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Updated: May 11, 2026
A 3D System for Culturing Human Articular Chondrocytes in Synovial Fluid
Published on: January 31, 2012
Histomorphometric Evaluation of Cartilage Degradation using Rabbit Articular Chondrocytes Cultured in Alginate Beads
K Nakatsuka1, K Kurita, Taro Hayakawa
1Department of Oral and Maxillofacial Surgery, School of Dentistry, Aichi-Gakuin Unuverity, Japan;
Objective:
A 3-dimensional alginate bead culturing method using rabbit articular chondrocytes was studied for the screening of the effectiveness of drugs for articular diseases.
Design:
The beads cultured with IL-1β, TGF-β, and Hyaluronan (HA) were evaluated histochemically with Alecian blue and immunohistochemically with CS-56 antibody. Chondrocytes in alginate beads were arbitrarily classified into four groups: 1) chodrocyte surrounded with cell-associated matrix (CAM) in which proteoglycan (PG) was positively stained (PG-possitive chondrocyte); 2) chondrocyte with PG-negative CAM; 3) PG-positive CAM alone, and 4) PG-negative CAM alone. Total sulfated GAG concentrations in the culture media were quantitated by dimethylmethylene blue (DMMB) assay. ProMMP-3, TIMP-1 and -2 concentrations in the culture media were determined by sandwich enzyme immunoassays.
Results:
Significant increase of PG-nagative cells were immunohistochemically found by IL-1β stimulation. The pretreatment with TGF-β almost fully suppressed those increase of PG-negative cells by IL-1β. Both GAG and proMMP-3 concentrations in the culture media were significantly increased after IL-1β stimulation. There were no significant differences in both TIMP-1 and TIMP-2 concentrations in the culture media with or without IL-1β stimulation. 800-kDa HA reduced significantly the number of PG-negative cells and proMMP-3 concentration in the culture media, but showed no effects on the concentrations of both TIMPs.
Conclusions:
Because this 3-dimensional chondrocyte culture in alginate beads is close to in vivo conditions, this method can be used for evaluation of the effectiveness of novel drugs for articular diseases.
