Intermittent mechanical loading of articular cartilage explants modulates chondroitin sulfate fine structure

SummaryObjectiveAlterations in the sulfation pattern of chondroitin sulfate (CS) chains of proteoglycans have been associated with aging and degeneration of articular cartilage. The purpose of the present study was to investigate systematically the effect of load amplitudes, frequencies and load durations of intermittently applied mechanical pressure on the sulfation of CS chains of cultured bovine articular cartilage explants.MethodsUsing a sinusoidal waveform of 0.5 Hz frequency, cyclic compressive pressure of 0.1–1.0 MPa was applied for 10 s followed by a period of unloading lasting 10–1000 s. These intermittent loading protocols were repeated for a total duration of 1–6 days. Newly synthesized as well as endogenous CS chains were isolated, depolymerized and subsequently quantitated after fractionation by high-performance anion-exchange chromatography.ResultsIncreasing the mechanical demands on cartilage explants by elevating either the duration or the frequency of loading can significantly alter the fine structure of newly synthesized CS in that less chains terminate on galNAc4,6S and, in that simultaneously the ratio of the internal disaccharides ΔDi6S to ΔDi4S is increased. Similar results were obtained with explants being slightly mechanically challenged by low magnitudes of loads.ConclusionOur data show for the first time that intermittent loading of articular cartilage explants can significantly alter the sulfation pattern of the terminal CS residues as well as of the internal disaccharides. Furthermore, our results indicate that explants possess a physiological window of stress in which they are able to produce also a normal extracellular matrix.