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Hard tissue engineering has emerged to compensate for the losses in the properties of damaged bones by using scaffolds and cells. In this study, natural polymers chitosan and gelatin were used as scaffold materials because of their great resemblance to extracellular matrix elements. To enhance the mechanical properties and osteoconductivity of structures hydroxyapatite was added. This study is about the characterization of physico-chemical properties and cell behavior on scaffolds to optimize chitosan-gelatin ratio and aims to assess the effects of hydroxyapatite addition into polymeric matrix. According to thermal test results, decomposition temperatures decreased with non-sintered hydroxyapatite (nsHA) addition. Mechanical test results of composite structures were similar to that of cortical bone. Chitosan-gelatin films prepared with same amount of polymers showed the highest swelling degree. While nsHA added pure chitoan films showed higher cell number with respect to chitosan films; on the contrry, chitosan-gelatin polymeric blends indicated an opposite trend after nsHA addition.