Fig. 4.

Chondrogenesis and osteogenesis in the tri-layered scaffold. A) Calcium deposition in the tri-layered scaffold in standard, osteogenic, chondrogenic and osteochondrogenic media. B) Alizarin Red staining for calcium at day 28 following culture in osteogenic media, chondrogenic media, osteochondrogenic media (Black ​= ​bone layer, green ​= ​intermediate layer, yellow ​= ​cartilage layer) C) sGAG deposition in the tri-layered scaffold over 28 days in standard, osteogenic, chondrogenic and osteochondrogenic media. D) Toluidine Blue staining of representative samples of the scaffolds at day 28 cultured in standard, osteogenic, chondrogenic and osteochondrogenic media (Black ​= ​bone layer, green ​= ​intermediate layer, yellow ​= ​cartilage layer).