Temporal regulation of chondrocyte metabolism in agarose constructs subjected to dynamic compression
Section snippets
Preparation of chondrocyte/agarose constructs
Full depth slices of cartilage were removed from the metacarpalphalangeal joints of 18-month-old steers. The cartilage slices were washed, diced finely using a scalpel, and incubated at 37 °C on rollers for 1 h in Dulbecco’s minimal essential medium supplemented with 20% foetal calf serum, 2 μM l-glutamine, 5 μg ml−1 penicillin, 5 μg ml−1 streptomycin, 20 mM Hepes buffer, and 0.85 μM l-ascorbic acid (DMEM + 20% FCS) + 700 U ml−1 pronase and for a further 16 h at 37 °C in DMEM + 20% FCS supplemented with 100 U ml−1
Effects of continuous compression on chondrocyte/agarose constructs
The effects of continuous dynamic compressive strain on 35SO4 incorporation, [3H]thymidine incorporation, and nitrite release within unstrained constructs and constructs subjected to dynamic compressive strain are summarised in Table 1. The corresponding normalised strained values, presented as a percentage of the unstrained controls, are illustrated in Fig. 2. In unstrained constructs, mean 35SO4 incorporation values measured from individual experiments ranged from 0.0248 to 0.0361 μmol/h/μg
Discussion
A number of studies, previously published by the authors, have demonstrated that 15% compressive strain amplitude, applied in a static or dynamic manner, will induce distinct metabolic responses, in full depth chondrocytes and cells isolated from surface and deep zones of articular cartilage, when applied continuously over a 48 h test period to chondrocytes seeded in agarose constructs [12], [18], [19], [27]. Previous studies have demonstrated that low amplitude dynamic compression-induced
Acknowledgements
This study was supported by a Framework V European grant, “Imbiotor,” Project No. GRD1-2000-25394.
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