RU1836905C - Method for conservation of demineralized bone transplants - Google Patents

Abstract

The invention relates to the field of medicine, namely to transplantology, and can be used in the preservation of demineralized bone grafts. The purpose of the invention is to preserve the osteinductive properties of bone grafts when stored without cooling. The method of preserving demineralized bone grafts consists in the fact that the grafts are pretreated with calcium detergent solution with the addition of sodium chloride, and then placed in a preserving solution of urea with the addition of detergent and calcium chloride, sodium dodecyl sulfate being used as a detergent. 2 IL:

Description

The invention relates to medicine, namely to transplantology, and can be used in the preservation of demineralized bone grafts for subsequent transplantation during osteoplastic operations in the treatment of injuries and orthopedic diseases.

The purpose of the invention is to preserve the osteoinductive properties of demineralized bone grafts when stored without cooling.

This goal is achieved in that the demineralized bone grafts are pretreated with a calcium detergent solution with the addition of sodium chloride and then placed in a preserving solution of urea with the addition of a detergent and calcium chloride, sodium dodecyl sulfate being used as a detergent.

Pretreatment of demineralized bone grafts in a calcium detergent solution with the ability to stabilize protein structures, followed by storage of the grafts in a preserving solution of urea with the addition of a stabilizing complex, makes it possible to maintain the osteoinductive activity of the grafts for three months at room temperature.

The method is carried out as follows: the bone graft is freed from soft tissues, periosteum and bone marrow and placed in 1.2 N. hydrochloric acid solution. After the demineralization process is completed, the grafts are washed with distilled water and poured with 5% sodium bicarbonate solution for 30-40 minutes to neutralize acid residues. After this demineralized bone

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the transplants are transferred to a sterile bed and filled with a calcium detergent solution with the addition of sodium chloride, having the following composition:

Sodium Chloride 0.9 g

Calcium Chloride 0.3 g

Sodium dodecyl sulfate 10.0 mg Distilled water Up to 100 ml. The grafts are poured with the solution so that they are completely immersed in the solution. Transplant dishes are tightly sealed and refrigerated at 4 ° C. After 16-20 hours, the dishes with transplants are removed from the refrigerator, the solution is drained and the transplants are poured with a preserving solution of the following composition;

Urea 20 g

Calcium Chloride 0.3 g

Sodium dodecyl sulfate 10 mg Distilled water Up to 100 ml. The dishes are sealed and stored at room temperature. The maximum shelf life during which osteoinductive properties are maintained is 3 months,.

An example of the implementation of the method: fragments of the femur and tibia of rats are isolated from soft tissues, periosteum, and bone marrow, filled with 1.2 N in a glass vessel, hydrochloric acid solution, and placed in a refrigerator at 4 ° C. After 16 hours, the demineralized bones were removed from the acid solution, washed with distilled water, and filled with 5% sodium bicarbonate solution. After 30 minutes, the solution of sodium bicarbonate was drained, the bone fragments were divided into two groups, one of which was filled with a 0.5% formalin solution and then stored at room temperature - control. The remaining transplant is filled with a stabilizing calcium detergent solution and refrigerated at 4 ° C. After 16 hours, the stabilizing solution is drained, the bone fragments are filled with a preserving solution of urea with the addition of a stabilizing complex. Subsequently, these grafts were stored at room temperature - an experiment.

After three months of storage, the grafts were transplanted into the thigh muscles of 8 white rats. Transplants stored in a 0.5% formalin solution were transplanted into the muscles of the right thigh, and those stored in the preservation solution we proposed were transferred to the muscles of the left thigh. One month after surgery, 4 rats were killed. X-ray0

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Figure 1 shows the shadow of a bone fragment in the left thigh. There is no transplant shadow in the muscles of the right thigh. The grafts are removed. The method of ashing in a muffle furnace determined the content of mineral substances in them. Before transplantation, the content of mineral substances in the transplants was 1.2 ± 0.3%. The mineralization of the grafts stored in a urea solution with a stabilizing complex was 10 ± 2.1%. The mineralization of the grafts stored in a 0.5% formalin solution was 0.9 ± 0.3%.

Two months after the operation, the remaining four rats were sacrificed. On radiographs of the hind legs of rats (figure 2) in the muscles of the left thigh, a clearly visible shadow of the bone fragment. There is no shadow in the muscles of the right thigh. The mineralization of grafts extracted from the left thigh was 21 ± 0.8%. The grafts in the right thigh are almost completely resorbed.

When transplanting grafts stored in a urea solution with a stabilizing complex at room temperature for 4 months, the degree of their mineralization two months after surgery was 8 ± 3.2%, which indicates a significant decrease in the osteoinductive properties of the bone matrix.

Thus, the method of preserving demineralized bone grafts by treatment with a stabilizing solution for 4–20 h at 4 ° С for 16–20 h, followed by storage of urea in a preserving solution with the addition of a stabilizing complex, allows the osteoinductive properties of the bone matrix to be maintained at room temperature for three months, which eliminates the difficulties in transporting bone grafts to medical facilities and storing them before surgery;

Claims (1)

The claims A method of preserving demineralized bone grafts by placing in a 20% urea solution, characterized in that, in order to preserve the osteoinductive properties of the bone graft at room temperature, the graft is pre-treated with a 0.3% solution of calcium chloride with the addition of sodium dodecyl sulfate and sodium chloride, and dodecyl sulfate and calcium chloride are added to the preservation solution. FIG. I figure 2.