RU2400240C1 - Method of obtaining biologically active tissue preparation - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to veterinary. Method of obtaining biologically active tissue preparation based on cows' placenta includes filtration, crushing, homogenisation, centrifugation, pregnant uterus being removed in the first half of pregnancy (4-5 months) in period of evisceration of incalvers at slaughterhouse, longitudinal cut of serous-muscular-mucous layer of pregnant uterine horn is performed, cotyledons are collected by removing them from caruncles, preserving integrity of villi, cutting cotyledons with curved sterile scissors from tissue of baby placenta, puncturing with a needle chorionically amniotic membrane by vacuuming amniotic fluid is collected from amniotic bubble, cut cotyledons are crushed by cutting, and after that in homogeniser, earlier collected from the same uterus amniotic fluid is added in amount 10:1 of homogeneous mass, mixed slowly and settled, after that centrifuged at 10 thousand rev/min for 10 minutes, conservation is performed, supernatant part is poured into vials and sealed.

EFFECT: invention ensures increase of preparation efficiency.

2 tbl

Description

The invention relates to veterinary medicine, in particular to a method for producing a biologically active tissue preparation from the placenta of cows.

A known method of preparing a drug from the placenta, including grinding raw materials, conducting weak acid hydrolysis, isolating the product by centrifugation, neutralizing the supernatant, isolating and purifying the drug, freeze-drying (1).

The disadvantage of this invention is that the extraction of biostimulating, therapeutic substances of natural origin under the influence of any degree of acid hydrolysis, as well as sublimation treatment leads to the extraction of a reduced amount or properties of the therapeutic factor.

There is also a known method for producing the PDE preparation (denatured emulsified placenta), in the manufacture of which the tissues of the infant placenta are used: amniota, chorion tissue (2).

The disadvantage of this method is that upon receipt of the drug, chemicals are added that lead to changes in the natural properties of protein molecules, nucleic acids, and when using dispersion and emulsification, a drop-liquid phase is isolated from solutions, which is usually accompanied by a break in chemical bonds. Therefore, the natural healing properties of the drug are changing, losing the healing bactericidal and stimulating effect.

Closest to the present invention according to the technical and biological nature of the achieved result when using is a method for producing medicinal substances from the placenta.

The proposed method for producing a biologically active tissue preparation from human placenta involves washing the placenta, homogenizing the placenta tissue without separation into its constituent parts, hemolysis of the tissues in purified water, loading the tissues into a rotary pulsation unit (RPU) of the UM-075 brand. After that, a 3% solution of hydrogen peroxide in a ratio of 2: 1 is poured into the tank capacity and can withstand for 50-60 minutes with constant stirring (3).

The main disadvantages of the known method are the following: a double-processed placenta in a rotary pulsating apparatus using a cavitation shock wave and a 60-minute homogeneous mass content in a 3% hydrogen peroxide solution destroy placental enzymes and stem cells in the resulting preparation, which leads to a change in biostructure physiological, regenerative, reparative functions.

The objective of the invention is to develop and create a method for producing a tissue preparation using the obtained cotyledon fractions within 1.5-2 hours after removing the fetus from the uterus for the treatment of cows with postpartum diseases of the genital organs.

The aim of the invention is to improve the healing properties of drugs made from placental tissues for the treatment of cows with genital diseases after difficult birth.

This goal is achieved by the fact that cotyledons taken from the baby placenta of a pregnant cow in the first half of pregnancy (4-5 months) are rich in medicinal substances, because during this period of development of the calf fetus in the amniotic fluid (the product of cotyledons and caruncles) there is a maximum number of bactericidal and bacteriostatic enzymes: lysozyme, compliment, properdine, etc., which are detrimental to microorganisms with one (gram-positive) and two (gram-negative, viruses, fungi ...) membranes.

Cotyledons (cows on a baby placenta from 90 to 120 pcs.) Were collected from the baby placenta from an isolated uterus with fetus taken from the abdominal cavity from pregnant cows in a slaughterhouse as a result of the bankruptcy of the slaughterhouse.

The sequence of the proposed method is as follows.

1. A pregnant uterus is removed in the first half of pregnancy (4-5 months) during the period of nutrition of pregnant cows in a slaughterhouse.

2. Perform a longitudinal section of the serous-muscular mucous layer of the pregnant uterine horn.

3. I collect cotyledons by removing them from the caruncles, preserving the integrity of the villi, cutting off the cotyledons with curved sterile scissors from the tissue of the baby placenta.

4. By puncturing the chorially amniotic membrane with a needle, the amniotic fluid is collected from the amniotic bladder by evacuation.

5. The cut cotyledones are crushed by cutting, and then the amniotic fluid collected from the same uterus is added in a mechanical homogenizer at the rate of 10: 1 of a homogeneous mass, slowly mixed and settled, then centrifuged at 10 thousand rpm for 10 minutes, preservation , the supernatant part is poured into bottles and corked.

The drug is a whitish liquid with a yellowish tint, which meets the requirements for medicinal products.

Practical test of the proposed drug

A comparative assessment of the methods of practical application of our proposed ointment 30% form of a biologically active tissue preparation prepared by the method of homogenization and fractionation of cotyledon and amniotic fraction with 30% ointment of a biologically active tissue preparation made by the method in the prototype (copyright certificate RU 2148408 C1, 10.05. 2000), which was prepared in accordance with the method proposed by Polkovnikov PM, who used mechanical and electrical for the homogenization of whole placenta from pregnant cows Odom apparatus.

Ointments were prepared on lanolin (3: 7), adding 30% of each test preparation by mixing. Stab-cut and burn wounds were lubricated daily for 7 days, and 50-100 ml of 30% ointment was injected into sick cows in the postpartum cervix.

The result of treatment of animals with stab and cut wounds with preparations from placental tissues are shown in table 1, 2,

Table 1 The results of the treatment of pigs with wounds and burns. Ointment 30% Animals were treated Of them recovered Terms of treatment (goals) 1 number number % Cotyledon Cut wounds four 80 5 Burn Wounds 9 7 77.7 9 Whole placenta of cattle Incised Wounds 4 3 75 8 Burn Wounds 9 5 71,4 10

From table 1 it is seen that burn wounds under the action of the cotyledon drug create the healing, relief and healing of burn wounds in 77.7% of patients 5 days earlier than in the treatment of burn wounds with an all-placental preparation, i.e. treatment of burn wounds with the proposed drug in the form of a 30% ointment after daily lubrication was 6.3% more effective than in the control group of animals.

Sick cows with postpartum cervicitis were injected daily with 100 ml of 30% cotyledon ointment into the cervical canal. The results of treatment of cows are shown in Table 2.

The control group of cows in the cervix was injected with 100 ml of 30% ointment from the whole placental fraction.

table 2 The effectiveness of the drug in the treatment of cows with cervical diseases. A drug Cows total Of them recovered Recovery time (days) number % Ointment 30%, after 24 hours cotyledon drug 5 four 80 10 Full placental ointment 30% 8 6 75

From table 2 it is seen that the cotyledon 30% ointment proved to be a curative positive effect, which was 5% more effective than in the control group of sick cows.

References

1. US patent No. 4054648, IPC 6 A 61 to 35/50.

2. Belyaev V.I., Nezhdanov A.G., Lobodin K.A., Seminikhin I.P., Tyurenkov I.T. “The biological activity of preparations from the placenta”, Veterinary practice, No. 7 2006, p. 58-61 Journal 3.

3. Colonels P.M. Patent No.RU 2148408 C1, 09/14/1999.

Claims (1)

A method of obtaining a biologically active tissue preparation based on cow placenta, including filtration, grinding, homogenization, centrifugation, characterized in that the pregnant uterus is removed in the first half of pregnancy (4-5 months) during the period of pregnancy of pregnant cows in a slaughterhouse, a longitudinal incision of serous the muscular-mucous layer of the pregnant uterine horn, collect cotyledons by removing them from the caruncles, preserving the integrity of the villi, cutting off the cotyledons with curved sterile scissors from the tissue of the baby placenta, by the amniotic fluid from the amniotic bladder is collected by vacuum with a needle of the chorionic amniotic membrane, the cut cotyledones are crushed by cutting, and then the amniotic fluid collected from the same uterus is added in a mechanical homogenizer at a rate of 10: 1 of a homogeneous mass, slowly mixed and settled, then centrifuged at 10 thousand .rpm for 10 minutes, preservation is carried out, the supernatant is poured into bottles and sealed.