Development of a novel tissue culture system for the intervertebral disc

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Purpose of the current study was to establish a reliable, cost effective and reproducible in vitro tissue culture system for the intervertebral disc to investigate the metabolism of the intervertebral disc as a whole organ, thereby obtaining information which may lead to the elucidation of possible mechanisms of intervertebral disc degenerationRabbit intervertebral discs were extirpated and embedded in alginate gel and were cultured for up to 4 weeks. At weekly intervals, the contents of DNA, sulfated proteoglycans, antigenic keratan sulfate,hyaluronic acid and collagen in the cultured disc tissues were measured to evaluate the tissue integrity, Incorporation rates of 3SS sulfate were also measured at each time point to investigate proteoglycan synthesis and newly synthesized radiolabelled proteoglycans were characterized by separation with sepharose CL2B column sieve chromatography. Intervetebral disc cells cultured in medium alone were used as controls throughout the experimentsThe annulus fibrosus and ucleus pulposus of the intervertebral disc cultured in alginate gel maintained a higher content of various extracellular matrix molecules and sustained a higher rate of proteoglycan synthesis than those cultured in medium alone at all time points. Newly synthesized proteoglycans in the discs embedded in alginate gel also maintained their molecular sizes close to intact discsIncorporation of whole disc tissues in alginate gel proved useful to maintain the integrity of intervertebral disc and may be useful for studying the metabolism of intervertebral disc as a whole organ