Biotechnological aspects of the working-out and manufacturing of living bone equivalent

Abstract

Objective. To handle biotechnological aspects in manufacturing processes of three-dimensional living bone equivalent for restoration of critical sized bone defects for innovative treatment of combat-related casualties. Methods. To fabricate living bone equivalent we used devitalized xenogeneic bone scaffolds (DBM chips) and autologous fibrin hydrogel seeded with autologous cultured bone marrow-derived multipotent mesenchymal stem/stromal cells (BM-MSCs). Quality/identity control of cell cultures was assured by donor and cell culture infection screening (IFA, PCR), flow cytometry (cell phenotype), karyotyping (GTG banding), functional assays (CFU assay, multilineage differentiation assay). Results. The BM-MSC cultures had a normal karyotype and appropriate phenotype, multilinear differentiation potential and functional properties, appropriate CFU frequency and hadn’t any signs of cell senescence. The FDA/PI combined staining showed the demineralized bone chips’ regular seeding with viable cells. Conclusions. An actual regenerative medicine approach to organ-saving transplantation of the three-dimensional living bone equivalent for combat-related casualties requires further preclinical and clinical approbation for thorough studies on the bone integrity restoration, forming new bone tissue in a site of bone defect, and duration of rehabilitation period compared to the gold standard of the conventional bone defect cure.