocytes sometimes remained alive in the portion where cavity formation was induced by quinolones. Chondroitinase depleted the matrix basophilia, and sometimes produced necrotic areas. DNA synthesis inhibitorsn‐ethyl‐maleimide, CPT‐11, and etoposide (VP‐16) caused chondrocyte necrosis, but never caused cavities in the articular cartilage. The DNA synthesis inhibitorsn‐ethylmaleimide, CPT‐11, and hydroxyurea were administered concurrently with po LVFX administration and significantly increased the incidence of LVFX‐induced cavity formation.n‐Ethylmaleimide was the most effective of all the inhibitors. The quinolone‐induced cavity formation is suggested to be site specific in the articular cartilage of rat femoral condyles. The depletion of matrix proteoglycans and chondrocyte necrosis may be necessary, although insufficient, to produce such lesions. Disruption of the collagen framework is suspected to contribute to their development. Involvement of altered DNA metabolism may play a role in the chondrocyte necrosis that occurs early in the specific sites.