FIELD: medicine.
SUBSTANCE: invention relates to medicine, namely to traumatology and orthopedics, and can be used for growing bone regenerate between fragments of an injured rabbit tubular bone. The surgical field is prepared and surgical access to the diaphysis of the tubular bone of the animal is performed. A segmental resection of the diaphysis of the lower third of the tibia 3 cm long is performed. A periosteum 0.5 cm wide is removed from the edges of the sawdust of the bone. A funnel bioreactor in the form of a truncated hollow cone made of bioinert natural latex is put on the ends of the hollow tube in vivo. First, sawdust of a fragment of the distal part of the tibia is inserted into the opening of the tube of the in vivo bioreactor. Sawdust of a fragment of the proximal part of the tibia with the fit of the cuffs of the funnel to the outer surfaces of the sawdust of the tibia. Silicone rubber tubes are additionally placed in the in vivo cavity of the bioreactor, the free ends of which are brought out through punctures to the surface of the skin. The free ends of the tubes of the in vivo bioreactor are fixed through a metal fitting to a Luer-Lock connector with a protective cap. Stabilization of bone fragments is performed using an external fixation device for transosseous osteosynthesis. The surgical wound is sutured with the application of an aseptic bandage. The animal is placed on its healthy side, the silicone rubber tubes brought out to the surface of the skin of the animal with a metal plug of the connector for a Luer-Lock type syringe with a protective cap are transferred to a vertical position. The protective cap is opened and the liquid composition is injected through the silicone tube. Fill in vivo bioreactor with control of the liquid level of the composition in another tube. As a liquid composition, a nutrient medium for cell cultures DMEM with glutamine, containing 1 g/l of glucose, with Na pyruvate with the addition of human vascular endothelial growth factor-A, isoform 165, recombinant rhVEGF-A165 in the amount of 0.25 ng per 1 ml of nutrient medium. The liquid composition is replaced twice a day in the first 2 days from the start of the operation. After draining the solution of the liquid composition in vivo, the bioreactor is washed once with a sterile isotonic sodium chloride solution. Once a day for 8 days, the space of the in vivo bioreactor is filled with a nutrient medium for cell cultures DMEM with glutamine containing 1 g/l of glucose, Na pyruvate with the addition of 10% rabbit platelet lysate, with the addition of human vascular endothelial growth factor-A, isoform 165, recombinant rhVEGF-A165 in the amount of 0.25 ng per 1 ml of medium. Then, once a day for 15 days, the space of the in vivo bioreactor is filled with a nutrient medium for F12 cell cultures with the addition of BMP-7 recombinant human morphogenetic protein-7 rhBMP-7 at a concentration of 25 nanograms per milliliter of nutrient medium on even days, and also filled with nutrient medium for cultures of F12 cells with the addition of BMP-2 recombinant human morphogenetic protein-2 rhBMP-2 at a concentration of 25 nanograms per milliliter on odd days. After 25-30 days, surgical removal of the in vivo bioreactor is performed with access through the suture from the previous operation to install the bioreactor, the wall of the in vivo bioreactor is exposed from the side of the silicone rubber tubes, the silicone rubber tubes are cut off at the point where the tube enters the wall of the hollow tube of the in vivo bioreactor, the metal fitting is removed, and funnels are removed from both ends of the hollow tube of the in vivo bioreactor. The walls of the funnel are cut along the entire length with pliers, the wall of the hollow tube of the in vivo bioreactor is split lengthwise with the pliers, and after the fragments are removed, the hollow tube of the in vivo bioreactor is rotated around its axis and the pliers are split lengthwise, the formed two parts of the tube are removed from the tissues by rotational movements. Remove the silicone rubber tubes by pulling. The wound is sutured in layers. The Ilizarov apparatus is dismantled 140-150 days after the mineralization and remodeling of the bone regenerate.
EFFECT: method provides restoration of a bone defect without the need for a secondary operation for transplantation of an artificially created bone graft and the ingrowth of vessels and nerves into it due to reliable supply of bone regenerate with biologically active and nutrients and ensuring orthotopic assembly of bones.
1 cl, 3 ex
