OBJECTIVE: Bone tissue engineering needs to mimic the key aspects of bone structure and composition in order to achieve the desired functional tissue.since, bone has a potential for regeneration, it is a good example for tissue engineering. Bone fractures related to conditions such as osteoporosis usually occur in cancellous bone regions and this type of bone is one of the commonly used material due to its natural porous structure and. Due to the fact that porosity has implications in tissue engineering. In this study, we aimed to create an acellular matrix from bovine s cancellous bone. The use of bovine cancellous bone has the advantage of being easily obtained in large amounts. In order to decellularize the tissue, were used physicochemical techniques with out use of enzymatic treatment to remove the cells. MATERIALS AND METHODS: Cancellous bone was obtained from young bovine epiphysial bone, aged about 6 months and divided into small cylindrical pieces ( 2 or 3 mm in diameter and 2cm in length). These pieces were washed with normal saline to remove blood cells and then transferred to -4°с for two weeks and exposed to 5 cycles of wet snap freeze-thaw. Specimens were immersed in phosphate buffer saline(PBS) for further treatment and then rinsed for different times in various concentrations of SDS at room temperature with gentle agitation.finally, the samples immersed in PBS and stored at -80°с. RESULTS: The efficiency of decellularization was evaluated by histotechniques such as H&E staining. The results of H&E staining showed that cancellous bone did not have any cells and all bone cells had been removed. The control sample still contained cells such as osteocytes and bone marrow cells. DAPI staining also proven that all cells had been removed. CONCLUSION: In this study, it was demonstrated that using freez- thaw cycles and SDS can decellulize cancellous bone which can serve as a scaffold to study the behavior and interation of cells with the matrix and could be used for future bone clinical therapies. Currently, we are trying to investigate this interaction.

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