High molecular weight hyaluronan promotes repair of IL-1β-damaged cartilage explants from both young and old bovines Gene A Homandberg, Ph.D., Vijay.

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High molecular weight hyaluronan promotes repair of IL-1β-damaged cartilage explants from both young and old bovines Gene A Homandberg, Ph.D., Vijay Ummadi, M.S., Hellan Kang, M.D. Osteoarthritis and Cartilage Volume 11, Issue 3, Pages 177-186 (March 2003) DOI: 10.1016/S1063-4584(02)00371-0

Fig. 1 Effect of removal of HA from cultures of Fn-f-damaged and PG-restored cartilage. (Panel A) Bovine cartilage was cultured in 10% serum/DMEM alone (Control) or with 100nM Fn-f in 10% serum/DMEM from day 0 to 7 to remove about half of the total cartilage PG. At day 7, the media of some of the untreated cultures were adjusted to HA alone (HA [d7–42]). The media of the Fn-f-treated wells was changed to 10% serum/DMEM alone (Fn-f [d0–7]) or to 1mg/m HA in 10% serum/DMEM (Fn-f [d0–7]; HA [d7–14 or d7–21 or d7–42]). PG content was measured every 7 days. (Panel B) Similar conditions as in panel A except the Fn-f treatment was for 14 days, followed by HA treatment from day 14 to 28 or from day 14 to 42. Osteoarthritis and Cartilage 2003 11, 177-186DOI: (10.1016/S1063-4584(02)00371-0)

Fig. 2 Visualization of Fl-HA in normal (C) and Fn-f (F)-damaged cartilage as a function of time—Fl-HA was incubated in 10% serum/DMEM bovine cartilage cultures from day 0 to 7 and then removed. Sections were analyzed by fluorescence microscopy at days 7, 14 and 28. Shown are control cultures (C) incubated with Fl-HA, but not with Fn-f and cartilage cultured with Fn-f and Fl-HA for days 0–7. The −HA corresponds to no addition of Fl-HA. Ph is phase image of −HA section. Osteoarthritis and Cartilage 2003 11, 177-186DOI: (10.1016/S1063-4584(02)00371-0)

Fig. 3 Effect of HA on blocking depletion of PG and promoting restoration of PG in IL-1β-treated explants. (Panel A) Conditions were similar to those in Fig. 1(A), except 10ng/ml IL-1β in 10% serum/DMEM were used to damage rather than 100nM 29-kDa Fn-f. (Panel B) Conditions were similar to those in Fig. 1(B), except repair was tested after treatment with 10ng/ml IL-1β in 10% serum/DMEM from day 0 to 7. Osteoarthritis and Cartilage 2003 11, 177-186DOI: (10.1016/S1063-4584(02)00371-0)

Fig. 4 Effect of removal of HA from cultures of IL-1β-damaged and PG-restored cartilage. (Panel A) Bovine cartilage was cultured in 10% serum/DMEM alone (Control) or with 10ng/ml IL-1β in 10% serum/DMEM from day 0–7 to remove about half of the total cartilage PG. At day 7, the media of some of the untreated cultures were adjusted to HA alone (HA [d7–28]). The media of some of the IL-1β-treated wells were changed to 10% serum/DMEM alone (IL-1 [d0–7]) or to 1mg/m HA in 10% serum/DMEM (IL-1 [d0–7]; HA [d7–14 or d7–28]). PG content was measured every 7 days. (Panel B) Similar conditions as in panel A, except the IL-1β-treatment was for 14 days, followed by HA treatment at days 14–28 or days 14–42. Osteoarthritis and Cartilage 2003 11, 177-186DOI: (10.1016/S1063-4584(02)00371-0)

Fig. 5 Effect of HA on decreasing IL-1β-mediated damage in 6-year-old bovine cartilage and promoting repair—To test blocking activity, cartilage from 6-year-old bovines was cultured in 10% serum/DMEM alone (Control) or with 10ng/ml IL-1β in 10% serum/DMEM from day 0 to 7 (IL [d0–7]) or with both IL-1β and 1mg/ml HA from day 0 to 28 (IL, HA [d0–28]). To test for repair, cartilage was cultured with 10ng/ml IL-1β from day 0 to 7, to remove PG and then incubated with 1mg/ml HA in 10% serum/DMEM from day 7 to 28 (IL [d0–7]; HA [d7–28]). Controls for continuous IL-1β for days 0–28 are not shown, but were similar to day 0–7 treatment. PG content was measured every 7 days. Osteoarthritis and Cartilage 2003 11, 177-186DOI: (10.1016/S1063-4584(02)00371-0)

Fig. 6 Comparison of HA and IGF-1 on promoting restoration of PG in IL-1β-mediated damage in 6-year-old bovine cartilage— Conditions similar to those in Fig. 5 were used to test for repair. Cartilage was first treated with 10ng/ml IL-1β for 7 days in 10% serum/DMEM and was then treated with 1mg/ml HA (HA) or 100ng/ml IGF-1 (IGF-1) or a combination of the two (Both) from day 7 to 32. A duplicate control curve for day 0 to 7 IL-1β treatment is included. Osteoarthritis and Cartilage 2003 11, 177-186DOI: (10.1016/S1063-4584(02)00371-0)