RU2755347C1 - Composition for the correction of age-related skin changes and the method for its preparation - Google Patents

Abstract

FIELD: cosmetology.SUBSTANCE: The composition for the correction of age-related skin changes includes a stromal-vascular fraction (SVF) 5-20% by volume, platelet autologous plasma (PRP) 5-75% by volume, 0.1–3% aqueous solution of hyaluronic acid 20-75% by volume. A method for obtaining a composition for correcting age-related skin changes and a method for correcting age-related skin changes are also disclosed.EFFECT: group of inventions provides a high potential for stimulating the regeneration of connective and cartilaginous tissues.3 cl, 1 ex

Description

The field of technology.

The invention relates to cosmetology and is intended for the correction of age-related skin changes.

State of the art.

In recent years, regenerative medicine has become an independent, rapidly developing discipline. Developments are being carried out in a variety of areas of medicine. In cosmetology, attention is also paid to preparations containing endogenous stem cells and other components that stimulate the restoration and rejuvenation of tissues.

So, from the prior art there is known a composition containing platelet-rich plasma and hyaluronic acid, and is intended to stimulate wound healing and use for cosmetic purposes by mixing with a cream and applying to the skin (patent RU 2667964, 11.03.2011).

It is known to use a biopolymer that can be injected subcutaneously or intradermally in the form of a monophasic hydrogel containing a gel obtained from crosslinked hyaluronic acid and one of its physiologically acceptable salts. The biopolymer is used to fill in wrinkles and stimulate epidermal cells and / or maintain the mechanical properties of skin firmness and elasticity. (patent RU 2448740, 06.12.2007).

It is known to use platelet-rich plasma for the preparation of a preparation for intradermal administration to stimulate reparative and trophic processes in the skin (patent RU 2470677, 19.10.2011).

Also known is a method of obtaining a composition containing stem cells derived from adipose tissue (ASC), primary culture (P0), grown ex-vivo, mixed with selected adipose tissue. for use as an agent for stem cell-enriched breast lipofilling or breast stem cell filing, cosmetic method for stem cell-enriched breast lipofilling or breast stem cell filing, cosmetic method for stem cell-enriched face lipofilling or facial stem cell filing and cosmetic method of administration agent into the skin. The invention provides correction of volumetric defects (patent 2660561, 06/26/2014)

The disadvantage of the known drugs designed to stimulate regenerative processes in tissues is that they are one- or two-component compositions with a limited set of stimulating factors.

The objective of the present invention is to develop a composition intended for the correction of age-related changes in the skin, containing a wider range of components stimulating regeneration, to develop a method for its production and use.

To solve this problem, we propose a composition for correcting age-related skin changes, including stromal-vascular fraction (SVF) 5-20% by volume, autologous platelet plasma (PRP) 5-75% by volume, 0.1-3% aqueous solution of hyaluronic acids 20-75% by volume.

We also propose a method of obtaining the claimed composition for correcting age-related changes in the skin, which consists in the fact that the stromal-vascular fraction (SVF) is preliminarily isolated from the patient's adipose tissue, autologous platelet plasma (PRP) is obtained from the patient's blood, then into a sterile disposable syringe No. 1 with a threaded needle tip, place the SVF, put the PRP into a sterile # 2 syringe with a threaded needle tip; syringe no. 3 with a threaded nozzle for a needle is filled with a 0.1-3% aqueous solution of hyaluronic acid, syringe no. 1 is connected in series to a four-end connector with a threaded connection with syringes through nozzles; syringe # 2; syringe # 3; and an empty syringe No. 4, the contents of syringes No. 1 and No. 3 are transferred into the syringe No. 4, then the liquids are alternately moved from the syringe No. 2 to the syringe No. 4 in both directions, after which the final product is kept in the syringe No. 4.

We also propose a method for correcting age-related changes in the skin using the proposed composition, including injections of the composition subcutaneously, papularly or bolus, the procedures are carried out once with an interval of 3-9 months.

The technical result of the claimed invention is to obtain a composition that has a high potential for stimulating the regeneration of connective and cartilaginous tissue and can be used in a wide range of medical and cosmetic procedures.

This effect is achieved through the use in the composition of the stromal-vascular fraction (SVF), autologous platelet plasma (PRP), hyaluronic acid solution.

SVF is a cell complex that includes mesenchymal stem cells (MSC), endothelial and smooth muscle cells of blood vessels and their precursors, pericytes, and fibroblasts. The main component of SVF are MSCs capable of self-renewal and multipotent differentiation. The mechanism of action of SVF is based on a heterogeneous cell population, leading to stimulation of cell differentiation, angiogenesis, immunomodulatory, anti-apoptotic effects and, accordingly, the restoration of damaged cells and tissues. The regenerative effect of SVF, realized by multipotent stem and progenitor cells, which are generally similar to bone marrow MSCs in morphology and immunophenotype, is associated with their ability to migrate to the damaged areas. The paracrine mechanism of action of MSCs is the main one.

PRP contains powerful factors that promote regeneration: PDGF (platelet-derived growth factor) - initiates the regeneration process, including the healing of bones and blood vessels, activates macrophages; VEGF (vascular endothelial growth factor) has marked angiogenic, mitogenic and vascular permeability-improving properties. It is known that TGF (transforming growth factor beta) promotes cellular mitosis and differentiation for tissue and bone fusion, affects mesenchymal stem cells, osteoblastic progenitor cells and fibroblasts, and inhibits the formation of osteoclasts. It is known that EGF (epidermal growth factor) induces epithelial development and promotes angiogenesis.

Hyaluronic acid (HA) injectable gels are widely used for cosmetic purposes, for filling or replacing biological tissues, as well as for rehydrating the skin using mesotherapy. Implants based on hyaluronic acid are used to replace the synovial fluid in the joints. Hyaluronic acid is a biodegradable biopolymer, hyaluronic acid monomers have active polar ends, which allows them to retain molecules of biologically active substances with which they interact through chemical bonds. Biodegradation of large-sized hyaluronic acid occurs under the action of the human body enzyme hyaluronidase, which separates monomers from the ends of polymer chains, the greater the mass of the large size, the longer the chain and the longer the biodegradation and decomposition of the polymer takes place. During the enzymatic decomposition of polymers, biologically active substances are released, including signaling molecules, which were retained due to chemical interaction after mixing with active factors from autologous platelet plasma and stromal-vascular fraction. The retention and prolonged entry into the tissue of biologically active factors and signaling molecules allows to significantly prolong the effect of the drug, at least for the period of resorption of hyaluronic acid. For an additional prolongation of the therapeutic effect, cross-linked hyaluronic acid is used, due to the cross-linking of large-sized ones into three-dimensional forms, hyaluronidase splits off monomers more slowly, while the biodegradation period increases, and the therapeutic effect is prolonged.

Experimentally, the authors have selected such ratios of these components, which provide the most pronounced effect.

In addition to the high efficiency of the proposed technique, it should be noted the simplicity and safety of the method for creating a biological product in the presence of a patient, since all active components intended for use in the composition of an autologous implant are obtained from the patient himself, the mixing of the constituent parts of the final product occurs under aseptic conditions thanks to the mixing technology using a cruciform connector with threaded adapters for syringes, with a luer-lock type outlet cannula, excluding uncontrolled disconnection and damage to components.

The essence of the invention.

The preparation of the claimed composition includes the following steps:

Stage 1: obtaining SVF. In the patient under local anesthesia, adipose tissue is taken from the lower third of the abdominal wall by liposuction. The resulting lipoaspirate is treated with collagenase for 15 minutes, then the resulting suspension is centrifuged at 600g for 5 minutes, the fat ring and supernatant are removed, and the stromal cell sediment is resuspended in saline so that the concentration of MSCs is about 5-7 million cells / ml. The specified suspension is filled with a syringe No. 1 - a syringe with an output cannula of the luer-lock type.

Stage 2: obtaining autologous platelet blood plasma involves drawing venous blood into a syringe with a pre-drawn anticoagulant. The syringe is closed, the contents are mixed and injected into a test tube. Centrifugation is carried out at room temperature with a rotation speed of 1800 rpm for 10 minutes. The supernatant layer of platelet-rich plasma is collected in a syringe No. 2 - a syringe with an exit cannula of the luer-lock type.

Stage 3. Preparation and heating in a thermostat to 37 ° C of a syringe pre-filled with a 0.1-3% aqueous solution of cross-linked hyaluronic acid (for example, the REVI trademark, manufactured by CLC International, Tyumen) - syringe No. 3 - a syringe with an exit cannula of the luer-lock type.

Stage 4 - connecting syringes 1-3 and an empty syringe No. 4 - a syringe with a luer-lock outlet cannula through a cross-shaped connector. For mixing, a four-end connector with a threaded connection is used, the connector is connected in series: syringe no. 1 with SVF; syringe # 2 with PRP; syringe No. 3 with HA; syringe No. 4 sterile with a threaded attachment for a luer-lock needle as a storage of the final product (CP). The connector is made of a material that allows the transportation of infusion solutions and products obtained from blood plasma.

Stage 5 - transferring the contents of syringes No. 1-3 to syringe No. 4 to obtain the final product. The movement is carried out in the following sequence: the syringe No. 4 is transferred to the set position, while a vacuum is created inside the syringe, the syringes No. 1 and No. 3 are transferred to the injection pressure position, while the process of fluid overflow from the area of increased pressure of syringes No. 1 and No. 3 to the area of reduced pressure occurs. pressure of syringe No. 4. After complete emptying of syringes No. 1 and No. 3, the liquid from them is in syringe No. 4. Then there is an alternating 10-fold movement of liquids from syringe No. 2 to syringe No. 4 in both directions. The final product remains complete and sufficiently mixed for 5 minutes in syringe # 4 in a thermostat at a temperature close to normal body temperature of 37 ° C.

The method of application makes it possible to use various injection techniques. Injections can be made in a papular and / or bolus manner. With the papular method of administration, the syringe needle is inserted into the skin at an angle of 30-40 (to a depth of about 1-2 mm, a micropapule is formed (its diameter is about 1 mm), then the next injection is made in the same way at a distance of 5-7 mm from the previous one. one puncture is injected with 0.01-0.02 ml of the drug. Injections are carried out along the wrinkles or along the lines of skin tension. With the bolus method of administration, the needle is immersed in the skin to the level of the periosteum, the drug is injected in a volume of 0.05-0.1 ml. are carried out along the lines of skin tension.

The procedures are performed once with an interval of 3-9 months.

The invention is used in cosmetology for the treatment of age-related changes in the skin of the face, décolleté and hands.

Implementation of the invention.

Example 1: Patient K.N., a 57-year-old woman, lean build, pronounced mimic wrinkles on the forehead, noticeable mimic wrinkles in the lateral corner of the eyes like crow's feet. Dry thinned skin with normal pigmentation. On the anterior surface of the neck, there are two transverse wrinkles over the entire surface, 6 mm wide and about 1 mm deep. In the decollete area, the skin is even, normally pigmented, dry, thinned. The skin of the hands is thinned, the patient complains of constant dryness of the skin of the hands. A woman has menopause for about 7 years, there are no chronic diseases, the hormonal profile corresponds to age, the lifestyle is mobile, and is engaged in fitness. He categorically refuses to use preparations containing botulinum toxin.

Before the procedure, the patient was photographed with a digital video camera, the level of hydration in various areas of the skin of the face and hands was measured by corneometry, and a U3 scan of the skin was performed.

Prescribed injections of the composition according to the claimed invention for the correction of facial wrinkles of the face and neck, containing 0.5 ml of SVF fraction; 1 ml of PRP and 1 ml of 0.1% aqueous solution of hyaluronic acid cross-linked trade mark REVI manufactured by CLC International, Tyumen. The drug was injected into the wrinkles on the forehead in a reciprocating manner, as if lifting the bottom of the wrinkles outward, into the area around the eyes and temples along the skin tension lines with a small papule distance between the papules of 5-7 mm. For one puncture, 0.01-0.02 ml of the drug is injected. To correct age-related changes in the skin of the hands and décolleté, injections of the composition according to the invention are prescribed in the form of a composition containing 1 ml of SVF; 2 ml of PRP and 1 ml of hyaluronic acid of the REVI trademark, injections of 0.07 ml were made bolus with an interval of 2 cm between injections along the lines of skin tension.

An examination was carried out 3 weeks after the procedure. The patient rated the result as "amazing", the frontal and mimic wrinkles were visually reduced. Objectively: the depth of wrinkles on the neck decreased, the number of wrinkles around the eyes decreased according to the results of photography, the level of hydration in various areas of the facial skin increased by 10-20%, on the skin of the hands the increase in the level of hydration was 30%. U3-scanning of the skin showed an increase in the echogenicity of the dermal layer, uniformity of signal distribution, which indicates the restoration of the structure of collagen and elastin fibers.

Example 2. Patient M., 43, Complaints about pronounced horizontal wrinkles on the forehead.

Prescribed injections are a composition according to the claimed invention containing 5% by volume of the SVF fraction; 75% by volume of PRP and 20% by volume of 1% aqueous solution of hyaluronic acid of the cross-linked trade mark REVI manufactured by CLC International, Tyumen. The drug was injected into the wrinkles on the forehead in a reciprocating manner, as if lifting the bottom of the wrinkles outward, with a distance between the injections of 5-7 mm. For one puncture, 0.01-0.02 ml of the drug is injected.

An examination was carried out 3 weeks after the procedure.

To objectify the results of exposure, an ultrasound study was carried out. Comparative images taken on the DUB SkinSkanner ultrasound device before and after the procedure clearly showed the leveling (smoothing) of the skin surface and an increase in the echo density of the dermal layer of the skin directly under the wrinkles.

Example 3. Patient A. woman, 48 years old. She complained of widespread occurrence of facial wrinkles, decreased skin elasticity, and dryness. On the anterior surface of the neck, there are two transverse wrinkles along the entire width with a depth of about 1 mm.

Prescribed injections of the composition according to the claimed invention for the correction of facial wrinkles of the face and neck, containing 20% by volume of the SVF fraction; 5% by volume of PRP and 75% by volume of a 3% aqueous solution of hyaluronic acid of the cross-linked trade mark REVI manufactured by CLC International, Tyumen. The drug was injected into the skin of the forehead, in the area around the eyes and temples along the lines of tension of the skin, in small papules with a distance between the papules of 5-7 mm. For one puncture, 0.01-0.02 ml of the drug was injected.

An examination was carried out 3 weeks after the procedure. The depth of wrinkles on the neck decreased, the number of wrinkles around the eyes decreased according to the results of photography, the level of hydration in various areas of the facial skin increased by 10-15%. U3-scanning of the skin showed an increase in the echogenicity of the dermal layer, uniformity of signal distribution, which indicates the restoration of the structure of collagen and elastin fibers.

The claimed composition was used to correct wrinkles in 15 patients. After the course of injections, an increase in skin moisture and elasticity, leveling of its relief was observed, which was confirmed both visually and by instrumental methods of skin examination.

Claims (3)

1. Composition for the correction of age-related skin changes, including stromal-vascular fraction (SVF) 5-20% by volume, autologous platelet plasma (PRP) 5-75% by volume, 0.1-3% aqueous solution of hyaluronic acid 20-75 % by volume. 2. A method of obtaining a composition for correcting age-related changes in the skin according to claim 1, comprising the fact that the stromal-vascular fraction (SVF) is preliminarily isolated from the patient's adipose tissue, autologous platelet plasma (PRP) is obtained from the patient's blood, then into a sterile disposable syringe # 1 with threaded needle bar is placed SVF, PRP is added to sterile # 2 syringe with threaded needle bar; syringe no. 3 with a threaded nozzle for a needle is filled with a 0.1–3% aqueous solution of hyaluronic acid; syringe no. 1 is connected in series to the four-end connector with a threaded connection with syringes through nozzles; syringe number 2; syringe number 3; and an empty syringe no. 4, the contents of syringes no. 1 and no. 3 are transferred into syringe no. 4, then the liquids are alternately moved from syringe no. 2 to syringe no. 4 in both directions, after which the final product is kept in syringe no. 4. 3. A method for correcting age-related changes in the skin using the composition according to claim 1, characterized in that the composition is injected subcutaneously, papularly or bolus, while performing one procedure at intervals of 3-9 months.