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The effects of hyaluronic acid (HA) on the proliferation and chondroitin sulfate (CS) synthesis of chondrocytes embedded in collagen gels were examined. Articular cartilage was isolated from the humerus, femur, and tibia of 21 10-week-old Japanese white rabbits. Chondrocytes isolated by collagenase digestion were embedded in type I collagen gels and cultured in Dulbecco's modified Eagle's medium (DMEM) with various doses of HA for 4 weeks. Histological and biochemical evaluations were performed at postculture weeks 1, 2, 3, and 4. For biochemical evaluations, isomers such as chondroitin 6-sulfate (delta(di)-6S) and chondroitin 4-sulfate (delta(di)-4S) synthesized by cultured chondrocytes were determined by high performance liquid chromatography (HPLC) combined with fluorometry. Morphological and histological studies demonstrated that HA-treated chondrocytes in collagen gel proliferated profusely while maintaining their phenotype. At postculture week 4, 0.1 mg/ml of HA induced an eightfold increase in cell counts compared with HA pretreatment values, or 1.5-fold more than control group. Synthesis of delta(di)-6S (delta(di)-6S content/cell) in groups treated with 0.01 and 0.1 mg/ml of HA significantly increased, while gel accumulation rates in groups treated with 0.1 and 1.0 mg/ml of HA scored significantly higher values than other groups. In collagen gel culture, HA enhanced the proliferation and delta(di)-6S synthesis of chondrocytes while maintaining their phenotype. In clinical application, since the supply of autologous chondrocytes for transplantation is not unlimited, the HA-treated culture method may be useful for increasing the number of chondrocytes and thus improving the quality of implants.

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The effects of cell density on the proliferation and chondroitin sulfate synthesis of chondrocytes embedded in Atelocollagen gel were examined. Chondrocytes of 21 10-week-old Japanese white rabbits isolated by collagenase digestion were embedded in Atelocollagen gel and cultured in Dulbecco's modified Eagles medium at cell densities of 2 x 105 cells/ml (105 group), 2 x 106 cells/ml (106 group), and 2 x 107 cells/ml (107 group) for 4 weeks. Chondrocytes in the 105 group gradually proliferated more than the other two groups. In contrast, most chondrocytes in the 107 group showed increased capability to produce chondroitin 6-sulfate. Cartilage-like tissue was produced from high-density cultures (107 cells/ml), although a decrease in cell number was seen. Even in three-dimensional cultures, the proliferation and chondroitin sulfate synthesis of chondrocytes were influenced by the cell density. These results are informative for the clinical application of chondrocyte transplantation in three-dimensional cultures for cartilage repair.

The purpose of this study was to examine the relationship between osteochondritis dissecans (OCD) of the lateral femoral condyle and lateral menisci. From 1993 to 2002, 38 knees (28 patients) were diagnosed with OCD of the lateral femoral condyle. OCD locations were graded by the Cahill and Berg classification. The types of lateral menisci were classified by Watanabe's classification. The relationship between OCD of the lateral femoral condyle and lateral menisci was examined. On the anterior-posterior view, 25 OCDs were located in zone 4 and 13 in zone 5. The types of lateral menisci were complete discoid in 19 knees, incomplete discoid in 15, and normal in 4. Ten of the 19 complete discoid menisci were damaged. Complete discoid menisci without tears were found in OCDs located in zone 4; incomplete discoid menisci were found in OCDs located in zone 5. The authors found a relationship between the type of OCD and the state of the lateral meniscus.