Tissue engineering is the promising therapeutic approach that use combines cells, biomaterials, and microenvironmental factors for tissue repair. Recently, the use of decellularized tissues has been considered in tissue engineering.These tissues can be used as a scaffold for cellular studies. It is important to study cellular behavior in the three dimensional culture compare to two dimensions, because this culture is similar to in vivo. Biological scaffolds are derived from the tissues and organs and can be used on human. In this study, we prepare a declassified 3D bone matrix from human epiphyseal bone. Firstly, we declassified this bone matrix with acid solutions and then decellularized with physical, Enzimatic and chemical methods. in physical method bone samples were excluded at -4°C a week, then used of snap melting–freezing for 5 times. In enzymatic method and chemical method, Tripsin and sodium dodecyl sulfate (SDS) were used as an ionic detergent. General staining such as Hematoxylin-Eosin and specific staining such as DAPI were shown successful removal of the cells from the prepared scaffolds. Therefore, declassified and de- cellularized 3D bone matrix from human epiphyseal bonecan easily be prepared and createda suitable scaffold for cell and tissue engineering research and can be used in the cellular studies and interaction between the extracellular matrix and cells.

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