Polyploidy. Types of fibroblasts

fibroblasts(fibroblastocytes) (from lat. fibra - fiber, Greek blastos - sprout, germ) - cells that synthesize components of the intercellular substance: proteins (for example, collagen, elastin), proteoglycans, glycoproteins.

In the embryonic period, a number of mesenchymal cells of the embryo give rise differon fibroblasts, which includes:

· stem cells,

semi-stem progenitor cells

unspecialized fibroblasts,

differentiated fibroblasts (mature, actively functioning),

fibrocytes (definitive forms of cells),

myofibroblasts and fibroclasts.

The formation of the ground substance and fibers is associated with the main function of fibroblasts (which is clearly manifested, for example, in the healing of wounds, the development of scar tissue, the formation of a connective tissue capsule around a foreign body).

Semi-specialized fibroblasts are low-growth cells with a round or oval nucleus and a small nucleolus, RNA-rich basophilic cytoplasm. The cell size does not exceed 20-25 microns. In the cytoplasm of these cells, a large number of free ribosomes are found. The endoplasmic reticulum and mitochondria are poorly developed. The Golgi apparatus is represented by clusters of short tubules and vesicles.
At this stage of cytogenesis, fibroblasts have a very low level protein synthesis and secretion. These fibroblasts are capable of mitotic reproduction.

Differentiated mature fibroblasts are larger in size. These are active cells.

In mature fibroblasts, intensive biosynthesis of collagen, elastin proteins, proteoglycans, which are necessary for the formation of the ground substance and fibers, is carried out. These processes are enhanced under conditions of reduced oxygen concentration. Stimulating factors of collagen biosynthesis are also iron, copper, chromium ions, ascorbic acid. One of the hydrolytic enzymes collagenase- splits immature collagen inside the cells, which regulates the intensity of collagen secretion at the cellular level.

Fibroblasts are mobile cells. In their cytoplasm, especially in the peripheral layer, there are microfilaments containing proteins such as actin and myosin. The movement of fibroblasts becomes possible only after their binding to the supporting fibrillar structures with the help of fibronectin- glycoprotein synthesized by fibroblasts and other cells, providing adhesion of cells and non-cellular structures. During movement, the fibroblast flattens, and its surface can increase 10 times.

The fibroblast plasma membrane is an important receptor zone, which mediates the impact of various regulatory factors. The activation of fibroblasts is usually accompanied by the accumulation of glycogen and increased activity of hydrolytic enzymes. The energy generated during glycogen metabolism is used to synthesize polypeptides and other components secreted by the cell.

According to the ability to synthesize fibrillar proteins, reticular cells of the reticular connective tissue of hematopoietic organs, as well as chondroblasts and osteoblasts of the skeletal variety of connective tissue, can be attributed to the fibroblast family.

Fibrocytes- definitive (final) forms of fibroblast development. These cells are spindle-shaped with pterygoid processes. [They contain a small number of organelles, vacuoles, lipids and glycogen.] The synthesis of collagen and other substances in fibrocytes is sharply reduced.

Myofibroblasts- cells similar to fibroblasts, combining the ability to synthesize not only collagen, but also contractile proteins in a significant amount. Fibroblasts can transform into myofibroblasts, functionally similar to smooth muscle cells, but unlike the latter, they have a well-developed endoplasmic reticulum. Such cells are seen in granulation tissue healing wounds and in the uterus during the development of pregnancy.

fibroclasts- cells with high phagocytic and hydrolytic activity, take part in the "resorption" of the intercellular substance during the period of organ involution (for example, in the uterus after the end of pregnancy). They combine the structural features of fibril-forming cells (developed granular endoplasmic reticulum, Golgi apparatus, relatively large but few mitochondria), as well as lysosomes with their characteristic hydrolytic enzymes. The complex of enzymes secreted by them outside the cell breaks down the cementing substance of collagen fibers, after which phagocytosis and intracellular digestion of collagen occur.

The following cells of fibrous connective tissue no longer belong to the differon of fibroblasts.

A polyploid is an organism derived from one or two parental forms by doubling the number of chromosomes. The phenomenon of increasing the number of chromosomes called. polyploidy. This doubling may be spontaneous or artificially induced. For the first time, the phenomenon of polyploidy was discovered by I.I. Gerasimov in 1890.

POLYPLOIDY is an increase in the number of sets of chromosomes in the cells of the body, a multiple of the haploid (single) number of chromosomes; type of genomic mutations. Sex cells of most organisms are haploid (contain one set of chromosomes - n), somatic - diploid (2n).

Organisms whose cells contain more than two sets of chromosomes are called polyploids: three sets are triploid (3n), four are tetraploid (4n), etc. The most common organisms with a multiple of two chromosome sets are tetraploids, hexaploids (6 n), etc. Polyploids with an odd number of sets of chromosomes (triploids, pentaploids, etc.) usually do not produce offspring (sterile), because the sex cells they form contain an incomplete set of chromosomes - not a multiple of the haploid one.

Polyploidy occurs when chromosomes do not separate meiosis. In this case, the germ cell receives a complete (non-reduced) set of somatic cell chromosomes (2n). When such a gamete fuses with a normal one (n), a triploid zygote (3n) is formed, from which a triploid develops. If both gametes carry a diploid set, a tetraploid is produced.

Polyploid cells can arise in the body with incomplete mitosis: after chromosome doubling, cell division may not occur, and two sets of chromosomes appear in it. In plants, tetraploid cells can give rise to tetraploid shoots whose flowers produce diploid gametes instead of haploid ones. Self-pollination may result in a tetraploid, while pollination with a normal gamete may result in a triploid. During vegetative propagation of plants, the ploidy of the original organ or tissue is preserved.

Polyploidy is widespread in nature, but among different groups of organisms it is represented unevenly. Great importance this type of mutations had in the evolution of wild and cultivated flowering plants, among which approx. 47% of the species are polyploids. A high degree of ploidy is inherent the simplest- the number of sets of chromosomes in them can increase hundreds of times. Among multicellular animals, polyploidy is rare and is more characteristic of species that have lost the normal sexual process - hermaphrodites (see. Hermaphroditism), e.g. earthworms, and species in which eggs develop without fertilization (see. Parthenogenesis), e.g. some insects, fish, salamanders. One of the reasons why polyploidy in animals is much less common than in plants is that plants can self-pollinate, and most animals reproduce by cross-fertilization, and, therefore, the resulting polyploid mutant needs a pair - the same mutant - polyploid of the opposite sex. The likelihood of such a meeting is extremely low. Quite often, animals have polyploid cells of individual tissues (for example, in mammals - liver cells).

Polyploid plants are often more viable and prolific than normal diploids. Their greater resistance to cold is evidenced by the increase in the number of polyploid species in high latitudes and high mountains.

Since polyploid forms often have valuable economic traits, artificial polyploidization is used in crop production to obtain initial breeding material. For this purpose, special mutagens(e.g., alkaloid colchicine), which violate the divergence of chromosomes in mitosis and meiosis. Productive polyploids of rye, buckwheat, sugar beet, and other cultivated plants have been obtained; sterile triploids of watermelon, grapes, banana are popular due to seedless fruits.

Application of remote hybridization in combination with artificial polyploidization allowed domestic scientists in the 1st half. 20th century for the first time to obtain fertile polyploid hybrids of plants (G.D. Karpechenko, hybrid tetraploid of radish and cabbage) and animals (B.L. Astaurov, hybrid tetraploid of silkworm).

(Polyploid series)

Distinguish:

-autopolyploidy(a multiple increase in the number of sets of chromosomes of one species), characteristic, as a rule, for species with a vegetative method of reproduction (autopolyploids are sterile due to a violation of the conjugation of homologous chromosomes during meiosis),

-allopolyploidy summation in the body of the number of chromosomes from different types), when cutting, a doubling of the number of chromosomes usually occurs in an infertile diploid hybrid, and it becomes fertile as a result.

- endopolyploedy - a simple increase in the number of chromosomes in one cell or in the cells of an entire tissue (tapetum).

As can be seen from the diagram, mitotic polyploidization occurs as a result of doubling the number of chromosomes in a somatic cell without the subsequent formation of a cell septa. With zygotic polyploidization, the formation of zygotes proceeds normally, but the first division according to the type of mitosis is not accompanied by its division into two cells. As a result, the cells of the resulting embryo will have a double set of chromosomes (4x). And finally, meiotic polyploidization takes place in the absence of a reduction in the number of chromosomes in generative cells (egg, sperm).

Spontaneous polyploidization- a very rare occurrence. In studies, heat shock and nitrous oxide were most often used to obtain polyploids. However, real progress in the study of polyploidy was made after the discovery by Blaxley et al. in 1937. colchocin alkaloid(C 22 H 26 O 6), obtained from colchicum. Since then, it has been successfully used to produce polyploids in hundreds of plant species. Colchicine acts on the spindle of division in the cell, preventing the divergence of chromosomes to the poles at the anaphase stage, thus contributing to the doubling of their number in the nucleus: see fig.

The apical meristems are exposed to colchicine, which makes it possible to obtain quite fertile forms of plants with a doubled number of chromosomes.

Polyploidy has importance in the evolution of cultural and wild plants(it is believed that about a third of all plant species arose due to P.), as well as certain groups of animals (predominantly parthenogenetic). Polyploids are often characterized by large size, high content of a number of substances, resistance to adverse external factors. environment, and other economically useful features. They represent an important source of variability and might. used as starting material for breeding (on the basis of P., high-yielding varieties of agricultural plants that are resistant to diseases have been created). In a broad sense, under the term "P." understand both multiple (euploidy) and non-multiple (aneuploidy) changes in the number of chromosomes in the cells of the body.

· Autopolyploidy- a hereditary change, a multiple increase in the number of sets of chromosomes in the cells of an organism of the same biological species. On the basis of artificial autopolyploidy, new forms and varieties of rye, buckwheat, sugar beet and other plants have been synthesized.

Autopolyploid An organism that has arisen by spontaneous or induced direct increase in the number of chromosomes by a factor of two. An increase in the number of chromium in the class of autopolyploids leads to an increase in the size of the nucleus and cells. generally. This entails an increase in the size of stomata, hairs, vessels, flowers, leaves, pollen grains, etc. The increase in the number of chromium is associated with the enlargement of the whole plant as a whole and its individual organs.

to the physiological characteristics autopolyploids include:

Deceleration of cell division

Growing period extension

Low osmotic pressure

Decreased resistance to abiotic environmental factors, etc.

As a rule, autopolyploids are characterized by reduced fecundity (this is due to the peculiarities of meiosis).

The inheritance of traits in autopolyploids and diploids is also different, since in the genome of the former, each gene is present in four doses. Therefore, for example, a heterozygous tetraploid AAaa with complete dominance forms the following gametes: 1AA + 4Aa + 1aa. The ratio (number) of gametes of a certain type depends on the probability of conjugation of chromium-m carrying genes A and a:

These five genotypes are named:

- quadriplex (AAAA)

- triplex (АААа)

- duplex (AAaa)

- simplex (ahhh)

- nullplex (aaaa)

According to the dose of dominant alleles. In general, the ratio will be 35:1, in contrast to the Mendelian splitting in monohybrid crosses in diploids, equal to 3:1.

In the wild, as well as in culture, autopolyploids are isolated from diploids by a barrier of inbreeding, usually determined by the absence of normal germination of pollen tubes on the stigma of pistils, and impaired development of the embryo and endosperm.

Increasing the size of plants, the size of flowers, seeds, etc. led to the use of autopolyploids in ornamental floriculture (varieties of chrysanthemums, asters, etc.) and the selection of field grain and fodder crops.

· Allopolyploidy- a multiple increase in the number of chromosomes in hybrid organisms. Occurs during interspecific and intergeneric hybridization.

Alloploid is an organism resulting from the combination of chromosome sets of different species.

One of the first such hybrids was obtained by G.D. Karpechenko when crossing radish with cabbage. Both species have a diploid number of chromium = 18 and belong to different genera. Usually the resulting plants are sterile, but in this case the gametes with unreduced chromium number spontaneously combined, resulting in a fertile plant with 2n=36 (18+18). It was called the rare-cabbage hybrid. With the discovery of colchicine, obtaining such hybrids does not present a problem.

ANEUPLOIDY.

Aneuploid is an organism with an increase or decrease, not a multiple of the haploid number of chromium. The most common types of aneuploids are:

Nullisomics 2n-2

Monosomy 2n-1

Trisomics 2n+1

Tetrasomics 2n+2

Monosomy, cat. One chromium is missing (2n-1), and nullisomics (2n-2) do not survive in most plants.

Nullisomics are obtained by self-pollination of monosomic. These plants lack both homologues of a particular chromosome.

Monosomics have reduced fertility. This is explained by the fact that male gametes (n-1) practically do not survive, and less than half of the eggs survive.

Trisomics (2n+1) are obtained by crossing triploids with diploids. At the same time, trisomics also survive in plants with a small amount of chromium, while monosomics in these plants are not completely viable.

Haploidy.

Haploid - an organism containing in somatic cells a complete set of non-homologous chromium-m (n) for a given species. In appearance, haploids correspond to diploid plants, but are much smaller, because. have small cells with small nuclei.

№ 52 REMOTE HYBRIDIZATION.

BUTthe 11th-century slave poet Al-Maarri once exclaimed bitterly: “It seems to us that youth does not wear out, but years roll like stones from a slope.” Centuries have passed since then. Scientists and doctors did not waste time: they worked to give humanity techniques that could slow down the aging process. One of the most advanced anti-aging technologies is fibroblast therapy - a reliable and safe procedure that provides amazing results. It allows you to return the spring of life - the time when we look great even after a sleepless night. If your skin requires true rejuvenation, and you want to look younger every day, modern cellular technologies will help you achieve the desired result.

P The leading clinics in Europe and the USA have long adopted a progressive method of rejuvenation with fibroblasts. Over the past 7 years, several thousand Americans have participated in clinical trials of this technology, which have shown striking effect rejuvenation, which occurs in some patients after a few weeks, in others - only after a few months. After the introduction of fibroblasts, patients notice a long-term improvement in skin quality, the positive effects of which accumulate up to 18-24 months and remain stable for 7 years or more. The results of the studies were so convincing that the procedure was approved by many reputable medical institutes(e.g. ISA (Medicine Control and Administration Agency)).

Euntil recently, our compatriots had to go to England, Switzerland or the United States and pay huge sums of money there in order to undergo a course of cell therapy. Today, procedures using autologous fibroblasts are also available in Russia.

AndMore than one doctoral dissertation is devoted to the study of fibroblasts in our country; many serious medical institutions are engaged in their study (for example, the A. V. Vishnevsky Institute of Surgery of the Russian Academy of Medical Sciences). Why are these cells human body cause such a strong interest of scientists? It's all about their unprecedented rejuvenating potential. They contain the same magical formula of eternal youth that people have been trying to bring out for many centuries.

What are fibroblasts and what are they for?

The word fibroblast contains two roots - "fibra", which means "fiber", and "blastos" - "sprout". Fibroblasts are connective tissue cells that have a nucleus and are characterized by a round or fusiform shape and many processes. These are the most valuable cells of the middle layer of the skin (dermis), which are part of the stromal-vascular fraction, which is fundamentally divided into 2 groups:

1. Vascular (vascular) cells: endothelial, pericytes, smooth muscle, circulating blood cells - erythrocytes, leukocytes, monocytes, macrophages, T-lymphocytes, preadipocytes.

2. Fibroblast-like cells, which directly include fibroblasts and their precursors - stromal (they are also multipotent, mesenchymal) stem cells.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

Based on the above classification, it becomes clear that fibroblasts are not stem cells, but their more mature and highly organized successors. Unlike stem cells, which can give rise to cells of any tissue in our body, fibroblasts can only turn into an inactive fibrocyte.

Without fibroblasts, the preservation of the structural integrity of the connective tissue would not be possible, so the role of fibroblasts cannot be overestimated - they are powerful factories that produce and constantly renew the structural components of the dermis and intercellular substance, as well as many biologically active substances that affect regeneration processes:

1. It is fibroblasts that synthesize the constituent components of connective tissue, for the reproduction of which the most modern and high-tech cosmetic procedures are being developed. We are talking about collagen, elastin and hyaluronic acid - natural substances of the dermis that provide its turgor, firmness, elasticity and moisture. Thanks to fibroblasts, proteoglycan, fibronectin, chondroitin sulfate, laminin and other elements of the intercellular matrix are also produced, which are responsible for the beauty and health of the skin.

2. Fibroblasts constantly renew the dermis and do not allow damaged fibers to accumulate in it. Enzymes that are secreted by fibroblasts destroy obsolete, old and damaged elastin, collagen and hyaluronic acid, while replacing them with new and healthy ones. The process of destruction-recovery takes place continuously, providing renewal of the intercellular substance. Especially intensive is the exchange of hyaluronic acid.

3. Fibroblasts are the unique healers of our body. In case of any damage, they “run” with the blood flow to the site of injury and provide the fastest possible restoration of damaged areas, wound healing and epithelialization (rapid restoration of the epidermis - the surface layer of the skin).

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

P The first task pursued by fibroblasts is to restore the barrier to maintain the constancy of the internal environment, i.e. "plug holes" Therefore, they begin to divide very actively and in an emergency mode produce connective tissue molecules, which in a hurry are formed large, coarse, immature, located randomly in the tissues. This is how the first scar appears - red, dense, inelastic, "weak".

F ibroblasts multiply much faster than epidermal cells, therefore, if the damage to the basement membrane is more than 5 mm, then the scar will come to the surface. If less, then full-thickness skin will be restored.

W Then fibroblasts begin to produce enzymes that destroy fibers and gradually replace them with mature, elastic, structural ones. And the scar turns pale, becomes elastic, thin, durable.

4. Fibroblasts are also responsible for skin regeneration (recovery, renewal), since it is they who produce very important growth factors - regulatory proteins (tissue hormones), the function of which is to stimulate the division and growth of cells in the dermis and epidermis, as well as the formation of new blood vessels. We list only some of the growth factors produced by fibroblasts:

The main fibroblast growth factor (bFGF) is responsible for the formation and development of all types of skin cells, causing fibroblasts to actively produce collagen and elastin fibers, hyaluronic acid.

Transforming growth factor (TGF-beta) is responsible for the rapid regeneration of the damaged dermis. It attracts fibroblasts to the site of injury and activates their production of collagen fibers and fibronectin - substances that ensure the restoration of injured skin.

Transforming growth factors (TGF-alpha, a-NGF) cause neoangiogenesis - the process of formation of new blood vessels in the skin.

Epidermal growth factor (EGF) accelerates the division and maturation of keratinocytes.

Keratinocyte growth factor (KGF) accelerates the healing and epithelialization of wounds, stimulating the reproduction and development of epidermal cells (keratinocytes).

5. Trauma is a kind of signal for fibroblasts, forcing them to divide at an accelerated pace and produce growth factors, which, in turn, attract fibroblasts and other cells to the site of damage, ensuring the restoration of damaged tissue.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

Unique properties of fibroblasts

1. The cells of our body cannot multiply indefinitely and their number is reduced by approximately 10-15% every 8-10 years. And the process goes exponentially. This is because with each cell division, a small piece of DNA is lost. At first, sections of DNA (telomeres) are lost that do not carry important information for the functioning of the cell. With each division, the length of telomeres decreases, and when they "end" and there is a threat of loss of DNA fragments that carry significant information for the cell, its division stops. The maximum possible number of divisions averages 50 ± 10 and is called the "Hayflick limit", in honor of the American scientist who discovered this phenomenon in 1961. The countdown of the number of divisions begins in the embryonic period and after the limit is exhausted, the aging of cells, tissues and the body as a whole begins.

2. Previously, it was believed that over time, fibroblasts lose their ability to divide and turn into fibrocytes - mature cells that are characterized by low activity. However, as a result of scientific research, it was found out that despite the fact that the number of fibroblasts decreases with age, they do not lose their functional qualities and are still able to divide, but for some reason they stop doing this, they simply “fall asleep” and, if necessary, can switch back to the active form. Apparently, the reason for this lies in the presence of the telomerase enzyme, which after each cell division restores the length of the telomere, thereby increasing the number of fibroblast divisions. For the first time, this mechanism, which provides the ability of infinite division, was discovered in stem cells.

E This discovery led to the development of a technique for culturing autologous fibroblasts with their subsequent transplantation into the patient's dermis. The procedure is, in fact, the embodiment of the dream of eternal youth, because it involves not only the elimination of signs of age, but also the impact on the very cause of skin aging.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

3. The age of the fibroblast donor does not matter for their lifespan, functional activity and ability to divide. This phenomenon is due to the fact that in the process of cultivation their rejuvenation occurs. Cristofalo et al. came to this conclusion after many years of research. In his opinion, in the laboratory, cells return to a state characterized by high functional activity and bringing them closer in properties to mesenchymal stem cells.

4. In the process of isolating fibroblasts from a piece of the patient's skin, a primary cell culture is obtained, containing both young and old cells. Further, all these cells are placed in a medium containing fetal serum, i.e. to conditions that are observed in the embryonic state. This stimulates the division of young cells that have retained a high ability to grow, and the dilution or washing out of the culture of old cells that have lost the ability to proliferate. Thus, the culture is rejuvenated, as it were. In addition, according to Makinodan, old cells are reactivated under such conditions and subsequently, when injected into the dermis, they populate it and intensively synthesize the entire complex of extracellular matrix components and growth factors necessary to maintain the patient's skin in an optimal physiological state.

It is important to note that we are talking about the patient's own cells, which, while growing up, will not be absorbed by macrophages, unlike transplanted donor cells.

5. In the process of cultivation, fibroblasts lose the alien gene, and they are also unable to cause oncology, which makes it possible to use “foreign” donor cells for therapy, which has already been proven by many years of clinical trials. For the first time, the fibroblast culture technique appeared in 1968 and was used to accelerate wound healing. In 1998, the FDA approved the first fibroblast-based cell product, Apligraf, for use in cambustiology (burn treatment). And only after that a new direction appeared in aesthetic medicine, namely, the treatment of age-related changes with fibroblasts, and in dentistry, the treatment of gingivitis. True, at first only donor fibroblasts were used.

The mechanism of action of this method is associated with the ability of fibroblasts to synthesize collagen, elastin, hyaluronic acid and other components of the intercellular substance, as well as growth factors, which accelerates the division and growth of the epithelium, and ultimately leads to the restoration of the surface and middle layers of the skin - the epidermis and dermis.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

6. With any thermal damage to the skin (burns or frostbite), skin damage occurs, and the degree of severityinflammation and duration (and sometimes the ability to recover) depends on the depth of its defeat:

I degree- redness, swelling of the skin (subside after 3-4 days) and pain (remains 1-2 days) due to reversible damage to the surface layers of the epidermis. In cosmetology, such skin damage is applied specifically with the help of superficial chemical or laser peels for the purpose of rejuvenation.

II degree- the formation of blisters filled with transparent contents as a result of the death of the layers of the epidermis (up to the basal, germ layer) and their detachment. At the site of the burn for some time kept severe pain and burning, but within 10-14 days there is a complete restoration of the integrity of the epidermis without scarring. Suitable for medium peels.

IIIa degree- incomplete skin necrosis with preservation of the dermis and its derivatives - sweat and sebaceous glands, hair follicles, from the epithelium of which an independent restoration of the epidermis occurs within 4–6 weeks, sometimes with the formation of skin scars with areas of hyper- and depigmentation.

IIIb degree- complete necrosis of the entire thickness of the skin.

IV degree- necrosis of the skin and tissues located under it. Epithelialization in such cases is possible only from the edges of the wound and it occurs very slowly. Only a small wound can heal on its own. the possibility of restoring the epidermis along the edges of the wound is no more than 5 mm.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

An important feature that distinguishes IIIa and IIIb degrees is the preservation pain sensitivity In the first case. In children before puberty, such burns often heal with the formation of hypertrophic scars. At this level, deep laser resurfacing or deep chemical peeling of the skin is performed. Perhaps this is only on the face, which is characterized by a very large number of skin appendages, a high ability to regenerate, a very active metabolism in cells and blood supply. In other areas of the skin of our body, such an aggressive effect inevitably leads to the formation of scars.

With superficial burns of I, II and IIIa degrees, fibroblasts are applied to complex treatment wounds of a large area in order to accelerate epithelialization. With deep ones - in combination with transplantation of one's own skin, which, at the same time, requires much less.

7. Autologous (own) and donor cultured fibroblasts do not cause allergic reactions or oncogenesis after transplantation. The body recognizes them as its own, not foreign cells, and therefore does not include a defense mechanism against them.

An important nuance is that the rejuvenating effect of our own cultured fibroblasts is much more prolonged than the similar effect of donor cells. The latter, over time, are still recognized and absorbed by the immune cells of our body, so the result remains stable for no more than 2 years.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

Features of aging

BUT American scientists published data according to which after 44 years for women (based on medium duration life, which is 78.8 years) and after 40 years for men (based on an average life expectancy of 72.6 years), a person inevitably begins to face diseases. In other words, for almost half of his life he is doomed to fade away, suffering from ailments and infirmities. The first signs of the destructive aging process appear already at the age of 30. The situation is aggravated by the modern rhythm of life, associated with mental overload, which most detrimentally affects the human body.

To As mentioned above, thanks to the activity of fibroblasts, the dermis is constantly updated due to the balance of two multidirectional processes: the destruction of obsolete, old fibers and the synthesis of new ones. BUT, at some point, for some reason (still not clear, because this phenomenon can begin in people in different ages) the ability of fibroblasts to divide and synthesize substances decreases. At the same time, the process of destruction of old fibers will continue for a long time, which will entail a decrease in the volume of connective, muscle, bone and other types of tissues. That is, the process of destruction begins to prevail over the process of creation.

Due to the reserve of cells provided by nature, the consequences of the imbalance remain not too noticeable for several years. Meanwhile, after 40-45 years, no one manages to avoid age-related changes, and sometimes they overtake us like an avalanche, and for many women this period is associated with the onset of menopause and the onset of hormonal aging. That is why Victor Hugo called this age "the old age of youth." After a while, the process of cell and tissue death stops, the balance between creative and destructive processes is restored, but by this age a person turns into a “shrunken” old man or old woman. In aging skin, the thickness of the dermis decreases, the moisture content in it decreases, as a result, the skin loses firmness and elasticity. The result is stretching of the skin and the formation of wrinkles.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

P The processes of tissue renewal and regeneration slow down, which entails unpleasant consequences:

- the basal (growth, regenerative) layer becomes thinner, less and less keratinocytes are formed;

The cells of the epidermis (horny scales) become thinner;

The process of removing horny scales from the surface of the skin slows down, as a result of which the stratum corneum becomes thicker;

The dermis rapidly loses thickness, the number and size of fibroblasts, macrophages, tissue basophils and other cells of the dermis decreases. They cease to cope with their functions, which sooner or later leads to a deficiency of collagen, elastin and intercellular substance. Starting at about 25 years of age, the synthesis of collagen and elastin - the fibers that make the skin look firm and healthy - is reduced by 1% annually.;

The structure of elastin and collagen fibers is deformed: they become thicker, more rigid than they should be in the norm, the order of their arrangement is disturbed;

The body produces less and less hyaluronic acid, which leads to a loss of moisture in the dermis, leads to overdrying of the skin, the formation of microcracks and wrinkles on it, a decrease in its elasticity and turgor;

The blood supply and the supply of nutrients to the cells of the dermis are deteriorating;

Recovery processes are slow.

The above changes may affect appearance skin cover. The feeling of dryness and tightness of the skin gradually increases, the skin becomes flabby, thin, inelastic, covered with fine wrinkles and age spots. Over time, all these signs of aging accumulate and become more pronounced. Open areas of the skin and folds age especially quickly.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

Fibroblast rejuvenation stops the aging process

Hsaturation of the dermis with young fibroblasts - the most natural, effective method rejuvenation and prevention of aging, as it allows you to revive the structure of the dermis, and in aged patients it is a replacement therapy.

At The unique procedure of cellular skin rejuvenation, based on the use of autologous fibroblasts, stops the process of depletion of the dermis' own cells. The technique not only corrects age-related changes, but affects them at the microtexture level: young fibroblasts rejuvenate the dermis from the inside, and also stimulate the activity of those fibroblasts that are present in the body. As a result, the rate of cell division increases, the surface layer of the skin is renewed faster, new young collagen and elastin fibers are formed, and the content of hyaluronic acid in the dermis increases. You again enjoy the look of radiant velvety skin, for a long time you forget about wrinkles, enlarged pores, age spots, peeling and dryness.

To flying therapy even copes with stretch marks - defects that are almost impossible to eliminate using other minimally invasive techniques. Fibroblasts do not just stop the biological clock, but make it run in the opposite direction. And if, after some time, their activity decreases and they fall asleep, then simple methods of physical trauma penetrating the dermis (such as diode and carbon dioxide laser skin rejuvenation) will wake them up again and make them work for a long time, multiplying the rejuvenating effect.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

ATintroduction of laboratory-cultivated fibroblasts into the dermis makes it possible to restore the elasticity and firmness characteristic of youth to the skin. Moreover, if in the future you will do cosmetic procedures or plastic surgery, you will get a much more pronounced effect than those who did not activate their own fibroblasts first and did not replenish their supply.

FROM The fabulous effect of using autologous fibroblasts has already been appreciated by many celebrities. After all, the results of the procedure are really amazing: the network of small wrinkles disappears without a trace, deep folds are smoothed out. Day after day, you watch in the mirror how the skin becomes more radiant and elastic, its tone improves, the network of fine wrinkles is smoothed out, and the complexion becomes healthy. The neck and arms no longer give out age - the skin of these parts of the body takes on a toned look and fullness. Transplantation of cultured fibroblasts increases the protective barrier properties of the skin, which means that adverse factors and stress will not be able to steal youth and beauty.

T Therapy with autologous fibroblasts is much more effective than Botox injections, which, with prolonged and frequent use, can cause nerve damage and malnutrition of the skin.

P In addition, the introduction of fibroblasts is more effective than replacement therapy hyaluronic acid, which rejuvenates the skin for a short period of time, and then they must be repeated. With frequent use and over time, the body begins to produce antibodies to artificial hyaluronic acid, and the destruction of the injected preparations occurs faster and faster. In addition, the introduction of an excess (especially under the age of 35!) of hyaluronic acid has an inhibitory effect on the synthesis of the structural components of the skin by fibroblasts, thereby indirectly accelerating aging.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

Indication for fibroblast therapy:

Prevention of aging - injections can be started from the age of 40, thereby performing replacement therapy;

Rejuvenation of the skin of the face, neck, decollete, hands eliminates the signs of aging: thinning, flabbiness, reduced turgor and elasticity, pigmentation, atrophy and fine wrinkling;

Improving the quality of the skin of the body: abdomen, back, thighs. Fibroblast therapy enhances elasticity and tone, thereby providing a lifting effect;

Elimination of pigmentation around the eyes;

Accelerating the "maturation" of young scars - at the age of up to 12 months;

Treatment of post-acne scars;

Stretch marks treatment;

Preparation for plastic surgery and quick recovery after them;

Acceleration of recovery after peels, laser procedures, etc.

Contraindications to fibroblast therapy:

Acute infectious diseases;

Exacerbation of chronic diseases;

Autoimmune diseases of the connective tissue;

Tendency to keloid and hypertrophic scars;

Oncological diseases;

Long-term steroid therapy;

Pregnancy, lactation.

Fibroblast therapy

E If we talk about therapy with autologous fibroblasts simply, then it consists of several stages:

1. Taking a piece of leather. It can be taken on any part of the body, it is only important to observe the size - about 5 * 1.5 cm. The amount of fibroblasts that will be received in the laboratory of the Pokrovsky Stem Cell Bank (with which our clinic cooperates) depends on the size of the taken skin area. In order to properly saturate the skin with young cells, in one procedure it is necessary (by a special method!) to introduce a sufficient amount of fibroblasts (about 2-3 million per 1 ml). Therefore, laboratory technologists ask for a larger piece of skin.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

Most often, we take it away by excising healthy skin along the existing scars on the body from previous operations or injuries, and in their absence from the groin area. After taking the required area, the wound is sutured in layers, ending with an intradermal suture, which we remove on the 7-10th day after the operation. As a result, a thin, inconspicuous thread-like seam will remain in this place, which is easily hidden even in the most open panties.

To complete this stage, it is required to pass blood tests: general clinical, biochemical (glucose, ALT, AST, bilirubin, urea, creatinine) and a coagulogram.

Skin removed during aesthetic operations (facelift, blepharoplasty, abdominoplasty, etc.) is suitable for isolating and cultivating fibroblasts. Many patients want to repeat the procedure in the future, and technologists immediately cultivate two portions of fibroblasts, one of which is stored in a cryogenic chamber until the right moment - so that they do not have to take a piece of skin next time.

2. Isolation and cultivation of fibroblasts in the laboratory of the Pokrovsky Stem Cell Bank: a piece of skin is crushed, processed with special enzymes, washed with saline. Then the cells released in this case are deposited in a centrifuge and sown on a special nutrient medium. They multiply to the required amount, removed from the substrate, cleaned of the remnants of the medium, and precipitated in a centrifuge.

3. The quality control system of fibroblasts obtained as a result of cultivation is very important. To do this, cells with oncogenic potential are monitored daily and removed from the culture: daily control of the shape, structure, activity of cell reproduction, as well as DNA testing and, if necessary, the level of expression (isolation) of oncogenic markers. In addition, quality control includes analysis for bacterial contamination and for the absence of HIV and hepatitis viruses. For each batch of cells, a fibroblast passport is provided, which indicates the name of the donor, date, time of manufacture, the number of cells per 1 ml, and negative results of tests for oncogenicity and infection.

Further options are possible - they are either introduced in saline by the method of mesotherapy (for rejuvenation), or placed on a special gel carrier (for healing wounds and burns). This whole process can take 4 to 6 weeks.

Not a public offer! There are contraindications. Before use, a specialist consultation is necessary.

4. Each portion of the cells is prepared not only for a specific date, but also for a predetermined time, because. fibroblasts outside of human tissues are vulnerable and must be injected within 6 hours, as after that they will die. Fibroblasts are injected 5 times with an interval of 2 weeks, using mesotherapeutic technique, into the upper layers of the skin. This technique ensures the stability of the result and a rejuvenating effect. At the same time, the rule works well: the larger the treatment area, the greater the rejuvenation.

R The result of therapy with own fibroblasts is cumulative, it appears at the level of sensations after 1-1.5 months from the start of the procedures (usually by the third) and then gradually increases up to 12-18 months, and then remains stable for 5 years or more. This is a method of natural skin rejuvenation, absolutely safe, highly effective and long-lasting. The introduction of fibroblasts is aimed at improving the quality of the skin, i.e. its turgor, tone, color, density, etc., BUT never leads to a lift!

E If you start noticing that fibroblasts decrease their activity, they can be easily stimulated by laser therapy (CO, diode, neodymium lasers, acting on the dermis), the effect of which will be much more pronounced and noticeable.

1. Production of all components of the intercellular substance (fibers and the main amorphous substance). Fibroblasts synthesize collagen, elastin, fibronectin, glycosaminoglycans, etc.

2. Maintaining the structural organization and chemical homeostasis of the intercellular substance (due to balanced processes of its production and destruction).

3. Regulation of the activity of other connective tissue cells and influence on other tissues. Production of cytokines (colony-stimulating factors of granulocytes and macrophages).

4. Wound healing. During inflammation and wound healing, fibroblasts are activated by macrophages.

Rice. 3.2. Loose and fibrous connective tissues - film preparation I - basic substance; II - collagen fibers; III - elastic fibers; IV - cells; V is a blood vessel. 1 - fibroblasts, 2 - fibrocyte, 3 - macrophages, 4 - mast cells, 5 - plasma cells, 6 - leukocytes, 7 - fat cell.

Fig.3.3. Electron diffraction pattern of fibroblast among collagen fibers
(x 18.500).

Ct - transverse,

Сl – longitudinal sections of collagen fibers;

N - the cell nucleus is displaced to the periphery;

ER, endoplasmic reticulum;

G is the Golgi complex.

Rice. 3.4. Actin microfilaments in the cytoplasm of myofibroblast (immunofluorescent method).

macrophages. Macrophages are in second place in quantitative terms among the cells of loose connective tissue. Macrophages are formed by differentiation and reproduction of monocytes that have entered the tissue from the blood. There are free and fixed macrophages. Compared to fibroblasts, they are smaller in size 10-15 microns. Have different shape- rounded, elongated or irregular. The basophilic cytoplasm of macrophages contains many lysosomes, phagosomes, and pinocytic vesicles. Mitochondria, ER, Golgi complex have moderate development. Macrophages are actively phagocytic cells rich in organelles for intracellular digestion of the absorbed material (lysosomes) and synthesis of antibacterial and other biologically active substances (pyrogen, antiferon, lysozyme, EPS). The nuclei contain more chromatin and stain more intensely than the nuclei of fibroblasts. The cytoplasm of macrophages forms deep folds and long microvilli that capture foreign particles. The macrophage surface has receptors sensitive to erythrocytes, T and B-lymphocytes, antigens and immunoglobulins. The latter provide the possibility of their participation in the body's immune responses.

BUT

B

Rice. 3.5. Macrophage ultrastructure. A – active form, B – macrophage surface (x11.600). Scanning electron microscopy. 1 - processes of the cell. Pp, 1 - pseudopodia; P, phagocytosed particles; M - mitochondria; L - lysosomes. Irregular shaped core.

Macrophages, along with the ability to phagocytosis, synthesize a number of substances that provide innate immunity (lysozyme, interferon, pyrogen, etc.). Macrophages secrete mediators - monokines, which promote a specific reaction to antigens and cytolytic factors that selectively destroy tumor cells.

Functions of macrophages:

1. phagocytosis: recognition, absorption and digestion of damaged, infected, tumor and dead cells, components of the intercellular substance, as well as exogenous materials and microorganisms.

2. participation in the induction of immune responses, tk. (play the role of antigen-presenting cells).

3. regulation of the activity of cells of other types (fibroblasts, lymphocytes, mast cells, endotheliocytes, etc.).

Macrophages develop from monocytes. A set of cells with one nucleus is called a monocular phagocytic system, and mononucleids that have the ability to phagocytosis: to capture foreign particles, dying cells, non-cellular structures, bacteria, etc. from the tissue fluid of the body. The phagocytosed material undergoes enzymatic cleavage inside the cell (“completed phagocytosis” ), due to which agents harmful to the body, arising locally or penetrating from the outside, are eliminated. Macrophages (histiocytes) of loose fibrous connective tissue, stellate cells sinusoidal vessels liver, free and fixed macrophages of hematopoietic organs (bone marrow, spleen, lymph nodes), lung macrophages, inflammatory exudates (peritoneal macrophages), osteoclasts, giant cells foreign bodies and glial macrophages of the nervous tissue (microglia). All of them are capable of active phagocytosis, have immunoglobulin receptors on their surface, and originate from bone marrow promonocytes and blood monocytes. In contrast to such “professional” phagocytes, the ability to facultative absorption can be expressed independently of these cytoreceptors in other cells (fibroblasts, reticular cells, endotheliocytes, neutrophilic leukocytes). But these cells are not part of the macrophage system.

I.I. Mechnikov (1845-1916) was the first to come to the conclusion that phagocytosis, which arises in evolution as a form of intracellular digestion and is fixed in many cells, is at the same time an important protective mechanism. He substantiated the expediency of combining them into one system and proposed calling it macrophage. The macrophage system is a powerful protective apparatus that takes part in both general and local defense reactions of the body. In the whole organism, the macrophage system is regulated both by local mechanisms and by the nervous and endocrine systems. In the 1930s and 1940s, this defense system was called reticuloendothelial. Recently, it has been called the system of mononuclear phagocytes, which, however, characterizes it inaccurately due to the fact that among the cells included in this system, there are also multinuclear (osteoclasts).

Plasma cells - plasma cells have a rounded shape. The size of plasma cells is from 7 to 10 microns. The nucleus is round or oval and usually lies eccentrically. Lumps of chromatin in it are located along the radii. They resemble pyramids, the base of which lies on the nuclear envelope. It seems that chromatin is arranged in the form of spokes in a wheel. This circumstance is one of diagnostic features in the determination of plasma cells.

BUT

B

AT

Rice. 3.6. Plasma cell. A - in a blood smear. B - scheme. B - electronogram .

The cytoplasm of cells is sharply basophilic, especially along the periphery. In the center in front of the core there is a small enlightenment - a "courtyard". It contains a mesh apparatus, centrioles, mitochondria. Cytochemically, a huge amount of ribonucleoproteins is found in plasma cells, which determine the basophilia of the cytoplasm. Among the proteins, a lot is found - γ-globulin. The main function of cells is associated with it - participation in the protective reactions of the body.

Mature plasma cells are characterized by high basophilia and an eccentric nucleus. Under an electron microscope, parallel membranes are determined. The presence of parallel membranes in the cytoplasmic reticulum is typical for cells synthesizing protein for export. The protein produced by the plasma cell can have a different composition and is determined by the quality of the protein of the stimulus or antigen. Therefore, we say that protein synthesis in plasma cells is a particular expression of the ability of these cells to take part in protein metabolism. Along with this, the cytoplasm of the cell secretes a small amount of glycosaminoglycans entering the intercellular substance.

Comparison of globulin concentration showed that it is less in mature cells than in immature ones. Recently, it is believed that a mature cell is a plasma cell at rest. When meeting with an antigen, an irritant, it can also intensively form globulin and, in its own way, morphological features approach that cell, which is called "immature". Plasma cells are called immunocompetent, because they retain a "memory" of antigenic stimuli and, when they encounter it again, block the antigen with a specific antibody.

One of the manifestations immune response in vertebrates, when a foreign agent enters the body, antibodies are released by plasma cells.

In the cytoplasm of plasma cells, crystalline inclusions may appear that perceive acidic dyes, the so-called Roussel bodies. It is believed that they are conglomerates of globulins synthesized earlier by this cell.

Plasma cells provide humoral immunity by producing antibodies. In 1 second, each plasma cell synthesizes up to several thousand immunoglobulin molecules (more than 10 million molecules per hour).

Tissue basophils (labrocytes, mast cells). mast cells- a permanent cellular component of loose fibrous connective tissue, which performs important regulatory functions. These cells have a granularity in the cytoplasm resembling granules of basophilic leukocytes. They are regulators of local connective tissue homeostasis.

BUT

B

Rice. 3.7. Mast cell structure A - Mast cells (M) in the connective tissue (x1200); B - relief of the cell surface.

Mast cell development occurs in tissues from a precursor believed to be of bone marrow origin. Their differentiation and growth are influenced by factors of the cellular microenvironment (fibroblasts, epithelial cells and their products). Unlike basophils, which after migration into tissues do not live long (from several hours to several days), mast cells have a relatively long lifespan (from several weeks to several months). During this period, under the influence of appropriate stimuli, mast cells are apparently able to divide.

Rice. 3.8. Electronogram of a mast cell (x12.000). G - large granules fill the entire cytoplasm; Mi - mitrchondria located between them, in the center is the nucleus.

Tissue basophils have a variety of shapes. In humans and mammals, their shape is more often oval. Dimensions 3.5x14 microns. The nucleus is small and rich in chromatin. There are binucleated cells.

Mast cell granules contain a variety of biologically active substances. Submicroscopically, they are irregularly shaped dense bodies with a diameter of 0.3-1.4 microns, stained metachromatically. Cells contain mitochondria, an intracellular network apparatus. The components of mast cells are different in different animals and in different parts of the connective tissue. in rabbits and guinea pigs mast cells are few, white mice have a lot. In humans and animals, mast cells are found in all places where there are layers of loose connective tissue. They are located in groups along the course of the blood and lymphatic vessels. The number of mast cells changes with different states body - during pregnancy, the number of mast cells in the uterus and mammary glands, in the stomach and intestines increases in the midst of digestion. Mast cells contain a variety of mediators and enzymes.

Structural and functional differences of mast cells. The mast cell population is formed by elements that have different morphofunctional properties and can differ qualitatively and quantitatively even within the same organ. It is suggested that individual subpopulations of mast cells perform different functions in the body.

Mast cell functions:

1. Homeostatic, which is carried out under physiological conditions by slowly releasing small amounts of biologically active substances that can affect various tissue functions - first of all, vascular permeability and tone, maintaining the balance of fluids in tissues.

2. protective and regulatory which is provided by local release of inflammatory mediators and chemotactic factors that provide (a) the mobilization of eosinophils and various effector cells involved in the so-called late phase reactions; (b) influence on the growth and maturation of connective tissue in the area of ​​inflammation.

3. Participation in the development of allergic reactions due to the presence of high-affinity receptors for class E immunoglobulins (IgE) on their plasmolemma and the functional connection of these receptors with the secretory mechanism. Involvement of mast cells in the development of allergic reactions, as well as basophilic granulocytes includes:

Ø binding of IgE to high-affinity receptors on their plasmalemma;

Ø interaction of membrane IgE with an allergen;

Ø activation and degranulation of mast cells with the release of substances contained in their granules and the production of a number of new ones.

Ø It is assumed that mast cells perform a magnetoreceptor function.

Degranulation can also be mediated by complement receptors or induced by neutrophil proteins, proteinases, neuropeptides (substance P, somatostatin), and lymphokines.

According to Walker's calculations, a complete change of mast cells in loose connective tissue can occur in 16 to 18 months. According to N.G. Khrushchev for 9 days.

Table 3.2.

Mediators and enzymes found in mast cells

Fat cells, lipocytes. There are two types of fat cells: white and brown fat cells. White fat cells are monovacuolar, having one fat vacuole. They are located in the loose connective tissue mainly along the course of the vessels, and in some parts of the body (under the skin, between the shoulder blades, in the omentum and other places) forming significant accumulations. This allows you to isolate a special adipose tissue built almost exclusively from fat cells. Fat cells are spherical in shape. They are larger than other connective tissue cells. Their diameter is 30-50 microns. The immediate precursors of fat cells are poorly differentiated connective tissue cells located mainly near the capillaries (pericapillary or adventitial cells). The formation of lipocytes from histiocytes that phagocytize fat droplets is possible. In the process of differentiation, small drops of neutral fat accumulate in the fat cell, which, by fusion, form larger ones. The main function of lipocytes is to store fat as a macroergic compound. When it breaks down, a large amount of energy is released, used by the body as a source of heat, as well as for phosphorylation of ADP with the formation of ATP. Fat serves as a source of water formation, performs a protective and supporting function. Fat cells synthesize biologically active substances - leptin, which regulates the feeling of fullness, estrogens, etc.

BUT

B

Fig.3.9. White fat cells (apudocytes, monovacuolar cells) A - a set of fat cells forms a fat lobule, equipped with a large number of blood vessels (C) x480); B – electron micrograph of the periphery of 2 apudocytes, L – fat vacuole; D - small droplets of fat; M - mitochondria; C-collagen fibers in the intercellular space. (x6.000).

Rice. 3.10. Electron micrograph of a brown fat cell: The nucleus is located in the center,

L - fat vacuoles,

M - mitochondria,

C - capillaries.

Fat cells, in addition to the role of an energy depot, perform the functions of an endocrine gland, the hormones of which regulate the volume and weight of the body. This hormone is leptin.

White adipose tissue makes up 15-20% of the body weight of adult males and 5% more in females. In a sense, it can be spoken of as a large metabolically active organ, since it is mainly involved in the absorption from the blood, the synthesis, storage and mobilization of neutral lipids (fats). (To mobilize fat means to make it mobile so that it can be used as a fuel” in other parts of the body.) In a fat cell at body temperature, fat is in the state of liquid oil. It consists of triglycerides containing three fatty acid molecules that form an ester with glycerol. Triglycerides are the most caloric type of nutrients, so fat in fat cells is a storehouse of "high-calorie" fuel, and relatively light. In addition, in the inhabitants of cold camps, fat is involved in the regulation of the temperature of the organs underlying it. And, finally, fat serves as an excellent filler for various “crevices” in the body and forms “pillows” on which certain internal organs can lie.

brown fat cells found in newborns and in some animals on the neck, near the shoulder blades, behind the sternum, along the spine, under the skin between the muscles. It consists of fat cells densely braided with hemocapillaries. Brown fat cells are polyvacuolar. The diameter of brown fat cells is almost 10 times smaller than the diameter of white fat cells. These cells take part in the processes of heat production. Brown adipose tissue adipocytes have many small fatty inclusions in the cytoplasm. Compared to white adipose tissue cells, many mitochondria are found here. Iron-containing pigments - mitochondrial cytochromes - give brown color to fat cells. The oxidative capacity of brown fat cells is approximately 20 times higher than that of white ones and almost 2 times the oxidative capacity of heart muscle. With a decrease in ambient temperature, the activity of oxidative processes in brown adipose tissue increases. In this case, thermal energy is released, heating the blood in the blood capillaries. The sympathetic system plays a role in the regulation of heat transfer. nervous system and hormones of the adrenal medulla - adrenaline and norepinephrine, which through cyclic adenosine monophosphate stimulates the activity of tissue lipase, which breaks down triglycerides into glycerol and fatty acid. The latter, accumulating in the cell, uncouple the processes of oxidative phosphorylation, which leads to the release of thermal energy that heats the blood flowing in numerous capillaries between lipocytes. During starvation, brown adipose tissue changes less than white.

Pigmentocytes ( pigment cells). contain melanin pigment in their cytoplasm. They have a process shape and are divided into two types - melanocytes that produce pigment, and - melanophores that can only accumulate it in the cytoplasm. In people of black and yellow races, pigment cells are more common than the color of the skin, which does not change depending on the season, is determined. Pigmentocytes have short irregularly shaped processes. These cells only formally belong to the connective tissue, as they are located in it. There is now strong evidence that these cells originate from neural crests and not from mesenchyme.

Table 3.3. Differences between white and brown fat cells

adventitial cells. These are unspecialized cells that accompany blood vessels. They have a flattened or fusiform shape with weakly basophilic cytoplasm, an oval nucleus, and underdeveloped organelles. In the process of differentiation, these cells, apparently, can turn into fibroblasts, myofibroblasts, and adipocytes. Many authors deny the existence of adventitial cells as an independent cell type, considering them to be fibroblast cells.

endothelial cells- lined with blood vessels, therefore their totality is called the vascular endothelium. The structure of the vascular endothelium is similar to the structure of the epithelial tissue. The endothelium has the following general features.

1. Border position of the integumentary epithelium and endothelium.

2. Continuity of the endothelial lining within all blood and lymphatic vessels in vertebrates.

3. The absence of the main intermediate substance around the entire circumference of endothelial and epithelial cells.

4. The presence of a basement membrane that performs the function of supporting and fixing endothelial cells. Its basis, as well as the basis of the basement membranes of the epithelium, is type IV collagen.

5. Heteropolarity in the structure of cells. In endotheliocytes, this manifests itself in the formation of microvilli on the luminal surface of cells (with a relative smoothness of the basal one), in the unevenness of the elements of the cytoskeleton and the concentration of micropinocytic vesicles in the cytoplasm of opposing cell surfaces.

6. Specialized contacts between endothelial cells of the trailing type, the fibrillar strips of which are located closer to the luminal surface of the cells, which emphasizes its polarity.

7. Barrier, secretory, transport functions in their ideal combination.

8. Growth of endothelium in tissue cultures in the form of a monolayer of polygonal cells with pronounced contact inhibition.

Because of this similarity, many researchers classify the endothelium as an epithelial tissue. However, the endothelium originates from the mesenchyme, on the basis of which it is referred to as a connective tissue.

Endothelial cells play an important role in the processes of transcapillary metabolism, take part in the formation of tissue mucopolysaccharides, histamine, fibrinolytic factors.

Functions of the endothelium:

1. Transport - selective two-way transport of substances between blood and other tissues is carried out through it. Mechanisms: diffusion, vesicular transport (with possible metabolic transformation of transported molecules).

2. Hemostatic - plays a key role in blood clotting. Normally forms an athrombogenic surface; produces procoagulants (tissue factor, plasminogen inhibitor) and anticoagulants (plasminogen activator, prostacyclin).

3. Vasomotor - participates in the regulation of vascular tone: secretes vasoconstrictor (endothelin) and vasodilator (prostacyclin, endothelial relaxing factor - nitric oxide) substances; participates in the exchange of vasoactive substances - angaotensin, norepinephrine, bradykinin.

4. Receptor - expresses on the plasmalemma a number of compounds that provide adhesion and, and subsequent transendothelial migration of lymphocytes, monocytes and granulocytes.

5. Secretory - produces mitogens, inhibitors and growth factors, cytokines that regulate hematopoiesis, proliferation and differentiation of T- and B-lymphocytes, attracting leukocytes to the site of inflammation.

6. Vascular - provides new formation of capillaries (angiogenesis) - both in embryonic development and during regeneration.

Pericytes- star-shaped cells adjacent to the outside to arterioles, venules and capillaries. Most numerous in postcapillary venules. They have their own basement membrane that fuses with the basement membrane of the endothelium, so that it appears that the pericyte is enclosed in a stratified basement membrane of the endothelium. Pericyte covers the vessel wall, which suggests their involvement in the regulation of the vascular lumen.

Pericytes have a discoid nucleus with small depressions, contain the usual set of organelles, multivesicular bodies, microtubules, and glycogen. In the area facing the vessel wall, contain bubbles. Near the nucleus and in the processes there are contractile proteins, incl. actin and myosin. Pericytes are covered by a basement membrane, but are closely associated with the endothelial cell, since the basement membrane between them may be absent. Gap and adhesive contacts were found in these places.

The functions of pericytes are not clearly established. We can talk about specific functions with varying degrees of probability.

1. Contractile properties. It is likely that pericytes are involved in the regulation of the microvessel lumen.

2. Source of smooth muscle cells (SMC). During wound healing and restoration of blood vessels, pericytes differentiate into SMCs within 3-5 days.

3. 3. Influence on endothelial cells. Pericytes control endothelial cell proliferation, as in normal growth vessels, and during their regeneration; modulate the function of endothelial cells by regulating the transport of macromolecules from capillaries to tissues.

4. Secretory function. Synthesis of components of the capillary basement membrane.

5. Participation in phagocytosis.

intercellular substance loose fibrous connective tissue consists of fibers and a basic amorphous substance. It is a product of the activity of the cells of this tissue, primarily fibroblasts.

Functions of the intercellular substance of loose fibrous connective tissue:

1. providing architectonics, physico-chemical and mechanical properties of the tissue;

2. participation in the creation of an optimal microenvironment for cell activity;

3. integration into a single system of all connective tissue cells and ensuring the transfer of information between them;

4. impact on numerous functions of various cells (proliferation, differentiation, motility, receptor expression, synthetic and secretory activity, sensitivity to the action of various stimulating, inhibitory and damaging factors, etc.). This effect can be carried out by the contact action of the components of the intercellular substance on the cells, as well as due to its ability to accumulate and release growth factors.

Collagen fibers in the composition of different types of connective tissue determine their strength. In loose, unformed fibrous connective tissue, they are located in different directions in the form of wave-like curved, spirally twisted, rounded or flattened strands with a thickness of 1-3 microns or more. Their length is different. The internal structure of the collagen fiber is determined by the fibrillar protein - collagen, which is synthesized on the ribosomes of the granular endoplasmic reticulum of fibroblasts.

Rice. 3.11. I. Scheme - levels of structural organization of collagen fibers. II. Electron micrograph - collagen fibril. There are four levels of organization of collagen fibers: tropocollagen molecules (1), protofibrils (2), fibrils (3) and fibers (4).\

Collagen fibers are distributed not only in the connective tissue itself, but also in bone and cartilage, where they are respectively called ossein and chondrin. These fibers determine the tensile strength of fabrics. In loose unformed connective tissue, they are located in different directions in the form of wave-like curved strands 1-3 microns thick. Collagen fibers consist of bundles of parallel microfibrils with an average thickness of 50-100 nm, interconnected by glycosaminoglycans and proteoglycans. Their thickness depends on the number of fibrils, which have transverse striation (black and light areas) with a repetition period of 64-70 nm. Within one period there are secondary bands 3–4 nm wide.

Collagen structures, which are part of the connective tissues of the human and animal body, are its most common components. Their main component is a fibrous protein - collagen.

Collagen is the main protein of connective tissue, which makes up over 50% of the body weight of humans and animals. At the same time, according to the calculations of the Swiss scientist F. Verzar, collagen accounts for about 30% of the total amount of protein in the body. Consequently, collagen in quantitative terms is among the proteins in the first place.

Deciphering the primary structure of collagen is the most important stage in the development of this knowledge. The importance of revealing the structure of collagen should be considered in view of the great interest shown in collagen in various areas knowledge. It underlies entire areas of technology. All leather production is essentially the processing of collagen. Denatured collagen-gelatin is an indispensable component of photo-film materials. Recycled collagen is used to make many materials used in veterinary and medical practice.

The collagen molecules extracted from the fibers are 200 nm long and 1.4 nm wide. They are called tropocollagen. Molecules are built from triplasts - three polypeptide chains that merge into a single helix. Each chain contains a set of three amino acids regularly repeated throughout its length. The first acid in such a set can be any, the second - proline or lysine, the third - glycine.

The arrangement of amino acids can vary, as a result of which four types of collagen are formed.

Type 1 - in the connective tissue proper, bone, cornea, sclera, dental ligament, etc.

Type 2 - in hyaline and fibrous cartilage, vitreous body.

Type 3 - in the dermis of the skin of the fetus, blood vessels, in reticular fibers.

Type 4 - in the basement membranes, in the lens capsule.

In 1973, one of the collagen polypeptide chains was deciphered, which seems to be an outstanding event. Collagen is significantly larger in molecular weight than other studied proteins. Difficulties in establishing the structure of collagen were due to the size of the molecule and the special monotony of its structure - the frequency of repetition of amino acid residues and their combinations, which greatly complicated the task of research.

Collagen molecules are about 280 nm long and 1.4 nm wide. They are built from triplets - three polypeptide chains, the precursor of collagen - procollagen, twisting into a single helix while still in the cell. it the first, molecular, level of organization of collagen fibers. Procollagen is secreted into the intercellular substance.

Second, the supramolecular level - the extracellular organization of the collagen fiber - represents tropocollagen molecules aggregated in length and cross-linked by means of hydrogen bonds, formed by cleavage of the terminal peptides of procollagen. First, protofibrils are formed, and 5-6 protofibrils, fastened together by lateral bonds, make up microfibrils with a thickness of about 5 nm.

With the participation of glycosaminoglycans, also secreted by fibroblasts, third, fibrillar and, the level of organization of collagen fibers. Collagen fibrils are transversely striated structures with an average thickness of 20-100 nm. The repetition period of dark and light areas is 64-67 nm. Each collagen molecule in parallel rows is believed to be shifted relative to the adjacent chain by a quarter of the length, which causes the alternation of dark and light bands. In dark bands under an electron microscope, secondary thin transverse lines are visible, due to the location of polar amino acids in collagen molecules.

Fourth, fiber, organization level. Collagen fiber formed by fibril aggregation has a thickness of 1-10 µm (depending on topography). It includes a different number of fibrils - from single to several tens. The fibers can be folded into bundles up to 150 microns thick.

Collagen fibers are characterized by low tensile strength and high tensile strength. In water, the thickness of the tendon as a result of swelling increases by 50%, and in dilute acids and alkalis - 10 times, but at the same time the fiber is shortened by 30%. The ability to swell is more pronounced in young fibers. During heat treatment in water, collagen fibers form a sticky substance (Greek colla - glue), which gave the name to these fibers.

Reticular (reticulin, argyrophilic) fibers. They are found in loose and some other types of connective tissue, in the stroma of hematopoietic organs, the liver, and the inner membranes of blood vessels. On preparations impregnated with silver, they are located in the form of a network.

Rice. 3.12. reticular fibers in lymph node by impregnation with silver nitrate. The fibers branch, forming a thin network. BV - blood vessel (x800).

The question of the nature of reticular fibers remains controversial. Most researchers believe that reticulin, the protein that forms the basis of these fibers, is a substance close to collagen, and the impregnation and histochemical differences between reticular fibers and collagen fibers are associated with the properties of glycosaminoglycans that stitch the fibers. Unlike collagen and elastin, reticulin contains more serine, oxylysin and glutamic acid.

elastic fibres. Elastic fibers give the fabric elasticity. They are less durable than collagen ones. In loose connective tissue, they form a looped network, anastomosing with each other. The thickness of the fibers is from 0.2 to 1 micron. Unlike collagen ones, they do not have microscopically visible fibrils and submicroscopic transverse striation.

BUT

B

Rice. 3.13. A - Elastic fibers in the connective tissue (x320). B - elastic fibers in the wall of a large artery (x400), E - thin elastic fibers, Sar - branched capillary, P - plasma cells, C - collagen fibers.

The basis of elastic fibers is a globular glycoprotein - elastin, synthesized by fibroblasts and smooth muscle cells (the first, molecular, level of organization). Elastin is characterized by a high content of proline and glycine and the presence of two amino acid derivatives - desmosine and isodesmosine, which are involved in stabilizing the molecular structure of elastin and giving it the ability to stretch and elasticity. Elastin molecules, having globules 2.8 nm in diameter, outside the cell are connected into chains - elastin protofibrils 3-3.5 nm thick (second, supramolecular, level of organization). Elastin protofibrils in combination with a glycoprotein (fibrillin) form microfibrils with a thickness of 8-19 nm (third, fibrillar, level of organization). The fourth level of organization is fiber. The most mature elastic fibers contain about 90% of the amorphous component of elastic proteins (elastin) in the center, and microfibrils along the periphery. In elastic fibers, unlike collagen fibers, there are no structures with transverse striation along their length.

In recent decades, in the field of professional cosmetology, the correction technique has become increasingly popular. skin through restorative biological technologies. These include, in particular, rejuvenation by injection of autologous fibroblasts.

Scientific Validity

This technique has a serious biological basis and is based on the body's natural ability to regenerate. Fibroblasts are fibrous cells found in every human body. Their goal is the constant production of the most valuable substances on which the healthy state of the human body directly depends.

First of all, these cells synthesize the structural components of proteins, as well as connective fibers and hyaluronic acid. The presence in the tissues of these elements in the required quantity and in the correct proportions ensures the stability of the hydrostatic pressure in the cells and gives them elasticity. During life, as a person approaches adulthood, the percentage of fibroblasts in the skin decreases. They lose their elasticity and under the influence of gravity become flabby and saggy.

At the end of the 20th century, cellular rejuvenation by fibroblasts was included in the number of classical surgical techniques. Feedback from the first patients to whom this technique was applied showed that in 100% of cases the use of injections took place without any negative consequences.

Sequencing

The collection of tissues for the preparation of the solution is carried out under local anesthesia. Samples are sent to the laboratory, from where, within a few weeks, ready-made materials are delivered to the clinic, which are necessary in order to carry out rejuvenation with fibroblasts. How the procedure takes place can be seen in the photo below.

The skin of the face, as well as the neck, décolleté and hands are extensively injected. Shortly before the start of therapy, the areas indicated by the doctor are carefully treated with an anesthetic cream. The drug is injected using special thin needles. Once in the layers of the dermis, active cells begin to produce the most important proteins for the body (collagen and elastin), as well as hyaluronic acid and other elements that are an integral part of the matrix.

The rest of the fibroblasts not used for injection, at the request of the patient, remain in the cryobank, where they are stored indefinitely at a low temperature in liquid nitrogen. They can be obtained at any time for repeated procedures.

Cellular rejuvenation with fibroblasts: the essence of the procedure

Renewal of connective regenerating cells not only accelerates the recovery processes in the structure of the skin, but also allows for their correction. Along with the folds, shallow scars and other aesthetic defects disappear.

Fibroblast rejuvenation is a complex of medical procedures, built taking into account individual characteristics patient and referred to as SPRS therapy. It is carried out strictly in clinical conditions.

For injection, the surgeon takes samples of the patient's skin and makes many copies of its structural elements in the laboratory. Since fibroblasts are human's own, and not foreign, cells, the procedure for their implantation occurs absolutely naturally. Natural recovery processes are launched in the body, which after a while becomes noticeable visually.

The injection procedure is no more painful than any of the so-called "beauty injections" and does not leave behind any visible traces, in addition to positive ones.

Rejuvenation course

Most often, the introduction of the required amount of fibroblasts is carried out in two short procedures. They are held for 12 weeks at regular intervals. However, this schedule may vary as SPRS therapy involves individual approach, depending on the particular characteristics of the patient's skin.

The result of the procedure is often obvious after the very first session, which indicates the amazing speed with which fibroblast rejuvenation occurs. The photo below clearly demonstrates the effect of ongoing recovery processes.

SPRS therapy does not side effects in the form of allergic reactions. Since fibroblasts are the main element of mesenchymal stem cells, the probability of their rejection by the body is excluded. Courses of therapy are perfectly combined with almost all other methods that currently exist in cosmetology.

Indications for the procedure

The introduction of cloned regenerating cells is indicated for people aged 40 years. However, this technique can be applied at earlier stages. In addition, it is worth remembering that the saturation of the skin with fibroblasts is also carried out in order to correct minor scars or defects.

The technology of introducing repair cells is recommended for people:

• with pronounced signs of aging;
• middle-aged (for the prevention of withering of the skin);
• with various kinds of defecation (scars, pockmarks, burns, etc.);
• wishing to start the formation of fibroblasts in order to improve and maintain tone.

Patients who have indications for rehabilitation measures after cosmetic procedures (peeling, polishing, plastic surgery), fibroblast rejuvenation may also be indicated. Feedback on this procedure suggests that collection of samples for cell expansion is best done at a younger age, when their ability to regenerate is highest.

The principle of operation of the introduced cells

Morphological studies of the dermis artificially saturated with fibroblasts testify to the extraordinary productivity of such technologies. Soon after the injection, the newly acquired cells are fixed in small groups. This is due to the dosed introduction of biological material, which is characterized by weak diffuse properties.

Synthesized substances begin to be observed inside the intercellular fine-grained substance, which is a direct consequence of active restoration work. Characteristic features last up to 18 months, after which the fibroblasts are completely integrated into the structure of the skin and become no more active than all its components.

After these processes, active cells can be introduced again according to an individually selected scheme. As a rule, the effect of the repeated procedure differs in a brighter result, since the regenerative processes in the skin are already running.

Benefits of restorative biotechnologies

Fibroblasts embedded in the skin retain their activity for at least one and a half years. The necessary proteins are produced in the dermis, resulting in natural renewal cells. The intensity of the rejuvenating effect during the entire period of action is parabolic, increasing and then gradually fading away. By the end of the period, the activity of the implanted cells begins to correspond as much as possible to the real age of the patient.

Signs of correction of age-related and other changes make up the following list:

• the number of folds and the depth of old scars are significantly reduced;
• skin tone is evened out, its elasticity returns;
• the regenerative abilities of cells are obviously enhanced;
• there is a pronounced rejuvenation.

Fibroblasts are cells responsible for the freshness of the skin and, ultimately, for the beauty of a person. Constituting the framework of the dermis among other elements, they produce and organize various components, maintaining its necessary physiological state.

• active stage of an infectious disease;
• the presence of malignant tumors;
• dysfunction of the immune system;
• rashes and other defects not associated with the action of infection.

In addition, this therapy is contraindicated in pregnancy and breastfeeding.

Fibroblast injections are quite a productive base for other procedures, the purpose of which is to restore the microstructure of the skin and correct its defects. The extensive practice of applying biological rejuvenation technologies shows that the effect of each cosmetic product applied to the SPRS therapy procedure is significantly enhanced.