Tissue engineering is founded on three principal compositions of biological tissues including scaffold, cell and growth factors. Biological scaffolds that were derived from decellularized tissues and organs, successfully have been used in tissue engineering. Decellularization studies have shown that natural scaffolds which maintaine their main structure and properties could be a suitable tool for studying cellular behaviors.Several technologies have been developed to fabricate such as artificial valves, blood vessels and dermis. Decellularization techniques are classified by the chemicals used, such as acid or alkaline treatment, detergent treatment, or enzymatic digestion, and the physical methods used, such as snap freezing and mechanical agitation. Among these, detergent treatment is the most widely used. The purpose of this study is to develop an appropriate method to decellularize of rat's calvaria bone tissue. In this experimental study, after isolation of rat's calvaria decellularization was performed using physical (snap freezing- thow),enzymatic (trypsin 0.25%) and chemical ( SDS2 %). Histological examination revealed the scaffolds that prepared by trypsin 0.25% for 18h and SDS 2% for 20h, successfully decellularized rat's calvaria. This scaffold can be used as an accessible and practical 3-D matrix to be used in study cell differentiation and behavior.

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