RU2344826C1 - Method of orgamax bioactive osteoplastic material preparation - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method of obtaining Orgamax multipurpose bioactive osteoplastic material involves grinding demineralised bone into bone powder with particle size of 0.1-0.5 mm and mixing it with prepared 20% collagen-containing solution at 1:1 ratio. For production of tissue engineering compositions sterilised material is enriched with growth factors or cell culture immediately before the operation.

EFFECT: osteoplastic material procurement for health institutions.

Description

The invention relates to medicine, namely to the procurement and preservation of bone tissue, and can be used in the work of "bone banks" to provide bone-plastic material for health facilities.

A known method of preparing bioactive bone-plastic material "Deprotex" (I. Kirilova, patent RU No. 2232585), in which deproteinized bone tissue is used as the main component. In addition to the obvious advantages, this method has certain disadvantages, it frees the material from morphogenetic proteins that stimulate osteogenesis in bone tissue.

Closest to the proposed method is a method of preparing a composite bioactive osteoplastic material "Kostma" (Patent RU No. 2211708). However, in this method, a native allosteal flour is used as the main component, in which the bone morphogenetic proteins are in a state associated with the mineral component, which impedes the formation of regeneration foci and the manifestation of osteoinductive properties of the resulting material.

The objective of the invention is to develop a method for the preparation of bioactive osteoplastic material with enhanced osteoinductive and prophylactic antibacterial properties, stimulating angiogenesis and the intensity of metabolic processes.

The solution of this problem allows one to obtain a multifunctional composite bioactive bone-plastic material that stimulates osteogenesis, ensures tissue growth and development, prevents purulent complications, stimulates angiogenesis and increases the intensity of metabolic processes in damaged bone tissues.

The technical result is achieved through the use of demineralized allogeneic bone meal as the main component, preserving bone morphogenetic proteins and potentiating a large number of foci of osteogenesis, inducing bone formation, and by introducing a collagen-containing component, which is the building material for the formation of regenerate, as a nutrient medium for cellular elements (for creation of tissue-engineering compositions) and a “carrier” of drugs of various pharmacological groups, ensuring their conductive sustained release that optimizes processes of reparative regeneration.

The problem is solved due to the fact that crushed demineralized allocity is used as the main component, and a 20% collagen-containing solution is used as a binder component, which is mixed with the main component in a 1: 1 ratio, the material is homogenized, sterilized, enriched with growth factors or cell culture to create tissue engineering compositions.

The method is as follows.

The first step is preparing the main component of the material - demineralized allogeneic bone meal.

1. Fragments of diaphysis of allogeneic tibia are cleaned mechanically from soft tissues.

2. They are sawn into smaller fragments of a given size.

3. Repeated mechanical cleaning of the obtained fragments is carried out with washing them with filtered filtered water.

4. The processed fragments are degreased in a solution of ethyl alcohol and ether in a ratio of 1: 1 for 30 minutes twice.

5. Decalcification is carried out in an alternating magnetic field with a strength of 15 Oe to 200 Oe and a frequency of 8.3-10 Hz. Bone fragments in a glass container are placed on a horizontal ceramic stand located at a distance of 4.5-5 cm from the surface of the magnet.

6. The container is filled with a solution of 0.6 N. hydrochloric acid in sterile physiological saline in a volume of 1:10 relative to the bone material and placed on an MM-3 magnetic stirrer with an average magnet rotation speed of 500-600 rpm (according to the technical conditions of the device). In this mode, the demineralizing solution is mixed without involving the bone material in rotation together with the magnet.

7. The process of demineralization and subsequent processing takes place at a temperature of 2-4 ° C, for which a magnetic stirrer and containers with bone material in a solution of hydrochloric acid are placed in a refrigerator.

8. A preliminary control of the degree of demineralization is carried out by a breakdown on the bending of bone material and by piercing the drug with a needle. The quality of decalcification is checked by the Kossa histochemical reaction.

9. Demineralized bone material obtained by the above method is washed for 2.5-3 hours with running filtered tap water until a pH of 7.0-7.2 is established.

10. Then the bone material is aged in a 10% solution of calcium chloride for 4 hours to recover.

11. Next, the bone material is sequentially placed in a 0.5 M solution of Trilon B (EDTA) for 1-2 hours and in a 0.8 M solution of lithium chloride for 4 hours.

12. Bone fragments are placed under the fan for 4 hours in a sterile box.

13. After that, the bone fragments are crushed to a size of 2-5 mm and crushed to the state of bone meal.

14. Bone meal is sieved through sieves to obtain uniform particles of 0.1-0.5 mm.

The second step is preparing a 20% collagen-containing solution.

1. Saline solution (0.9% NaCl) is supplemented with drugs of various pharmacological groups (antibacterial, vascular agents that affect metabolism, etc.), depending on needs;

2. Then, the prepared solution is combined with a collagen-containing component and kept for 1-1.5 hours at room temperature to saturate the collagen-containing component with preparations of various pharmacological groups.

3. After a specified time, the flask with the solution is placed in a water bath and heated to a temperature of + 40 ° C.

4. During heating, the contents of the flask are periodically mixed by shaking until the solution is homogenized.

In the third stage, demineralized flour and a 20% collagen-containing solution are combined in a 1: 1 ratio, the material is homogenized and cooled at room temperature.

The prepared bioactive osteoplastic material "Orgamax" is dosed, packaged, sterilized and preserved by freezing. If necessary, immediately before implantation, the sterilized material is enriched with growth factors or cell culture to create tissue-engineering compositions.

Claims (1)

A method of preparing a bioactive osteoplastic material by combining several components, characterized in that crushed demineralized allost is used as the main component, and a 20% collagen-containing solution is used as a binder component, which is mixed with the main component in a 1: 1 ratio, the material is homogenized, sterilized, enriched with growth factors or cell culture to create tissue-engineering compositions.