Figure 8

Biochemical analysis and gene expression of chondrocyte-specific phenotypes induced by herbal extracts. A, ATDC5 cells were seeded in 96-well cell culture plates and allowed to differentiate. Thereafter, cells were cultured in the absence (Control) or presence of 10 μM AD, or 1, 10 and 100 μg/ml C. atratum (Atratum) M. azedarach (Azedarach) and C. turtschaninovii (Turtschaninovii) extracts. After 3 days, cells were subjected to MTT assay. B, C and D, ATDC5 cells were seeded in 24-well cell culture plates and allowed to differentiate. Thereafter, cells were cultured in the absence (Control) or presence of 10 μM AD, or 1, 10 and 100 μg/ml C. atratum (Atratum) M. azedarach (Azedarach) and C. turtschaninovii (Turtschaninovii) extracts. After 7 days, the cell layer was lysed with 300 μl of 0.02% Triton-X 100 in saline. The aliquot was then subjected to DNA (B), ALPase activity (C) and sulfated GAG (D) measurement. Data are means ± standard deviation of 3 sets of cultures, and each value (except for DNA) is standardized to DNA content (μg) in the cell layer. E, F and G, ATDC5 cells were seeded and cultured, as described above, and then total cellular RNA was purified, and real-time RT-PCR for ALPase (E), aggrecan (F) and type II collagen (G) was carried out. Data are means ± standard deviation of 3 sets of cultures and each value is standardized to β-actin. *, p < 0.05 vs control.