Objective：To explore a method to prepare deproteinized osteo-articulation（DPOA） with hydrogen peroxide and ether as well as to discuss its physicochemical characteristics and biological safety. Methods：Distal femur semi-joint cancellous bones of adult hybrid canine were made into bio-derived osteo-articulation materials with hydrogen peroxide and ether by degreasing and deproteinizing；outsaide surface，pore texture of interal surface，pore size and traffic situation among pores of DPOA were observed by stereology microscope and scanning electron microscope；eliminated cell composition of DPOA was observed by HE staining；culture solution pH values of DPOA were detected by instrument；protein contents of DPOA were detect by micro Kjeldahl method；stress weight was detect by biomechanics test；biological safety in vitro was tested by cytotoxicity test and hemolysis test. Results：Results of stereology microscope and scanning electron microscope reveal that DPOA had three-dimensional and porus-grid structure system of original bone tissue. Results of computer picture analytical system reveal that porosity was （84.76±4.63）%，maximum aperture was （318.64±16.35） μm and minimum aperture was （112.13±15.42） μm. HE staining revealed that cell composition of DPOA was eliminated entirely. pH value of DPOA culture solution was 7.382±0.009. Protein content of DPOA was （0.017±0.003）%. Maximum stress weight of biomechanics test was （226.00±7.65） N，approaching mechanics loading of normal osteo-articulation.Cytotoxicity was 0 grade. Result of hemolysis test was < 5%. Conclusions：Allogeneic natural derived DPOA made by hydrogen peroxide and ether method retains pore net structure of original bone tissue and possesses the intrinsic anatomical configuration. Its physicochemical characteristics and biological safety are qualified and can be used as an osteo-articulation grafting trestle material.