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


METHODS
Using a sinusoidal waveform of 0.5 Hz frequency, cyclic compressive pressure of 0.1-1.0 MPa was applied for 10s 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.


RESULTS
Increasing 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 DeltaDi6S to DeltaDi4S is increased. Similar results were obtained with explants being slightly mechanically challenged by low magnitudes of loads.


CONCLUSIONS
Our 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.
Sauerland
K
K
Orthopaedic Research Laboratories, Department of Orthopaedic Surgery, University Clinics Giessen and Marburg GmbH, Paul-Meimberg-Strasse 3, 35385 Giessen, Germany.
Steinmeyer
J
J
D000818:Animals; D001696:Biomechanical Phenomena; D002358:Cartilage, Articular / Q000378:metabolism*; D002417:Cattle; D002809:Chondroitin Sulfates / Q000096:biosynthesis* / Q000648:ultrastructure; D002852:Chromatography, Ion Exchange; D004187:Disaccharides / Q000378:metabolism*; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism*; D066298:In Vitro Techniques; D011509:Proteoglycans / Q000096:biosynthesis / Q000737:chemistry; D015238:Sulfotransferases / Q000378:metabolism
D004187:Disaccharides; D011509:Proteoglycans; D002809:Chondroitin Sulfates; D015238:Sulfotransferases