Hyaluronic acid secretion by synoviocytes alters under cyclic compressive load in contracted collagen gels

作者：Uehara Kazuki; Hara Masao; Matsuo Toshiki; Namiki Go; Watanabe Mutsuto; Nomura Yoshihiro^*

来源：Cytotechnology, 2015, 67(1): 19-26.

DOI：10.1007/s10616-013-9669-9

• Knee osteoarthritis
• Hyaluronic acid
• Synoviocytes
• Three-dimensional collagen gel culture
• Gel contraction
• Cyclic compressive load

Knee osteoarthritis is a degenerative disease of diarthrodial joints. Biomechanical factors are considered as risk factors for the disease, the knee joint being normally subject to pressure. Some studies have examined the biomechanical environment of the knee joint in vitro. The aim of this study was to establish a culture model to mimic the knee joint environment. As a first step, synoviocytes induced contraction of three-dimensional collagen gels. Next, contracted collagen gels containing synoviocytes underwent cyclical compression ranging from 0 to 40 kPa at a frequency of 1.0 Hz for 1.5, 3, 6 and 12 h using the FX-4000C (TM) Flexercell(A (R)) Compression Plus (TM) System. RNA in collagen gels was extracted immediately after compression and mRNA expression levels of HAS genes were analyzed by quantitative RT-PCR. Culture medium was collected 48 h after compression and analyzed by agarose gel electrophoresis and cellulose acetate electrophoresis. Synoviocytes in contracted collagen gels were stimulated by cyclic compressive load. Long-term compressive stimulation led to the production of higher molecular weight hyaluronic acid, whereas, short-term, compressive stimulation increased the total amount of hyaluronic acid. Furthermore, mRNA expression levels of both HAS-1 and HAS-2 were significantly higher than without compression. Taken together, using this gel culture system, synoviocytes synthesized higher molecular weight hyaluronic acid and produced large quantities of hyaluronic acid through up-regulation of HAS gene expression. Therefore, the contracted collagen gel model will be a useful in vitro three-dimensional model of the knee joint.

• 出版日期2015-1