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Pulsed electromagnetic fields preserve proteoglycan composition of extracellular matrix in embryonic chick sternal
1Department of Veterinary Basic Sciences, The Royal Veterinary College, London, UK. hliu@rvc.ac.uk
Abstract:
The influence of pulsed electromagnetic fields (PEMF) on proteoglycan composition in cartilage extracellular matrix has been investigated. Day 16 embryonic chick sternal cartilage was explanted to culture and exposed for 3 h per day for 2 days to a repetitive single-pulse PEMF with frequency of 15 Hz and peak magnetic field of 1.25 G. PEMF treatment did not affect cell proliferation, as indicated by [3H]thymidine incorporation, but significantly stimulated the retention of glycosaminoglycans in the explants and reduced the release of glycosaminoglycans into the media. Determination of incorporation of [35S]sulfate and [3H]N-acetylglucosamine into proteoglycans in vitro and breakdown of in ovo labelled [35S]sulfated proteoglycans in vitro showed that PEMF treatment significantly suppressed the synthesis of proteoglycans and the degradation of both newly synthesized and pre-existing proteoglycans. Sepharose CL-2B chromatography demonstrated that PEMF did not affect either the size distribution of newly synthesized and pre-existing [35S]sulfated proteoglycans or their ability to aggregate with hyaluronate. Sepharose CL-6B chromatography followed by cellulose acetate electrophoresis revealed that the chain length and degree of sulfation of [35S]sulfated glycosaminoglycans were identical in control and PEMF-treated cultures. It is concluded that PEMF treatment preserved extracellular matrix integrity of cultured cartilage explants by down-regulating proteoglycan synthesis and degradation in a co-ordinated manner without affecting their gross structural nature.