media containing 10 fcs was added to the cell suspension to neutralize the activity of trypsin the cells were counted and the volume adjusted to give a concentration of cells of 250 000 cells per 500 u03bcl of media the discs prepared in step 1 were individually placed into single wells in a non tissue culture treated 2 ml deep polypropylene 96 well plate aliquots of the media containing 250 000 cells were added to the individual wells the plates were centrifuged at 400 g for 5 minutes to facilitate contact between the cells and the felt step 3 culture of cell seeded primary scaffolds in the presence of a chondrogenic agent the cell seeded discs was incubated in media supplemented with 20 ng ml tgf u03b2 3 for up to 18 days with regular media changes fig5 illustrates the staining of the scaffolds with safranin o after 18 days in culture the arrows indicate different levels of staining intensity as follows 1 very strong safranin o staining 2 strong safranin o staining 3 moderate safranin o staining 4 non specific safranin o staining of pga fibres 5 no staining the high levels of safranin o staining indicate chondrogenic differentiation and the secretion of a cartilage extracellular matrix the scaffold has been resorbed by the chondrocytes polyglycolic acid pga non woven felt is reinforced with poly l lactide co glycolic acid pllga by dipping the felt in a solution of pllga and dried discs of between about 0 5 mm u00d7 1 mm.