Oksymets V. Cell and tissue technologies for treatment of the reparative osteogenesis disorders and bone defects: theoretical basis and possibilities for clinical use (experimental and clinical study)

Object: violations of reparative osteogenesis which occur as a result of diaphyseal fractures of the extremities. The aim: to improve the effectiveness of the treatment of patients with outdated nonunion fractures and bone defects by introducing cellular and tissue technologies developed on the basis of new results of clinical and experimental studies on the cellular mechanisms violations callus formation and mechanisms of the influence of mesenchymal stromal cells on reparative osteogenesis. Methods: Clinical, experimental, histomorphological, biotechnology, immunocytochemistry, immune-enzyme, instrumental (radiography, tomography) and statistics. For the first time the relationship terms of healing of diaphyseal fractures of the lower limbs with energy traumatic agent was established on the basis of retrospective analysis. For the first time in the experiment condition of cellular sources osteoreparation with low- and high-energy injuries were studied. For the first time on the basis of experimental studies the cellular mechanisms of violations of osteoreparative processes in high-energy trauma were described. For the first time in the experiments in vitro morphology, some membrane receptors, secretion of cytokines, alkaline phosphatase and osteopontin of uncommitted and osteogenic committed human mesenchymal stromal cells were investigated. For the first time in the experiment histomorphological analysis of nonunion area after transplantation of cultured mesenchymal stromal cells was conducted. Firstly the inductive properties of cell delivery system based collagen gel, hydroxyapatite and bone xenografts and allografts were studied experimentally in vitro. For the first time in the experiments in vitro the effect of the periosteum cells on mesenchymal stem cells in the co-cultured process was studied. It was found that in the co-cultured process the cells periosteal have osteogenic induction on mesenchymal stem cells. For the first time a theoretical concept of the use of cell-tissue technologies in the complex treatment of reparative osteogenesis disturbances was justified. The obtained research results facilitate understanding of mechanisms of reparative osteogenesis disturbances at the cellular level, allow us to understand how the transplanted mesenchymal stem cells promote restoration of disturbed processes of osteoreparation, and supplement the theoretical knowledge in this field. Developed and patented technology of biological osteogenic differentiation of mesenchymal stem cells in vitro (patent of Ukraine №15689) allows to obtain osteogenically directed cell culture, preserving their proliferative activity and capable for integration into the recipient tissue. Developed and patented technology of creating three-dimensional osteoprogenitor graft (patent of Ukraine №15680) allows obtain under in vitro conditions required amount of graft, which morphologically resembles bone and can be considered as an alternative to autologous bone graft. The developed technology of transplantation of autologous mesenchymal stem cells allows determine the best way to cell transplantation depending on the bone pathology, to create conditions for vascularization and engraftment of transplanted cells, restoration of reparative osteogenesis and / or the formation of new bone (resolution of the Coordination Center of transplantation of organs, tissues and cells of the Ministry of Health of Ukraine for the manufacture of cell / tissue transplants, the code 04.2008). Results of the study are introduced in the practical work of the Research Institute of Traumatology and Orthopedics of Donetsk National Medical University named M.Gorky Ministry of Health of Ukraine and the "National Institute of Emergency and Reconstructive Surgery named K. Husak NAMS of Ukraine". Traumatology and Orthopedics