ИЗОЛИРОВАННЫЙ ЖИЗНЕСПОСОБНЫЙ СЕГМЕНТ ПЕРЕДНЕЙ БРЮШНОЙ СТЕНКИ СВИНЬИ КАК ЗАМЕНИТЕЛЬ ЦЕЛОГО ЖИВОТНОГО, ОБЕСПЕЧИВАЮЩИЙ ОЦЕНКУ ЭФФЕКТИВНОСТИ ОТБЕЛИВАТЕЛЕЙ КРОВОПОДТЕКОВ

Ураков А.Л.

ORCID: 0000-0002-9829-9463, доктор медицинских наук, профессор, Институт термологии, Ижевская государственная медицинская академия, Ижевск, Россия, Институт механики Уральского отделения РАН

ИЗОЛИРОВАННЫЙ ЖИЗНЕСПОСОБНЫЙ СЕГМЕНТ ПЕРЕДНЕЙ БРЮШНОЙ СТЕНКИ СВИНЬИ КАК ЗАМЕНИТЕЛЬ ЦЕЛОГО ЖИВОТНОГО, ОБЕСПЕЧИВАЮЩИЙ ОЦЕНКУ ЭФФЕКТИВНОСТИ ОТБЕЛИВАТЕЛЕЙ КРОВОПОДТЕКОВ

Аннотация

Результаты исследований показывают, что оценить эффективность средств-отбеливателей кровоподтеков можно в лабораторных условиях без использования целых животных, если использовать для этого изолированные сегменты передней брюшной стенки, полученные сразу после забоя животных на мясо. Показано, что производимые в лабораторных условиях инъекции крови животных в кожу изолированных сегментов обеспечивают создание искусственных синяков, а динамика цвета кожи в области таких синяков позволяет оценивать эффективность отбеливающих средств. В связи с этим, сегмент передней брюшной стенки  свиньи с   искусственным синяком предложен в качестве новой биологической модели, которая позволяет в лабораторных условиях проводить скрининг отбеливателей кровоподтеков и оценивать локальную отбеливающую эффективность средств. Показано, что использование такой биологической модели позволяет проводить качественные фармакологические исследования при отсутствии живых животных и вивария,  поэтому существенно упрощает и удешевляет процесс изыскания и изучение новых лекарств.

Ключевые слова: свиньи, свиная кожа, модель болезни, лекарства.

Urakov A.L.

ORCID: 0000-0002-9829-9463, MD, Professor, Institute of Thermology, Izhevsk State Medical Academy, Institute of Mechanics of the Ural Branch of the Russian Academy of Sciences

ISOLATED VIABLE SEGMENT OF ANTERIOR ABDOMINAL WALL OF PIG AS A SUBSTITUTE OF THE WHOLE ANIMAL PROVIDING THE EVALUATION OF THE EFFECTIVENESS OF BLEACHER OF BRUISES

Abstract

The results of the studies show that it is possible to evaluate the effectiveness of bleacher of bruises in the laboratory without the use of whole animals if isolated segments of the anterior abdominal wall obtained immediately after slaughtering the animals for meat are used for this purpose. It is shown that the injections of animal blood into the skin of isolated segments produced in the laboratory ensure the creation of artificial bruises, and the color dynamics of the skin in the area of such bruises makes it possible to evaluate the effectiveness of bleaching agents. In this regard, the segment of the anterior abdominal wall of a pig with an artificial bruise is proposed as a new biological model that allows screening bleacher of bruises in the laboratory and assessing the local bleaching efficacy of the agents. It is shown that the use of such biological model makes it possible to carry out qualitative pharmacological studies in the absence of live animals and vivarium, so it greatly simplifies and cheapens the process of researching and studying of new drugs.

Keywords: pigs, pig skin, model of disease, medicines.

Introduction

In pharmacology in the search for new pharmacological tools in the study of their mechanisms of action, scientists begin their research with experiments on animals. However, as a rule, the experiments conducted on mice. And the first series of experiments has traditionally focused on the study of acute toxicity and dependence of effect on dose. The fact that the main part of researchers conducting studies for detection of drugs are suitable for shared action on the entire body. However it doesn't account for local operation of medicines, because it most depends of physical-chemical factors of local interaction with a specific tissue, e.g., with the venous blood, with a bruise, with a bloody crust, pus, sputum, skin, brain etc. [1, P. 106], [2, P. 466], [3, P. 1], [4, P. 6], [5, P. 2].

At the same time, any drug can have not only general but also local action. It is shown that the medications can have some local effect only when used in certain dosage forms and with the direct introduction of specific organs and tissues in which there is a well-defined pathology [6, P. 95], [7, P. 216]. So, ulcerogenic action of drugs on the wall of the stomach is manifested when they are introduced in the form of compressed tablets in an empty stomach [8, P. 51].

In this regard, the search for new drugs can begin with the study of not only general, but their local action on the selected part of the body, the organs and tissues of animals. In particular, for screening of bleach bruises and evaluation of their effectiveness, it was proposed to use clothing and gauze pads soaked with the blood of donors or domestic animals, and viable isolated body parts of pigs [2, P. 466], [9, P. 1], [10, P. 1], [11 P. 127].  However, this proposal was not noticed.

The aim of this study was to investigate the features using the cut-off heads and segments of the anterior abdominal wall of the pig to simulate bruising and study of bleaching effectiveness of the means in experiments.

Material and methods

In experimental conditions the experiments were conducted on 5 waking piglets of Landrace. The piglets were aged 2 months and weighing 7 – 8 kg. The dynamics of the local temperature in the area of intradermal blood were examined with a thermal imager Thermo Tracer TH9100XX (NEC, USA).

In the laboratory conditions were investigated the dynamics of the viable color of the skin in isolated segments of the anterior abdominal wall 10 adult pigs and in one cut off the head of adult pigs. Severed pig's head had a mass of 9.5 kg. Experiments were conducted in 2 – 3 hours after slaughter. The slaughter of animals was carried out in a special shop in the factory in the village Vavozh of the Udmurt Republic. Before the experiments, the bristles on the skin of animals are not burned with an open flame. Sometimes the bristles was shaved off with a razor, applying cosmetics shaving foam. We used pigs with white skin and colorless bristles.

Artificial bruises were created using intradermal injections of animal blood. Blood was injected into the skin in amounts of 0.5 – 1 ml.  The blood, accidentally gets on your skin, not washed off. Waiting for its drying and simultaneously the stabilization of the sizes of bruises. Then we began to explore the effectiveness of leaching bruising. Monitoring of the dynamics of skin color were performed by eye and by photography in the range of visible spectrum radiation.

To assess the effectiveness of skin discoloration to the area of bruising medicines used in the form of solutions, which were introduced in the cutaneous form of application for up to 10 minutes and/or were driven into the skin by injection. Injection was made by means of ordinary syringes and injection needles, and an application was made using swabs. In the interaction region supported a local temperature between 42 and 45°C with the help of a medical hot-water bottles.

Statistical processing of the results was performed using the program BIOSTAT according to standard methods.

Results

The obtained results confirm the assumption that the skin in isolated and viable body parts of pigs can be used to simulate bruises with a view to using it to search for new bleaching agents and/or evaluation of the effectiveness of bruises bleach. It is established that in the normal skin color in the segments of the abdomen and in isolated the head in animals is a physiological color slightly and remains unchanged at least during the first day after slaughter. It is shown that intradermal injection of venous blood produce intradermal bruises, over which the skin takes on a blue color. Artificially created on the skin blue spot (i.e. artificial bruises) applied to the skin of those of biological objects more than a day without significant changes of skin color. It is established that local application of remedies, bleaching the skin, may reduce the intensity of the color of the skin.

Initially experiments were conducted on isolated segments of the anterior abdominal wall pigs. Artificial bruises were created by introducing into the interior of the skin of 0.5 ml venous blood of animals. Infusion of blood immediately caused the formation of blue spots in the skin. The spots were round in shape with average diameter of 7.5 ± 0.7 mm (n = 20, P ≤ 0.05). Then made the injection of 1 ml of the investigated solution into the skin in the Central part of the spots on the bruise and began the registration of dynamics of skin color. As an example, present the results of one series of experiments (Table 1).

 

Table 1 – The efficiency and speed of skin discoloration in artificial bruises caused by intradermal injections of animal blood 0.5 ml, with subsequent single intradermal injection in the place of the solution of certain drugs in a volume of 1 ml at a temperature of +25°C

The investigated tools	The effectiveness of the bleaching	Speed of onset of skin whitening (sес.)
A solution of 3% hydrogen peroxide	Significant discoloration	65 ± 8 (n = 5, P ≤ 0,05)
A solution of 3% hydrogen peroxide and 10% of sodium sulfacyl	Significant discoloration	55 ± 5 (n = 5, P ≤ 0,05)
A solution of 3% hydrogen peroxide and 10% aminophylline	Significant discoloration	50 ± 4 (n = 5, P ≤ 0,05)
A solution of 3% hydrogen peroxide and 10% sodium bicarbonate	Complete discoloration	43 ± 4 (n = 5, P ≤ 0,05)

 

As follows from the presented results, a complete discoloration of the skin bruise is only the injection of a solution of 3% hydrogen peroxide and 10% sodium bicarbonate. The skin over the bruise becomes colorless completely in 43 ± 4 (n = 5, P ≥ 0,05) seconds after injection of a solution of hydrogen peroxide and sodium bicarbonate.

The results of studying the dynamics of the local temperature in awake piglets showed that the injection of blood does not cause local hyperthermia. Thus, the blood has a local irritant when injected.

Then we investigate the features of an intradermal injection of 1 ml of blood and solutions. Found that intradermal injection of 1 ml of blood or solution of the drug on the face of the pig requires more than 60 seconds, and on the front wall of the abdomen - less than 3 seconds. This forms an artificial bruise (Figure 1).

Fig. 1 – The creation of artificial bruise in the pig's severed head  (a). The color of the skin in the area of the bruise within 5 minutes after intradermal injection of 1 ml of bleach bruise (b)

 

It is shown that skin and subcutaneous fat on the face in piglets and adult pigs have a very durable properties. It turned out that when trying to pierce the skin injection needles, they are always bent, often broke and always demanded to attach to the injection needle significant physical effort. Moreover, blunted injection needle designed for repeated intramuscular injections, virtually were inserted into the skin and very much bent. In the attempt of piercing the skin of the cheeks from waking piglets one needle of 4 used broke. Even more difficult to stick the needle into the skin under the eyes in adult pigs.

The results of the experiments showed that the skin on the face in piglets and in pigs is a very solid. Therefore, the pig's face is very difficult to puncture the skin of the injection needles, is very difficult to enter into the skin the blood and solutions of medications. In this regard, it is difficult to simulate shadows under the eyes in these animals by intradermal injection of blood. However, the severed head can be used to search for new bleaching agents and evaluation of efficiency of the known brighteners of bruising.

Then, experiments were conducted on the isolated skin segments of the abdominal wall of animals. For the artificial formation of experimental bruising in the skin, the isolated segment of the abdominal wall of pigs were injected with 0.5 ml cadaveric pig's blood. Punctures on the skin made by the injection needle at the distance of 8 – 10 cm from each other. Thus formed colored spots that had the shape of a circle with diameter 8 – 11 mm each. The skin over these spots first had blue and cherry colour, and 10 minutes later skin color became blue and the blue color remained unchanged.

It was conducted a series of experiments in which was studied the dynamics of color of skin over bruises after the application for 10 minutes, warm wash with a solution of bleach bruises. It is shown that the application of bleach solution may completely bleached blood stains on the skin and significantly discolored the skin in the field of artificial bruises (Figure 2).

Fig. 2 – An isolated segment of the anterior abdominal wall of the pig before (a) and 10 minutes after application of the bleach solution bruise, consisting of 1% hydrogen peroxide, 2% lidocaine hydrochloride solution and 10% sodium bicarbonate at +45°C (b)

 

The results showed that the skin in the abdomen in young pigs and in adult pigs is a very delicate and subtle. The skin of this part of the body of pigs on their properties significantly different from the skin on the face and is very reminiscent of animal skin from people. Therefore, the abdominal wall of the pig is the most attractive biological target for simulation of bruises and assessment of whitening agents.

Conclusion                                                                                                                        

The results of our experiments indicate that isolated segments of the anterior abdominal wall of pigs can be used to simulate bruising and to develop new methods and tools for whitening of the skin over the bruises. The fact that the injection of blood into the skin of the isolated segment allows you to easily and quickly model a bruise, and the dynamics of color of the skin over the bruise used to evaluate the effectiveness of bleach bruises and methods of their use.

A new biological model allows for pharmacological studies in the laboratory without the use of live animals and without vivarium. In connection with this  new biological model greatly simplifies and cheapens the process of finding and studying new drugs.

I express my gratitude to the students of A.O.Bannikov for assistance in conducting the experiments.

The study was conducted with the support of a grant from the Russian Fund to Support  Innovation:  project N 24398, application C1-19369 "Development tools for bleaching facial skin with bruises under their eyes".

Список литературы / References

1. Габдрафиков Р.Р. Новое косметическое средство и новый способ экстренного обесцвечивания подногтевой гематомы. /Р.Р.Габдрафиков, Д.Р.Габдрафиков, А.А.Гадельшина// Журнал научных статей Здоровье и образование в ХХI веке. - 2017. - Т. 19. - № 4. - С. 104-108.
2. Уракова Н.А. Изобретено новое лекарство – разрыхлитель высохшей крови, который обеспечивает размягчение бинтов, прилипших к ране. /Н.А.Уракова, А.А.Гадельшина// Международный журнал прикладных и фундаментальных исследований. – 2016. – № 11-3. – С. 464-467.
3. Уракова, Н.А. Способ экстренного отбеливания и удаления кровавой корочки с кожи на месте выдавленного угря./ Н.А.Уракова, А.Л.Ураков, Т.В.Уракова, Р.Р.Габдрафиков, Д.Р.Габдрафиков, А.А.Гадельшина.// RUS Патент № 2631593. - 2017. - Бюл. № 27.
4. Urakov A.L. The change of physical-chemical factors of the local interaction with the human body as the basis for the creation of materials with new properties./A.L.Urakov// Epitőanyag – Journal of Silicate Based and Composite Materials. - 2015. - V. 67. - N. 1.- P. 2–6.
5. Urakov A.L. Development of new materials and structures based on managed physical-chemical factors of local interaction./A.L.Urakov// IOP Conf. Ser.: Mater. Sci. Eng. - 2016. - V. – 012008.
6. Уракова Н.А. Синяки под глазами. Способы и средства их выведения. / Н.А.Уракова// Международный журнал прикладных и фундаментальных исследований. – 2017. – № 7-1. – С. 94-96.
7. Уракова Н.А. Создание нового косметического средства для отбеливания кожи под глазами при наличии синяков. /Н.А.Уракова// Международный журнал прикладных и фундаментальных исследований. – 2017. - № 3-2. – С. 213 – 218.
8. Ураков А.Л. Неспецифические свойства таблеток, влияющие на перемещение и действие лекарств в ротовой полости, желудке и кишечнике./ А.Л.Ураков, Н.А.Уракова, Н.А.Михайлова, А.П.Решетников.// Медицинская помощь. - 2007. - № 5. - С. 49-52.
9. Решетников А.П. Способ экспресс удаления пятен крови с одежды./А.П.Решетников, А.Л.Ураков, Н.А.Уракова, Н.А.Михайлова, М.В.Серова, И.В.Елхов, В.Б.Дементьев, Н.А.Забокрицкий, Ю.С.Сюткина.// RUS Патент №  - 2008. - Бюл. № 30.
10. Ураков А.Л. Отбеливатель кровоподтеков./А.Л.Ураков, Н.А.Уракова, Л.В.Чернова, Е.Л.Фишер.// RUS Патент № 2539380. - 2015. - Бюл. № 2.
11. Уракова Н.А. Лекарства для отбеливания кожи под глазами при наличии синяков / Н.А.Уракова, А.Л.Ураков, А.О.Банников // Журнал научных статей Здоровье и образование в ХХI веке. - - Т. 19. - N 5. -  С. 125 – 128.

Список литературы на английском языке / References in English

1. Gabdrafikov R.R. Novoe kosmeticheskoe sredstvoi novyj sposob jekstrennogo obescvechivanija podnogtevoj gematomy. [New cosmetic product and a new way of emergency discoloration subungual hematoma] /R.R.Gabdrafikov, D.R.Gabdrafikov, A.A.Gadel'shina// Zhurnal nauchnyh statej Zdorov'e i obrazovanie v XXI veke. [Journal of scientific articles health and education in the twenty-first century].- 2017. - V. 19. №4. P. 104-108. [in Russian]
2. Urakova N.A. Izobreteno novoe lekarstvo – razrihlitel’ visohshej krovi, kotorij obespechivaet razmiagchenie bintov, prilipshih k rane. [Invented a new drug - a tool that dissolves the dried blood, a tool that dissolves the dried blood, soften the bandages sticking to the wound] /N.A.Urakova, A.A.Gadelshina// Mezhdunarodnij zhurnal prikladnyh i fundamental’nyh issledovanij [International journal of applied and fundamental research].– 2016. – № 11-3. – P. 464-467. [in Russian]
3. Urakova, N.A. Sposob jekstrennogo otbelivanija i udalenija krovavoj korochki s kozhi na meste vydavlennogo ugrja. [Method for emergency bleaching and blood crust removal from skin in place of squeezed out acne] / N.A.Urakova, A.L.Urakov, T.V.Urakova, R.R.Gabdrafikov, D.R.Gabdrafikov, A.A.Gadel'shina.// RUS Patent № 2631593. - 2017. - Bul. № 27. [in Russian]
4. Urakov A.L. The change of physical-chemical factors of the local interaction with the human body as the basis for the creation of materials with new properties./A.L.Urakov// Epitőanyag – Journal of Silicate Based and Composite Materials. - 2015. - V. 67, - N. 1.- P. 2–6.
5. Urakov A.L. Development of new materials and structures based on managed physical-chemical factors of local interaction./A.L.Urakov// IOP Conf. Ser.: Mater. Sci. Eng. - 2016. - 123. - 012008
6. Urakova N.A. Sinjaki pod glazami. Sposobi i sredstva ih vivedenia. [Bruising under the eyes. The ways and means of their elimination] / N.A.Urakova// Mezhdunarodnij zhurnal prikladnyh i fundamental’nyh issledovanij [International journal of applied and fundamental research].– 2017. – № 7-1. – P. 94-96. [in Russian]
7. Urakova N.A. Sozdanie novogo kosmeticheskogo sredstva dlja otbelivania kozhi pod glazami pri nalichii sinjakov. [The creation of a new cosmetic product for whitening of the skin under the eyes while bruises] /N.A.Urakova// Mezhdunarodnij zhurnal prikladnyh i fundamental’nyh issledovanij [International journal of applied and fundamental research].– 2017. - № 3-2. – 213 – 218. [in Russian]
8. Urakov, A.L. Nespecificheskie svojstva tabletok, vlijajushhie na peremeshhenie i dejstvie lekarstv v rotovoj polosti, zheludke i kishechnike. [Nonspecific properties of tablets, affecting the transport and action of drugs in the oral cavity, stomach, and intestine]/ A.L.Urakov, N.A.Urakova, N.A.Mihajlova, A.P.Reshetnikov.// Medicinskaja pomoshh'. [Medical care] - 2007. - № 5. - P. 49-52. [in Russian]
9. Reshetnikov, A.P. Sposob express-udalenia pjaten krovi s odezhdi. [Method the Express remove blood stains from clothes] /A.P.Reshetnikov, A.L.Urakov, N.A.Urakova, N.A.Mikhailova, M.V.Serova, I.V.Elhov, V.B.Dement'ev, N.A.Zabokritski, Ju.S.Syutkina.// RUS Patent № 2371532. 2008. Bul. № 30. [in Russian]
10. Urakov, A.L. Otbelivatel' krovopodtekov. [Bruise bleacher] /A.L.Urakov, N.A.Urakova, V.Chernova, E.L.Fisher.// RUS Patent № 2539380. - 2015. - Bul. № 2. [in Russian]
11. Urakova N.A. Lekarstva dlja otbelivanija kozhi pod glazami pri nalichii sinjakov [Medications for whitening skin under the eyes in the presence of bruises] / N.A.Urakova, A.L.Urakov, A.O.Bannikov // Zhurnal nauchnyh statej Zdorov'e i obrazovanie v XXI [Journal of scientific articles Health and education in the twenty-first century] - 2017. - V. 19. - N 5. -  P. 125 – 128. [in Russian]