Hydrating gel (bioscaffold) containing exosomes derived from serum, plasma, and blood platelets to cleanse and rejuvenate the skin.

Abstract

] Improving wrinkles, reconstructing and removing skin scars is one of the problems and needs of society. In this regard, exosomes, which contain proteins and growth factors related to cell proliferation and reconstruction and do not carry risks such as transplant rejection and cancer, are of interest. One of the problems with previous treatments with cell-derived exosomes, such as stem cells, is that harmful agents such as antibiotics and compounds such as choline chloride, sodium hypoxanthine salt, thymidine, putrescine dihydrochloride, iron nitrate, and L-glutamine required for cell culture as cosmetic and health raw materials are harmful and prohibited. The present invention overcomes the disadvantages of previous methods by directly accessing serum, plasma, and platelet-derived exosomes through ultracentrifuge methods and laboratory kits containing buffers such as phosphate saline, which are consistent with physiological conditions without the need for cell culture. In this invention, transferring exosomes to a substrate of polymers such as carbomer, which is water-soluble and has no toxic effect on the skin, allows the contents of the exosomes to be preserved for a longer period of time and to be properly transferred to the epidermis of the skin.]

Description

In the name of God Description of the invention Title of the invention Hydrating gel (bioscaffold) containing exosomes derived from serum, plasma, and blood platelets to cleanse and rejuvenate the skin. Technical background of the relevant invention This invention relates to medical-pharmaceutical and cosmetic products based on exosomes that can rejuvenate and repair skin tissue and, unlike most cell-based products, eliminate transplant rejection reactions and severe allergies. Technical problem and statement of invention objectives Aging of the human body is inevitable with age and is first observed in the skin, which is the largest and most superficial organ of the human body. The reasons for this can be attributed to endogenous factors (heredity and gene expression) as well as exogenous factors (various effects of skin exposure to the external environment). The effects of both endogenous and exogenous factors cause skin aging in terms of appearance or phenotype, function and structure. Sunlight, which is an important external factor and the UVB rays contained in it, plays a fundamental role in wrinkles caused by skin aging. This type of skin aging, also called photoaging, appears in the form of wrinkles, skin roughness and irregular spots. Histologically, uneven thickening of the epidermis or the outermost surface of the skin, disruption of the skin capillary network, decreased type I collagen content in the skin, relative increase in type III collagen content, increased reticular fibers, denaturation of elastic fibers and accumulation appear in skin aging. Furthermore, with age, the balance between collagen synthesis and degradation, which is crucial for maintaining youthfulness and skin regeneration, changes.Type I collagen and type III collagen constitute the largest amount of total skin collagen, respectively. The balance of collagen maintenance over time is decreasing for type I collagen and slightly increasing for type III collagen. Various anti-aging treatments, especially cosmetics, peels and needling, which are used to improve cell function to stimulate collagen and elastin synthesis, are usually short-lived, and the various chemical products used in these products may have side effects in the long term. Even herbal ingredients used in the manufacture of cosmetic products, due to their low solubility compared to chemical compounds, limit the widespread use of these products in treatments. In recent years, research and treatment of skin aging have progressed, and the use of stem cells has become more widespread. With the development of stem cell transplantation for similar treatments such as clinical wounds, shortcomings in their use have gradually become apparent.For example, stem cells must be transplanted within a short period of time after extraction, because the necessary conditions for their survival are not present, and their long-term use and storage cause them to differentiate into different cell lineages, which is inconvenient and harmful for clinical use. Transplantation of stem cells from allogeneic or xenogeneic sources may also cause graft rejection and immune system sensitivity, and increase the risk of viral infection for the host. In addition, relevant studies have shown that direct differentiation and proliferation of stem cells for wound healing does not improve conditions and reduce clinical symptoms. In contrast, the main mechanism through which progress in damaged tissue repair is made is the paracrine secretion of a number of active substances by the damaged tissue, of which exosomes are an important component. Exosomes have a similar function to stem cell transplantation in wound repair and treatment, except that they do not cause immune rejection and graft rejection.Exosomes, which are membrane (phospholipid) vesicles with a size of about 30-150 nm, are biocompatible because they are derived from cells and their uptake rate into cells is also very good. Exosomes also have potential for diagnostic-therapeutic applications due to the long-term preservation of their contents, stable physiological properties, wide range of sources and large-scale extraction. They mainly contain proteins, microRNA, RNA, DNA and lipids. However, few studies have reported the effects of exosomes on skin aging. Transdermal products are discrete dosage forms that deliver drugs through the skin at a controlled rate to epidermal and dermal cells. In the past, skin rejuvenation and regeneration by releasing some drugs at the skin surface was associated with a decrease in the intensity of the drug's effect in the long term and the occurrence of allergic side effects. An essential prerequisite for the development of this mechanism is that the drug must be able to cross the skin quickly enough to reach therapeutic concentrations.However, the outermost layer of the skin, or stratum corneum, forms a major barrier to most exogenous substances, including large-sized drugs. This barrier is due to the skin's sebum, pores, and enzymes that are present to combat infectious and external agents. The discovery of this method is a major advance in the field of targeted delivery systems for skin regenerative and rejuvenating structures and is non-invasive compared to methods such as Botox, fat injections, and other similar methods that are based on periodic injections with many side effects. Therefore, there is no need for a separate process of encapsulating the culture fluid on liposomes, as is the case with the conventional method, and it has the advantage of being easily applied to the skin. Invention objectives Improving the duration of the effect of these products compared to previous methods and the ability to use them for a long time. These products do not cause transplant rejection reactions, unlike methods such as cell therapy. These products do not cause severe allergic reactions due to the immune system's tolerance for these vesicles based on their surface markers, which are derived from the individual's own cells. High absorption power and low toxicity of the products, due to their nanometric size and passage through the epidermis layer as a delivery system for cell growth factors to help regenerate the skin and remove scars and wrinkles. Producing products with antioxidant agents naturally enclosed in exosomes that reduce free radicals and protect the skin from sunlight and other damaging factors. Creating scaffolds in products to maintain the integrity and activity of exosomes and prevent their structural disruption. This invention will be a suitable alternative to common cosmetic and invasive methods for treating eczema, spots and freckles, sagging skin and wrinkles, and improving skin scars, and a new step towards increasing the level of health and well-being of societies. Description of the state of prior art and history of developments related to the invention [The following describes several inventions using exosomes: In the patent number CN110869033A published in 2020, a cosmetic-hygienic composition for preventing, suppressing, reducing, improving or treating itching is provided, which comprises stem cell-derived exosomes as an active ingredient. Due to the effect on various cytokine targets in this composition (e.g., IL-4, IL-31, and TSLP) that cause itching, the compositions of this invention are therefore useful for a wide range of itching caused by various causes and can be effectively used. Patent No. CN114306194A, published in 2022, discloses a cosmetic composition as a freckle remover containing umbilical cord blood extract, a freckle removal cream, and a method for preparing the freckle removal cream. In this product, it is explained that exosomes extracted from pomegranate, which have strong effects on removing freckles and are non-irritating and have high product safety, are used as a synergistic role. Patent No. KR101663912B1, published in 2016, describes a cosmetic composition for skin whitening, wrinkle improvement or regeneration, which contains exosomes extracted from stem cells as an active ingredient. The exosomes according to this composition contain genes and proteins related to cell proliferation, differentiation and regeneration of stem cells and can be used as a scar treatment for cosmetic purposes. Patent number WO2019231133A1, published in 2018, provides a cosmetic composition for reducing pores, comprising stem cell-derived exosomes as an active ingredient using this composition. In the patent number CN111148520A published in 2020, a cosmetic composition for preventing, reducing or treating dermatitis is provided, which comprises adipose stem cell-derived exosome as an active ingredient. The composition can be applied to a wide range of dermatitis caused by various etiologies by acting on the targets of various cytokines that cause dermatitis, and can effectively inhibit or reduce dermatitis. A 2016 study by Shiqi Hu showed that human fibroblast cells, which make up most of the dermis, lose their ability to produce collagen with age and the aging process. In this study, they showed that exosomes derived from bone marrow mesenchymal stem cells increased the expression of procollagen type I and were used to treat skin aging. In 2021, Shi showed that platelet-derived exosomes enriched with fibrin gel induced chronic wound healing and complete skin regeneration in rabbit ears.] Providing a solution to an existing technical problem [As previously mentioned, one of the problems associated with the use of cell-based therapies is the provision of sufficient cells and the toxic effects of culture media. In the case of stem cells, cells are often extracted from tissues such as bone marrow and fat, which are obtained by invasive methods and repeated sampling is often problematic. In the present invention, serum, plasma and platelet samples obtained by non-invasive methods and the possibility of sampling from each individual repeatedly have been used. Another problem associated with cell therapy is the unknown side effects of the treatment, including the uncertain fate of the cells in the body, and in this invention, in order to enhance the safety of the treatment, engineered vesicles (exosomes) from platelet cells, serum and plasma have been used instead of the cells themselves.] Accurate, sufficient and integrated description of the invention According to the embodiment of the present invention, exosomes not only carry genetic information, proteins and cell growth factors, but the exosome itself can also act as a carrier. The cosmetic composition of the present invention can also be a skin lotion, a skin softener, a skin toner, an astringent, a milk lotion, a nourishing lotion, a massage cream, a nourishing cream, a moisturizing cream, a hand cream, a foundation, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion, a body cleanser, a serum, an ointment and a gel, and may even be in the form of a conventional cosmetic composition not limited to a specific formula. In addition, additives are not limited and additives common in the field of cosmetics may be added thereto. Examples of common additives in the cosmetic field include binders, disintegrants, diluents, lubricants, stabilizers, preservatives, fragrances, oils, water, surfactants, humectants, lower carbon number alcohols, thickeners, pigments, and preservatives, and the like.The pharmaceutical compositions of the present invention may also be used in the form of pharmaceutically acceptable salts and may be used alone or in combination with other pharmaceutically active ingredients as well as in a suitable combination. The salt is not limited as long as it is pharmaceutically acceptable and includes hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid, acetic acid, tartaric acid. It is also possible to induce skin regeneration without other additives such as various factors. Since the exosome is a purified component that does not contain harmful factors such as antibiotics, serum and culture medium, it can overcome the problems of cosmetic products and works well due to its cell-derived and phospholipid nature in cellular penetration and effective efficiency in factor delivery. Accordingly, the present invention can be used for functional cosmetic-hygienic compositions for the purposes of skin whitening, improvement or reconstruction of wrinkles, collagen formation, treatment of eczema and spots, sagging, stratum corneum water retention, and scar treatments.Also, the products of this invention are not derived from stem cells, and the problems of cell culture and the contents of the culture media that were previously explained have been eliminated in this method. Obtaining exosomes is very fast and is obtained with high concentration and excellent quality. Finally, the exosomes extracted from serum, plasma and blood platelet samples were combined with cosmetic and hygienic ingredients and examined after creating skin scratches on Wistar rats with diabetic ulcers and also on a human model with acne and skin rashes, wrinkles and sagging skin. The method of obtaining these vesicles, the steps of synthesizing the desired products, as well as examining the quality of the products and measuring their effect are described below: - Sample preparation, extraction and confirmation of exosomes Initially, 2 cc of blood is collected in a tube containing 0.2 ml of sodium citrate, which also keeps platelets active, and centrifuged for 20 minutes at 3000 rpm to separate the plasma containing platelets. To separate the serum, 2 cc of blood is poured into a glass laboratory tube and centrifuged for 8 minutes at 2800 rpm. After centrifugation, the supernatants are removed and stored at -20°C. Now, in order to extract exosomes from samples obtained from biological kits such as (ExoCIB-AnaCell, Iran), we proceed according to the manufacturer's protocol below: 1. Centrifuge platelet-containing plasma samples for 10 minutes at 3000 rpm to remove contamination and excess material. 2. Vortex and heat solution A from the kit at 37°C, which contains polyethylene glycol (PEG) weighting agent to precipitate exosomes. 3. Add the plasma supernatant from the first step in a ratio of five to one to solution A (50 μL plasma sample + 250 μL solution A) and vortex the tube for 5 minutes for better mixing. 4. Incubate the tube from the previous step for 12 hours at 4°C (for better results, we shook the tube every few hours) 5. Vortex the samples from the previous step for 1 minute and then centrifuge for 10 minutes at 3000 rpm and 4°C. 6. Remove the supernatant and add 100 microliters of solution B containing phosphate buffered saline (PBS) to the pellet at the bottom of the tube. 7. Storage of extracted exosomes at -20°C The resulting exosomes may be at a concentration of 1 to 100 μg / mL, specifically at a concentration of 1 to 90 μg / mL, but this ratio is changed and not limited by adding different amounts of the centrifuge exosomal sediment dissolving solution. The size, shape and percentage of exosomes are evaluated using transmission electron microscopy (TEM) and dynamic light scattering (DLS) measurement methods. For electron microscopy analysis, 5 μL of plasma-extracted exosomes suspension should be diluted with 10 μL (PBS) and fixed with 1% glutaraldehyde. After that, 10 μL of the desired diluted sample is placed on carbon-coated grids at room temperature. The grids should be washed twice with sterile PBS at 5 min intervals. Finally, staining with 1% uranyl acetate is performed for 5 min and the samples are examined using a transmission electron microscope such as the company (Carl-Zeiss, USA) at 80 kV. Dynamic light scattering measurement method is also used to determine the size and percentage of exosomes.For this purpose, 20 microliters of the suspension of exosomes extracted from plasma are diluted with 100 microliters of PBS in a ratio of 1 to 5 and after preparation, the assay is performed with the relevant device such as the company (100-Z-Horiba-SZ, Japan). Figure 1 shows the electron microscope examination of the extracted exosomes, which is part of the supplementary information of this invention. Figure 2 also shows the concentration and size of the extracted exosomes using the DLS technique. -How to make products and formulate some cosmetic ingredients Formulation and preparation of hydrating gel Pour 40 ml of double distilled water into a sterile container and heat to 70°C. Slowly and with stirring, add 4% glycerin by weight and 2% carbopol-940 by weight to it and bring to a volume of 45. Stir with a propeller stirrer and slow speed until a uniform mixture is formed and all the polymer dissolves and a uniform opaque liquid is obtained. Now, place the resulting mixture at room temperature and sterile conditions for about half an hour to equilibrate with the environment. While stirring, slowly add 0.1% triethanolamine by weight to obtain a consistent and uniform mixture. Now, place the resulting mixture at room temperature and sterile conditions for about half an hour to equilibrate with the environment. Measure the pH value, which should be in the range of 5-5.5. Now we add exosomes at a rate of 1% by weight along with essential oil and preservative to a uniform mixture and store the contents of the container containing the exosomes at 4°C to achieve greater consistency, and then package them in airtight containers that prevent sunlight. How to use [Take a small amount of the product introduced with your index finger, which has been previously washed with soap and water, and gently massage it on the cheeks and forehead from the bottom to the top of the cheek and also between the eyebrows to the sides. For the area around and under the eyes, massage from one centimeter below the eye outwards to near the ears. On the chin, massage from under the lower lip to the end of the chin. In all the above steps, massage should be done gently to enhance blood circulation. The best time to use is in the morning after waking up and at night before going to bed. Before using the product, wash your face and body thoroughly with lukewarm water and a neutral shampoo. Note that it should not be used on the eyelids and under the eyebrows.] Proof of exosomes being active after 20 days from product preparation One of the most important things to consider is the activity of exosomes in products. The ultimate goal of this invention is to deliver growth factors and skin regenerative and scar repair stimuli to the epidermal and dermal layers of the skin. Therefore, examining whether the synthesized product is active in terms of the desired factors can be done in various ways. One of the most important ways is to examine the amount of macromolecules carried by these vesicles (exosomes). For this purpose, we need to extract RNA from creams, lotions, gels, and other cosmetic products. After extraction, by examining the absorption of this biomacromolecule in the sample containing exosome after 20 days of product preparation compared to the control sample without exosome, we can determine the existence and activity of the product. But why did we choose RNA among the macromolecules?!! In terms of stability, RNA is vulnerable compared to other macromolecules at ambient temperature due to the free OH group at carbon number 2 of the ribose sugar, and it quickly becomes unstable and decomposes.Therefore, the proof of the presence of RNA indicates that this biomacromolecule is protected in exosomes and the final morphology is preserved. The steps for extracting RNA from exosomes and examining its concentration at a wavelength of 260 / 280 nm, which is the absorption range of nucleic acids, are given below: The extraction process was performed using the RNX-PLUS kit (SinaClon, Iran). According to the manufacturer's protocol, first, 1 mL of cold RNX-PLUS solution was mixed with 100 μL of the exosome suspension contained in the cosmetic-hygiene product in 2 mL microtubes. Then, vortex for 10 seconds to mix them, and then incubate on ice for 5 minutes. Then, 200 μL of chloroform was added to the previous mixture, and after 5 minutes of re-incubation on ice, centrifuged for 15 minutes at 12,000 rpm and 4°C. After centrifugation, the supernatant was added to new 2 mL microtubes containing 600 μL of cold ethanol (Merck, Germany) and incubated on ice for 15 minutes. After that, centrifuge for 15 minutes at 12000 rpm and 4°C. Then, discard the supernatant and add 1 ml of 75% ethanol to the microtubes and centrifuge for 8 minutes at 7500 rpm and 4°C.Then the supernatant in the tube is discarded and the plate in the tube is dried for 10 minutes at room temperature. Finally, the plate is dissolved in 40 μl of RNase and DNase-free water (BioGene, Iran). Finally, the concentration and purity of the extracted RNA are checked using Nanodrop in Figure 4. Proving the existence of biologically active factors related to wrinkle improvement in exosomes After extracting RNA and examining its concentration using the Nanodrop device, it is necessary to examine biological factors related to wrinkles, which are examined by the Real Time PCR technique and gene expression examination. Important biological factors include important ones such as Platelet-derived growth factor (PDGF) and EGF (Epidermal Growth Factor). For this purpose, the expression level of these two factors in products containing exosome is examined. The steps of this examination and the expression graphs related to it are mentioned below. In this invention, Real-time PCR by the Cybergreen method, which is the most common and least expensive method for examining gene expression, is used to measure the expression of PDGF and EGF derived from exosomes using the device (ABI STEP ONE real-time PCR system Applied Biosystems-USA). To synthesize cDNA using the Pars Toos kit, we proceed according to the manufacturer's protocol. After that, the synthesized cDNA is stored at -20°C until the Real-time PCR reaction is performed. To perform the real-time reaction, 6.5 μL of Amplicon's ROX-containing master mix, 4.25 μL of nuclease-free water, 0.5 μL of forward primer and 0.5 μL of reverse primer, and 1 μL of cDNA are placed in the special strips of the device, for a total volume of 13 μL, and inserted into the device. The device's time program is a step of 95°C for 5 minutes, followed by 40 steps of 95°C for 5 seconds and 64°C for 30 seconds, and in the final step, a step of 72°C for 5 minutes. After the reaction, the final fold change is calculated and an expression graph is drawn using Graphpad software and is shown in Figure 5. Investigating the effect of topical gel containing exosome on skin scars caused by diabetic wounds in diabetic rats with heart disease using histological examination and gene expression analysis In order to investigate the effect of exosomes on improving collagen synthesis and repairing skin lesions, laboratory studies are needed. For this purpose, one of the best laboratory models is diabetic rats that have skin scars and wounds. In this model, animals with scars are divided into two treatment groups with topical gel containing exosome and a control group that receives topical gel without exosome. The product is used daily and massaged once in the scar area to a size of 1 cm. On day 21, a sample is taken from the scar area and sent to the laboratory in paraffin form. Pathological examination with hematoxylin and eosin staining in the two groups is shown in Figure 6. In addition, the gene expression of collagen type I and type III along with elastin in these two groups is shown in Figure 7 with a significant difference of 0.01. Explanation of shapes, maps and diagrams 1. Figure 1 shows exosomes derived from serum, plasma and platelet samples by transmission electron microscopy. White arrows indicate exosomes, scale bar = 100 nm. The spherical morphology of the exosomes indicates that their membrane has maintained its integrity and cohesion during the separation processes and the spherical structure has not been damaged. Also, in this image, the diameter of the exosomes is 100 nm or less, indicating that contamination with other things such as apoptotic bodies is minimal. 2. Figure 2 shows the size of exosomes examined using the DLS technique. According to this image, more than 80% of the extracted vesicles have a size of about 100 nm, which indicates the presence of extracted exosomes. 3. Image number 3 shows a laboratory model made of a hydrating gel containing exosomes. 4. Figure 4 shows the concentration graph of RNA extracted from exosomes using the Nanodrop device in two groups: control and exosomes. According to the image, in the cosmetic-health product group, we have an absorption peak in the range of 260 / 280 nm, which was obtained with a concentration of 157.7 ng / μl. In the control group, which did not have any exosomes, no absorption peak was observed and the concentration of the measured macromolecules was reported as a negative number, in fact, without concentration. 5. Figure 5 shows the expression graph of exosome-derived PDGF and EGF genes. 6. Figure 6 shows H&E staining for histological examination of the scarred area before and after treatment with topical gel containing exosome. According to Figure 6, in the areas indicated by white arrows, the depth of the scar in the groups receiving topical gel containing exosome is reduced. Also, the marginal areas indicated by black arrows are healing compared to the control group, which had a deeper scar area, and also the marginal areas that had less healing compared to the treatment group. 7. Figure 7 shows the expression graph of Collagen I, III, and Elastin genes in the control and treatment groups. The amount of collagen types I and III was significantly increased in the treatment group, and elastin in the treatment group increased significantly compared to the control, but was not significant. A clear and precise statement of the advantages of the claimed invention over prior inventions. 1. Reducing costs compared to foreign products and even many domestic products and the ability to earn foreign exchange for the country. 2. Reduction of side effects due to the absence of harmful factors in the culture medium, antibiotics, and carcinogenic chemical compounds. 3. Biodegradability and lack of reactions such as graft rejection and immune sensitivities compared to cell-based products. 4. Possibility of personalization for each individual according to their serum, plasma, and platelet samples and better adaptation of each product to the physiological conditions of the individuals. 5. Increased duration of effect compared to other products and the possibility of long-term use. 6. Effective on a wide range of skin problems such as wrinkles, scars, eczema, acne and rashes, and other similar issues. 7. Relatively quick response after using the products due to their dynamism compared to purely chemical products. 8. Increasing the synergistic effect of the compounds used in these products, which helps to achieve faster effectiveness. Description of at least one implementation method for implementing the invention The product is produced in ready-made, waterproof packaging that is sterilized with 70% ethanol and can be used at home and outdoors. To produce the invention in the industrial phase, we can follow the following steps: Providing sterile conditions in the laboratory environment, such as isolation rooms or clean rooms, to isolate exosomes. Preparing some laboratory equipment such as centrifuges, nanodrops, and other equipment needed in different stages of work. Need for equipment and raw materials to make lotions, creams, tonics, masks, scrubs, etc. The need for appropriate equipment for final packaging and preventing contamination at various stages. Explicit mention of the industrial application of the invention and how it will be produced in the industrial phase. This product can be used in industrial, commercial and medical phase production by packaging it in waterproof packages and can be used in homes and beauty clinics. Also, this invention can be used in the medical, pharmaceutical and cosmetic industries for auxiliary and beauty treatment. According to the study, this invention provides evidence that exosomes derived from serum, plasma and blood platelets can be used in improving skin lesions and collagen production. These biomolecules, given that they are carriers of growth and tissue repair factors and unlike stem cell transplantation and injection that may transmit toxic properties from the culture medium and serums, do not have toxic effects and have a more convenient preparation process. It should be noted that due to the lipid and bilayer nature of these vesicles, the need for encapsulation and carriers similar to many nanoproducts is not felt in these bags. In addition, they themselves can also act as a platform for drug delivery.Finally, according to the studies and investigations conducted, this invention can be a new way to improve skin lesions, wrinkles, eczema, rejuvenate and remove scars, help improve blemishes and acne, reduce pores, help maintain cell water and maintain elasticity, brighten and whiten skin color, and improve wounds from diseases such as diabetes. For production in the industrial phase, a clean room or isolated rooms with a hood are required to separate exosomes. Other required equipment such as centrifuges, Erlenmeyer flasks and beakers, dry baths, mixers, refrigerators and samplers are also required. It should be noted that the large-scale preparation process is the same as mentioned in the formulation section. This means that for production in higher volumes, each of the required materials increases by the amount of production. It should be noted that the steps in this section are exactly the same as the production method in the formulation section.

Claims

Claims What is claimed: Claim 1) Hydrating gel (bioscaffold) containing exosomes derived from serum, plasma and blood platelets for cleansing and rejuvenating the skin, which is obtained by dissolving 4% glycerin by weight and 2% carbopol-940 by weight in 40 ml of double-distilled water and bringing to a volume of 45 ml at a temperature of 70 degrees Celsius. To obtain the final product, 0.1% triethanolamine by weight is used at ambient temperature and sterile conditions to obtain a mixture with consistency and uniformity, and changes in the stated values ​​​​take the elasticity and consistency out of optimum conditions. Claim 2) According to claim number 1, each particle of 2% by weight of Carbopol-940 biopolymer exhibits a high concentration property and swells ten times its initial volume, creating the necessary porosity for exosome placement.

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