Fibroblasts

Skin regenerator potential

SPRS therapy® – the only technology approved in Russia for the correction of age-related skin changes based on the use of its own cultured fibroblasts.

The technology is protected by Russian and international patents. The main technology "Use of autologous dermal fibroblasts for the correction of age and scar skin defects" has a permission of Roszdravnadzor for use (FS No. 2009/308) issued to the Human Stem Cell Institute in July 2010 - the only organization with official permission to use this technology in Russia.

The processes that develop in the skin during aging are based on fundamental changes associated with the main cell population of the dermis - fibroblasts: their number, morphology, proliferative potential, functional activity.

They not only support the homeostasis of the intercellular matrix of the dermis, providing its remodeling and renewal (due to the degradation of "depleted" and the synthesis of new components of the dermis - collagen, elastin and the main substance), but also play a significant role in maintaining the physiological state of other layers of the skin, representing a key link in its biology.

Study by J. Varani et al. [2006] showed that in old people, compared with young people, the total number of fibroblasts in the skin is 35% less on average. According to G. Fisher et al. [2002], collagen production in the skin of old people is reduced by an average of 75%. A natural consequence of these processes is a decrease in the thickness of the skin, a decrease in its elasticity, the formation of wrinkles.

Therefore, the process of age-related skin changes comes down to a decrease in the population of fibroblasts and a decrease in their proliferative and biosynthetic activity, which naturally manifests itself in a decrease in the quantitative and qualitative composition of the intercellular matrix of the dermis. In this regard, it becomes quite clear that dermal fibroblasts are the main point of application of therapeutic effects in the correction of age-related skin changes.

The purpose of applying a number of methods of modern cosmetology (mesotherapy, biorevitalization, peels, fractional photothermolysis, radio wave therapy, dermabrasion, etc.) is to stimulate the functional activity of dermal fibroblasts. And this goal is being realized, including through damage to the skin tissue.

Before performing a traumatic procedure, an important question must be answered that will determine the success of exposure and minimize the risks of adverse events. Namely: what intensity will be most optimal for the skin of a particular person in terms of achieving the desired effect and safety?

This question can be answered by determining the regenerative and proliferative capabilities of the main cell population of the skin - fibroblasts. To date, the Institute of Human Stem Cells (ISCH) has developed the "Passport of Skin ®" technology (based on the method of cloning stromal progenitor cells developed by A. Ya. Fridenstein et al. in 1970), which makes it possible to assess the effectiveness of colony formation of skin fibroblasts (ECOf) in vitro and the degree of proliferation of cell colonies.

The technology protected by both Russian and international patents makes it possible to determine with a high degree of reliability the regenerative and proliferative potentials of the population of human skin fibroblasts, on the basis of which it is possible to conclude on the regenerative potential of the skin of a particular patient.

Fibroblasts are known to be a heterogeneous cell population comprising all fibroblastic differon from multipotent mesenchymal stem cell, progenitor cells (fibroblast progenitor cells), and differentiated (mature) fibroblasts to finely differentiated fibrocyte.

This means that stem/progenitor cells are present in the dermis of each person, the main task of which is to maintain the cell population of tissue. These cells can be detected in culture by observing the formation of colonies - clones consisting of 50 or more cells, each representing offspring of a single stem/progenitor cell. For this, clonal analysis is used - a specific method for determining the effectiveness of colony formation of fibroblasts (ECOf), which consists in the following.

A primary fibroblast culture is obtained from a 4 mm biopsy of the patient's skin in a specialized HSCI laboratory, from which a strictly determined number of cells are taken (arbitrarily). These cells are explanted into petri dishes and cultured under strictly defined (and always identical) conditions. After 14 days, using a special computer program, the formed colonies are counted and the value of ECOf is determined, which is the ratio of grown colonies (cell clones consisting of 50 or more cells) to the number of explanted cells.

The ECOf value, as shown by the results of clonal analysis of dermal fibroblast cultures, does not depend on age - this is an individual parameter for each person. ECOf is characterized by rather high values, averaging 45,0±9,5%. By determining the value of ECOf and counting it on the mass of the biopsy, one can get an idea of ​ ​ the content of stem/progenitor cells in the skin of each person. Thanks to the functioning of the latter, dermis recovery/remodeling is carried out. Based on ECOf values, a conclusion can be made about the individual regenerative potential of the dermis, that is, the ability of a population of skin fibroblasts of a particular person to maintain tissue homeostasis and restore it if damaged.

Moreover, the higher the ECOf, the more fibroblast progenitor cells in the skin, which means more differentiated (mature) functionally active fibroblasts, therefore, the regenerative potential of the tissue is higher; conversely, the less ECOf, the lower the regenerative potential of the skin tissue. Regenerative potential correlates with the ability of tissue to repair after injury. It is genetically determined and independent of age.

Along with investigating the regenerative capabilities of a patient's dermal fibroblasts, it is important for a beautician to know what their proliferative potential is - the number of divisions that cells can pass before they die. It is determined by percentage ratio of dense, diffuse and mixed colonies in skin fibroblast culture of patients

Obviously, the higher the proliferative potential of the cells constituting this tissue, the more mitotically active cells in it and, therefore, the faster the regenerative processes will be carried out.

It is known that the progenitor cells of the dermis (according to the results of the study of their cytomorphology, proliferative potential and ability to synthesize specific cytokines/growth factors) are divided into three types of cell populations: MFI, MF2 and MF3 (MF - mitotically active fibroblasts). In this case, the MF1 cell pool has the highest proliferative potential, its cells pass about 25-30 cell divisions before differentiation into the MF1 cell population. In turn, cells of the MFII pool perform about 15-20 divisions before switching to the MF3 pool, and cells of the MF3 pool perform only about 5-8 divisions before differentiation into PMF (postmitotic fibroblasts). Accordingly, in culture, mitotically active fibroblasts form three kinds of colonies

1. dense, consisting of hundreds and even thousands of fibroblasts (which seems to correspond to the differentiation stage of MA1);
2. diffuse, consisting of 50-100 cells (which apparently corresponds to the differentiation stage of MA3);
3. mixed, consisting of fibroblasts in the stages of differentiation of MA1 and MA3.

According to the quantitative ratio of cell colonies of different types in culture (using the formula derived by E. B. Vladimirskaya et al. for stromal progenitor cells) and conclude on the proliferative potential of skin fibroblasts of each patient. Moreover, the denser colonies in the culture, the higher the patient's proliferative potential of the dermal fibroblast population. And accordingly, the larger the diffuse colony culture, the lower the proliferative potential of the patient's skin fibroblast population.

Modern cosmetological methods that allow remodeling of the dermis microstructure can be divided into two groups:

1. the first includes technologies that stimulate resident (i.e. present in the dermis) fibroblasts, for example laser and radio wave technologies, injections of platelet-rich PRP plasma, etc.;
2. to the second - methods from the field of regenerative medicine, the main goal of which is to replenish the dermis cell population decreasing with age by introducing functionally active fibroblasts (SPRS- therapy).

Accordingly, the variety of modern methods of aesthetic correction poses a number of pressing questions for each doctor practicing in this field of medicine related to the implementation of certain procedures in a particular patient:

1. what procedures are shown?
2. how often should they be carried out?
3. what are the safe parameters of their use in a particular case (for example, hardware procedures - laser or radio waves)?
4. At what age can the patient can be recommended to carry out interventions for aesthetic indications? etc.

Today, in our opinion, these questions can be answered with greater accuracy if we take into account the individual indicators of the regenerative and proliferative potentials of the skin obtained using the original technology "Passport of Skin ®," based on the method of cloning fibroblasts.

For example, at low rates of these parameters, any cosmetic procedures should be carried out with a certain caution so as not to deplete an already small pool of fibroblast progenitors. These patients need to undergo SPRS therapy before any skin effects can be applied to fill the deficiency of functionally active dermal cells. At the same time, if the patient has high or normal regenerative/proliferative potentials, any cosmetological methods can be used (according to the instructions).

SPRS - therapy (use of autologous fibroblasts to correct age and scar changes in the skin) personalized complex of diagnostic and therapeutic procedures for skin repair based on application of the patient's own skin cells - fibroblasts. Fibroblasts are the main cells of the skin, on which its youth and beauty depend, since it is these cells that produce collagen, elastin, hyaluronic acid and other substances important for the skin.

With age, the number of functionally active fibroblasts in the skin decreases, their biosynthetic activity is impaired, i.e. the ability to produce collagen and elastic fibers - the main framework of the skin, as well as hyaluronic acid, which retains water in the skin. As a result, the thickness of the skin decreases, its hydration and elasticity decreases, wrinkles form.

The use of own skin fibroblasts grown in special laboratory conditions increases the number of these functionally active cells in those areas of the skin that require correction. In the process of cultivation, only functionally active fibroblasts are selected and stimulated, i.e. cells that have maintained a high ability to divide and biosynthesis. These cells, after introduction into the skin, actively synthesize collagen, elastin and other skin-important components. As a result, the thickness of the skin increases, its elasticity increases, the number and depth of wrinkles decreases. I.e. this medical technology allows not only to correct visual defects of the skin, but also to restore the skin at the level of its microstructure.

The doctor takes a biopsy sample from a patient from the ear area - a small piece of skin with a diameter of 4 mm. The wound after a biopsy does not require special care, it heals on its own. The biopsy is placed in a container with a special medium and delivered to a specialized laboratory of the ISC (Human Stem Cell Institute), where fibroblasts are isolated from it and increased to the required amount. Part of obtained cells is used for preparation of SPRS - preparation, and part is frozen according to special technology and stored in cryovessel in individual cell (Cryobank of patient's skin fibroblasts). At any time, the cells can be extracted from Cryobank and used to prepare SPRS, a drug for treating the skin of the face, neck, neckline, hands. Prepared SPRS - the drug is delivered to the clinic, where it is administered intradermally using mesotherapy needles. An anaesthetic cream is applied to the skin 30-40 minutes before the procedure.

Improvement in the skin condition (increase in its elasticity and thickness, decrease in the number and depth of wrinkles, improvement in color), patients begin to notice as early as 10-14 days after the procedure. The clinical effect has a gradually increasing and long-term effect, since the own, functionally active skin cells are used.

- age-related skin changes (skin thinning, decreased elasticity, presence of fine and medium wrinkles);

- rehabilitation after peels and plastic surgery

Age: 18 - 65

- acute and chronic (in the stage of exacerbation) skin diseases

- acute and chronic (in the stage of exacerbation) systemic diseases

- autoimmune diseases of connective tissue

- oncological diseases

SPRS therapy is the only method that allows you to fight not only the external manifestations of skin aging, but also the causes of aging at the level of its microstructure. The technology makes it possible to replenish the deficiency of fibroblasts that develops in the skin with age by introducing millions of young and active own fibroblasts into the skin, which, after introduction into the skin, fully take root and actively produce all substances important to the skin.

All other cosmetological methods only stimulate the skin fibroblasts, the number of which decreases significantly with age.