RU2682763C1 - Method of treating androgenetic alopecia - Google Patents

Abstract

FIELD: medicine.SUBSTANCE: invention relates to medicine, namely to dermatology, trichology and cosmetology. Blood is taken by venipuncture into a sterile vacuum tube with an anticoagulant at a ratio of 10:1. First centrifugation of the tube is carried out for at least 5 minutes at 1,800 rpm. Then the plasma, leukocyte layer and the upper part of the erythrocyte layer is taken from the tube with a syringe of 5 ml with a needle of not more than 18G. This mixture is transferred to another vacuum sterile tube, which is then centrifuged for at least 8 minutes at 2,500 rpm. After that, 2 ml of plasma is taken into a 2–3 ml injection syringe containing 0.1 ml of 10 % calcium chloride solution in the ratio of calcium chloride:plasma 1:20 with a needle of no more than 18G placed by the lower end of the cut into the plasma layer 1 mm above the leukocyte film. Subsequent manual suspension is performed by inverting the syringe at least 10 times. Obtained autologous activated, platelet-rich plasma by the linear retrograde technique is injected in the form of injections over the entire surface of alopecia at the level of the lower layers of the dermis, 0.15 ml per injection. Such injections are carried out 1 time a month for 4 months with the possibility of repeating a similar course of treatment in the event of a relapse.EFFECT: method provides a high and fast effective effect in terms of hair density, thickness of hair rods and intensity of hair loss with preservation of the clinical result up to 16 months.1 cl, 2 dwg, 1 tbl, 2 ex

Description

The invention relates to medicine, namely to dermatology, trichology, cosmetology, and may find application in the treatment of androgenetic alopecia.

Androgenetic alopecia (AHA) is the most common type of hair loss and affects up to 80% of men and 40% of women. A decrease in hair length and hair diameter, as well as a decrease in scalp hair density, is the result of a combination of changes in the hair growth cycle (premature onset of the catagen phase and a gradual decrease in the duration of the anagen phase) and follicular miniaturization. During the development of baldness, the relative contribution of these two components varies between patients.

The main role in the pathogenesis of AHA is played by the androgen-converting enzyme 5a-reductase, which catalyzes the conversion of testosterone to a more active form - 5 alpha-dihydrotestosterone. In the presence of dihydrotestosterone, androgen receptors undergo conformational changes, resulting in an “activated complex”. This hormone-receptor complex, after interacting with a nuclear acceptor, passes into the cell nucleus and binds to specific nuclear chromatin receptors. The expression of specific genes is activated, the global process of changing the primary morphology and physiology of the hair follicle, in which the androgenic effect is realized, is launched.

The result of the implementation of the pathogenetic mechanisms of AHA is a progressive thinning of the hair rods to a state of vellus and miniaturization of the hair follicles up to complete loss of hair in the androgen-dependent areas of the scalp.

Being a pronounced cosmetic defect, the AHA creates a number of social (limiting the choice of profession, device of personal life, etc.) and economic (high costs of treatment) problems. In addition, baldness affects the psycho-emotional state of a person, which significantly reduces the quality of life. Some authors believe that this process is a normal biological phenomenon, and not a disease. Regardless of how their condition is classified, people with AHA perceive it as a deviation from the norm, a phenomenon that is unacceptable from a psychosocial point of view, and, therefore, consult a doctor for treatment.

Despite the high prevalence of AHA, the first European recommendations for its treatment were developed only in 2011.

Minoxidil for external use is a first-line drug for the treatment of AHA in many countries and is included in the standard of care for patients with AHA in the Russian Federation. Minoxidil prolongs the stage of anagen (hair growth), increases the size of the hair follicle by activating potassium channels and prostaglandin endoperoxide synthase-1, which increases the level of prostaglandin E2 (PGE2). The drug has a number of significant drawbacks: the lack of treatment efficacy desired by the patient, the presence of side effects, the need for continuous use, a pronounced withdrawal symptom, and inefficiency in the case of the patient's minoxidil resistance.

For specific systemic treatment of androgenetic alopecia in men, the European Recommendations prescribe one of the strongest 5a-reductase inhibitors with proven efficacy in AHA, blocking the conversion of testosterone to dihydrotestosterone at the target organ level - finasteride at a dose of 1 mg per day. Antiandrogen therapy helps stop hair loss, improve hair quality. The disadvantages are the lack of restoration of hair density, the presence of contraindications and side effects. Finasteride is registered in the Russian Federation as a drug for use in urology and andrology in a dosage of 5 mg, finasteride (in a recommended dose of 1 mg) for the treatment of AHA in Russia is not used.

With severe baldness, the surgical correction is the method of choice, which consists in transplanting androgen-resistant follicles into the alopecia zone from the skin of the scalp not affected by baldness to completely or partially cover the bald areas of the head. Hair transplantation is a very successful treatment method, but involves many different aspects: the patient’s fear of surgery and the risks associated with it, its high cost, the need to continue using medications to prevent the loss of preserved androgen-sensitive hair. According to our observations, patients, especially those who are in the early stages of baldness, prefer conservative drug therapy over surgery.

The number of studies proving the positive effect of other drugs and methods is very small, as well as the amount of reliable data and evidence showing that they improve the condition of the hair with AGA or prevent its progression. Left untreated, baldness is steadily progressing. Therapy should be carried out for a long time, discontinuation of treatment leads to the resumption of hair loss. The most effective treatment of AHA is possible at an early stage of the disease (before replacing long hair with fluffy). Drug treatment has 2 goals, one of them is to increase the area of scalp hair growth in case it decreases with the course of AHA, the second is to stop the progressive thinning and / or hair loss.

The treatment of AHA at the present stage has several disadvantages associated with contraindications for use, side effects, the need for lifelong use of systemic and topical drugs, the absence of the patient's expected treatment results, patients' fear of surgical treatment and the risks associated with it, and to this day remains one of the relevant dermatology problems.

Currently, injection therapy is used in the complex treatment of AHA. Platelet-rich plasma therapy (Platelet Rich Plasma, PRP) is being discussed as one of these methods.

The present invention relates to solving the problem of AHA using PRP.

PRP is an autologous plasma with a higher platelet concentration than in whole blood, obtained by centrifuging the patient’s own blood, during which the separation of blood cells occurs due to their difference in weight. The clinical and molecular biological effects of PRP have been studied since the 1970s, and the method has been used in many areas of medicine, such as traumatology and orthopedics, maxillofacial surgery, plastic surgery, and dermatology.

Until 2011, only a few publications on the use of PRP in the treatment of alopecia related to pilot studies could be found in the literature. Since 2011, the number of studies, including randomized, placebo-controlled, during which data were obtained confirming the positive effect of PRP injections on hair growth and their morphometric parameters in AGA.

The main clinical effect obtained during the studies was manifested in a decrease in hair loss and an increase in hair density. However, as noted in the publications, the effect obtained largely depends on the method of preparing PRP, the number of procedures with its use and the interval between them. The methods for evaluating the results obtained are also excellent.

Traditionally, the PRP-therapy procedure consists of several stages: patient’s blood sampling, centrifugation, subsequent plasma activation and its introduction to the patient.

The key point at all stages of PRP preparation is the preservation of the integrity and viability of platelets, since blood centrifugation at high speeds is traumatic and platelets can undergo mechanical destruction. In the process of activation, platelets go through the stage of degranulation with the release of growth factors and other bioactive substances into the surrounding intercellular space. Most growth factors stored in platelets in an inactive state interact with certain structures on the cell membrane, as a result of which platelets transform into an active form and become bioavailable. If the platelet is grossly destroyed by inadequate preparation technology, then inactive substances enter the environment, which means that the expected clinical effect will not be. With an increase in centrifugation speed, the morphological changes and the reactivity of platelets, which characterize their ability to activate, decrease.

Platelets contain various growth factors, the main targets of which are blood vessel cells, fibroblasts, mesenchymal stem cells, "directed" to the differentiation pathway to replenish the pool of functionally active cells. The interrelation of the reparative and hemostatic functions of platelets ensures the formation of new skin tissue during wound healing and structural changes in the tissue, expressed in cell growth, cell differentiation, and growth of blood vessels, which are the main effects in response to injection of PRP.

The final stage in the preparation of the drug is the activation of platelets with the subsequent degranulation of bioactive substances. Under natural conditions, platelet activation occurs under the action of thrombin after they are attached to a fibrin clot. In many commercial PRP technologies used in trichology and cosmetology, the option of producing a fibrin matrix on which activated platelets are adsorbed (they, in turn, will serve as a source of growth factors and other bioactive substances) consists in adding calcium gluconate or calcium chloride to PRP. Calcium ions stimulate the release of own fibrin from platelets with all the ensuing consequences - the formation of a fibrin gel, platelet adsorption on it, their activation and degranulation. The physiological concentration of platelets in the blood is in the range of 150-300 thousand / μl of whole blood. In PRP, this concentration increases from 2 to 9 times, and this figure varies depending on the technology of preparation of the drug. It matters the initial volume of whole blood, single or double centrifugation of blood, the number of revolutions of the centrifuge, the initial platelet content in whole blood.

Currently, 4 basic options for preparing PRP are known in world medical practice.

1. Using a single centrifugation and involves the activation of platelets in a natural way under the action of thrombin with a low amount of released growth factors and other bioactive substances. In this case, the platelet count in the plasma rises no more than 2-2.5 times.

2. Using a single centrifugation with platelet activation by adding calcium ions, which implies a greater number of released growth factors and other bioactive substances than in the version of claim 1. The number of platelets in the plasma also increases by no more than 2-2.5 times.

3. Using double centrifugation, which increases the number of platelets by 5 or more times, and involves the activation of platelets in a natural way under the action of thrombin.

Medical devices for obtaining PRP according to the first three options are registered in the Russian Federation and are used in maxillofacial surgery, traumatology and cosmetology.

4. Using double centrifugation with activation by adding calcium ions.

According to the technical nature and the achieved result of the treatment of AHA using PRP obtained by this method, the closest to the proposed product is a medical device for the preparation of enriched plasma "GLO PRP", which we took as a prototype. A medical device for the possibility of obtaining PRP in this way (GLO PRP, Korea) is registered and approved for use in medical practice on the territory of the Republic of Kazakhstan, but is not available on the territory of the Russian Federation (Reg. Certificate RK-IMN-5 No. 011357 of 05/03/2013)

According to the prototype, PRP is prepared as follows: in a 10 ml syringe containing an anticoagulant - 3.8% sodium citrate solution 1.5 ml - by venipuncture and the use of a “butterfly needle”, the patient’s blood is taken in the amount of 8.5 ml, mixed . The ratio of anticoagulant: blood is approximately 1: 5.5. For centrifugation, a removable piston is provided in the same syringe. The first centrifugation is carried out at a rotation speed of 1800g (about 3200 rpm) for 3 minutes. Next, the erythrocyte layer is dumped vertically down into the container with a disinfectant solution by pressing on the piston to the border with the plasma, visually determined in the syringe. The second centrifugation is performed in the same syringe at the same speed - 3200 rpm - for 6 minutes. The PRP is transferred from the syringe directly to the syringe for injection using a vertically connecting special syringe coupling included in the kit. As PRP, 1, 2, 3 or 4 ml of the lower plasma layer with different platelet concentrations in each milliliter can be used - from a high 6-8 fold concentration in the first milliliter of PRP to a low 1.5-2 fold concentration in the fourth milliliter. The GLO PRP product involves only receiving plasma enriched with platelets, its further use - the purpose of use, activation of PRP, treatment regimen - remains at the discretion of the doctor without a specific protocol. The use of PRP prepared by this method is used in orthopedics, wound healing, cosmetology, ophthalmology. The developer's practical recommendations for the treatment of AHA show a papular technique for introducing PRP into the patient’s skin without specifying a treatment regimen. The technique consists in the formation of papules of various sizes in order to improve the deposition of medicinal substances, PRP is introduced to a depth of 2-3 mm, the angle of the needle is 45 degrees, the diameter of the papules is 2-4 mm, approximately 0.1 ml per injection. In the case of administration of 4 ml of PRP, about 40 injections are required.

Practical recommendations and clinical results of the use of “GLO PRP” in the treatment of AHA are clearly reflected and are available at http://www.glofinn.com/applications/hair-loss (Appendix 1).

The publication of PRP cooking is available at https://www.youtube.com/watch?v=XJrOrCm8h2w

The advantages of the prototype are the simplicity of the method of preparing enriched plasma, which is carried out in the same container (syringe) without plasma transfer between 1 and 2 centrifugation, which ensures minimal possibility of infection, as well as the possibility of obtaining a high concentration of platelets in PRP under the condition of platelet activation.

A significant disadvantage of the prototype is the centrifugation of blood and subsequently plasma with a centrifuge speed of more than 3000 rpm, as is known from other sources and proved by us, is traumatic for platelets and significantly reduces the number of viable cells, which reduces the therapeutic potential of PRP. The developer stated that the technique allows you to collect more than 60% of platelets. This amount is enough to obtain a clinical result, but subject to the use of platelet activation. Although the authors do not emphasize the possibility of activation, our repetition of this method of obtaining PRP, but with its activation confirmed a good clinical result in the treatment of AHA. Without activation, the effect was slightly lower. Nevertheless, we introduced plasma activation into the restrictive part of our formula as a common one with the prototype, since it was mentioned in other sources of the same developer. We believe that we have enough other differences from the prototype.

Currently, you can purchase GLO PRP KIT kits in the country of the developer company (Korea) or in the EU countries, there is no representative office in the Russian Federation. The cost of one syringe container is 50 euros, excluding delivery and payment of state duty. To carry out only one course of treatment of AHA with the help of plasma obtained in this way often 2 syringes are required. Obviously, obtaining PRP in one container is convenient, but given the high cost of such syringes and the need for repeated courses of treatment for AHA, their use significantly increases the cost of treatment and makes it inaccessible to most patients. In addition, the use of such a syringe, in our opinion, is not essential for obtaining the necessary and sufficient plasma quality, which provides an effective and safe treatment of AHA.

The technical result of the present invention consists in achieving a result of the treatment of AHA, not inferior to the effect obtained in other known methods of treating AHA, and above the effect of the prototype due to the use for treatment of plasma obtained under conditions ensuring the preservation of the necessary and sufficient number of viable platelets, as well as cheaper way.

This result is achieved in that in the known method for the treatment of AHA by intradermal administration of autologous blood plasma, obtained in a sterile container containing the patient’s blood and 3.8% sodium citrate solution as an anticoagulant, followed by double centrifugation and separation of platelet-rich plasma, according to of the invention, blood sampling is carried out by venipuncture in a vacuum sterile tube with an anticoagulant at a ratio of 10: 1, the first centrifugation of the tube is carried out for at least 5 minutes at 1800 rpm n, then plasma, leukocyte layer and 1-2 ml of the upper part of the erythrocyte layer are slowly taken from a 5 ml syringe with a needle of no more than 18G and this mixture is transferred to another sterile tube, which is then centrifuged for at least 8 minutes at 2500 rpm, then with a needle no more than 18G placed in the plasma layer 1-2 mm higher than the white blood cell film, 2 ml of plasma is taken into a 2-3 ml injection syringe containing 0.1 ml of a 10% solution of calcium chloride in the ratio of calcium chloride / plasma 1:20, followed by manual suspension by gentle replay syringes at least 10 times and the obtained autologous activated platelet-rich plasma, linear retrograde technique is injected over the entire surface of alopecia at the level of the lower layers of the dermis at 0.15 ml per injection to a depth of 2-4 mm and such injections are carried out 1 time per month for 4-6 months.

The first centrifugation at 1800 rpm. and the second - at 2500 rpm allows you to exclude the mechanical destruction of platelets and, most importantly, to maintain their viability. We have previously shown that centrifugation at speeds above 3000 is traumatic, destroying more than 20% of platelets and, accordingly, reducing their viability. It is known that for the high biological activity of the resulting plasma, it is important not only the number of platelets, which in itself cannot be a sufficient marker for predicting the biological activity of the plasma, their viability is also important.

The time of the first centrifugation for at least 5 minutes and the second - at least 8 minutes, as we have shown, is necessary and sufficient to maintain the viability of more than 80% of platelets.

Slow sampling from the test tube after its first centrifugation of the plasma, leukocyte layer and 1-2 ml of the upper part of the erythrocyte layer into another sterile tube with a syringe with a needle no more than 18G, and after the second centrifugation - the PRP layer located in the test tube above the leukocyte film, an injection syringe with a needle no more than 18G, placed 1-2 mm above this film, provides a plasma containing a high platelet content. At the same time, the selection of 1-2 ml of the upper part of the erythrocyte plasma layer for the second centrifugation, as we have shown, is mandatory, because after a soft back there is also a certain amount of platelets that add to the platelets from the plasma and increase their total number in PRP.

The use of no more than 18G needles in either case is necessary for the additional exclusion of vortices, as in the case of plasma retraction by a syringe with a needle of a narrower diameter, and in vitro mixing of the lower layer with PRP and the upper layer with a platelet-poor plasma. In the case of mixing, the upper platelet-poor portion of the plasma will “dilute” the PRP and the platelet count in the PRP will ultimately be reduced.

The intake of 2 ml of plasma in an injection syringe containing 0.1 ml of a 10% solution of calcium chloride in a ratio of 1 part calcium chloride to 20 parts of plasma, followed by manual suspension by gentle inversion of the tube at least 10 times, ensures uniform mixing of the activator and plasma and obtaining PRP suitable for intradermal administration to patients.

The ratio of activator and plasma 1:20 provides a sufficient amount of time (about 20 minutes) for the procedure to run until the cascade of coagulation processes starts, which makes the procedure impossible. Adding a larger amount of activator, for example, in a ratio of 1:10, leaves for medical procedures about 10 minutes, which may be insufficient due to coagulation in the syringe and obstruction of the plasma through the needle. The use of a linear retrograde introduction technique promotes a more uniform distribution of PRP in all skin layers due to the formation of a linear channel in the dermis (in the growth zone of the hair follicles) and its uniform filling in the reverse course of the needle. In addition, it is known that the use of linear retrograde technology can almost 1.5 times reduce the number of injections into the scalp due to the larger treatment area and more plasma per injection, which is significant from the point of view of the pain of the procedure.

The use of the linear retrograde PRP injection technique in our method of treating AHA in the area of alopecia at the level of the lower layers of the dermis to a depth of about 2-4 mm, in the volume of 0.15 ml per injection, 1 time per month for 4-6 months, we found experimentally and provide, along with its other advantages, a clinical result that exceeds the prototype and other known to us from a number of publications.

The essence of the method is as follows.

0.8 ml of 3.8% sodium citrate is added to a 9 ml sterile vacuum tube as an anticoagulant, 8 ml of the patient’s blood is taken into the same tube by venipuncture, the tube is inverted 6-8 times to mix the blood and anticoagulant. Next, the tube is placed in a centrifuge with a horizontal rotor and the first centrifugation is carried out for at least 5 minutes at a speed of 1800 rpm, using the "soft start" function. After the first centrifugation, all the obtained plasma located in the upper half of the tube, as well as the leukocyte and upper erythrocyte layers, are taken and transferred to another sterile vacuum tube without fillers, which is carried out with a 5 ml syringe with a needle of no more than 18G. This tube is placed in a centrifuge, and a second centrifugation is carried out at a speed of 2500 rpm for at least 8 minutes. As PRP, 2 ml of plasma located immediately above the white blood cell film is used. PRP sampling is carried out slowly, in order to avoid mixing the fractions, into the injection syringe, with a volume of 2.5-3 ml with a needle, a thickness of not more than 18G, placed 1-2 mm above the white blood cell film. Then, a 10% calcium chloride solution is added to this syringe as an activator at the rate of 0.1 ml per 2 ml of PRP and the mixture is manually suspended by gentle inversion of the tube at least 10 times.

PRP-therapy is carried out in a treatment room in compliance with all sanitary standards. After suspending, the 18G needle on the syringe is replaced with an injection needle 0.3 × 13 mm in size. The scalp is treated with a standard chlorhexidine solution. Injections are carried out in the field of alopecia with linear retrograde technique at the level of the lower layers of the dermis, a depth of about 2-4 mm, depending on the thickness of the skin, in a volume of 0.15 ml per injection. The procedures are carried out once a month, for a course of 4 procedures in the case of stage 1-3 of the AHA on the Norwood-Hamilton scale. For more severe stages, 6 treatments with the same interval are recommended.

Example 1. Patient S., a man of 40 years, stage V of the AHA, the duration of the disease is about 10 years. Examined to exclude endocrinopathies that affect hair growth. He began treatment in January 2017.

Blood sampling was carried out in 2 sterile vacuum tubes of 9 ml each, since the area of baldness was large enough, an anticoagulant was added to each tube - 3.8% sodium citrate in an amount of 0.9 ml, the tubes were turned upside down 6-8 times to mix blood and anticoagulant. The first centrifugation for 5 minutes was carried out at a speed of 1800 rpm, all the obtained plasma in the upper half of the tube, as well as the leukocyte and upper erythrocyte layers, were taken and transferred to other sterile vacuum tubes without fillers using a 5 ml syringe needle 18G. These tubes were placed in a centrifuge, and a second centrifugation was carried out at a speed of 2500 rpm for 10 minutes. As PRP, 2 ml of plasma was used from each tube located immediately above the white blood cell film formed above the erythrocyte layer.

PRP sampling was carried out slowly in syringes for injection, with a volume of 2.5-3 ml, a needle thickness of 18G, placed 1-2 mm above the white blood cell film. After that, 0.1 ml of a 10% solution of calcium chloride was added to each syringe as an activator, and the mixture was manually suspended by gentle inversion of the tubes 10 times. A total of 4 ml of PRP was obtained from the two tubes, 2 ml each.

The resulting autologous activated PRP was used within 15 minutes after activation. The scalp was treated with a standard chlorhexidine solution. Injections were performed in the area of alopecia (parietal and frontal zones) using linear retrograde technique, at the level of the lower layers of the dermis, depth 2-4 mm, depending on the thickness of the skin, in the amount of 0.15 ml per injection. In one procedure, 26 injections were performed. In total, 4 PRP procedures were performed with an interval of 1 month.

The results of therapy were evaluated 1 month after the last procedure, in June 2017. The evaluation method is phototrichograms obtained before and after the course of treatment (Fig. 1a and Fig. 1b), which are attached to the description and performed using a specialized computer program for evaluating the parameters hair growth TrichoSciencePro v1.3. As can be seen from the images, the patient’s hair density after the first course of treatment increased from 300 to 512 / cm2 (+ 70%), the proportion of vellus hair decreased from 33 to 24% (-27%), the intensity of hair loss decreased, which was reflected in a decrease in the proportion of telogen hair from 32 to 10% (-69%).

Conclusion: the treatment of AHA in a patient with severe baldness made it possible to obtain a high and quick effective effect in terms of hair density, hair shaft thickness and hair loss intensity. Phototrichogram data obtained 10 months after the start of treatment (November 2017) showed the stability of the result. The patient was recommended dynamic monitoring with a phototrichogram after another 6 months (in May 2018) and, if necessary, and the patient's desire to repeat the course of treatment.

Example 2. Patient M., 35 years old, stage 2 AHA, parietal zone, disease duration of about 2 years. Examined to exclude endocrinopathies that affect hair growth. Got a similar course of treatment.

After the first course of treatment from May to August 2015 and control in September 2017, the hair density changed from 501 to 546 per cm2 (+ 9%), the average hair diameter changed from 55 to 58 μm (+ 5.5%), the proportion of vellus and telogen hair remained unchanged, at 18%, which is acceptable in men.

Conclusion: treatment at the initial stage of AHA in a patient allowed to obtain a positive result on the density and diameter of hair with a prolonged effect. Slight positive dynamics after treatment is due to the initial stage of AHA and not pronounced deviations in the quality of the hair. The control measurement of morphometric parameters of hair growth was carried out a year after the start of treatment - in May 2016, another 10 months - in March 2017, hair growth indicators remained stable. In January 2018, there was a slight deterioration in the average hair diameter, which was regarded by us as a relapse. The patient is recommended a repeated course of 4 procedures with subsequent monitoring of the treatment result one month after the last procedure.

For comparison, we conducted a course of treatment for volunteers using PRP obtained by the method of the prototype on one half of the head, and the present invention on the other half. The increase in the amount of hair on half the head using PRP obtained by the proposed method was 15.4% higher than the increase in the amount of hair using PRP obtained by the prototype; the increase in the average diameter of all hair was 13.2% higher, terminal hair - 15.3% higher; the decrease in the proportion of telogen hair is 14.4% more intense than that of the prototype. The difference in the dynamics of the proportion of wellus hair was 2.3%, which was not statistically significant. These data are shown in table 1, indicating the advantages of our method for the treatment of AHA.

From 2013 to date, the proposed method has been treated in more than 100 men with AHA of 2-4 severity and disease duration from 1 to 12 years with a good clinical result that lasted from 4 to 16 months. Since AHA is a chronic progressive process with inevitable relapses after the end of any treatment, maintaining a clinical result for such a long time is a very good indicator of the prolongation of the effect.

To develop the proposed method, we were prompted by the well-known data on the possibility of using PRP therapy with a high positive result of the treatment of AHA. However, as noted above, at the present time it is possible to purchase GLO PRP KIT kits in the country of development (Korea) or in the countries of the European Union. The cost of one syringe is 50 euros. Since more than one syringe is required for a full course of treatment of AHA using PRP obtained using such syringes, the cost of treatment increases several times and amounts to a significant amount, which makes treatment inaccessible to most patients. This prompted us to develop a domestic method for obtaining PRP using medical devices available in any medical institution of this profile. As the examples of treatment of AHA using the devices (test tubes) widely used in medical practice show, we were able to create a method that is not inferior to or even superior to all known methods both in obtaining PRP and in the method of its administration.

We used standard VACUETTE sterile vacuum tubes (manufacturer Greiner Bio-One, Austria), available throughout the Russian Federation. The cost of one test tube for January 2018 is about 13 rubles. The cost of one procedure, subject to the use of two tubes, taking into account syringes, needles and other consumables, does not exceed 100 rubles.

In accordance with the foregoing, the proposed method in comparison with the known has several significant advantages, the main of which is that it is not inferior in effectiveness to other known methods of treating AHA, higher than the effectiveness of the prototype and at the same time is affordable for a wide range of patients.

The method was developed in LLC Trichological Center for Health and Hair Treatment (Russia, St. Petersburg) and has been clinically tested to date in more than 100 patients with a positive result.

Claims (1)

A method for the treatment of androgenetic alopecia by intradermal administration of autologous blood plasma obtained in a sterile container containing the patient’s blood and a 3.8% sodium citrate solution as an anticoagulant, followed by double centrifugation and separation of platelet-rich plasma, characterized in that the blood is taken by venipuncture in a sterile vacuum tube with an anticoagulant at a ratio of 10: 1, the tubes are first centrifuged for at least 5 minutes at 1800 rpm, then from the syringe tube Using a 5 ml needle with a needle of no more than 18G, the plasma, the leukocyte layer and the upper part of the erythrocyte layer are taken, this mixture is transferred to another sterile vacuum tube, which is then centrifuged for at least 8 minutes at 2500 rpm, after which the needle is placed no more than 18G the end of the cut into the plasma layer 1 mm higher than the white blood cell film, take 2 ml of plasma into a 2-3 ml injection syringe containing 0.1 ml of a 10% solution of calcium chloride in the ratio of calcium chloride: plasma 1:20, followed by manual suspension by flipping the syringe e less than 10 times, and the obtained autologous activated platelet-rich plasma by linear retrograde technique is administered as injections over the entire surface of alopecia at the level of the lower layers of the dermis of 0.15 ml per injection and such injections are carried out once a month for 4 months with the possibility of repeating a similar course of treatment in case of relapse.

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