The use of cellular technologies to improve the appearance of scars

Modern science is characterized by the rapid development of a number of related disciplines, united under the common name of "biotechnology." This section of science, based on recent advances in biology, cytology, molecular genetics, genetic engineering, transplantology, aims to use the enormous potential inherent in plant and animal cells - the basic structural units of all living things. "A living cell is a ready biotechnological reactor in which not only the processes leading to the formation of the final product are realized, but also a number of others that contribute to maintaining the catalytic activity of the system at a high level," John Woodward, 1992. The beginning of the science of the cell was laid in 1665, when the English physicist R. Hooke created the first microscope and in the plug found cells - cellulae ("cells"). In 1829, M. Schleiden and T. Schwann substantiated the "cellular theory", which proved that all living things consist of cells. R.Virkhov in 1858 has proved, that in a basis of all illnesses infringement of the structural organization and a metabolism of cells lays. He became the founder of "cellular pathology." The fundamental contribution to the science of the cell was made in 1907-1911. R. Garrison and AA Maksimov, proving the possibility of culturing cells outside the body. Their work showed that for the cultivation of cells, animal tissues and parts of plants should be mechanically disconnected to small pieces. To isolate cells, the tissues are cut with a sharp knife or microtome in thin sections, approximately 0.5-1.0 mm. Physical division of cells is called immobilization. Isolated cells are obtained by enzymatic dispersion of pieces of plants or tissues. After crushing with sharp scissors, the pieces are treated with trypsin or collagenase to obtain a suspension - suspension of individual cells or their microaggregates in a special medium. Alginate gels (calcium alginate) are widely used to immobilize plant cells. It is proved that the immobilized plant and animal cells retain the ability to biosynthesis. Products of cellular biosynthesis accumulate in cells, their expression occurs either spontaneously, or with the help of special substances that promote increased permeability of cell membranes.

Cultivation of animal cells is a much more complicated process than the cultivation of plant cells, which requires special modern equipment, high technologies, the presence of various media, growth factors designed to preserve the viability of cells and maintain them in a state of high functional activity. It has been found that most cells of hard tissues, such as kidney, liver, and skin tissues are surface-dependent, so in vitro they can only be cultured in the form of thin layers or monolayers directly attached to the substrate surface. The longevity, proliferation and functional stability of cells obtained by enzymatic tissue dispersal depends to a large extent on the substrate on which they are grown. It is known that all cells obtained from vertebrate tissues have a negative surface charge, so that positively charged substrates are suitable for their immobilization. Isolated cells obtained directly from whole tissues can be maintained in the primary culture in the immobilized state while maintaining high specificity and sensitivity for 10-14 days. Immobilized, surface-dependent cells play an important role in biology today, especially their role is important for clinical studies. They are used to study the cycles of cell development, the regulation of their growth and differentiation, functional and morphological differences between normal and tumor cells. Immobilized monolayers of cells are used in biotests, for quantitative determination of biologically active substances, as well as for studying the effects of various drugs and toxins on them. A great interest in the cell, as a medical device, has been shown by doctors of all specialties for several decades. In this direction, cell technologies are rapidly developing.

With the onset of tissue and cell therapy, the name of the famous Russian scientist V.P. Filatova, who in 1913 laid the foundations of the doctrine of tissue therapy, studying the results of corneal transplants of healthy donors to patients with cataracts .. In the process of working with corneal transplantation, he found that the cornea preserved in the cold for 1-3 days at -2 -4 degrees C gets better than fresh. Thus, it has been found that the property of cells secrete in unfavorable conditions some substances that excite life processes in transplanted tissues and regenerate in the tissues of the recipient. The tissue and cells separated from the body are in a state of experience, that is, a slowed down life. In them, blood circulation stops, and consequently nutrition. Tissue breathing is extremely difficult, innervation and trophic are upset. Being in a new qualitative state, adapting to the new conditions of existence of the cell, they produce special substances possessing medicinal properties. These substances of non-protein nature were named by V.P. Filatov as biogenic stimulants. He established, together with V.V. Skorodinskaya, that the material from animals and plants can be freely autoclaved at t 120 degrees C for an hour after aging under unfavorable conditions, while not only did they lose their activity, but rather intensified, which was explained by the yield biological stimulants from canned tissues. In addition, they lost antigenic properties, which significantly reduced the possibility of rejection. The canned sterile material was injected into the body by implantation (replanting) under the skin or in the form of injections of extracts, with adequate results. It has also been found that fetal tissues contain significantly more biologically active substances than adult tissues, and some factors are found only in embryos. Inoculated fetal tissues are not perceived by the recipient organism as foreign due to the absence of proteins in the cytoplasmic membrane structure responsible for specific, tissue and individual specificity (proteins of the main histocompatibility complex). As a result, inoculation of human fetal tissues into the human body does not trigger the mechanisms of immunological defense and the reaction of incompatibility and rejection. VP Filatov in the medical practice widely used the human placenta and skin. Treatment courses consisted of 30-45 injections of tissue extracts and 1-2 implants of autoclaved tissues.

Having begun research with human and animal tissues and cells, he transferred his generalizations to the plant world. Making experiments with living parts of plants (aloe, plantain, agave, beet tops, St. John's Wort, etc.), he created unfavorable conditions for them, placing the cut leaves in a dark place, since for life the plant needs light. From the muddy peat and peat they were also given biogenic stimulators, due to the fact that mud and peat are formed with the participation of microflora and microfauna.

A new stage in the development of tissue therapy was received in the late 1970s, when the knowledge and experience accumulated over decades allowed the use, on a qualitatively new level, of animal and plant tissues and cells to treat a person and prolong his active longevity. So, in some domestic clinics and a number of foreign women in physiological menopause with a climacteric syndrome or against the background of ovariectomy, tissue therapy with fetal tissues of the placenta, hypothalamus, liver, ovaries, thymus and thyroid glands. In one of the most prestigious geronto-cosmetology clinics in Western Europe, for several years already, injections of extracts derived from fetal tissues of the sexual glands of rams have been used for the same purposes.

In our country, biostimulating treatment has also been widely used. Patients with various diseases until recently were actively prescribed injections of placenta extracts, aloe, collant, large (biodoside), FBS, peloidodistillate, peloidin, peat, humisol prepared according to the method of VP Filatov. Currently in pharmacies it is practically impossible to buy these highly effective and inexpensive domestic tissue preparations of animal, vegetable and mineral origin.

At the basis of obtaining various biogenic preparations from tissues and human organs of import production, such as rumalon (from cartilage tissue and bone marrow), actovegin (from calves' blood), solcoseryl (extract of cattle blood), and also domestic preparations - vitreous from the vitreous body of the eye of the cattle), kerakol (from the cornea of cattle), splenin (from the spleen of cattle), epithalamin (from the epithelial-epiphyseal region) also belong to VP Filatov. The common property for all tissue preparations is the overall effect on the whole organism. Thus, the "Tissue Therapy" of Academician VP Filatov formed the basis for most modern developments and directions in surgery, immunology, obstetrics and gynecology, gerontology, combustiology, dermatology and cosmetology associated with the cell and products of its biosynthesis.

The problem of transplantation of tissues has worried humanity since ancient times. So in Ebers' papyrus, dated 8,000 BC, there is already a mention of the use of tissue transplantation in order to compensate for defects in individual parts of the body. In the "Book of Life" of the Indian scientist Sushruta, who lived for 1,000 liters. BC. There is a detailed description of the restoration of the nose from the skin of the cheeks and forehead.

The need for donor skin grew in proportion to the increase in the number of plastic and reconstructive surgeries. In this regard, began to use cadaveric and fetal skin. There was a need to conserve donor resources and find ways to replace human skin with animal tissues, various skin modeling options. And it was in this direction that scientists worked, when in 1941 P.Medovar first showed the principal possibility of keratinocyte growth in vitro. The next important step in the development of cellular technologies was the work of Karasek M. And Charlton M., who in 1971 carried out the first successful transplantation of autologous keratinocytes from the primary culture to rabbit wounds, using collagen gel as a substrate for the cultivation of CC, which allowed to improve cell proliferation in the culture. J.Rheinvvald. H Green. Have developed a technology for serial cultivation of large quantities of human keratinocytes. In 1979, Green and co-authors opened the perspectives of the therapeutic use of the cellular culture of keratinocytes in the restoration of skin with extensive burns, after which this technique, being constantly improved, was used by surgeons of burn centers abroad and in our country.

In the process of studying a living cell, it was revealed that the cells produce not only biogenic stimulators of non-protein origin, but also a number of cytokines, mediators, growth factors, polypeptides that play an important role in regulating the homeostasis of the whole organism. It was found that, in various cells and tissues, peptide bioregulators are contained. Which have a wide range of biological effects and coordinate the development and functioning of multicellular systems. The era of the application of cell culture as a therapeutic agent began. In our country, the implantation of a suspension of fibroblasts and multilayered cellular keratinocyte seams in recent decades has been adopted in combobustology. Such an active interest in the transplantation of skin cells burned due to the need for rapid closure of large burn surfaces and a shortage of donor skin. The ability of a small piece of skin to isolate cells capable of covering the wound surface in 1000 and even 10,000 times larger than the area of donor skin turned out to be very attractive and important for combustiology and burn patients. The percentage of engraftment of keratinocyte seams varies depending on the burn area, age and health condition of the patient from 71.5 to 93.6%. Interest in the transplantation of keratinocytes and fibroblasts is associated not only with the ability to quickly close the defect in the skin, but also because these transplants have a potent biologically active potential for improving the type of tissues obtained as a result of transplants. Vascular neovascularization, hypoxia removal, trophic improvement, acceleration of maturation of immature tissue - this is the morpho-functional basis of these positive changes that occur due to the release of growth factors and cytokines transplanted by cells. Thus, due to the introduction of advanced cellular technologies for the transplantation of multicellular seams of autologous and allogenic keratinocytes and fibroblasts into extensive wound surfaces, the combobustiologists were able not only to reduce the percentage of deaths burned with a high percentage of skin lesions, but also to improve qualitatively the scar tissue that is unavoidable occurs on the site of burns IIb and IIIa and 6 degrees. The experience of combobustologists, obtained in the treatment of wound surfaces in burn patients, suggested the use of the already modified Green method in dermatological surgery with various skin and cosmetic pathologies (trophic ulcers, vitiligo, nevuses, bullous epidermolysis, removal of tattoos, age-related skin changes, as well as to improve the appearance of scars).

The use of allogenic keratinocytes in surgery, combustiology and dermatocosmetology has several advantages over the use of autologous keratinocytes, since the cellular material can be prepared in advance in unlimited quantities, canned and applied if necessary. It is also known that allogeneic KCs have a reduced antigenic activity, since Langerhans cells, carriers of antigens of the HLA complex, are lost in vitro cultivation. In favor of the use of allogeneic KC, it is also said that they are replaced by autologous ones after transplantation, according to the data of different authors in terms of 10 days to 3 months. In connection with this, cell banks have been created in many countries, thanks to which cell transplants can be produced in the right amount and at the right time. Such banks are in Germany, the US, Japan.

Interest in the use of cellular technologies in dermatocosmetology is due to the fact that "cellular compositions" carry a powerful bioenergetic and information potential, thanks to which it is possible to obtain qualitatively new results of treatment. The autokines released by the transplanted cells (growth factors, cytokines, nitric oxide, etc.) primarily act on their own fibroblasts, increasing their synthetic and proliferative activity. This fact is especially attractive for researchers, since fibroblast is the key cell of the dermis, the functional state of which depends on the state of all layers of the skin. It is also known that, after traumatizing the skin with a cauter, laser, needle and other tools, bone marrow, adipose tissue and pericyte capillaries replenish the skin with fresh stem precursors of fibroblasts, which helps to "rejuvenate" the pool of own cells. They actively begin to synthesize collagen, elastin, enzymes, glycosaminoglycans, growth factors and other biologically active molecules, which leads to an increase in hydration and vascularization of the dermis, improve its strength,