RU2580621C1 - Biocidal composition for aseptic storage of preserved prosthetic material from tissues of animal origin - Google Patents

Abstract

FIELD: pharmaceutics.

SUBSTANCE: invention relates to biocidal composition for aseptic storage preserved prosthetic material from tissues of animal origin. Biocidal composition for aseptic storage preserved prosthetic material of animal origin tissues, including the antimicrobial agent from mixture of diatomic alcohols fatty row, phenoxyethanol and sorbic acid, buffer and osmolar aqueous solutions, as buffer aqueous systems are buffer systems, providing pH within range 5.0-7.0, as component of isotonicity buffer solutions of sodium chloride solution, providing osmolarity of composition in range 260-310 mosm/l, and to improve solubility of antimicrobial agent simplest mono-atomic spirits in certain proportions are added.

EFFECT: composition described above provides reliable protection against contamination, allows to reduce time of cleaning of bioprostheses of preserving solution before implantation and does not contain toxic and potentially toxic components.

9 cl, 1 tbl, 2 ex

Description

The invention relates to medicine, namely to the manufacturing technology of prosthetic material from allo- and xenogenic tissues, and can be used to prevent contamination of biological prostheses during their long-term storage until implantation.

The earliest methods for the aseptic storage of bioprostheses involved the use of solutions of a crosslinking agent, which is the main preservative of biomaterial. The sterilizing effect of cross-linking agents is based on cross-linking of bacterial proteins, which leads to the death of microorganisms.

Thus, a known method of sterilization and storage of xenoprostheses in a 0.5% solution of glutaraldehyde (Guidelines for sterilization of xenobioprostheses with a solution of glutaraldehyde. USSR Ministry of Health. N 28-6 / 26 of 09/19/1986). The disadvantage of this method is the long washing of the canned biomaterial from the sterilizing solution - for 90 minutes with vigorous stirring in 500 ml of a 0.9% sterile sodium chloride solution with a change of solution every 15 minutes.

A known method of preservation and sterilization of xenogenic prostheses of heart valves and blood vessels in a buffer solution of diglycidyl ether of ethylene glycol (DEE) (RF patent No. 20088767, class A01N 1/00, publ. In 1994, bull. No. 5). Preservation and sterilization of biological material is achieved by treating with a 2-5% DEE solution. Until the implantation of the product is stored in the same solution. This method has the following disadvantages:

1) before implantation, it is necessary to wash bioprostheses from a preservative in a sterile 0.9% sodium chloride solution for a long time (within 1 hour with a 3-fold change of solution every 20 minutes). This leads to an increase in the duration of the operation, which is especially critical for cardiovascular bioprostheses in open heart surgery;

2) epoxides in aqueous solutions are unstable, in contrast to glutaraldehyde. Oxyran rings easily open, and chemical compounds lose their crosslinking and, therefore, biocidal activity. Thus, the long-term preservation of the sterility of products in epoxide solutions is doubtful.

In order to reduce the intraoperative pre-implantation washing of bioprostheses to 2-10 minutes, methods for aseptic storage of chemically cross-linked, sterile material have been developed. For this, compounds have been proposed that are used for other purposes in the cosmetic and pharmaceutical industries.

So, there is a method of storing biological prostheses in a solution containing methyl and propyl parabens (US patent 5188834, CL A61L 27/36, publ. 02.23.1993). The disadvantage of this method is that parabens have only a bacteriostatic effect and are partially deactivated in the presence of proteins, which increases the likelihood of loss of sterility during prolonged storage of biological prostheses. In addition, the potential carcinogenic and allergenic effects of parabens are still widely debated.

There is also a method of preimplantation storage of bioprostheses in a water-buffer (0.05-2.0 M phosphate buffer at pH 4.0-8.0) solution of 2-bromo-2-nitropropane-1,3-diol with a concentration of 0.01 -2.0% (RF patent No. 2357766, class A61N 1/02, A61L 27/00, priority dated July 6, 2007). The disadvantages of this method include the following negative aspects regarding the main biocidal agent:

1) 2-bromo-2-nitropropane-1,3-diol in aqueous solutions in the presence of proteins (which form the basis of all biological prostheses) decomposes with the formation of highly toxic and carcinogenic products - formaldehyde and nitrosamines, as evidenced by the staining of the biomaterial in intensely yellow color over the course of time (1-2 months);

2) 2-bromo-2-nitropropane-1,3-diol is among the 15 most common allergens. Both reasons led to the prohibition of the use of 2-bromo-2-nitropropane-1,3-diol in the cosmetic and pharmaceutical industries of several countries (for example, Canada).

Thus, long-term storage for subsequent implantation of bioprostheses in patients with potential toxic, allergenic and carcinogenic properties cannot be considered optimal.

At the same time, in recent years, a number of non-toxic biocidal mixtures have been proposed for cosmetic products, including mixtures consisting of fatty dihydric alcohols (1,2-pentanediol, 1,2-hexanediol, 1,2-octanediol), phenoxyethanol and co-biocidal components (US patent No. 7935732 B2, class A01N 31/00 (2006.01), publ. 05/03/2011; US patent No. 8519010 B2, class A01N 31/00 (2006.01), publ. 08.27.2013) . The mixture contains either all three alcohols: 1,2-pentanediol, 1,2-hexanediol and 1,2-octanediol in a ratio of 1: 1: 1 in a total mass fraction of 40-60%, or only 1,2-octanediol ( caprylyl glycol) in a similar mass fraction; the next component is phenoxyethanol (40-60%); 0-10% of the mixture can be represented by sorbic or benzoic acids, dibromo-dicyanobutane, 1,2-benzo-isothiazoline or a mixture thereof. To achieve a biocidal effect, it is recommended to introduce no more than 2% (by weight) of this mixture into the composition of the preserved product; the optimal concentration is 0.5-1.5%.

The disadvantage of these mixtures is their poor solubility in water due to dihydric fatty alcohols. As a result, the mixture is intended to be used as emulsions in gels, shampoos, creams, but not in preparations based on aqueous solutions. At the same time, the storage of prosthetic material from biological tissues in suspensions and emulsions is unacceptable due to the formation of hard-to-remove formations on the surface of the biomaterial, which can lead to a violation of the bio- and hemocompatible properties of bioprostheses.

The solubility of these mixtures in an aqueous medium can be increased by using several approaches and their combinations:

1) by reducing the mass fraction of sparingly soluble dihydric alcohols of the fat series;

2) due to the introduction of low molecular weight monohydric alcohols into the composition of the biocidal composition, the most preferred of which is ethyl alcohol, which is widely used for medical purposes; non-toxic in small concentrations and having its own biocidal activity;

3) by increasing the ionic strength of the aqueous solution by introducing a salt component into the composition, the most preferred of which is sodium chloride. When using a 0.9% aqueous solution of sodium chloride, among other things, iso-osmolarity of the composition is achieved. Maintaining osmolarity close to plasma osmolarity prevents impaired structural integrity of the biological tissue, which is manifested by edema under conditions of hypoosmolarity or dehydration under conditions of hyperosmolarity.

A biocidal composition is proposed for the aseptic storage of canned prosthetic material from tissues of animal origin, containing several components:

1) an antimicrobial mixture with a mass concentration of 2-atomic fatty alcohols of not more than 30%, a phenoxyethanol content of 60-65%, and a sorbic acid concentration of 6-10% of the antimicrobial mixture. The mass fraction of the antimicrobial mixture in the composition can be brought up to 5% by weight;

2) low molecular weight monohydric alcohol (ethyl or isopropyl) in a concentration of 5-20% by weight;

3) an aqueous solution of saline buffer, providing not only pH, but also the osmolarity of the solution, close to the osmolarity of blood plasma. In this case, it is preferable to use phosphate, citrate or phosphate-citrate buffers prepared on a 0.9% sodium chloride solution, because they provide both a wide pH range - from 3.0 to 8.0, and osmolarity in the range of 300-320 mosm / l.

The technical result of the invention is to obtain a biocidal composition for the aseptic storage of canned prosthetic material from tissues of animal origin that does not contain toxic and potentially toxic components, which provides reliable protection against contamination and reduces the time it takes to wash bioprostheses from a preservation solution before implantation.

The following non-limiting examples illustrate the present invention.

Example 1. A biocidal composition was prepared as follows: 5 g of an antimicrobial agent (1 g of 1,2-octanediol + 3.7 g of phenoxyethanol +0.3 g of sorbic acid) was dissolved in 20 g of ethanol (96%) and the volume was adjusted to 100 ml of 0.1 M phosphate buffer (pH 5.8) prepared in a 0.9% sodium chloride solution, thus obtaining a 5% solution of the antimicrobial agent.

By diluting this 5% solution with the above buffer, working solutions of the biocidal composition with an antimicrobial agent concentration of 2%, 1%, 0.5% and 0.1% were obtained.

In tubes containing 4.5 ml of biocidal composition with a concentration of antimicrobial agent of 0.1%, 0.5%, 1%, 2% and 5%, 0.5 ml of a suspension of microorganisms was added: E. coli (strain 1257) , Ps.aeruginosa (ATCC strain 27853), St. aureus (strain 906), Ent. faecalis (hospital strain), Candida albicans (strain 15) and Aspergilla niger (museum strain NIID) at a concentration of 2 × 10 ⁹ microbial bodies / ml according to the optical turbidity standard No. 20.

Incubation was carried out for 1, 3, 7 and 10 days, after which seeding was carried out on the appropriate medium. The results are presented in table 1.

The test results show that the proposed composition has a good biocidal effect - a 0.1-5.0% solution of the antimicrobial agent inhibits the growth of all test microorganisms - both bacteria and fungi.

Example 2. Biological prostheses of vessels from the internal thoracic artery of cattle (4 pcs.) And xenopericardial flaps (7 pcs.) Were produced under standard conditions: the biomaterial was preserved with ethylene glycol diglycidyl ether for 14 days, after which at all stages of aseptic production bioprostheses were stored in the proposed biocidal composition with a concentration of an antibacterial agent of 1%. Upon completion of all production manipulations, bioprostheses were packed under aseptic conditions in standard sealed containers containing a biocidal composition with an antimicrobial agent concentration of 0.5%. After 6 months of storage, all bioprostheses were examined for sterility. Result: all samples are sterile.

Claims (9)

1. Biocidal composition for aseptic storage of canned prosthetic material from tissues of animal origin, including antimicrobial agents, buffered and osmolar aqueous solutions, as well as simple monohydric alcohols, characterized in that a mixture of fatty dihydric alcohols, phenoxyethanol and sorbic is used as antimicrobial agent acids, as buffer water systems - buffer systems providing a pH in the range of 5.0-7.0, as a component of the isosmolarity of buffer solutions - a solution of chlorine sodium chloride, providing osmolarity of the composition in the range of 260-310 mOsm / l, and to increase the solubility of the antimicrobial agent is added simple monohydric alcohols in the following ratio, wt.%:
- antimicrobial agent - 0.5-5;
- aqueous buffer solution - 74.1-93.8;
- isosmolar component is 0.7-0.9,
- low molecular weight monohydric alcohol - 5-20. 2. The composition according to p. 1, characterized in that the antimicrobial agent includes, as fatty alcohols, a mixture of 1,2-pentanediol, 1,2-hexanediol and 1,2-octanediol in a ratio of 1: 1: 1 in a mass fraction of 0 , 05-1.5% based on the weight of the solution. 3. The composition according to p. 1, characterized in that the antimicrobial agent includes, as the dihydric alcohols of the fatty series, 1,2-pentanediol in a mass fraction of 0.05-1.5% with respect to the weight of the solution. 4. The composition according to p. 1, characterized in that the antimicrobial agent includes, as the dihydric alcohols of the fatty series, 1,2-hexanediol in a mass fraction of 0.05-1.5% with respect to the weight of the solution. 5. The composition according to p. 1, characterized in that the antimicrobial agent includes 1,2-octanediol as a diatomic fatty alcohol in a mass fraction of 0.05-1.5% with respect to the weight of the solution. 6. The composition according to p. 1, characterized in that the antimicrobial agent comprises phenoxyethanol in a mass fraction of 0.39-2.5% relative to the weight of the solution. 7. The composition according to p. 1, characterized in that the antimicrobial agent comprises sorbic acid in a mass fraction of 0.01-0.5% relative to the weight of the solution. 8. The composition according to p. 1, characterized in that the buffer solution is selected from a phosphate, citrate or phosphate-citrate buffer system. 9. The composition according to p. 1, characterized in that to increase the solubility of the antimicrobial agent use ethyl or isopropyl alcohols.