FIELD: medical science.
SUBSTANCE: invention can be used in treating bone defects caused by various reasons related to loss of bone material volume. Method of producing 3D scaffolds involves washing allogenic or xenogenic bone material, sizing and shaping as required, followed by processing the bone material: 6% hydrogen peroxide solution at ratio of 1:3 to 1:5 for 36-72 hours at room temperature; with a mixture of chloroform and diethyl ether in ratio 1:1 for 24-48 hours; hypotension with 0.075 M potassium chloride solution in ratio 1:1 for 24-48 hours; washing with purified water; treatment at temperature 2-4 °C 0.3 M hydrochloric acid solution for 4-6 hours, followed by neutralization and treatment with 0.05% trypsin solution for 2-3 hours. When processing the material, the solution is replaced with a fresh solution every 12 hours. Treated material is then frozen in liquid nitrogen with 1-3% DMSO for not less than 2 days with subsequent defrosting and holding for 12 hours in 2-3 M sodium hydrophosphate solution and phosphate-salt buffer solution. That is followed by lyophilisation, packaging and sterilisation. That is followed by aseptic addition of a conditioned medium from the cultivation of allo- or xenogenic mesenchymal stem cells of at least 3rd passage, pre-differentiated in the osteogenic direction by means of an acceptable inductor, in ratio 1:1, and repeated lyophilisation in aseptic conditions. Influence of 3D scaffolds of each batch on adhesion and proliferation of mesenchymal stem cells during co-cultivation and evaluation of cytotoxic properties in vitro are also monitored.
EFFECT: method enables to obtain 3D scaffolds capable of osteoinduction and osteoconduction, having high biocompatibility.
4 cl, 4 dwg, 3 ex
