The surface of the skin into which the cells are injected is treated with sterilizing solutions before performing the procedures, and all the manipulations are performer strictly in sterile rubber gloves and in a gauze bandage covering the respiratory organs.
The results of the application of autogenous fibroblasts
According to one of the leading experts in the application of autologous fibroblasts, the PhD of biological Sciences A. A. Zgursky, the procedure has several attractive aspects for cosmetologist and, accordingly, for the patient (Zgursky, 2004):
it uses the patients own cells;
a single biopsy is usually performed;
there is only a slight discomfort in the process of applying procedures for the patient;
the effect lasts for a long time;
easiness of application;
a small number of procedures are performed to obtain the effect;
there are no complications and side effects.
Autologous fibroblasts are used for the correction of wrinkles, atrophic scars, atrophic areas of the skin, disorders of skin structure (Boss et. al., 2000).
The application advantages are the following:
it slows down skin aging and improves the color of the skin;
deep wrinkles are smoothed out and small wrinkles disappear;
it improves firmness and elasticity of the skin connective tissue;
there is a significant effect in scar correction and nasolabial folds in 7 and 12 months.
During the first hours after administration of cells local effects in the form of hyperemia (local increase in blood flow) and a small edema are observed, disappearing 23 hours after the procedure. A positive effect is seen 68 hours after the procedure, resulting in increased skin turgor and the elasticity of the skin, as well as its blood supply. There was a positive effect, that consisted of the disappearance of small wrinkles and reduce large one (Kricheli et al., 2004). Skin improvement occurs within a few months. The positive effect observed for 12 months. An example of the use of autogenous fibroblasts for the treatment of nasolabial folds is shown in figures 10a, 10b. Figure 11 shows an example to improving the surface of the skin near the eye: fig. 11 a the area of the eye before the procedure, fig. 11 b a few months after the application of therapy. It can be seen that the number and depth of wrinkles after the procedures are significantly decreased. Fig. 12 shows the result of treatment of frontal wrinkles.
Some experts came to the conclusion that an anti-wrinkle treatment using autologous fibroblasts is the least laborious and time consuming and most effective method of correction of wrinkles in clinical practice.
This method appears to be the only one that effectively removes scar changes after acne (Fig. 13). According to Dr. Arnold Klein (United States), the effectiveness of this method reaches 95%.
The basic properties of fibroblasts are preserved by cultivating them outside the body. These cell:
retain the diploid karyotype during the whole life;
have a limited lifespan;
possess the property of contact inhibition of divisions and require attachment to the substrate (bottom of the culture flask) for proliferation;
have no oncogenic potency, i.e. there is a safety of autologous fibroblasts (Keller et al., 2000), there are no tumors in athymik (non-immune) mice after 2 months after the administration of 40 million cells;
have normal contractility as referred to collagen;
maintain in vitro an active synthesis of type I collagen after in vitro cultivation for 6 passages.
Specialists in one of USA firms claims that they have already executed thousands of similar procedures, and thousands of patients are treated. With the help of similar techniques in cosmetology, it is possible to correct age-related changes in the skin, wrinkles, skin scars. The use of persons own cells is justified by the fact that there is no rejection reaction, often arising from the use of another persons cells, both of embryonic origin and even more so, when received from adult donors. In addition, there are no ethical problems arising from the use of embryonic cells.
Examples of application of techniques are also shown in other figures. Thus, in figures 14a, 14b, 14c shows the face of
a patient with nasolabial fold (wrinkle) before the procedure with autologous fibroblasts (a), during cell therapy (b) and several months after it (c). It can be clearly seen that wrinkles are significantly reduced (smoothed) and almost disappeared. This was due to the fact that the administered autologous fibroblasts successfully adapted and began to produce new, young collagen and elastin.
In the fig. 15a one can see part of the patients face before the procedure of autologous fibroblasts and in fig. 15b 46 months after the therapy. The arrow shows a defect on the skin before and after therapy. A defect is a scar on the surface of the cheek after removal of warts (Watson et al., 1999).
The corrections of other such senile changes in the skin are also possible (Weiss et al., 2007).
CRYOSTORAGES
After the use of cell therapy, some cells may be still not utilized. These cells should be preserved in order to repeat the procedure in the distant future.
To preserve your own cells, you need to place them in a special cryostorages. It is a duralumin or steel container, made like a type of thermos, with double walls, between which a vacuum is created. It is filled with liquid nitrogen having a temperature of minus 1960C (Figures 16a, 16b).
Before freezing, the cells are washed from the growth medium and kept in a protective solution that prevents them from damage at low temperature.
Cell suspension is poured into special sterile plastic vials, closed hermetically. These ampoules with cells inside are first subjected to gradual smooth cooling to minus 70800С, after which they are immersed in liquid nitrogen in the cryostorage, where they remain for many years and decades. Each of the cryovials is assigned a unique identification number, which is entered into a computer database. This number is strictly consistent with the person from whom the cells were taken. The database contains the following data: name and surname of the person, date of birth, blood characteristics, information about hereditary diseases, the date of taking a biopsy, some other information.
The database exists not only in the computer, but the information is also duplicated on CDs or USB flash drives which are stored in a dedicated place. This eliminates the loss of information. The cryopreservation itself is constantly under the supervision of the personnel on duty, it is excluded from the access of unauthorized persons, and all manipulations with containers are made only by carefully trained personnel who are personally responsible for the safety of cellular material.