SU995718A1 - Method of obtaining biological envelopes for closure of burn-caused wounds - Google Patents

Description

(54) METHOD FOR OBTAINING A BIOLOGICAL SHELL FOR CLOSING BURNS

one

The image of ognosigs to medicine, namely to transplantation, can be used in the sterilization and preservation of the amniotic membrane to cover the burn wounds.

A known method of obtaining a biological membrane to close the burn wounds of the path is placed in a weak formalin solution (1 3However, this method does not allow jg to achieve a sufficient reduction in its immunospecificity.

The closest to the present invention is a method of obtaining a biological membrane for covering burn wounds by means of treating fetal membranes in solutions of a cationic detergent and antiseptic t2.

However, in the process of preserving the amesotoxic shell, there is no 20 decrease in its immunogenic properties, which leads to the development of a rejection reaction in terms of 14-3 ° C. after applying it to the burn surface, and

on the antisepticity of this method does not provide the transplant, at all stages of preservation.

The aim of the invention is to reduce the immunospecificity of the amniotic membrane and impart antibacterial activity to it.

: The goal is achieved by the fact that according to the method of obtaining a biological membrane for closing burn wounds by abrogating fetal membranes in solutions of cationic babies {) hevant n antiseptics, the blood is removed from fetal membranes obtained from rozhenpvp in non-sterile conditions with simultaneous sterilization , by shaking in an airtight container with a solution of M I containing, wt. including:

EthanolMurhlorvdl, O-t, 5

Cetylpirtzhy hpo-g

RID1.0-1.5

The remainder of the water is the ratio of the weight of hydrochloric acid to the volume of the solution (1: 15-20) at 18-25 ° C 6-t2 h until the end of staining the new portion of the solution in green color, then sterilized and canned in a sealed container with a solution containing, weight. h .: Ethanolmerkurhlorid0.4-0.6 Cetylpnridinium chloride0, 4-0.6 Sodium chloride7.2-7.8 Water bidistilled Ostae at the same ratio of tissue weight to solution volume of the same TavinepaType 5.0-8.5 h with the continuation of its storage in the same solution 1FI 2-4 ° C 20O24O h, then the amnion is separated from the chorionac in a cuvette with solution No. I at 20-25 o C, followed by sterilization of the amnion in the same solution at 18-25C 6 -12 h, after which the sterilized amniotic membrane is placed in a solution containing, by weight. h: FuratsilinO, 15-O, 2O Magnesium sulphate 99, 0-1O1, O Water is bad distilled The rest is at the same ratio of the weight of the fabric to the volume of the solution at 2-4 ° C where it is stored until use 480-48OO hours. Example I, A fetal shell, prepared for the woman from the mother in non-sterile conditions, is placed in a sealed container with solution number I, containing, by weight. h .; E tanolmerkurhlorid1,0 Cetylpyridinium chloride1, O Water bidistilled 9 97,0 when the ratio of the weight of tissue to the volume of 1:15 weight. The treatment of the graft was performed at 18 ° C by shaking with eight shakes per minute. After 8 minutes, the solution was changed. After cessation of the sixth portion of the solution in the greenish color, the allograft continues to be stored in this position of solution No. I 6h. Then, in the vial with the allograft of the fetal membrane, solution No. I is replaced with solution No. 2, containing (ratio of tissue weight to solution volume 1:15), weight. including: 3 tanolmercurlchloride, 0 4% ylpyridinium chloride 0, 4 Sodium chloride7,2 Bidistilled water 991, about which it is stored for TPR 18 C h and subsequent fananisia in the same solution at 2 ° C 2OO h. After that, the fetal swamp allograft is removed from vial with solution number 2 and placed in a cuvette filled with solution no. I, where the amnion is separated from the chorion at 2 ° C, then the separated amniotic membrane is sterilized in the bottle with solution no. I at 18 ° C. 6h. After that, the NI solution is replaced with the N 3 solution containing (the ratio of the weight of the tissue to the volume of the solution is 1:15) weight. h: Furacilin .0.15 Magnesium sulphate 99.0 Water was bidisglylated 898, 80 in which the amniotic membrane allograft is stored at 2 480 480 h. When examining an extract from this allograft with a disc electrophoresis method, one protein fraction is detected (before treatment by the proposed method, this amniotic eight fractions were revealed). Condensation of the total protein in the extract from this allograft is 18% compared to its content in the untreated amniotic membrane. A single precipitation arc was detected by the immunoelectrophoresis method in an extract from the amlotic membrane allograft (in the extract from this amniotic membrane before treatment by this method, six precipitations were detected). Antimicrobial activity is studied by placing a disc with a diameter of I cm, cut from this graft, on the surface of an agar seeded with Staphylococcus aureus. The zone of growth inhibition of microbes around the 8 mm allograft. Around the disc taken from the amniotic membrane before the treatment with the proposed method, the zone of growth inhibition of Staphylococcus aureus was not detected. PRI mme R 2. A fetal shell prepared from a woman in non-sterile conditions is placed in an airtight vessel with solution No. I containing, by weight.h: Ethanol-mercurcloride 1.5 Cetylpyridinium chloride 1, 5

Water distillirovanna 998, 0

when co-weighing with a volume of 1: 2O, it was done at 25 ° C with a five-shake regime per minute shaking per minute. After 5 microns, the solution was changed. After the staining of the eighth portion of the solution in a greenish color ceased, the allograft continues to be stored in a portion of solution no. I 12h. Then, in the vial with the apotransplantation of the fetal membrane, solution N) I is replaced with solution td 2 containing (coot-carrying the weight of the tissue to the solution 1:20), weight, hours:

Ethanol-Mercurcloride 0.6

Cetylpyridinium Chloe: Read0, 6

Sodium chloride 7.8

BidistihLi water. 992, O

in which it is stored at 25 8.5 hours with subsequent storage in the same solution at 4 ° C for 240 h. After that, the fetal membrane allograft is removed from the vial with solution No. 2 and placed in a cuvette filled with solution No. I, where amnion is separated. from chorion at 25 ° C, then the separated amniotic membrane is sterilized in a bottle with solution No. I at 25 ° C for 6 hours. After this, solution No. I is replaced with solution No. 3, containing (ratio of tissue weight to area of solution: 1: 2O), weight h:

Furacilin, 2O

Magnesium sulfate lOl.O

The water is bidistilled-.

IA 899.0

in which the allograft amniotic membrane is stored at 4 ° C 48OO hours.

In the study of the extract from this allograft using the d-sclerotherosis, two protein fractions were detected (nine fractions were detected in the amniotic membrane before the treatment by the proposed method). The concentration of total protein in the extract from this allotransport is 16% compared to its content in the untreated amniotic membrane. One precipitation precipitate was detected in the extract from the amloigrasion membrane allograft (in the extract from this amnnotic membrane prior to treatment using the proposed method, seven were found: precipitation precipitation). :

Antimicrobial activity has been studied by placing a disk with a diameter of t cm, cut out of this adlograft {| tata, on the surface of agar, seeded with Staphylococcus aureus. The area of microbial growth around the allograft is 9 mm. The ekrug disc cut from the amniotic membrane before processing by the proposed method, the zone of growth inhibition of Staphylococcus aureus is not detected.

The resulting biological membrane, sterilized, preserved and saturated with antiseptics, was used in a patient D, U years, diagnosis: burn I - HI And the degrees of the neck, chest, back,

an area of 10%. The burn wound was about but momentarily closed on an area of about, 5OO cm 2. The patient's treatment was wound up: with recovery, with complete epithelization of the burn surface, eleven days after application of the biological membrane on it, treated using the proposed method.

The proposed method allows to obtain a biological membrane with reduced immunospecificity, as evidenced by a decrease in the content of sheathed protein in extracts from it, depletion, its antigenic structure, with antibacterial activity, saturated with antiseptics.

Claims (2)

Invention Formula I. The method of obtaining a biological coating to cover burn wounds by treating fetal membranes in solutions of cationic detergent and antiseptics, characterized in that, in order to reduce the immunospecificity of the amniotic membrane and to give an anesthetic activity, the fetal membranes obtained from unborn women are washed from the blood x, with simultaneous sterilization by shaking in an airtight container with solution number I, containing, by weight. h: Eganolmerkurkhlorid1, O-1,5 Cetylpyridium Chloride1, O-1.5 ; The water is double distilled Else 1FI the ratio of the weight of the tissue to the volume of the solution (1: 15-2О) at 18–25 ° C for 6–12 h until the fresh portion of the solution is stopped in a greenish color, then grow to conserve in a 57 megic bottle with a container containing , weight. h: Eganolmerkurkhlorid0,4-O, 6 yetichirshny chloride, 4-0,65 Sodium chloride7,2-7,8 Water bidistilled The rest 1FI the same ratio of fabric weight to solution volume and the same temperature 5.0-10.10 h with continued storage in the same solution at 2-4 ° C 2OO-24O h, then the amnion is separated from the chorion in a cuvette with solution No. 1 at 20-25, followed by sterilization of 15 amnions in the same solution at 18-25 ° C 6-12 h, after which the sterilized amniotic membrane is placed in a solution containing, by weight. h: 188 FurvtsilinO, 15-O, 20 Magnesium sulphate 99.0-101.0 Water bididnlopirovana The rest at the same cohesion weight psani to the volume of the solution at 2-4 ° C 48O-48OO hours. Sources of information taken into account in the examination of 1 "Kozlova-Lavrinenko T, E., Circle °" The use of weak formalin solutions for storing skin grafts, - In the book: Burn disease, pathogenesis, clinic, treatment. Kiev, l975, p. IP-3O. 2. R. Klen. Ln Research fn burUs (Eds), R. Hotter. T.L.Barclay, L.Kon skara, Bern, Stutgart. Vienna, Hans Huber, 1971, p. 289-292,