Method for obtaining thromboresistant polymer materials

FIELD: medicine.

SUBSTANCE: invention refers to the field of polymer chemistry and medicine, namely to method for obtaining thromboresistant polymer materials which have widespread application in medical industry for manufacturing workpieces on blood contact, for example blood-vessels prostheses, parts of bioartificial organs implanted into living body, bloodlines for artificial blood-circulation apparatus, storages for storage and blood transfusion etc. Method for obtaining thromboresistant polymer materials implies mixture polymer with extender, and is inert to blood coagulation water-soluble compounds in amounts of 0.3-3.5 wt % used as extender.

EFFECT: invention enables to produce thromboresistant polymer materials with lowered tendency to adhesion of platelets and lowered ability to formation of fibrinous thrombs on the surface of material in the absence of influence of whole blood coagulation system, as evidenced by increase of buildup time of fibrinous clod from 60-80 seconds to 110-240 seconds.

3 tbl, 36 ex

 

The invention relates to the field of polymer chemistry and medicine, namely to a method for thromboresistant polymer materials, which find wide application in the medical industry for the manufacture of contact with blood products, such as artificial blood vessels, parts implanted in a living organism, artificial organs, arteries of artificial blood circulation, storage and transfusion of blood, etc.

All presently used in contact with blood materials are not truly thromboresistance. Blood contact with any foreign material, including polymers, leads to immediate its collapse, which is a natural protective reaction of the body. Positive results of implantation of some polymeric materials are either relatively small size of the implants (heart valves), or the fact that in conditions of intensive blood products blood clots constantly washed away from the surface of the hydrolyzed polymer and fibrinolytic enzymes blood. In both cases, a protective system able to cope with anticoagulation.

It is known that the process of blood coagulation on the surface of polymers involves several steps [Smurova E.V., Dobrova NB Creating polymer materials with trom resistentie properties. Chemistry and technology of macromolecular compounds, Moscow, VINITI, 1976, 10, p.30-60]. The first step is the adsorption of proteins, which is favourable and unfavourable. So the adsorption of serum albumin inhibits several further stages of coagulation of the polymer surface, and the adsorption of fibrinogen, on the contrary, accelerates the process of blood clotting. The second stage is the adhesion and aggregation of platelets with their destruction and the release of substances that cause further platelet aggregation. The third step is the activation of coagulation factors and the conversion of soluble fibrinogen into insoluble fibrin, which, together with the jam in it shaped blood cells and is a blood clot.

In this regard, the most common approach to improving thromboresistant polymers is their modification of biologically active compounds that affect the different stages of thrombogenesis [Biocompatibility, edited Viewactive. The Moscow state unitary enterprise "Information centre of Vniigeosistem", 1999, s-305]. Typically the modification is subjected to a surface already prepared polymer product, as in the modification of the feedstock in the process of its transformation into a product is the destruction of the modifying agent with the loss of their biological activity.

A method of obtaining trombones santih polymers by radiation graft copolymerization on their surface unsaturated derivative of a natural anticoagulant blood - heparin or its mixture with a hydrophilic monomer [Ed. St. USSR №1120679, C08F 291/00, 1979].

The disadvantage of this method is the low thromboresistant polymer (a relative measure of the adhesion of platelets equal to the ratio of the number of platelets per unit area of the sample to the number of platelets per unit area of standard glass, for the original and the modified polymer is 1.0±0.2)and also an active effect of heparin on blood coagulation (clotting time in contact with the polymer increases blood 7 times).

A method of obtaining thromboresistant polymers by radiation graft copolymerization on the surface of acrylic acid chloride or methacrylic acid followed by treatment of the grafted copolymer with a solution of heparin [Ed. St. USSR №1120680, C08F 291/00, 1979].

The disadvantage of this method is the active influence of the polymer on blood coagulation (clotting time in contact with the polymer increases blood 6 times).

The closest in technical essence and the achieved results is the method of obtaining thromboresistant polymeric materials by mixing the polymer with the modifying agent [Kim S.W., Ebert CD., Lin J., J.C. McRea, Nonthrombogenic polymers: pharmaceutical approaches, Journal of American Society Artificial Internal Organs, 1983, v.6, p.76-87]. As the modifying agent is sportsouth natural anticoagulant blood - heparin in the amount of less than 0.1% wt. During implantation into the bloodstream these materials act as a depot of heparin and increased thromboresistance of these materials due to the leaching of heparin in the blood from the surface of the implant and prevent them from clotting by the usual mechanism of action of the anticoagulant. To maintain thromboresistance the release rate of heparin in the blood flow should not be less than 4×10-2ág/cm2a minute.

The disadvantage of this method is the low thromboresistant received due to loss anticoagulatory activity of heparin in contact with blood (in the blood plasma of a person is more than 20 proteins that are able to neutralize the anticoagulant activity of heparin [Biocompatibility, edited Viewactive. The Moscow state unitary enterprise "Information centre of Vniigeosistem", 1999, s]), and reduce clotting of the whole blood under the action of released heparin, which can lead to bleeding [Biocompatibility, edited Viewactive. The Moscow state unitary enterprise "Information centre of Vniigeosistem", 1999, s-305].

The objective of the invention is to increase thromboresistant polymeric materials and the elimination of the impact modifying agent in blood clotting.

The technical result achieved when use is lovanii of the invention, is to increase thromboresistant polymeric materials and the elimination of the impact modifying agent in blood clotting.

The technical result is achieved in that in the method of obtaining thromboresistant polymeric materials by mixing the polymer with the modifying agent, as the modifying agent is an inert relative to the blood coagulation system is a water-soluble compound in the amount of 0.3 to 3.5 wt.%.

The mechanism of action of the modifying agent is in a continuous leaching it from the surface of the polymer with the surface renewal and destruction of platelets and blood proteins without affecting the whole blood clotting. Unlike polymer obtained by the method prototype, whose thromboresistant caused by the release of the anticoagulant and lower total blood clotting, the resulting polymer is true thromboresistant.

The following examples illustrate the invention but in no way limit its scope.

Example 1

5 g of polyurethane based on aliphatic polyester with a molecular weight of 40,000 is dissolved in dimethylformamide and to the solution was added 0.015 g of NaCl powder (to 0.3% wt.). After stirring the reaction was poured onto the glass surface and the dimethylformamide removed by evaporation PR is the temperature of 90°C and a pressure of 20 mm Hg.

Evaluation of thromboresistance obtained film is carried out by two methods: by measuring the time of the formation of a fibrin clot on the surface of the film and measuring the number of adhered platelets. In the first method, on the surface of the film put a drop (~50 µl) solution of fibrinogen with a concentration of 1.5 mg/ml, which is added to a solution of thrombin and measure the time of the formation of a clot of fibrin.

Evaluation of the adhesion of platelets carried out using the method of electron microscopy. On the surface of the film put a drop (~50 µl) platelet-rich plasma of human blood. Film and the plasma is maintained at 20°C for 15 minutes. Then the film was washed with 0.9% NaCl to remove leatheireannach platelets and proteins. The film is treated with 2.5% solution of glutaraldehyde for 1 hour and washed successively with water, 20%, 70%, 90% and 100% ethanol. The film is dried and sprayed with copper. On the surface choose 20 field size (28×28 μm), which measures the number of platelets. Evaluation of the impact of polymers on the coagulation is carried out by measuring the clotting time of blood after contact with the polymer. The results are shown in table 1.

Examples 2-14

The process is conducted according to example 1, using various modifying agents and different number of them. The results are shown in table 1.

Example 15 (a counter is local)

The process is conducted as in example 1, but without the use of the modifying agent. The results are shown in table 1.

Table 1
# exampleThe modifier and the number*During the formation of a fibrin clot, with ±10The number of adhered platelets on 100 μm2The clotting time after contact with the polymer, min ±8%
1NaCl, and 0.3%1301.8±0.36,3
2NaCl, 1,9%170of 1.3±0.36,0
3NaCl, 3,5%2101.0±0.25,8
4Glucose, 1,4%1201.9±0.25,8
5LiCl, 1,7%1401.0±0.2 6,2
6PAK, 0,3%1101.6±0.36,0
7PAK, 2,0%1201.5±0.36,3
8PAK, 3,5%1801.3±0.26,1
9PEG, 1,0%1201.1±0.26,0
10PEG, 3,0%1600.9±0.25,8
11PEG, 3,5%2001.0±0.25,9
12PVP, 0,3%1201.3±0.26,3
13PVP, 0,9%1701.2±0.26,0
14 PVP, 3,5%1900.6±0.25,9
15, the controlno702.6±0.26,1
*PAK - polyacrylic acid with a molecular weight of 42000,
PEG - polyethylene glycol with molecular weight of 12000,
PVP - polyvinylpyrrolidone with a molecular weight of 25000.

Examples 16-26

The process is conducted according to example 1, using polyurethane with a molecular weight of 35,000-based aromatic polyester. The results are shown in table 2.

Table 2
# exampleThe modifier and its quantityDuring the formation of a fibrin clot, with ±10The number of adhered platelets on 100 μm2The clotting time after contact with the polymer, min ±8%
16NaCl, and 0.3%1601.4±0.3 6,3
17NaCl, 3,5%2000.9±0.25,9
18LiCl, 1,3%1701.1±0.26,5
19PAK, 0,3%1301.5±0.36,3
20PAK, 3,0%1501.4±0.36,4
21PEG, 1,0%1701.0±0.26,4
22Glucose, a 1.8%2100.9±0.26,6
23PEG, 3,5%230<0,56,1
24PVP, 0,3%1801.2±0.26,3
25 PVP, 3,5%200<0,56,2
26, the controlno603.1±0.26,4

Examples 27-35

The process is conducted as in example 1, as the original polymer using a polydimethylsiloxane with a molecular weight of 50,000, and as a solvent using toluene. The polymer film is prepared by evaporation of toluene at 60°C and a pressure of 20 mm Hg.

The results are given in table 3.

Example 36

The process is conducted according to example 15, using a polyurethane-based aromatic polyester. The results are shown in table 3.

Table 3
# exampleThe modifier and its quantityDuring the formation of a fibrin clot, with ±10The number of adhered platelets on 100 μm2The clotting time after contact with the polymer, min ±8%
27NaCl, and 0.3%2401.6±0.3 6,3
28NaCl, 3,5%2601.1±0.26,8
29Glucose, 2,4%1901.0±0.27,0
30PAK, 0,5%1900.9±0.36,4
31PAK, 3,5%200<0,56,5
32PEG, 1,6%2101.1+0.26,9
33PEG, 3,5%230<0,55,9
34PVP, 0,3%2201.1±0.26,0
35PVP, 3,0%270<0,55,9
36, the control no802.3±0.26,8

It is evident that the modification of polymers of water-soluble compounds leads to a decrease in the number of adhered polymer surface of platelets, which is accompanied by a significant increase in thromboresistant polymers, as evidenced by increased time of formation of the fibrin clot on the polymer with 60-80 seconds before 110-240 seconds. When released into the blood water-soluble compounds, in contrast to the prototype method, have no effect on blood clotting (coagulation time after contact with the polymer does not change), which excludes the possibility of bleeding after implantation of the polymeric products.

Thus, the present invention allows to obtain thromboresistant polymeric materials with low tendency to adhesion of platelets and reduced ability to form a fibrin clot on the surface of the material in the absence of effects on the coagulation system of whole blood.

Limit the number of reactive compounds used in the preparation of polymeric materials, are defined as. When the concentration of the modifying compounds less than 0.3% wt. there is no effect of the appreciation is of thromboresistance, and when the concentration of the modifying compounds above a 3.5% wt. a deterioration of the mechanical properties of the original polymer.

The method of obtaining thromboresistant polymeric materials by mixing the polymer with the modifying agent, characterized in that the modifying agent is an inert relative to the blood coagulation system is a water-soluble compound in the amount of 0.3 to 3.5 wt.%.



 

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