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Hyaluronic acid enhances proliferation and chondroitin sulfate synthesis in cultured chondrocytes embedded in collagen gels
Author(s) -
Kawasaki Kenzo,
Ochi Mitsuo,
Uchio Yuji,
Adachi Nobuo,
Matsusaki Masahiko
Publication year - 1999
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/(sici)1097-4652(199905)179:2<142::aid-jcp4>3.0.co;2-q
Subject(s) - hyaluronic acid , chondroitin sulfate , collagenase , chemistry , chondrocyte , cartilage , chondroitin , type ii collagen , microbiology and biotechnology , biochemistry , andrology , glycosaminoglycan , in vitro , biology , anatomy , medicine , enzyme
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 (Δdi‐6S) and chondroitin 4‐sulfate (Δdi‐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 Δdi‐6S (Δdi‐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 Δdi‐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. J. Cell. Physiol. 179:142–148, 1999. © 1999 Wiley‐Liss, Inc.