RU2612260C1 - Ointment for treating superficial thermal burns - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to pharmaceutical industry, namely to ointment for treatment of superficial thermal burns, which contains essential oils of above-ground part of common yarrow, Pontic wormwood, camomile and blue tansy, collected in the phase of blooming, as well as petrolatum and anhydrous lanolin, with specified ratio of components. Said ointment demonstrates expressed anti-inflammatory, wound-healing, antimicrobial effects.

EFFECT: invention reduces terms of treatment of skin superficial thermal burns of IIIa degree to 3 weeks.

6 dwg, 5 tbl

Description

The invention relates to medicine (experimental and clinical) and veterinary medicine, in particular to medicines for external use, and can be used to treat superficial thermal burns in humans and animals.

A number of ointments are known for the treatment of superficial thermal burns, some of which are already used in medical practice, while others are proposed for use:

1. “Panthenol” 5% [Ostrovsky N.V., Petrov V.V., Bystrova A.S., Musatskova M.V. Comparative evaluation of the effect of drugs for local treatment of wounds on the healing of thermal burns of the II-III degree in the experiment // Basic research. - 2014. - No. 6 (part 3). - S. 512-515]. The active substance of the ointment is dexpanthenol, auxiliary components are lanolin, petrolatum, liquid paraffin, isopropyl myristate, mityl parahydroxybenzoate, cholesterol, propyl parahydroxybenzoate, water (http://www.neboleem.net/pantenol.php).

2. “Levomekol” [Pavlenko O.Yu., Bgatova NP, Panichev A.M., Gulkov AN Features of the ultrastructural organization of granulation tissue fibroblasts when using the “LITOPLAST” wound cover after thermal skin burn // Modern problems of science and education. - 2006. - No. 3 - S. 28-29]. Levomikol ointment contains two active ingredients (40 mg of methyluracil and 7.5 mg of chloramphenicol in 1 g of ointment) and two excipients that ensure uniform application of the ointment and optimal penetration into tissues (polyethylene oxide-400 and polyethylene oxide-1500).

3. “Methyluracil” 10% [The effect of the preparation of ionized silver on the reparative regeneration of the skin and underlying tissues in modeling thermal and chemical burns in rats // N.S. Ponomar, Yu.S. Maklyakov, D.P. Khloponin, A.O. Revyakin. - Biomedicine. - 2012. - No. 1. - From 143-148.]. As auxiliary substances, the ointment contains soft white paraffin, anhydrous lanolin, and purified water.

The composition of many ointments for the treatment of burns includes synthetic chemical compounds that have a number of undesirable side effects. As a rule, ointments created on the basis of plant materials, the range of which is currently limited, are deprived of this drawback [I. Percev. Pharmaceutical and biological aspects of ointments NFU Publishing House: Golden Pages, Kharkov, 2003].

4. RF patent 2256441 (11/25/2003) describes an ointment for treating burns, purulent and infected wounds of various etiologies and a method for treating burns, purulent and infected wounds. The ointment contains, by weight. %: beeswax 20-30, butter 30-40, vegetable oil 15-25, rosin 15-25. The ointment is applied with a layer of 0.5-3 mm on a dense cotton cloth, which is applied to the wound and fixed with a bandage or plaster. The dressing is changed 1-3 times a day for 3-14 days, depending on the degree of skin damage.

It is known that the use of dressings prevents the access to the wound of light and oxygen, which are necessary for the involvement of restoration mechanisms in the healing process [Peauetenvironnement / Dubertret Louis // C.r. Ser. 2 - 200. - Vol. 323. - No. 7. - R. 629-632]. When using this ointment, the wound is conducted in a closed manner, a possible complication of which is the danger of anaerobic infection.

Increasing the therapeutic effectiveness of the ointment, accelerating the rejection of necrotic tissue of wounds, accelerating the maturation of young granulation tissue, enhancing the proliferation of epithelium, reducing the healing time of wounds, which are described in the description of this patent, is not confirmed by comparison with any control or with another ointment. The indication that the ointment “has a multifaceted effect: antimicrobial, analgesic” is not supported by either a microbiological study or a study of pain sensitivity and wound healing speed. Only two isolated examples with burns are given.

In example No. 4, the authors describe a clinical case of the treatment of a burn, the signs of which are blackening of the skin, necrosis of the skin and internal organs. The described burn refers to deep burns of IIIb-IV degree, is subject to surgical treatment and cannot heal within 14 days. Burns of the IIIb degree are known to require longer conservative therapy. The healing process is 3-6 weeks [Loginov L.P. Modern principles of local treatment of thermal burns / L.P. Loginov // Russian Medical Journal. - 2001. - T. 9. - No. 34. - S. 123-125].

5. Known ointment for the treatment of burn wounds [RF patent 2287324, author - Bvzaev TV (29.04.2005)], containing vegetable oil, wax, rivanol, tannin, glycerin, in the following ratio of components, wt. %: wax 13-15; rivanol solution 1.2-1.3; tannin 3-5; glycerin 15-20; vegetable oil - the rest. The ointment is applied with a sterile medical spatula in a thin layer on a sterile napkin and applied to the wound surface.

The authors do not indicate which vegetable oil is used to make the ointment. Increasing the effectiveness of the healing effect of the ointment, which is described in the description of the invention, is not confirmed by comparison with any control or other ointment.

The formation of a “soft scar tissue” at the site of the burn wound (Example 4) suggests that the ointment was apparently insufficient [Peauetenvironnement / Dubertret Louis // C.r. Ser. 2 - 200. - Vol. 323. - No. 7. - R. 629-632].

The composition of the ointment includes 1.2-1.3% rivanol. According to the manufacturer's instructions (LLC Himaks Pharma, Bulgaria), a rivanol solution in a concentration of 0.1% has side effects and causes the development of contact dermatitis, photosensitivity, local reactions in the form of skin irritation. It is also known that for the treatment and treatment of fresh and infected wounds, aqueous solutions of rivanol are used at a concentration of 0.05% (1: 2000), 0.1% (1: 1000), 0.2% (1: 500).

6. Known wound healing, anti-inflammatory ointment described in the patent of the Russian Federation 2326683 (10/06/2005) (prototype). The ointment contains, by weight. h .: St. John's wort perforated (grass and color) - 2, yarrow (grass and color) - 2, plantain big (leaf, seed) - 2, chamomile - 1, wormwood and bitter - 1, large celandine - 1, calendula (color) - 1, burdock (roots) - 2, pine resin (resin) - 0.5, propolis - 0.1, arnica (color) - 2, horsetail (grass) - 1, Japanese Sophora - 0 ,5.

As a fat base, the ointment includes interior unsalted pork fat or ghee. The ratio of plant material to fat base is 1:10.

Example 3 refers to the “complete epithelization of the entire wound surface” after a single use of the ointment in the form of a compress for a IIIa degree burn. It is known that IIIa degree burns need longer conservative therapy and the healing process is 3-6 weeks [Loginov L.P. Modern principles of local treatment of thermal burns / L.P. Loginov // Russian Medical Journal. - 2001. - T. 9. - No. 34. - S. 123-125]. The data on the timing of wound epithelization indicated by the authors cast doubt on the reliability of the results.

The creation of new drugs in the form of ointments based on plant materials with anti-inflammatory, wound healing and antimicrobial effects is relevant.

Disclosure of invention

The objective of the invention is to develop a composition and technology for the production of ointments intended for the treatment of thermal burns that do not contain chemical or special synthetic additives, which has a wide range of anti-burn effects, reducing the time of conservative treatment of surface thermal burns of III degree.

The proposed ointment for the treatment of superficial thermal burns contains essential oils of yarrow, Pontic wormwood, pharmacy chamomile and blue tansy, as well as petrolatum and anhydrous lanolin in the following ratio of components, wt. %: yarrow essential oil (Achillea millefolium L.) - 1.0, Pontic wormwood essential oil (Artemisia pontica L.) - 1.0, pharmacy chamomilla essential oil (Matricaria chamomilla L.) - 2.0, tansy essential oil blue (Tanacetum annuum L.) - 2.0, petrolatum - 84.6, anhydrous lanolin - 9.4.

Essential oils for the inventive ointment are obtained from the aerial parts of Pontic wormwood, yarrow, pharmacy chamomile and blue tansy, collected in the flowering phase.

Essential oils of yarrow and Pontic wormwood, which are part of the ointment, the authors obtained independently by steam distillation in accordance with the known method [RF patent 2356567. Essential oil from wormwood and the method of its production / Khanina MA, Makarova DL, Kim N.E., Hanina M.G. (10.31.2007)]. Essential oils from the aerial parts of the pharmacy chamomile and blue tansy were prepared (purchased), manufactured by FLORAME (France) and obtained by water-steam distillation.

List of figures illustrative material

FIG. 1. A - type of burn wound. B - skin area in the area bordering the area of primary necrosis. 1st day of the experiment in animals of the 1st group. Dilation of blood vessels, increased vascular permeability. Picrofuchsin stain according to Van Gieson. SW 600.

FIG. 2. A - type of burn wound. B - skin area in the area of paranecrosis on the 3rd day of the experiment in animals of the 1st group. Severe neutrophilic infiltration, fragmentary scab rejection. Picrofuchsin stain according to Van Gieson. SW one hundred.

FIG. 3. A - type of burn wound. B - skin area in the area of paranecrosis on the 7th day of the experiment in animals of the 1st group. Plots of dystrophy and necrosis of the epidermis and dermis, severe edema and leukocyte infiltration. Picrofuchsin stain according to Van Gieson. SW 400.

FIG. 4. A - type of burn wound. B - skin area in the bottom of the burn wound on the 10th day of the experiment in animals of the 1st group. The formation of connective tissue. Picrofuchsin stain according to Van Gieson. SW one hundred.

FIG. 5. A - type of burn wound. B - skin area in the bottom of the burn wound on the 14th day of the experiment in animals of the 1st group. The formation of the epithelium over the newly formed connective tissue. Picrofuchsin stain according to Van Gieson. SW 600

FIG. 6. A - type of burn wound. B - skin area in the bottom of the burn wound on the 21st day of the experiment in animals of the 1st group. Complete epithelization of a burn wound. Picrofuchsin stain according to Van Gieson. SW 400.

EXPERIMENTS FOR THE DEVELOPMENT OF THE DECLARED OINT

1. Modeling standardized burns

Surface thermal burns of III degree to assess the therapeutic effectiveness of essential oils of Pontic wormwood, yarrow, chamomile and tansy blue, as well as ointments in general, were modeled in accordance with the "Method for experimental modeling of thermal burns in laboratory animals" (application for invention No. 2014152729 from 12.24.2014) developed by the authors.

In accordance with the purpose of the experiment (studying the therapeutic effect of essential oils), it was decided that the severity of the burn should correspond to grade IIIa (subject to conservative treatment), and the burn area should not exceed 1% of the total body surface area of the animal.

On the eve of thermal burns on the outer thigh of the animals, shave was shaved under sevoran anesthesia, and the skin was treated with 70% ethyl alcohol. A day after depilation, the integrity of the skin was evaluated and, in the absence of signs of skin damage, it was considered ready for thermal burns.

A thermal burn was applied under sevoran anesthesia. An infrared heater of an infrared soldering station of the brand YaXunYX865D was placed at a distance of 15 mm from the skin of the animal. The radiation was directed perpendicular to the surface of the skin of the animal.

Burns were applied to the outer surface of the animal’s thigh, close to flat, on which the burn was perpendicular to the infrared radiation of the soldering station, falling on it, in the form of a circle. Therefore, the required burn area was set by a circle diameter of 20 mm (314 mm ² ).

The heating temperature of the skin of animals in the experiment was set at 60 ° C, based on literature data, since at this heating temperature infrared radiation penetrates into the tissues to a depth of 5 mm, warming them to 50-60 ° C [Burns: (Manual for doctors) / Ed. B.S. Vikhrieva, V.M. Burmistrova. - 2nd edition revised and supplemented. - L. Medicine, 1986. - 272 p., Ill.]. A heating temperature of 60 ° C made it possible to obtain surface thermal burns of degree IIIa, which was confirmed by data from morphological studies of skin preparations of experimental animals.

On the body of animals in the heating zone, a K-type thermocouple was fixed using KPT-8 heat-conducting paste. After the heating temperature of 60 ° C and the radiation power of 100 W were established on the panel of the IR soldering station, the skin of animals was heated from the initial body temperature of 37 ° C to a predetermined temperature of 60 ° C, which was recorded with a thermocouple. The temperature indicator on the panel of the IR station first shows the set temperature of 60 ° C, and after five seconds - the actual temperature on the skin of the animal. By setting the temperature control to position 3 (60 °), the temperature in the heating zone was limited.

Tissue heating in the heating zone from 37 ° C to 60 ° C took place over 23 seconds. Upon reaching 60 ° C for 23 seconds, heating was stopped by turning off the YaXunYX865 soldering station.

The results of morphological studies of skin preparations of experimental animals showed that heating in this mode provides a third-degree burn.

A visual observation revealed that in all experimental animals, a site of dry coagulation necrosis was formed at the site of a thermal burn within 10 minutes after its application. The surface of the burn was dense, yellowish in color and clearly demarcated from the surrounding healthy tissue. Thus, the effect of infrared radiation at a temperature of 60 ° C and a power of 100 W at a distance of 15 mm from the skin surface for 23 seconds caused the development of coagulation necrosis in rats, spreading to all layers of the skin.

Dilution of essential oils

To conduct experiments with essential oils, they had to be applied to a wound, and for this, diluted in some kind of oil. As a solvent, sweet almond oil (Sweet Almondoil), obtained by mechanical pressing from the kernels of seeds of cultivated sweet almonds (manufactured by Desert Whale Jojoba Co. Ltd., USA), was chosen.

Sweet almond oil provides easy absorption and resorption of the medicinal substances that make up the essential oils, does not interfere with the heat and gas exchange of the skin, while softening it [RF patent 2160092 Antimicrobial agent for treating infected wounds / Chichkun-Krikova AD, Seliverstov D .V., Isakov S.A., Gausman B.Ya. (02.16.1998)].

Essential oils of Pontic wormwood, yarrow, pharmacy chamomile and blue tansy were bred in sweet almond oil in the ratio, wt. h: essential oil - 1, sweet almond oil - 20.

Experiments conducted to evaluate the wound healing, anti-inflammatory and antimicrobial effects of essential vegetable oils and finding their ratio in the composition of the claimed ointment

Experiments of the series No. 1. Evaluation of the wound healing, anti-inflammatory and antimicrobial effects of essential oils

The information in the next two paragraphs applies to all animal experiments described below.

The experiments used male Wistar rats aged 2.5 months old, obtained from the vivarium of the Central Research Laboratory of SBEI HPE Novosibirsk State Medical University of the Russian Ministry of Health. The animals were kept under standard vivarium conditions (under natural light, air t + 20-22 ° С and humidity 55-60%) in plastic cages 55 × 45 × 15 cm in size, with litter from sawdust, on a normal diet and free access to 5 animals per cage in water.

The experimental study was conducted in accordance with the requirements of the National Standard of the Russian Federation “Principles of Good Laboratory Practice”, adopted in March 2010, and providing for preclinical studies of animal medicines in accordance with international rules [National Standard of the Russian Federation GOST 53434-2009 “Principles of Good laboratory practice ”].

In the experiment of series No. 1, 30 rats were used. The choice of this type of animal as an object of study is determined by their omnivorous nature and a wide range of existence, which makes the reactions of various organs and systems of rats similar to those in humans [Zapadnyuk I.P. Laboratory animals. - Kiev, Science, 1974. - 303 p.].

The information in the next two paragraphs applies to all animal experiments described below.

The minimum weight of the rats of the experimental groups was 237 g, the maximum was 340 g, the average weight of the rats in the groups was 285.2 ± 5.1 g. The minimum body surface area of the animals in the groups was 436.59 cm ² , and the maximum was 555.34 cm ² , the average - 493.94 cm ² . The area of thermal burn caused in the experiment (314 mm ² ) amounted to 0.72% of the minimum surface area of the rat body, 0.56% of the maximum surface area of the rat body, and 0.64% of the average surface area of the rat body.

Meeubner's formula for calculating body surface area by weight (mass) of an animal:

,

,

where W is the mass of the animal in grams, S is the surface of the body in square centimeters [Kochetygov N.I. Burn disease. L. "Medicine", 1973].

Before the experiment, the animals were divided into 6 groups of 5 rats each.

The first group (experimental) - animals that were daily applied to the burned surface with a 5% solution of yarrow essential oil, diluted in sweet almond oil (1:20).

The second group (experimental) - animals that were daily applied to the burned surface with a 5% solution of essential oil of wormwood Pontic, diluted in sweet almond oil (1:20).

The third group (experimental) - animals that were daily applied to the burned surface with a 5% solution of essential oil of chamomile, diluted in sweet almond oil (1:20).

The fourth group (experimental) - animals that were daily applied to the burned surface with a 5% solution of blue tansy essential oil diluted in sweet almond oil (1:20).

The fifth group (control) - animals that did not receive treatment after a burn.

The sixth group (control) - animals that daily applied sweet almond oil to the burned surface.

Every day, starting from 1 day after modeling a superficial thermal burn, the animals of the experimental groups were once applied with a 1-channel microdoser 50 μl Light Micro (Lenpipet) to the burn wound 50 μl of a 5% solution of essential oils using single tips, and With a sterile glass rod, solutions of essential oils were evenly distributed over the surface of the burn wound.

To assess the wound healing and anti-inflammatory effects of essential oils of Pontic wormwood, yarrow, pharmacy chamomile and blue tansy daily, the area of the burn wound was measured before applying the solutions of essential oils. The measurement of the area of the burn wound in this and subsequent experiments was carried out using the “Device for measuring the surface surface defects of the skin” developed by the authors [RF patent 153724. Device for measuring the surface area of skin defects / Pakhomova A.E., Pakhomova Yu.V., Pakhomova E .E. (07/03/2015)] according to the method described in this patent.

The area of burn wounds was measured in all animals on the 1st, 3rd, 7th, 14th and 21st days from the beginning of the experiment (the experiment should be considered the 1st day after modeling the wound).

An estimate of the epithelialization rate was calculated by the L.N. index Popova [Effect of ionized silver preparation on the reparative regeneration of the skin and underlying tissues in modeling thermal and chemical burns in rats // N.S. Ponomar, Yu.S. Maklyakov, D.P. Khloponin, A.O. Revyakin. - Biomedicine. - 2012. - No. 1. - From 143-148.]:

,

,

where ΔS is the desired value, S is the size of the wound area in the previous measurement (mm ² ), S _n is the size of the wound area at the moment (mm ² ), t is the number of days between measurements [Kuzin M.I. Wounds and wound infection. / M.I. Cousin // M .: "Medicine". - 1990. - 258 p.]. The results of measuring the area of burn wounds are presented in table. one.

From table 1 it is seen that the most pronounced wound healing and anti-inflammatory effects have essential oils of yarrow and Pontic wormwood. Wound healing and anti-inflammatory effects of essential oils of chamomile pharmacy and blue tansy are less pronounced. So, the area of burn wounds on the 21st day of the experiment was 24 ± 0.28 mm ² (-92.36%) in the 1st group, and 28 ± 0.16 mm ² (-91.8 in the 2nd group) %), in the 3rd group - 32 ± 0.19 mm ² (-89.81%), in the 4th group - 48 ± 0.11 mm ² (-84.71%)), in the 5th group - 112 ± 0.20 mm ² (-64.33%) and in the 6th group - 148 ± 0.19 mm ² (-52.87%).

The anti-inflammatory effect was evaluated by the results of visual observation of a burn wound and a change in the phases of the course of the wound process [Butko Ya.A., Tkacheva O.V. Burns of the skin: classification and treatment methods // Pharmacist. - 2008. - No. 2. - http://www.provisor.com.ua/archive/2008/N02/butko_og_028.php]. On the 1st day after applying thermal burns in animals of all experimental groups, an intense local inflammatory reaction was observed. Visual inspection of the wound around the heat affected zone revealed the formation of a demarcation line of the damaged epidermis 2 mm in size. Marked hyperemia and swelling of the skin tissue at the border with the wound surface were noted. By the 3rd day of the experiment, animals of all experimental groups had a burn scab on the surface of the burn wound. In animals of the 1st and 2nd groups, fragmentary rejection of the burn scab was observed. In animals of other experimental groups, a close fit of the scab to the wound surface was noted. On the 7th day of the experiment, in animals of the 1st and 2nd groups, during visual observation at the bottom of the burn wound, intensive formation of young connective tissue in the form of granulations was noted. In animals of the 3rd and 4th groups, the formation of granulations at the bottom of the burn wound was also noted. On the 14th day from the time of burn injury in animals of the 1st, 2nd groups, 3rd and 4th groups, wound epithelization was observed, which almost completely ended in animals of the 1st and 2nd groups by the 21st days.

Experimental data showed that essential yarrow and Pontic wormwood have the greatest anti-inflammatory effect.

Microbiological examination

To evaluate the antimicrobial effect of essential oils of Pontic wormwood, common yarrow, pharmacy chamomile and blue tansy, a microbiological study of burn wounds was performed daily before applying the solutions of essential oils. It is known that all burns are either initially infected, or bacterial contamination of the wound occurs in the first hours after a burn injury [Biryukova VV, Aseptic method for treating burns, author. dis., L., 1951]. Conditionally pathogenic microflora is found on the surface of a burn scab as early as 1 day after the injury [Kuzin MI, Sologub V.K. Yudenich V.V. Burn disease, M. Medicine, 1982, 61 S.].

In the control terms of the experiment: the 1st, 3rd, 7th, 14th and 21st days, all animals underwent microbiological examination of burn wounds using contact cups "bactotest" (single-use backplates) (manufactured by Medpoliprom LLC ”, Russia) intended for taking an imprint of microbial bodies from the surface of a wound onto a nutrient medium (agar) and their subsequent investigation by germination under conditions of a thermostat at a temperature of plus 37 ° C. On the day the microbial bodies were imprinted from the surface of the burn wound, the Bactotest cups were poured to the top with Endo medium and stored in a refrigerator at a temperature of + 4-6 ° С. To take an imprint from the "bacterotest" the cover was removed and the surface of the medium was pressed to the surface of the burn wound. Then, the Bactotest cups were closed again and incubated in a TS-1/80 SPU thermostat (Russia) to germinate at + 37 ° С during the day, after which the number of microbial bodies was counted [Terekhova MV, Gurevich K .G., Zaborov V.A., Arzumanyan V.G., Skotnikova Yu.V. Comparative characteristics of the state of skin microbiocenosis in athletes of various sports // Materials of the international scientific-practical conference "East - Russia - West" Modern problems and innovative technologies in the development of physical culture and sports, Irkutsk, 2011. - P. 255-256].

According to microbiological research, in all groups on the 1st day of the experiment there was a continuous growth of opaque microbial colonies of white color. Microbial colonies had a smooth S-shape, even edges, uniform structure, soft consistency, and a convex profile. The diameter of the colony was 1 mm. On the 7th day of the experiment, in the 5th and 6th groups there was a continuous growth of microbial colonies (they cannot be counted). In the 1st and 2nd groups, there was a continuous growth of microbial colonies, forming shapeless clusters in the form of grapes. In the 3rd and 4th groups, moderate growth of microbial colonies was observed (50-55). On the 21st day of the experiment, in the 5th and 6th groups there was a meager growth of microbial colonies (20-25). In the 1st and 2nd groups, a single growth of microbial colonies was noted (up to 7–9). In the 3rd and 4th groups, the cessation of the growth of microbial colonies was observed.

Experimental data showed that blue tansy essential oil has the greatest antimicrobial effect. A good antimicrobial effect has essential oil of chamomile pharmacy.

Creation and evaluation of the effectiveness of the composition of essential oils. Finding the ratio of essential oils in the claimed ointment

Taking into account the results of the evaluation of the wound healing, anti-inflammatory and antimicrobial effects of essential oils of Pontian wormwood, common yarrow, pharmacy chamomile and blue tansy in the treatment of superficial thermal burns of III degree in order to potentiate the effect, work was carried out to select the ratios of essential oils and create their compositions, the following composition :

- composition of essential oils No. 1: essential oil of wormwood Pontic: essential oil of yarrow: essential oil of chamomile pharmacy: essential oil of blue tansy, wt. hours, respectively: 1.0: 1.0: 2.0: 2.0;

- composition of essential oils No. 2: essential oil of wormwood Pontic: essential oil of yarrow: essential oil of chamomile pharmacy: essential oil of blue tansy, wt. hours, respectively: 1.0: 1.0: 1.0: 2.0;

- composition of essential oils No. 3: essential oil of wormwood Pontic: essential oil of yarrow: essential oil of chamomile pharmacy: essential oil of blue tansy, wt. hours, respectively: 1.0: 1.0: 2.0: 1.0.

The composition of essential oils No. 1, No. 2, No. 3 was diluted in sweet almond oil in the ratio, wt. h: composition of essential oils - 1, sweet almond oil - 20.

Experiments series No. 2. Evaluation of the effectiveness of the composition of essential oils

Superficial thermal burns of the IIIa degree to evaluate the therapeutic effectiveness of the composition of essential oils No. 1, No. 2 and No. 3 were modeled as described above. The determination of the area of thermal burns caused in the experiment, relative to the surface area of the body of animals was carried out according to the method described above.

In the experiment, 30 rats were used.

Before the experiment, the animals were divided into 6 groups of 5 rats in groups.

The first group (experimental) - animals, which on the burn surface daily applied a composition of essential oils No. 1.

The second group (experimental) - animals, which daily on the burn surface was applied a composition of essential oils No. 2.

The third group (experimental) - animals, which on the burn surface were daily applied a composition of essential oils No. 3.

The fourth group (control) - animals that were daily applied to the burned surface with sweet almond oil.

The fifth group (control) - animals that did not receive treatment after a burn.

Every day, starting from 1 day after modeling a superficial thermal burn, the animals of the experimental groups were once applied with a 1-channel microdoser 50 μl of Light Micro (Lenpipet) 50 μl of the composition of essential oils were applied to the burn wound using single tips and with a sterile glass rod evenly distributed compositions of essential oils on the surface of the burn wound.

In the control terms of the experiment: 1st, 3rd, 7th, 14th and 21st days, the area of burn wounds was measured in all animals.

The experimental results are presented in table. 2.

Experimental data indicate that the composition of essential oils No. 1 has the greatest wound healing and anti-inflammatory effect. So the area of burn wounds on the 21st day of the experiment was 8 ± 0.33 mm ² (-97.45%) in the 1st group, 16 ± 0.27 mm ² (-94.9% in the 2nd group ), in the 3rd group - 12 ± 0.16 mm ² (-96.18%), in the 4th group - 48 ± 0.11 mm ² (-84.71%), in the 5th group 112 ± 0.20 mm ² (-64.33%).

In the control terms of the experiment: the 1st, 3rd, 7th, 14th and 21st days, all animals underwent microbiological examination of burn wounds using contact cups "bactotest" (single-use backplates) (manufactured by Medpoliprom LLC ”, Russia), intended for imprinting microbial bodies from the surface of a wound.

According to the microbiological study, in all groups on the 1st day of the experiment, abundant growth of white opaque microbial colonies was noted. Microbial colonies had a smooth S-shape, even edges, uniform structure, soft consistency, and a convex profile. The diameter of the colony was 1 mm. On the 7th day of the experiment, in the 4th and 5th groups there was a continuous growth of microbial colonies (they cannot be counted). In the 2nd and 3rd groups, abundant growth of microbial colonies was noted, forming shapeless clusters in the form of grapes. In the 1st group, moderate growth of microbial colonies was observed. On the 21st day of the experiment, moderate growth of microbial colonies was observed in the 4th and 5th groups (52-58). In the 2nd and 3rd groups, a single growth of microbial colonies was noted (up to 5-7). In the 1st group, the cessation of the growth of microbial colonies was observed.

Experimental data indicate that the composition of essential oils No. 1 has the greatest antimicrobial effect: Pontic wormwood essential oil: ordinary yarrow essential oil: pharmacy chamomile essential oil: blue tansy essential oil in a ratio of 1.0: 1.0: 2.0: 2.0, respectively.

Development and manufacture of ointment base

When creating a soft dosage form, it is very important to choose the right ointment base, which would not only contribute to the manifestation of the pharmacological activity of essential oils, but also have a number of positive properties. The ointment base should provide a manifestation of the specific activity of the ointment; provide the necessary mass of ointment and the concentration of medicinal substances; Do not disturb the physiological functions of the skin; do not cause allergic reactions, do not have a toxic, irritating, sensitizing effect on the body; be chemically indifferent and do not interact with medicinal substances, do not change under the influence of environmental factors (light, oxygen, moisture); provide the necessary consistency, optimal rheological properties (easy to apply on the skin or mucous membranes, do not exfoliate, easily squeezed out of the tube); provide optimal pharmacokinetics; it is easy to include medicinal substances and release them upon contact with the skin and mucous membranes; do not undergo microbial contamination; maintain stability; easy to remove from skin, hair, linen; be affordable and economically feasible; have a good presentation [Pharmaceutical technology. The manufacture of drugs: a textbook / A.S. Gavrilov. 2010. - 624 p.: Ill.].

Taking into account the biomedical requirements for ointments for use in the conservative treatment of superficial thermal burns [Yarnykh T.G., Garkavtseva O.A., Chushenko V.N. Development and standardization of ointment "DERMALIK" // Chemical and Pharmaceutical Journal. - Volume 45. - No. 12. - 2011. - S. 41-44] were selected 3 ointment base, the following composition:

Ointment base number 1. Heated to a liquid state and filtered vaseline (84.6 g) is thoroughly mixed with anhydrous lanolin (9.4 g) and heated in a water bath to 50-60 ° C. After dispersing the components to a homogeneous mass, the obtained ointment base is poured in a hot state into the prepared container. The ointment base is a uniform thick viscous mass with a pleasant smell. The melting point of the obtained ointment base is 50 ° C. The obtained ointment base refers to the basis of the diphilic type.

Ointment base number 2. Sunflower oil (87.0 g) is heated to a temperature of 50 ° C and mixed thoroughly with Aerosil (7.0 g) in a water bath at a temperature of 50 ° C. After dispersing the components to a homogeneous mass, the obtained ointment base is poured in a hot state into the prepared container. The ointment base is a uniform thick viscous mass with a pleasant smell. The obtained ointment base refers to the basics of the hydrophobic type.

Ointment base number 3. Distilled water (30.0 g) is heated to a temperature of 50 ° C and mixed thoroughly with T-2 emulsifier (4.0) in a water bath at a temperature of 50 ° C. Distilled water (30.0 g) is heated to a temperature of 50 ° C and mixed thoroughly with Na-CMC (1.0) in a water bath at a temperature of 50 ° C. The resulting mixture is thoroughly mixed with each other in a water bath at a temperature of 50 ° C. Glycerin (2.0) is added to the resulting mixture with thorough mixing. After dispersing the components to a homogeneous mass, the obtained ointment base is poured in a hot state into the prepared container. The ointment base is a homogeneous very thick viscous mass with a pleasant smell. The obtained ointment base refers to the basis of the emulsion type.

The thermostability of ointment bases was evaluated by a change in organoleptic properties and a change in pH. The thermal stability of ointment bases was determined by acting on samples of ointment bases at high and low temperatures. To determine the thermal stability, a sample of ointment bases 3.0 g was placed in a bottle, closed and placed in a thermostat at Т = 60 ° С for 6 hours, a sample of ointment bases 3.0 g was placed in a bottle (the bottle was not closed) and frozen at T = 10 ° C for 6 hours. At elevated temperatures (+ 60 ° C), all ointment bases were stable. At low temperatures (-10 ° C), all ointment bases also showed stability (table. 3). The results were evaluated by changing the organoleptic properties and pH of the ointment base samples.

From table 3 it can be seen that all the ointment bases studied showed thermal stability both at elevated temperature (+ 60 ° C) and at low temperature (-10 ° C).

The pH of ointment base No. 1 is in the range from 8.22 to 8.24 (slightly alkaline medium), which can be considered the optimal value, since the use of an ointment base with an acidic pH value for conservative treatment of burns will enhance the acidosis of damaged tissues, worsening the course of the inflammatory process. The pH of ointment base No. 1 after accelerated aging did not statistically significantly change (Table 4).

Considering the indicators of thermal stability and pH of the ointment base during long-term storage, it was found that the ointment base No. 1 (ratio of components, wt.%: Petrolatum - 84.6%; anhydrous lanolin - 9.4%) is a priority for the manufacture of the developed ointment.

The manufacture of the claimed ointment

The experiments carried out allowed us to establish the optimal content of the components in the inventive ointment, ensuring its maximum clinical effectiveness.

The preparation of ointment base No. 1 was described above. The addition of the composition of essential oils No. 1 to the ointment base No. 1 is carried out after cooling the ointment base to a temperature of 37 ° C.

The ability of the ointment to release essential oils was evaluated by bioavailability test by direct diffusion in vitro agar. The release of essential oils from the ointment base was evaluated by the diameter of the stained area, which after 7 minutes was 7.8 mm, after 1 hour - 8.1 mm, after 2 hours - 8.4 mm, after 3 hours - 8.6 mm, which indicates about the high rate of release of components of essential oils and a high degree of bioavailability of the ointment.

The resulting ointment is heated 10-15 ° C above room temperature and poured into the form of the desired configuration and volume (despenser, jars, tubes, etc.) for subsequent cooling naturally.

The result is a packaged ointment. The ointment is stored in a cool dark place for up to 10 years without loss of initial activity. Ointment at room temperature is a homogeneous composition of a solid yellow consistency. Upon contact with the body or heating above 25 ° C, the ointment softens and becomes plastic. It does not dissolve in water and alcohol. Non toxic Contraindication to use is individual intolerance (allergy).

Experiments series number 3. Comparative evaluation of the effectiveness of the ointment

Preclinical studies have allowed to evaluate the therapeutic efficacy of the claimed ointment compared to traditional ointments in the conservative treatment of thermal burns of the skin of the IIIa degree.

Grade IIIa superficial thermal burns were applied as described above. The determination of the area of thermal burns caused in the experiment, relative to the surface area of the body of animals was carried out according to the method described above.

In the experiment, 100 male Wistar rats were used.

Before the experiment, the animals were divided into 5 groups of 20 rats per group.

The first group (experimental) - animals that daily applied the claimed ointment to the burned surface.

The second group (experimental) - animals that daily applied 10% methylurocyl ointment to the burn surface.

The third group (experimental) - animals that daily applied ointment levomekol on the burn surface.

The fourth group (experimental) - animals that were daily applied to the burned surface with 20% ichthyol ointment.

The fifth group (control) - animals that did not receive treatment after a burn.

, Гладух

Вивчення

активностi мазевих основ на третiй фазi ранового процесу / Фармацевтичний журнал. - Киiв, 2006. - N2. - С. 70-73].The inventive ointment was applied evenly daily to the entire surface of the burn wound by the application method in the form of a layer 2 mm thick using a sterile spatula. Throughout the treatment, the burn wound was conducted in an open way, since the wound dressing seals the wound and creates conditions for the growth of anaerobic infection, and thereby worsens the course of the reparative process [Armless

Gladuh

Vivchennya

activity of ointment bases on the third phase of the wound process / Pharmaceutical journal. - Kyiv, 2006. - N2. - S. 70-73].

In the control terms of the experiment: 1st, 3rd, 7th, 10th, 14th, 21st, 28th and 35th days, the area of burn wounds was measured in all animals.

Experimental data indicate that the claimed ointment has the greatest wound healing, anti-inflammatory and antimicrobial effects.

In the control period of the experiment, animals were taken out of the experiment in accordance with the requirements of the National Standard of the Russian Federation “Principles of Good Laboratory Practice” [National Standard of the Russian Federation GOST 53434-2009 “Principles of Good Laboratory Practice”] by overdose of ether anesthesia. Burn wounds were totally excised along with a section of healthy skin, fixed in a 10% formalin solution, embedded in paraffin, made sections that were stained according to Van Gieson (acid fuchsin and picric acid) and subjected to morphological examination using the microscopic method [O. Volkova, V.V. Fundamentals of histology with a histological technique / O.V. Volkova, Yu.K. Yeletsky. - M .: Medicine, 1971. - 272 p.]. Microscopic examination of the obtained sections was performed on a Mikromed binocular microscope with a magnification of 400 times. Using the eyepiece micrometer, the sizes of the newly formed regenerate structures were determined: the height of the scab, granulation tissue, border zone of the epithelium, leukocyte shaft and the length of the epithelial wedge.

The studies showed that during the 1 day of the experiment in animals of all groups a standardized thermal skin burn of the III degree was formed and an increase in the area of burn wounds was observed in all groups. The skin on the site of burn wounds was dense, motionless, not sensitive to pain stimuli, and a loose, uneven thickness scab was formed (Fig. 1).

The maximum values of the area of burn wounds in animals of all groups reached the 3rd day. At the same time, the minimum size of burn wounds was noted in animals of the 1st group and amounted to 384 ± 0.34 mm ² (+ 22.3%) relative to the area of the initial (standardized) burn of 314 mm ² . The maximum values were noted in the 4th and 5th groups and amounted to 440 ± 0.37 mm ² (+ 40.13%) and 468 ± 0.17 mm ² (+ 49.04%), respectively (Table 5) . On the 3rd day of the experiment in animals of the 1st group (the claimed ointment), the scab became dense and rose above a healthy area of the skin, fragmentary scab rejection began (Fig. 2).

Further, throughout the observation period (up to 21 days), a decrease in the area of burn wounds was noted in all the studied groups. On the 7th day of the experiment in animals of the 1st group, the size of the burn wound was 228 ± 0.34 mm ² (-27.4%), while in animals of the 4th group, the area of the burn wound exceeded the initial values by 17, 2% (368 ± 0.15 mm ² ), which is comparable with the results of animals of the 5th group who did not receive treatment (+ 26.1%) (Table 5). In animals of the 1st group, scab rejection continued on the 7th day of the experiment, areas of dystrophy and necrosis of the epidermis and dermis were noted in the area of thermal burn application, pronounced edema and leukocyte infiltration were observed in the dermis (Fig. 3).

On the 10th day of the experiment, in animals of the 1st group (the claimed ointment), the area of the burn wound decreased by half compared with the initial size of the burn wound (-57.3%), the bottom of the burn wound was cleaned of scab remnants and filled with granulations, signs of transition appeared granulation tissue into the connective tissue, vascular sprouting was noted (Fig. 4). In animals of the 2nd and 3rd groups, the area of burn wounds decreased in comparison with the initial size by 8.3% and 3.4%, respectively, while in animals of the 4th and 5th groups, the area of burn wounds was 10 day of the experiment exceeded the initial indicators by 10.8 and 14.6% (table. 5).

In animals of the 1st group (the claimed ointment) on the 14th day of the experiment, the area of the burn wound decreased by 91.0% and amounted to 28 ± 0.24 mm ² . In animals of other groups, the area of burn wounds decreased in comparison with the initial values by 60.51% (2nd group), by 52.87% (3rd group), by 22.93% (4th group) and 22.29% (5th group) (table. 5). In animals of the 1st group, the newly formed epithelium actively grew on granulation tissue from both sides of the burn wound towards each other, under the epithelium a young connective tissue formed, the main cellular elements of which were fibroblasts (Fig. 5).

By the 21st day of the experiment in animals of the 1st group (the claimed ointment), burn wounds were completely epithelized, the connective tissue had a typical structure characteristic of healthy skin (Fig. 6). Complete epithelization of burn wounds in animals of the 2nd group occurred on the 28th day of the experiment, in animals of the 3rd and 4th groups on the 35th day of the experiment, and in animals of the control group on the 40th day.

From table 5 it is seen that the claimed ointment has the most pronounced wound healing and anti-inflammatory effects. By the 21st day of the experiment, in the animals of the 1st group, the burn wounds were completely epithelized, the connective tissue had a typical structure characteristic of healthy skin.

The inventive ointment is characterized by the following properties:

- It has a pronounced anti-inflammatory, wound healing, antimicrobial effects;

- does not contain chemical or special synthetic additives, the active components of the ointment are made from plant materials,

- does not contain toxic components (see above on rivanol),

- contains inexpensive, widespread ingredients for creating an ointment base, easy to manufacture;

- the ointment reduces the treatment time for superficial thermal burns of the IIIa degree skin to 3 weeks, which is 2 weeks less than in the treatment with 10% methyluracil, 20% ichthyol ointments, Levomikol;

- makes it possible to lead a wound in an open way (without dressing);

- in the treatment of the claimed ointment, no scars formed.

The effectiveness of the claimed ointment is shown both in comparison with the control, and in comparison with other anti-burn ointments.

Claims (2)

Ointment for the treatment of superficial thermal burns, characterized in that it contains essential oils of the aboveground part of the yarrow, Pontic wormwood, pharmacy chamomile and blue tansy collected in the flowering phase, as well as petrolatum and lanolin anhydrous in the following ratio of components, wt.%: essential oil of the aerial part of the yarrow 1,0 essential oil of the aerial part of wormwood Pontic 1,0 essential oil of the aerial parts of chamomile pharmacy 2.0 blue overhead essential oil 2.0 petroleum jelly 84.6 anhydrous lanolin 9,4

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