Hyaluronic acid enhances proliferation and chondroitin sulfate synthesis in cultured chondrocytes embedded in collagen gels.
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.
Kawasaki
K
K
Department of Orthopaedics, Shimane Medical University, Shimane-ken, Japan.
Ochi
M
M
Uchio
Y
Y
Adachi
N
N
Matsusaki
M
M
D000818:Animals; D002455:Cell Division / Q000187:drug effects; D019902:Chondrocytes / Q000187:drug effects*; D002809:Chondroitin Sulfates / Q000096:biosynthesis*; D003094:Collagen; D005782:Gels; D006025:Glycosaminoglycans / Q000096:biosynthesis; D006820:Hyaluronic Acid / Q000494:pharmacology*; D007150:Immunohistochemistry; D011817:Rabbits
D005782:Gels; D006025:Glycosaminoglycans; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates; D003094:Collagen