The method of forming anticoagulant surface

 

(57) Abstract:

The invention relates to biology, in particular biochemistry. The formation method of the surface is consistent application of water or water-alcohol solutions of dyes acridine series and heparin on the surface, is able to adsorb or chemically bind the dye molecules. Effect: method provides a quick and easy formation of the surface with atrombogenity properties. table 1.

The invention relates to the field of biology, in particular biochemistry.

Progress angiosurgery has made significant progress in the development and implementation of reconstructive operations on the vessels. However, thrombosis of the reconstructed vessels that occur both in the near future mainly in the late postoperative periods, significantly reduce the effectiveness of surgical interventions. In this regard, despite the numerous studies conducted in this direction, the problem of formation of anticoagulation, particularly heparinised surfaces of vascular prostheses, vessels with acetaminophenol the intima, ALLO - and xenotransplantation material, etc. is not completely solved in the least formation of anticoagulant (heparinised) surface:

a) local treatment of blood vessels and anastomoses with a solution of heparin during surgery (Operative surgery /Ed. And. of Littmann.-Budapest:Publishing house of the Academy of Sciences of Hungary, 1985,-s.). This measure is short-term: most of the anticoagulant is not delayed when restoring blood flow;

b) physico-chemical and chemical methods of surface treatment of blood vessels or ALLO - and xenografts. For example, in order to prevent thrombosis wall endarterectomies arteries exposed to ultrasound in the presence of heparin for 30 sec. Heparin retained in the vessel wall is not more than 12 hours (Antushev A. F. Prevention of infection of vascular prostheses and thrombosis endarterectomies arteries /Diss. for obtaining the academic degree of Cand. Sciences.-1985). The umbilical vein or the carotid artery of dogs, pigs, proposed for use as vascular grafts, chemically modified heparin using carbodiimide for at least 5 hours, reaching concentrations of heparin to 31.5 μg per cm2surface (E. S. Venkataramani, Senatore F., M. Feola, et al. Nonthrombogeni small-caliber human umbilical vein vascular prosthesis //Surgery.-1986.-V. 6.-P. 735-741);

C) surface heparinization of different synthetic (for example, polio is C. S., McRea J. C. et al. Heparinized polyurethanes: in vitro and in vivo stadies //J. of Biomedical Materials Research. -1985. -V. 19. -P. 419-436), impregnation of synthetic grafts heparin-collagen copolymer (Shankar N., Senatore F., Wu D. R. et al. Co-immobilisation and interaction of heparin and plasmin on collageno-elastic tubes //Biomaterials, Artificial Cells and Artificial Organs.-1990. - V. 18.-#1.-P. 59-73), etc., These methods heparinisation or are in the process of scientific development, or have not received practical application because of some disadvantages.

Also there is a method of binding heparin, which is a polyanionic electrolyte, with cationizing surface. For example, heparin or heparansulfate (endothelium bovine aorta), immobilized on a partially cationizing using 3-chloro-2 - hydroxypropyltrimethylammonium the surface of the pulp, possess anticoagulant activity and can be used as biomaterials, membranes with atrombogennoe surface similar to the surface of blood vessels (Baumann N., Keller R., Ruzicka E. Partially cationized cellulose for non-thrombogenic membrane in the presence of heparin and endothelial-cell-surface - heparansulfate //J. Membr. Sci.-1991. -V. 61.-P. 253-268) prototype. The same principle binding of heparin with a Quaternary ammonium salt used in uninominal chromatography. Also describes the interaction of heparin with Paul M., Fazal M. S., Wan K. S. Heparin binding on poly(L - lysine)-immobilized surface //J. Colloid Interface Sci.-1991.- V. 147.-#1.-P. 251-261). On the smooth surface of the polymer for 30 minutes from blood or plasma at a speed of 100 ml/min heparin concentration reaches a value of 0.52 ág/cm2on porous was 1.69 mg/cm2. For the manifestation of the surface of the anticoagulant properties of heparin concentration should not be less than 5 ág/cm2(E. S. Venkataramani, Fred Senatore, Mario Feola, et al. Nonthrombogenic small-caliber human umbilical vein vascular prosthesis //Surgery.-1986.-V. 6.-P. 735-741).

The authors propose a method of formation of anticoagulant surface using dyes acridine series, such as acridine orange (AO), ethacridine lactate (EL) and Any other surface that can either adsorb or chemically bind the dyes initially processed water or a water - alcohol solution of the dye, and then an aqueous solution of heparin. Heparin as a polyanionic electrolyte, interacting with aggregated dye molecules, gives the surface anticoagulant properties. The advantages of this method are speed and ease of formation atrombogennoe surface, and the ability in a wide range to change the surface concentration of heparin in a time-dependent vozdeistviyah in the quantitative determination of glycosaminoglycans in various biological fluids (G. P. Diakun at al. A simple purpose-built fluorimeter for the titrimetric assay of glycosaminoglycanes //Anal. Biochem.- 1979.-V. 94.-P. 378-382), as well as in the quality of medicines (M. D. Mashkovsky. Drugs.-Kharkov: Torsing, 1997.- So 2,-s.). For the formation of anticoagulant surfaces any tissue of a living organism, containing glycosaminoglycans, can be treated sequentially with solutions of dyes acridine series and heparin.

The invention is illustrated by the following example.

Aqueous solutions of AO or E put on a surface area equal in fragments artery or aorta with acetaminophenol the intima within a certain time. Excess dye after staining of tissue is removed with filter paper and treated surface with a solution of heparin (5000 u/ml) for 5 minutes. Unreacted with the surface of the copper dye and heparin washed with running water for 10-15 minutes. For the quantitative determination of heparin dye on the surface of the fragments restored with zinc in hydrochloric acid environment to leucosolenia. After bleaching of the dye heparin, passed into the solution, determine carbazolyl method by the reaction of the DISHA on uronic acid Carbohydrate analysis /Ed. by M. F. Chaplin and Kennedy J. F.-ia dyes acridine series fabric Dura, autovenous graft well procrasinate that is promising in terms of prevention of thrombosis during the reconstruction of blood vessels by using grafting materials.

The method of forming anticoagulant surface by applying the heparin solution, characterized in that the surface of the fabric is pre-processed water or a water-alcohol solution of the dye acridine number within 1 to 10 min, and then applied to the heparin solution.

 

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