Dynamic compression inhibits cytokine-mediated type II collagen degradation

Abstract

ObjectiveTo examine the effect of dynamic compressive loading applied intermittently on bovine cartilage explants stimulated with proinflammatory cytokines over 21 days. DesignCartilage explants were cultured for 21 days with Oncostatin M and TNFα (O ​+ ​T) [10/5 ​ng/mL] or in culture medium alone (w/o). The explants were either left free-swelling or subjected to dynamic compressive loading of 20 ​min, at 1 ​Hz, with loads ranging between 0.1 and 1 ​MPa, 5 times weekly. Metabolic activity was measured once weekly using Alamar Blue and cartilage turnover was assessed with biomarkers targeting degradation fragments of aggrecan (AGNx1) and type II collagen (C2M). Glycosaminoglycan degradation was quantified was the DMMB assay. Furthermore, explant weight and histological analysis was used to assess the cartilage degradation. ResultsDynamic compression of cartilage explants attenuated the O ​+ ​T-mediated C2M release on day 21 with 40% (p ​= ​0.0068) compared to the unloaded explants. Additionally, the change in explant weight from day −1 to day 21 showed that O ​+ ​T stimulation alone mediated a cartilage loss of 11%, whereas O ​+ ​T-stimulated explants subjected to compressive loading demonstrated a decreased cartilage weight loss of 6%, which was supported by the histological analysis. However, loading had no effect on aggrecan degradation. ConclusionIn cartilage explants cultured in a proinflammatory milieu, dynamic compressive loading confers anti-catabolic effects, inhibiting type II collagen degradation and reducing explant cartilage loss. These results demonstrate that compressive loading alters cartilage tissue turnover and enforces the need to include mechanical loading in a translation ex vivo cartilage model.