Anti-hair-loss hair-growing microneedle, preparation method thereof and anti-hair-loss hair-growing essence

Abstract

The invention relates to the field of hair care agents, and particularly discloses an anti-hair-loss hair-growing microneedle, a preparation method thereof and anti-hair-loss hair-growing essence. The anti-hair-loss hair-growing microneedle comprises a microneedle substrate and an anti-hair-loss hair-growing component loaded in the microneedle substrate, and the raw material of the microneedle substrate is selected from at least one of chitosan, sodium hyaluronate, I-type and II-type collagen complexes, silk fibroin, ganoderan, sodium alginate and PLGA (poly (lactic-co-glycolic acid)); the hair loss preventing and hair growing component is prepared from the following raw materials: walnut shell vinegar liquid, black-silk fructus viticis, zinc-loaded eggshell membrane powder and lactic acid bacteria fermentation liquid in a mass ratio of 1: (0.7-1.2): (0.3-0.6): (0.1-0.5). The anti-hair-loss hair-growing microneedle can be used for preparing anti-hair-loss hair-growing essence and has the advantages of being painless, efficient, high in hair growing speed and good in hair growing effect.

Description

Technical Field

[0001] The application relates to the field of hair care technology, and more specifically, it relates to a hair loss prevention and hair growth microneedle, its preparation method, and a hair loss prevention and hair growth essence. Background Technology

[0002] Hair loss is a common and frequently occurring skin condition characterized by abnormal and excessive hair loss and thinning hair. With increasing social pressure, a faster pace of life, and unhealthy dietary habits, the proportion of people experiencing hair loss is rising year by year and is showing a trend towards affecting younger people. Hair loss not only affects a patient's appearance, social life, and employment, but also brings psychological burden and mental stress, seriously impacting quality of life and even leading to emotional problems such as sleep disorders, depression, and anxiety.

[0003] Hair loss includes alopecia areata, stress-related hair loss, endocrine-related hair loss, and androgenetic alopecia (seborrheic alopecia). Among these, more than 90% of hair loss cases are androgenetic alopecia. The main mechanism is that testosterone is converted into dihydrotestosterone (DHT) with higher biological activity by 5α-reductase. This competitively inhibits the binding of testosterone to its receptor. This process inhibits the metabolic process, prevents dormant hair follicles from entering the growth phase, and causes the hair follicles to gradually shrink until they disappear, resulting in hair loss.

[0004] In the treatment of hair loss, commonly used Western medicines include minoxidil and finasteride. However, both drugs are prone to relapse after discontinuation and can cause side effects such as skin irritation, scalp itching, hirsutism on the face and hands, dizziness, anorexia, and tachycardia. In addition, while hair transplantation has some therapeutic effect, its clinical application is limited by the large surgical trauma, high cost, and the need for multiple surgeries. Furthermore, non-drug treatments such as acupuncture are not ideal in their effectiveness.

[0005] Currently, there is an increasing number of studies on the use of plant extracts, such as traditional Chinese medicine, to treat hair loss. For example, Chinese invention patent application number CN2023115367031 discloses the preparation of a traditional Chinese medicine essential oil composition using Polygonum multiflorum, mulberry bark, soapberry, arborvitae leaf, Ligusticum chuanxiong, ginger, and camphor. Another example is Chinese invention patent application number CN2023101519486, which discloses the use of supercritical extraction to remove 6-gingerol from ginger, and then extracting alkaloids, polysaccharides, and flavonoids from ginger, arborvitae leaf, Drynaria fortunei, and coffee seeds to make a hair loss prevention and hair growth composition. Plant-derived ingredients are relatively safe, effective, and convenient to use, but treatment with plant-based ingredients alone has a long treatment cycle and slow onset of action. Summary of the Invention

[0006] In order to achieve better hair growth and hair regrowth effects and effectively reduce hair loss, this application provides a hair loss prevention and hair growth microneedle, its preparation method, and a hair loss prevention and hair growth essence.

[0007] Firstly, this application provides a microneedle for preventing hair loss and promoting hair growth, employing the following technical solution: A hair loss prevention and hair growth microneedle includes a microneedle matrix and hair loss prevention and hair growth ingredients loaded in the microneedle matrix. The raw material of the microneedle matrix is ​​selected from at least one of chitosan, sodium hyaluronate, type I and type II collagen complex, silk fibroin, Ganoderma lucidum polysaccharide, sodium alginate and PLGA. The ingredients for preventing hair loss and promoting hair growth include walnut shell vinegar, *Ulmus pumila*, zinc-loaded eggshell membrane powder, and lactic acid bacteria fermentation liquid in a mass ratio of 1:0.7-1.2:0.3-0.6:0.1-0.5.

[0008] By employing the above technical solution, since the outermost layer of the skin, the stratum corneum, is the main barrier for hair growth, the active ingredients in anti-hair loss and hair growth products can only remain on the epidermis and cannot reach the dermis where the hair follicles are located. However, the tips of microneedles can form micron-level channels on the skin surface, directly penetrating the stratum corneum and allowing the active ingredients to penetrate to the area around the hair follicles in the dermis, improving the scalp efficiency of the active ingredients and directly acting on atrophied or dormant hair follicles. Using chitosan, sodium hyaluronate, gelatin, etc., as the microneedle matrix satisfies the requirements of being biodegradable, biocompatible, and capable of loading active ingredients. It also meets the requirements of having a certain mechanical strength and supporting puncture; microneedles can gradually degrade in the body, and the anti-hair loss and hair growth ingredients are slowly released as the microneedle matrix degrades, rather than being released all at once. This slow release allows the anti-hair loss and hair growth ingredients to maintain a stable concentration around the hair follicles, avoiding the problem of rapid loss of ingredients and the need for frequent reapplication in traditional application, thus prolonging the nourishing time for the hair follicles. Moreover, the anti-hair loss and hair growth ingredients enter the deep layers of the skin through the microneedles, without accumulating in large quantities in the epidermal layer, avoiding prolonged contact with the skin and reducing the loss of ingredients caused by sweat, friction, etc.

[0009] The anti-hair loss and hair growth ingredients include walnut shell vinegar and *Ulva prostrata* (a type of herb). Walnut shell vinegar, obtained through high-temperature carbonization and distillation of walnut shells, is rich in active ingredients such as phenols, aldehydes, and organic acids. Phenolics stimulate the proliferation of dermal papilla cells in hair follicles, awakening dormant follicles and promoting their entry into the growth phase. Organic acids balance the pH of the scalp, inhibit the growth of harmful bacteria, reduce oil production and dandruff, and provide a healthy growth environment for hair follicles. *Ulva prostrata* contains flavonoids and saponins, which promote scalp blood circulation, provide nutrients, strengthen hair, reduce breakage and loss, and improve dry and frizzy hair, thus enhancing hair quality. Silver-loaded eggshell membrane contains nano-silver and eggshell membrane. The eggshell membrane contains collagen, hyaluronic acid, chondroitin sulfate, etc., while the silver ion loading can disrupt bacterial cell membranes. To achieve antibacterial effects, the natural components of eggshell membrane can regulate the balance of scalp flora. The dual antibacterial effect can reduce oily hair loss caused by flora imbalance from the source. Moreover, the collagen in eggshell membrane has a similar structure to hair protein, which can penetrate into the gaps between hair cuticles, fill the cuticle damage, and provide the amino acids needed for hair follicle repair, thus improving hair follicle atrophy. The lactic acid bacteria fermentation liquid contains small molecule peptides, polysaccharides, vitamins, organic acids, etc. Small molecule peptides and polysaccharides can promote the proliferation of scalp epidermal cells, enhance the integrity of the stratum corneum, reduce the damage of external stimuli to hair follicles, and improve hair loss. Organic acids and active peptides can inhibit scalp inflammatory factors and alleviate hair follicle damage caused by inflammation. In addition, the active ingredients in the fermentation liquid are small molecules, which are more easily penetrated into the hair follicles through microneedles. At the same time, the pH value of the fermentation liquid is close to that of the scalp, which can reduce irritation to the scalp.

[0010] Using microneedles as a delivery carrier for anti-hair loss and hair growth ingredients breaks through the barrier of the stratum corneum while avoiding the risks of injection. Multiple natural ingredients activate hair follicles, condition the scalp, and repair damage. Combining microneedle scalp technology with natural active ingredients solves the problems of poor scalp absorption, slow onset of action, and long treatment time of traditional hair growth products. It has the advantages of high-efficiency delivery, safety and gentleness, and targeted hair growth.

[0011] Optionally, the zinc-loaded eggshell membrane powder is prepared by the following method: The eggshell membrane was soaked in alkaline solution, washed with water, and then soaked in a mixture containing zinc gluconate, ε-polylysine and water. The mixture was then freeze-dried to obtain the carrier material. The carrier material was immersed in a treatment solution containing adipose-derived mesenchymal stem cell exosomes and a film-forming agent, then filtered, dried, and pulverized to obtain zinc-loaded eggshell membrane powder.

[0012] By adopting the above technical solution, using the eggshell membrane as a base, its collagen fiber network provides a good adhesion and load-bearing foundation for zinc gluconate and ε-polylysine. Moreover, it is rich in collagen, hyaluronic acid, etc., which are friendly to the scalp and have certain moisturizing and repair-promoting effects. ε-polylysine (ε-PL) has a significant inhibitory effect on the growth and reproduction of bacteria and fungi. ε-PL is a homopolymer amino acid linked by amide bonds formed by α-carboxyl ε-amino groups of lysine monomer molecules. It has the advantages of being safe and non-toxic, having broad-spectrum antibacterial properties, good water solubility, and good thermal stability. Zinc gluconate has the advantages of fast absorption rate, good safety, and high utilization rate. Using zinc gluconate and ε-polylysine as antibacterial agents can effectively enhance the antibacterial ability of the eggshell membrane, reduce scalp micro-inflammation and excessive microbial proliferation, create a healthy scalp, and reduce hair loss caused by inflammation such as folliculitis.

[0013] Eggshell membranes loaded with zinc gluconate and ε-polylysine were freeze-dried to obtain a highly porous carrier material with increased specific surface area. However, its mechanical strength was not as good as the original membrane. Therefore, it was soaked in a treatment solution containing a film-forming agent. During soaking, the carrier material could quickly absorb the treatment solution due to its porous structure. The film-forming agent and adipose-derived mesenchymal stem cell exosomes could be quickly absorbed into the porous structure of the carrier material or adhered to the surface. The filling and adhesion of the film-forming agent could improve the strength of the carrier structure. The adipose-derived mesenchymal stem cell exosomes could activate hair follicle stem cells, prolong the growth phase, stimulate the generation of new capillaries around the hair follicles, and increase the supply of nutrients to the hair follicles. Moreover, the anti-inflammatory factors in the exosomes could regulate the local immune response, inhibit the release of inflammatory cells and pro-inflammatory factors, reduce the inflammatory attack around the hair follicles, inhibit hair follicle cell apoptosis, and enhance their proliferation and metabolic capacity, thus repairing cell function from the inside and achieving hair loss prevention and hair growth. Adipose-derived mesenchymal stem cell exosomes were filled into eggshell membranes loaded with zinc gluconate and ε-polylysine, and then added to a microneedle matrix. After the microneedles were inserted into the skin, they gradually dissolved and released zinc-loaded eggshell membrane powder. The zinc-loaded eggshell powder released adipose-derived mesenchymal stem cell exosomes, zinc ions, and ε-polylysine around the hair follicles. Through the mechanisms of inhibiting inflammation, promoting angiogenesis, and activating hair follicle stem cells, it promoted hair follicle regeneration.

[0014] Optionally, in the zinc-loaded eggshell membrane, the mass ratio of eggshell membrane, zinc gluconate, and ε-polylysine is 1:0.01-0.05:0.02-0.1; The mass ratio of the carrier material, adipose-derived mesenchymal stem cell exosomes, and film-forming agent is 1:0.1-0.3:0.5-1.

[0015] By employing the above technical solution, the combined use of ε-polylysine and zinc gluconate improves the antibacterial and anti-inflammatory effects of eggshell membrane. ε-polylysine, a natural cationic polypeptide, can bind to anionic sebum on the scalp surface, helping to regulate scalp sebum secretion, reducing sebum blockage of hair follicles, and enhancing the efficacy of the ingredients. It also prevents active ingredients from becoming ineffective due to oxidation, extending shelf life and improving the overall effect. Zinc gluconate contains zinc, an essential trace element for hair follicle growth, which replenishes the scalp and hair follicles, activating resting hair follicles into the growth phase, promoting hair growth. Furthermore, by influencing the activity of 5α-reductase, it reduces the conversion rate of testosterone to dihydrotestosterone (DHT), inhibiting the effects of DHT on hair follicles, reducing follicle atrophy, promoting normal renewal of the scalp stratum corneum, enhancing the scalp barrier function, reducing irritation from the external environment or chemical components, and indirectly protecting hair follicle health.

[0016] Using the eggshell membrane as a carrier, zinc gluconate nourishes hair follicles, while ε-polylysine protects the scalp. This precise targeting prevents ingredient loss and provides dual protection and nourishment. ε-polylysine addresses scalp microecological issues such as bacteria and oiliness, zinc gluconate addresses hair follicle nutrition, and the eggshell membrane replenishes basic moisture. Together, these three ingredients target inflammatory hair loss, nutritional hair loss, and oily hair loss. Furthermore, the combination of the eggshell membrane with gentle zinc gluconate and ε-polylysine reduces the potential side effects of traditional anti-hair loss ingredients like minoxidil, such as scalp irritation and dryness, making it suitable for a wider range of skin types.

[0017] Optionally, the water in the blend and the treatment liquid is Yulongkash River water.

[0018] By adopting the above technical solution, the Yulongkash River contains a large amount of minerals such as Hetian jade, mainly from glacial meltwater, containing macroelements such as calcium and magnesium, as well as trace metals such as iron, manganese, nickel, and chromium, which can provide certain nutrients for hair follicles.

[0019] Optionally, the mass ratio of zinc gluconate to ε-polylysine is 1:1-2.

[0020] By adopting the above technical solution, zinc gluconate and ε-polylysine can be used in appropriate proportions to achieve synergistic compatibility, balance the synergistic effects of antibacterial and oil-controlling and nutritional repair, and avoid excessive use of a single component.

[0021] Optionally, the film-forming agent comprises xanthan gum, chitosan, and hyaluronic acid in a mass ratio of 1:0.5:0.5.

[0022] By adopting the above technical solution, xanthan gum, which is natural, mild, and breathable for the scalp, is used as the main material of the film-forming agent, and chitosan is added to give it antibacterial effect, while hyaluronic acid improves the moisturizing properties of the film-forming agent.

[0023] Optionally, the raw materials of the microneedle matrix include chitosan and sodium hyaluronate in a mass ratio of 6:4 to 7:3.

[0024] By adopting the above technical solution, sodium hyaluronate exhibits excellent water solubility, rapidly dissolving in the scalp environment to release the loaded components. It also possesses strong natural moisturizing ability, alleviating scalp dryness after microneedling. Furthermore, it exhibits high compatibility with scalp tissue and is non-irritating. Chitosan, with its high mechanical strength, enhances the puncture force of the microneedles, ensuring effective penetration of the scalp stratum corneum. It also possesses mild antibacterial properties, reducing the risk of infection after microneedling. Moreover, its good sustained-release effect prolongs the duration of action of the active ingredients on the scalp. Chitosan supplements the mechanical strength of sodium hyaluronate, while sodium hyaluronate improves the solubility and irritation of chitosan, simultaneously meeting the requirements of rapid dissolution, ensuring puncture force, and gentle, non-irritating properties.

[0025] Secondly, this application provides a method for preparing anti-hair loss and hair growth microneedles, using the following technical solution: A method for preparing anti-hair loss and hair growth microneedles includes the following steps: Mix *Usnea indica* and bamboo shoots in a 1:1 mass ratio, extract the juice, filter to remove impurities, inoculate with 0.05-0.08% of compound bacteria, ferment at 35-38℃ for 46-48 hours, filter, and obtain mixed enzyme liquid. Mix the mixed enzyme solution with walnut shell vinegar solution, zinc-loaded eggshell membrane powder and lactic acid bacteria fermentation broth evenly, and sonicate for 20-30 minutes to obtain the active ingredient solution. The raw materials for the microneedle matrix are dissolved to prepare a matrix solution; The active ingredient solution and the matrix solution are mixed evenly at a mass ratio of 0.5-1:3 to obtain the precursor solution; The precursor solution was added to the microneedle mold, defoamed under negative pressure, dried at 30-35℃ for 24-28 hours, and then demolded to obtain anti-hair loss and hair growth microneedles.

[0026] By employing the above-mentioned technical solution, bamboo shoots and *Ulmus pumila* are juiced, filtered, and impurities removed before fermentation. After fermentation, *Ulmus pumila* contains flavonoids, polysaccharides, alkaloids, and other components, which can improve local blood circulation in the scalp, provide sufficient nutrition to hair follicles, and regulate the microenvironment of hair follicles. Moreover, the antioxidant components in *Ulmus pumila* can reduce oxidative damage, and the anti-inflammatory components can relieve scalp inflammation, indirectly protecting hair follicles. The carbohydrates and proteins in bamboo shoots can serve as a nutrient source for the complex bacteria, promoting microbial growth and metabolism, thereby enabling microorganisms to produce more bioactive small molecule peptides, organic acids, and polysaccharide degradation products, which can help enhance hair follicle activity, improve scalp barrier function, and indirectly synergize with the hair growth effect of *Ulmus pumila*.

[0027] Optionally, the compound bacteria include Lactobacillus plantarum, Lactobacillus paracasei, and Lactobacillus rhamnosus in a mass ratio of 4-5:2-4:0.5-1. The viable bacteria count in the mixed enzyme solution was (3-5) × 10⁻⁶. 11 cfu / ml.

[0028] By adopting the above technical solution, Lactobacillus plantarum, Lactobacillus paracasei, and Lactobacillus rhamnosus are mixed in a certain proportion as a compound bacteria. Through fermentation, the macromolecular components in the Chinese artemisia and bamboo shoots are degraded into small molecules, improving the skin permeability and bioavailability of the components, making them easier to be absorbed by hair follicles. Moreover, the beneficial components such as organic acids produced by the compound bacteria during metabolism can not only regulate the pH value of the scalp, maintain the microecological balance, and reduce hair loss caused by flora imbalance, but also assist in anti-inflammatory and moisturizing effects, providing a healthier growth environment for hair follicles.

[0029] Secondly, this application provides a hair loss prevention and hair growth essence, which adopts the following technical solution: A hair loss prevention and hair growth serum containing hair loss prevention and hair growth microneedles.

[0030] By adopting the above technical solution, microneedles are added to the essence. Through application, massage, and other actions, the microneedles pierce the stratum corneum of the scalp, improving the absorption rate of effective ingredients, accelerating the hair growth process, shortening the hair growth time, and improving the hair growth effect.

[0031] Optionally, the hair loss prevention and hair growth essence contains the following ingredients by weight percentage: 10-20% hair loss prevention and hair growth microneedles, 3-6% glycerin, 2-4% fucoidan, 1-3% niacinamide, 1.5-2% eggshell membrane polypeptide, 0.1-3% betaine, 0.1-1% PEG-40 hydrogenated castor oil, 0.2-1% preservative, 0.05-0.2% citric acid, 0.1-0.6% fragrance, and the balance being water.

[0032] By adopting the above technical solution, ingredients such as fucoidan, graphene oxide, and eggshell membrane peptides are added to the essence. All of these ingredients have good biocompatibility. Graphene oxide has a large number of hydrophilic groups and a large specific surface area, giving it good liquid absorption capacity. Eggshell membrane peptides have strong antioxidant capacity, which is conducive to the migration of fibroblasts and accelerates the healing of microneedle puncture wounds. In addition, the strong antibacterial properties of graphene oxide, under the adhesive force of fucoidan, can effectively adhere microneedles, graphene oxide, eggshell membrane peptides, and other ingredients to the scalp, reducing volatilization, increasing penetration, and improving the anti-hair loss and hair growth effect.

[0033] Optionally, heat glycerin, fucoidan, nicotinamide, betaine, and water to 70-80℃, stir well, add PEG-40 hydrogenated castor oil, glycerin, preservative, and citric acid, homogenize at 60-65℃ for 1-2 minutes, then add anti-hair loss and hair growth microneedles, eggshell membrane peptides, and fragrance, homogenize at 60-65℃ for 1-2 minutes, and cool to room temperature to obtain the anti-hair loss and hair growth essence.

[0034] In summary, this application has the following beneficial effects: 1. Since this application uses microneedles as delivery carriers for anti-hair loss and hair growth ingredients, the effective ingredients penetrate quickly, avoiding the risks of injection, reducing scalp irritation, and using zinc-loaded eggshell membrane, wormwood, walnut shell vinegar and lactic acid bacteria fermentation liquid as anti-hair loss and hair growth ingredients, it can provide a healthy growth environment for hair follicles, reduce the awakening of dormant hair follicles, promote their entry into the growth phase, regulate the balance of scalp flora, strengthen hair, reduce hair breakage and loss, and improve hair quality.

[0035] 2. In this application, eggshell membrane is preferably loaded with ε-polylysine and zinc gluconate, which is then freeze-dried and soaked with adipose-derived mesenchymal stem cell exosomes and a film-forming agent. ε-polylysine has broad-spectrum antibacterial properties, and zinc ions have anti-inflammatory effects, working together to build a healthy scalp. Moreover, the new element, as an enzyme cofactor, can support hair follicle cell metabolism and keratin synthesis. Adipose-derived mesenchymal stem cell exosomes stimulate hair follicles to transition from the resting phase to the growth phase, improve microcirculation, and enhance nutrient supply. The eggshell membrane and film-forming agent constitute a carrier system, prolonging the action time of the active ingredients, ensuring that the active ingredients do not evaporate quickly, and acting on the hair follicle target for a long time to promote hair regeneration.

[0036] 3. In this application, a mixed enzyme is preferably prepared by juicing a mixture of purslane and bamboo shoots and fermenting with compound bacteria. This enzyme can promote the regeneration of hair follicles, increase the activity of hair follicles, increase the accumulation of collagen, and promote the growth of hair by promoting cell proliferation proteins in subcutaneous tissue, thereby alleviating seborrheic alopecia. Attached Figure Description

[0037] Figure 1 Before and after photos of volunteer 1 using the anti-hair loss and hair growth essence prepared in Application Example 1 (left: before test, right: after test).

[0038] Figure 2 Before and after photos of Volunteer 2 who used the anti-hair loss and hair growth essence prepared in Application Example 1 (left: before test, right: after test).

[0039] Figure 3 Before and after photos of volunteer 3 who used the anti-hair loss and hair growth essence prepared in Application Example 1 (left: before test, right: after test).

[0040] Figure 4Before and after photos of volunteer 4 who used the anti-hair loss and hair growth essence prepared in Application Example 1 (left: before test, right: after test).

[0041] Figure 5 Before and after photos of volunteer 5 who used the anti-hair loss and hair growth essence prepared in Application Example 1 (left: before test, right: after test).

[0042] Figure 6 Before and after photos of volunteer 6 who used the anti-hair loss and hair growth essence prepared in Application Example 1 (left: before test, right: after test). Detailed Implementation

[0043] The following embodiments provide a further detailed description of this application.

[0044] Preparation of zinc-loaded eggshell membranes: Examples 1-7

[0045] In the preparation example, the adipose-derived mesenchymal stem cell exosomes were selected from Nanjing Wanmuchun Biotechnology Co., Ltd., specifically human umbilical cord mesenchymal stem cell exosomes, with endotoxin less than 0.25 EU / ml and BCS protein concentration of 0.5-1 mg / ml.

[0046] Preparation Example 1: 5g of zinc gluconate, 10g of ε-polylysine and Yulongkash River water were mixed evenly to prepare a blend solution with a zinc gluconate concentration of 5g / l and an ε-polylysine concentration of 10g / l. 100g of eggshell membrane was soaked in alkaline solution (0.1mol / L sodium hydroxide solution) for 72h, cleaned to remove impurities, then soaked in a blend solution for 24h, and then freeze-dried at -56℃ for 24h to obtain the carrier material. 5g of film-forming agent was dissolved in Yulongkash River water to prepare a solution with a concentration of 0.5wt%. 30g of adipose-derived mesenchymal stem cell exosomes were added and mixed evenly to prepare a treatment solution. The film-forming agent included xanthan gum, chitosan and hyaluronic acid in a mass ratio of 1:0.5:0.5. 100g of carrier material was soaked in the treatment solution and soaked at room temperature for 48 hours. After filtration, the material was dried at 40℃, pulverized, and ground to the nanoscale to obtain zinc-loaded eggshell membrane powder.

[0047] Preparation Example 2: 1g of zinc gluconate, 2g of ε-polylysine and Yulongkash River water were mixed evenly to prepare a blend solution with a zinc gluconate concentration of 1g / L and an ε-polylysine concentration of 2g / L. 100g of eggshell membrane was soaked in alkaline solution (0.1mol / L sodium hydroxide solution) for 72h, cleaned to remove impurities, then soaked in a blend solution for 24h, and then freeze-dried at -56℃ for 24h to obtain the carrier material. 10g of film-forming agent was dissolved in Yulongkash River water to prepare a 1wt% solution. 10g of adipose-derived mesenchymal stem cell exosomes were added and mixed evenly to prepare a treatment solution. The film-forming agent included xanthan gum, chitosan and hyaluronic acid in a mass ratio of 1:0.5:0.5. 100g of carrier material was soaked in the treatment solution and soaked at room temperature for 48 hours. After filtration, the material was dried at 40℃, pulverized, and ground to the nanoscale to obtain zinc-loaded eggshell membrane powder.

[0048] Preparation Example 3: The difference from Preparation Example 1 is that ε-polylysine was not added.

[0049] Preparation Example 4: The difference from Preparation Example 1 is that zinc gluconate was used in an equal amount to replace ε-polylysine.

[0050] Preparation Example 5: The difference from Preparation Example 1 is that no adipose-derived mesenchymal stem cells were added.

[0051] Preparation Example 6: The difference from Preparation Example 1 is that the carrier material was crushed and ground to the nanoscale without soaking in the treatment solution, and zinc-loaded eggshell membrane powder was obtained.

[0052] Preparation Example 7: 5g of zinc gluconate and Yulongkash River water were mixed evenly to prepare a blend solution with a zinc gluconate concentration of 5g / L; 100g of eggshell membrane was soaked in an alkaline solution (0.1mol / L sodium hydroxide solution) for 72 hours, cleaned to remove impurities, and then soaked in a blended solution for 24 hours. After soaking, it was dried at 40℃, pulverized and ground to nanoscale to obtain zinc-loaded eggshell membrane powder.

[0053] Example 8: Preparation of eggshell membrane peptides: 5g of eggshell membrane was mixed with 100ml of buffer solution (0.1mol / L Na2PO4·12H2O and 0.1mol / L NaH2PO4·2H2O solutions were mixed in an appropriate ratio to make the final pH 8), 40mmol / L reducing agent Na2SO3 and alkaline protease (Anzame AP-200A, 200,000 U / g). After stirring and mixing evenly, the mixture was placed on a magnetic stirrer and enzymatically hydrolyzed for 4h (heating temperature was 55℃). After 4h of enzymatic hydrolysis, the enzyme was inactivated at high temperature (temperature was 80℃). The supernatant was collected by centrifugation and dialyzed with a small molecule retention dialysis bag for 24h. The mixture was then freeze-dried to obtain eggshell membrane peptides (stored at -20℃ for later use). The amount of alkaline protease added was 2%. Example

[0054] In the following examples, chitosan was selected from Xi'an Xinfengda Pharmaceutical Excipients Co., Ltd., catalog number 10055; sodium hyaluronate was selected from Xi'an Fenghe Biotechnology Co., Ltd., catalog number FH-563214, with a molecular weight of 20-40W; Lactobacillus plantarum (ATCC8014) was selected from Shanghai Xuanya Biotechnology Co., Ltd.; Lactobacillus rhamnosus was selected from Shandong Pingao Pharmaceutical Co., Ltd., catalog number PG-LRT109; Lactobacillus paracasei was selected from Xi'an Dongchi Biotechnology Co., Ltd., catalog number DC21081323; and Lactobacillus plantarum was selected from Xi'an Tianhe Pharmaceutical Co., Ltd., catalog number TH-KMHUY, with an activity of 10 billion CFU / g.

[0055] Example 1: A hair loss prevention and hair growth microneedle, comprising a microneedle matrix and hair loss prevention and hair growth components loaded within the microneedle matrix. The raw materials of the microneedle matrix include chitosan and sodium hyaluronate in a mass ratio of 7:3. The raw materials of the hair loss prevention and hair growth components include walnut shell vinegar, *Ulmus pumila*, zinc-loaded eggshell membrane powder, and lactic acid bacteria fermentation broth in a mass ratio of 1:1.2:0.6:0.5. The walnut shell vinegar is obtained by crushing walnut shells, pressing them into rods, carbonizing them at 400°C for 3 hours, cooling the flue gas to obtain liquid, adding 2 times the liquid mass of purified water and 0.1 times the liquid mass of ginger powder, sealing and storing for 3 months, filtering and separating the pigments. The zinc-loaded eggshell membrane powder is prepared according to Example 1. The lactic acid bacteria fermentation broth is obtained by fermenting *Lactobacillus plantarum* (ATCC8014) at an inoculum of 1% on MRS liquid medium at 37°C for 24 hours, inactivating it at 65°C for 30 minutes, homogenizing it three times at 1000 bar, and filtering.

[0056] The preparation method of the above-mentioned anti-hair loss and hair growth microneedles includes the following steps: S1. Juice the Usmania grass, filter to remove impurities, add compound bacteria at an inoculum rate of 0.08%, ferment at 38℃ for 46 hours, filter, and obtain a viable count of 5×10⁻⁶. 11 The mixed enzyme solution contains cfu / ml of bacteria, including Lactobacillus plantarum, Lactobacillus paracasei, and Lactobacillus rhamnosus in a mass ratio of 5:4:1. S2. Mix the mixed enzyme solution with walnut shell vinegar solution, zinc-loaded eggshell membrane powder and lactic acid bacteria fermentation liquid evenly, and sonicate at 200W power for 30 minutes to obtain the active ingredient solution. S3. Dissolve chitosan in 1% (v / v) acetic acid, stir at 50°C, add sodium hyaluronate, mix and stir evenly to obtain a matrix solution with a concentration of 20wt%. S4. Mix the active ingredient solution and the matrix solution at a mass ratio of 1:3 to obtain the precursor solution. S5. Add the precursor solution to the PMDS microneedle mold. The microneedle mold has a needle length of 800 μm, a bottom diameter of 340 μm, a needle tip distance of 600 μm, a needle tip array of 10×10, and a groove depth of 2 mm. Then, place the mold with the precursor solution into a vacuum hood and defoam at room temperature under negative pressure three times, 5 minutes each time. Gently stir with the needle tip to remove air bubbles. Then, place it in a vacuum drying oven and dry at 35°C for 24 hours. Demold to obtain anti-hair loss and hair growth microneedles.

[0057] Example 2: A hair loss prevention and hair growth microneedle, comprising a microneedle matrix and hair loss prevention and hair growth ingredients loaded within the microneedle matrix. The microneedle matrix comprises chitosan and sodium hyaluronate in a mass ratio of 6:4. The raw materials for the hair loss prevention and hair growth ingredients include walnut shell vinegar, *Ulmus pumila*, zinc-loaded eggshell membrane powder, and lactic acid bacteria fermentation liquid in a mass ratio of 1:0.7:0.3:0.1. The walnut shell vinegar is obtained by crushing walnut shells, pressing them into rods, carbonizing them at 400°C for 3 hours, and cooling the flue gas. The body was prepared by adding 2 times the liquid mass of purified water and 0.1 times the liquid mass of ginger powder, sealing and storing for 3 months, and then filtering and separating the pigments. The zinc-loaded eggshell membrane powder was prepared by Preparation Example 2. The lactic acid bacteria fermentation broth was prepared by fermenting Lactobacillus plantarum (ATCC8014) at an inoculum of 0.8% on MRS liquid medium at 38°C for 24 h, inactivating at 70°C for 25 min, homogenizing 5 times at 1000 bar, and filtering.

[0058] The preparation method of the above-mentioned anti-hair loss and hair growth microneedles includes the following steps: S1. Juice the Usmania grass, filter to remove impurities, add compound bacteria at an inoculum rate of 0.05%, ferment at 35℃ for 48 hours, filter, and obtain a viable count of 5×10⁻⁶. 11 The mixed enzyme solution contains cfu / ml of bacteria, including Lactobacillus plantarum, Lactobacillus paracasei, and Lactobacillus rhamnosus in a mass ratio of 4:2:0.5. S2. Mix the mixed enzyme solution with walnut shell vinegar solution, zinc-loaded eggshell membrane powder and lactic acid bacteria fermentation liquid evenly, and sonicate at 250W power for 20 minutes to obtain the active ingredient solution. S3. Dissolve chitosan in 1% (v / v) acetic acid, stir at 50°C, add sodium hyaluronate, mix and stir evenly to obtain a matrix solution with a concentration of 30wt%. S4. Mix the active ingredient solution and the matrix solution at a mass ratio of 0.5:3 to obtain a precursor solution. S5. Add the precursor solution to the PMDS microneedle mold. The microneedle mold has a needle length of 800 μm, a bottom diameter of 340 μm, a needle tip distance of 600 μm, a needle tip array of 10×10, and a groove depth of 2 mm. Then, place the mold with the precursor solution in a vacuum hood and defoam at room temperature under negative pressure three times, 5 minutes each time. Gently stir with the needle tip to remove air bubbles. Then, place it in a vacuum drying oven and dry at 30°C for 48 hours. Demold to obtain anti-hair loss and hair growth microneedles.

[0059] Example 3: A hair loss prevention and hair growth microneedle, comprising a microneedle matrix and hair loss prevention and hair growth components loaded within the microneedle matrix. The microneedle matrix comprises chitosan and sodium hyaluronate in a mass ratio of 7:3. The raw materials for the hair loss prevention and hair growth components include walnut shell vinegar, *Ulmus pumila*, zinc-loaded eggshell membrane powder, and lactic acid bacteria fermentation broth in a mass ratio of 1:1:0.5:0.3. The walnut shell vinegar is prepared by crushing walnut shells, pressing them into rods, carbonizing them at 400°C for 3 hours, cooling the flue gas to obtain liquid, adding 2 times the liquid mass of purified water and 0.1 times the liquid mass of ginger powder, sealing and storing for 3 months, filtering, and separating the pigments. The zinc-loaded eggshell membrane powder is prepared as in Example 1. The lactic acid bacteria fermentation broth is prepared by fermenting *Lactobacillus plantarum* (ATCC8014) at an inoculum of 0.8% on MRS liquid medium at 38°C for 24 hours, inactivating it at 70°C for 25 minutes, homogenizing it 5 times at 1000 bar, and filtering.

[0060] The preparation method of the above-mentioned anti-hair loss and hair growth microneedles includes the following steps: S1. Juice the Usmania grass, filter to remove impurities, add compound bacteria at an inoculum rate of 0.07%, ferment at 35℃ for 48 hours, filter, and obtain a viable count of 5×10⁻⁶. 11 The mixed enzyme solution contains cfu / ml of bacteria, including Lactobacillus plantarum, Lactobacillus paracasei, and Lactobacillus rhamnosus in a mass ratio of 5:3:0.5. S2. Mix the mixed enzyme solution with walnut shell vinegar solution, zinc-loaded eggshell membrane powder and lactic acid bacteria fermentation liquid evenly, and sonicate at 250W power for 20 minutes to obtain the active ingredient solution. S3. Dissolve chitosan in 1% (v / v) acetic acid, stir at 50°C, add sodium hyaluronate, mix and stir evenly to obtain a matrix solution with a concentration of 40wt%. S4. Mix the active ingredient solution and the matrix solution at a mass ratio of 0.8:3 to obtain a precursor solution; S5. Add the precursor solution to the PMDS microneedle mold. The microneedle mold has a needle length of 800 μm, a bottom diameter of 340 μm, a needle tip distance of 600 μm, a needle tip array of 10×10, and a groove depth of 2 mm. Then, place the mold with the precursor solution into a vacuum hood and defoam at room temperature under negative pressure three times, 5 minutes each time. Gently stir with the needle tip to remove air bubbles. Then, place it in a vacuum drying oven and dry at 35°C for 48 hours. Demold to obtain anti-hair loss and hair growth microneedles.

[0061] Example 4: A hair loss prevention and hair growth microneedle, which differs from Example 1 in that the zinc-loaded eggshell membrane powder is made from Preparation Example 3.

[0062] Example 5: A hair loss prevention and hair growth microneedle, which differs from Example 1 in that the zinc-loaded eggshell membrane powder is made from Preparation Example 4.

[0063] Example 6: A hair loss prevention and hair growth microneedle, which differs from Example 1 in that the zinc-loaded eggshell membrane powder is made from Preparation Example 5.

[0064] Example 7: A hair loss prevention and hair growth microneedle, which differs from Example 1 in that the zinc-loaded eggshell membrane powder is made from Preparation Example 6.

[0065] Example 8: A hair loss prevention and hair growth microneedle, which differs from Example 1 in that the zinc-loaded eggshell membrane powder is made from Preparation Example 7.

[0066] Example 9: A hair loss prevention and hair growth microneedle, differing from Example 1 in that the mixed enzyme solution also includes bamboo shoots. The mixed enzyme solution is prepared by the following method: Usmania ulmoides and bamboo shoots are mixed in a 1:1 mass ratio, juiced, filtered to remove impurities, and compound bacteria are added at an inoculum rate of 0.08%. Fermentation is carried out at 38°C for 46 hours, followed by filtration to obtain a live bacteria count of 5 × 10⁻⁶. 11 The mixed enzyme solution contains cfu / ml of bacteria, including Lactobacillus plantarum, Lactobacillus paracasei, and Lactobacillus rhamnosus in a mass ratio of 5:4:1. Comparative Example

[0067] Comparative Example 1: A hair loss prevention and hair growth microneedle, which differs from Example 1 in that the raw materials for the hair loss prevention and hair growth ingredients include walnut shell vinegar, *Usnea indica*, eggshell membrane powder, and lactic acid bacteria fermentation liquid in a mass ratio of 1:1.2:0.6:0.5. The eggshell membrane powder is prepared by the following method: 100g of eggshell membrane is soaked in alkaline solution (0.1mol / L sodium hydroxide solution) for 72h, cleaned to remove impurities, dried, pulverized, and ground to nanoscale at 40℃.

[0068] Comparative Example 2: A hair loss prevention and hair growth microneedle, which differs from Example 1 in that the raw materials for the hair loss prevention and hair growth ingredients include walnut shell vinegar, sage, and lactic acid bacteria fermentation liquid in a mass ratio of 1:1.2:0.5.

[0069] Comparative Example 3: A hair loss prevention and hair growth microneedle, which differs from Example 1 in that the raw materials for the hair loss prevention and hair growth ingredients include walnut shell vinegar and black vine in a mass ratio of 1:1.2.

[0070] Comparative Example 4: A hair loss prevention and hair growth microneedle, which differs from Example 1 in that the raw materials of the hair loss prevention and hair growth ingredients include *Usnea indica*, zinc-loaded eggshell membrane powder and lactic acid bacteria fermentation liquid in a mass ratio of 1.2:0.6:0.5. Application examples

[0071] Application Example 1: A hair loss prevention and hair growth essence, comprising the following raw materials in parts by weight (g): 20g of hair loss prevention and hair growth microneedles prepared in Example 1, 6g of glycerin, 4g of fucoidan, 3g of niacinamide, 2g of eggshell membrane polypeptide prepared in Preparation Example 8, 3g of betaine, 1g of PEG-40 hydrogenated castor oil, 0.5g of preservative, 0.2g of citric acid, 0.6g of lemon flavor, and water to make up to 100g. The fucoidan is selected from Xi'an Tianguangyuan Biotechnology Co., Ltd., product number TGYHZ654301.

[0072] The preparation method of the above-mentioned anti-hair loss and hair growth essence includes the following steps: Glycerin, fucoidan, nicotinamide, betaine, and water are heated to 80°C and stirred evenly. PEG-40 hydrogenated castor oil, glycerin, preservatives, and citric acid are added. After homogenization at 60°C for 2 minutes, anti-hair loss and hair growth microneedles, eggshell membrane peptides, and fragrance are added. After homogenization at 60°C for 2 minutes, the mixture is cooled to room temperature to obtain the anti-hair loss and hair growth essence.

[0073] Application Examples 2-9: A hair loss prevention and hair growth essence, which differs from Application Example 1 in that it uses hair loss prevention and hair growth microneedles prepared in Examples 2-9.

[0074] Application Examples 10-13: A hair loss prevention and hair growth essence, which differs from Application Example 1 in that it uses hair loss prevention and hair growth microneedles prepared according to Comparative Examples 1-4. Performance testing

[0075] Prepare the anti-hair loss and hair growth essence according to the method in the application example, and conduct performance testing according to the following methods. Record the test results in Table 1.

[0076] 1. Detection of 5α-reductase inhibition rate: (1) Preparation of 5α-reductase: Male SD males were fasted overnight but allowed to drink water, and then euthanized. The prostate was removed and cut into pieces on an ice plate. The prostate was homogenized at 0℃ using a glass homogenizer with buffer (containing 0.32mol / L sucrose, 0.1mmol / L dithiothreitol, 1mmol / L EDTA and 0.2mol / L phosphate buffer). After centrifugation, the upper white fat layer was removed, and the supernatant was added to buffer to make up to 20mL. The mixture was then aliquoted into EP tubes and stored at -80℃. (2) Determination of 5α-reductase inhibition rate: 0.5mL phosphate buffer, 0.1mL test essence, 200μL 300mg / L testosterone solution and 200μL 0.8g / L NADPH (reduced nicotinamide adenine dinucleotide phosphate) solution were added to 0.5mL of the solution. 5α-reductase extract was reacted at 37℃ for 30 min, then 2.5 mL of dichloromethane was added to stop and extract testosterone. 0.25 mL of 100 mg / L propylparaben was added as an internal standard. After centrifugation, the upper aqueous phase was removed, and approximately 1 mL of the organic phase was removed and evaporated to dryness. The residue was dissolved in 1.5 mL of methanol, and the residual testosterone was determined using high-performance liquid chromatography. 5α-reductase reaction tubes and 5α-reductase blank tubes were prepared, with finasteride (0.05 mg / L) as the control group. The inhibition rate was calculated using the following formula: 5α-reductase inhibition rate (%) = (Rsample - Rreaction) / (Rblank - Rreaction) × 100%.

[0077] 2. Number of hair follicles: (1) Mix rosin and paraffin in a 1:1 ratio, heat and melt, and apply evenly to the back of the mouse. The application area is 2cm×3cm. After cooling and solidification, remove the hair to remove the back hair and confirm that the hair growth phase is in the resting phase; (2) On the day after hair removal, select mice with smooth hair removal area and no skin damage, and randomly divide them into 15 groups, with 5 mice in each group. Groups 1-13 are applied with the essence prepared in Examples 1-13. Group 14 is the negative control group and does not apply the product. Group 15 is the 2% minoxidil control group; (3) Apply 3 times a day at regular intervals, with an application amount of 0.1ml each time. Massage for 5-10 minutes after application and apply continuously for 20 days; (4) Sacrifice the mice, use HE staining, observe the mouse skin tissue under a microscope, count the number of hair follicles in three non-overlapping areas under 200x magnification, and take the average value to represent the number of hair follicles in the mouse. The test results are the average value of 5 mice in each group.

[0078] 3. Hair growth: (1) Modeling: Shave the hair on the back of the mouse (2cm×3cm), remove the short hair with depilatory cream, apply 0.1ml of 0.05% propionate testosterone injection to the hair removal area, and inject 5mg / (kg·d) of testosterone propionate injection subcutaneously in the back of the neck once a day for 30 consecutive days. The mouse back is smooth and the hair no longer grows, and the hair falls out. The hair is fine and brittle, thus becoming a seborrheic alopecia model; (2) Grouping and administration: SPF grade 5-7 week male C57BL / 6 mice weighing 20±2g were grouped into groups. Ninety mice were randomly divided into 15 groups of 6 mice each. Except for the blank group, the mice in the other groups continued to be injected subcutaneously with testosterone propionate injection at 5 mg / kg·d once a day. Then, the test essence was applied three times a day, 0.1 ml each time, and massaged for 5-10 minutes. The control group was treated with the same volume of minoxidil. The administration was continued for 30 days. (3) Evaluation index: 20 days after administration, 10 new hairs were randomly taken from the experimental area, and the hair length and weight were measured. The average value was calculated to evaluate the hair growth. The test results were the average value of 6 mice in each group.

[0079] 4. Hair density: 90 male volunteers aged 25-45 years were selected to be troubled by hair loss. The inclusion criteria were: mild to moderate hair loss, hair loss grade Norwood-Hamilton II to III, or Ludwig I to II, no other diseases related to hair loss recently, and exclusion of (1) those with atrophied hair follicles; (2) those with serious systemic diseases, immunodeficiency or autoimmune diseases; (3) those with dermatological diseases in the test area or those who are receiving drug treatment, such as scalp infection, seborrheic dermatitis, psoriasis and alopecia areata; (4) those who have used topical or systemic hair growth products or used topical or systemic drugs that affect hair growth or loss acid production in the three months prior to the start of the experiment; (5) those who have received hair transplantation. The volunteers were randomly divided into groups 1-15, Experiment 1-1 Three groups used the serum prepared in Examples 1-13. The negative control group used 2% minoxidil, and the blank group received no treatment. Apply 0.1ml once daily before bedtime, massaging for 5-10 minutes after application. Continue for 20 days. Hair density was measured before and after use as follows: ① Test area selection: To ensure accuracy, a classic, even scalp area was selected (avoiding the hairline, balding areas, or areas with excessively dense hair). The most commonly selected area was the central area of ​​the top of the head: A 2cm diameter circular area (approximately 3.14cm in area) was drawn centered on the Baihui acupoint on the top of the head. 2 Or, take a square area 2cm behind the hairline on the forehead: 2cm wide and 2cm long (area 4cm²). 2); ② Operating steps: Clean the scalp, wash the hair with neutral shampoo and dry it to avoid oil and dandruff interfering with observation; Fix the hair: Gently comb the hair in the measurement area with a fine-tooth comb to ensure there are no knots or frizz; Microscopic observation: Use a dermatoscope (magnification 10~100x) to aim at the measurement area and take a high-resolution image; Count the hairs: Mark and count all visible hairs in the area on the image (Note: Under the dermatoscope, "terminal hairs" (coarse and stiff hairs) and "vellus hairs" (fine and soft lanugo) are visible. Usually, the number of terminal hairs is counted because vellus hairs do not affect density assessment); Calculate the density: Hair density (roots / cm²) 2 = Number of hairs in the measurement area / area of ​​the measurement area. For example, if 400 terminal hairs are counted in a circular area with a diameter of 2cm (area ≈ 3.14cm²), the density is: 400 / 3.14≈127 hairs / cm² (close to the normal range for Caucasians). The test result is the average of 6 people in each group.

[0080] Table 1 Performance Tests of Anti-Hair Loss and Hair Growth Essence project 5α-reductase inhibition rate Number of hair follicles (per field of view) 20-day hair weight / mg 20d hair length / mm <![CDATA[Total hair density before use (roots / cm 2 ).]]> <![CDATA[Total hair density after use (roots / cm 2 )]]> Application Example 1 91.5 32.50 30.47 9.55 134 154 Application Example 2 91.1 31.32 30.12 9.34 133 153 Application Example 3 90.7 30.67 29.98 10.11 134 154 Application Example 4 87.1 27.38 28.61 7.98 132 151 Application Example 5 88.6 29.21 29.88 8.42 133 152 Application Example 6 85.2 25.54 27.43 7.48 132 150 Application Example 7 84.3 23.12 26.31 7.24 131 148 Application Example 8 80.2 21.85 25.13 6.71 132 147 Application Example 9 91.4 33.41 33.02 9.84 133 155 Application Example 10 78.6 20.42 24.97 6.28 131 146 Application Example 11 78.1 20.13 23.12 5.94 133 147 Application Example 12 77.2 19.21 22.21 5.75 131 145 Application Example 13 78.8 20.23 23.35 6.08 132 146 Minoxidil / 21.20 25.77 6.56 131 143 Blank group / 17.32 21.50 5.29 131 142 In Application Examples 1-3, the anti-hair loss and hair growth microneedles prepared in Examples 1-3 were used respectively. It can be seen that they can significantly inhibit 5α-reductase, making it difficult for male hormones to be converted into the more active dihydrotestosterone, preventing hair follicles from entering the resting phase, thus playing a good role in preventing hair loss. Moreover, the number of hair follicles in mice increased and entered the growth phase. After applying the anti-hair loss and hair growth essence to mice in the lipophilic alopecia model, the hair growth rate was fast, and the hair density after growth was high, and it was not easy to break or fall out. This shows that the anti-hair loss and hair growth essence prepared in this application can make hair follicles enter the growth phase, promote hair growth and nourish hair follicles, and achieve the effects of preventing hair loss and strengthening hair.

[0081] In Application Example 4, the anti-hair loss and hair growth microneedles prepared in Example 4 were used. In Example 4, the zinc-loaded eggshell membrane powder prepared in Preparation Example 3 was used. Compared with Example 1, no ε-polylysine was added. It can be seen that after continuous use of the anti-hair loss and hair growth essence prepared in Application Example 4, the 5α-reductase inhibition rate decreased, the hair follicle growth rate decreased, and the hair growth promotion effect weakened.

[0082] In Application Example 5, the zinc-loaded eggshell membrane powder prepared in Preparation Example 4 was used. Compared with the zinc-loaded eggshell membrane powder in Application Example 1, zinc gluconate was used to replace ε-polylysine. As can be seen from the data comparison in Table 1, compared with Application Example 1, the hair loss prevention and hair growth essence prepared in Application Example 5 had a reduced effect on promoting hair growth and preventing hair loss, a reduced number of hair follicles, and a slower hair growth rate.

[0083] In Application Example 6, the zinc-loaded eggshell membrane powder prepared in Preparation Example 5 was used without the addition of adipose-derived mesenchymal stem cells. Compared with Application Example 1, after continuous use, the hair loss prevention and hair growth microneedles prepared in Application Example 6 showed a decrease in the rate of increase in the number of hair follicles, a slowdown in hair growth, and a decrease in the total hair density. This indicates that the addition of adipose-derived mesenchymal stem cells can significantly enhance the anti-hair loss, growth promotion, and hair strengthening effects of the essence.

[0084] In Application Example 7, the zinc-loaded eggshell membrane powder soaking solution was used. The eggshell membrane loaded with zinc gluconate and ε-polylysine was freeze-dried and then pulverized. The treatment solution formed by adipose mesenchymal stem cells and polyvinyl alcohol was not used for soaking. Compared with Application Example 1, the hair growth-promoting effect of the essence was weakened after use.

[0085] In Application Example 8, the zinc-loaded eggshell membrane powder prepared in Preparation Example 7 was used. In this example, the eggshell membrane was soaked in zinc gluconate, dried, and pulverized. Compared with Application Example 1, the essence prepared in Application Example 8 had a more significant effect on preventing hair loss and promoting hair growth. This indicates that using zinc gluconate and ε-polylysine loaded onto the eggshell membrane, freeze-drying it, and then soaking it in a treatment solution containing adipose-derived mesenchymal stem cells and polyvinyl alcohol can effectively improve the hair growth promotion, hair strengthening, and hair loss prevention effects of the essence.

[0086] Compared with Application Example 1, Application Example 9 also added bamboo shoots when preparing the mixed enzyme solution. Table 1 shows that after using the anti-hair loss and hair growth essence prepared in Application Example 9, the number of hair follicles in mice increased, the amount of hair increased, and the growth rate accelerated. This indicates that using the mixed enzyme with added bamboo shoots can effectively improve the hair growth promoting effect of the essence.

[0087] Compared to Application Example 1, Application Example 10 used eggshell membrane powder instead of zinc-loaded eggshell powder, while Application Example 11 did not add zinc-loaded eggshell membrane powder. The data in Table 1 show that the anti-hair loss and hair growth essence prepared in Application Examples 10 and 11 had a reduced inhibition rate of 5α-reductase, slower hair growth rate, and reduced total hair density after use. The effect in Application Example 11 was even worse, indicating that using only eggshell membrane powder did not significantly improve the hair growth promotion effect and was less effective than using zinc-loaded eggshell membrane powder.

[0088] Compared with Application Example 1, Application Example 12 only used walnut shell vinegar and black vine as raw materials for preventing hair loss and promoting hair growth. Compared with Application Example 1, Application Example 13 did not add walnut shell vinegar. As shown in Table 1, the hair loss prevention and hair growth essence prepared in Application Example 12 and Application Example 13 were not as good as Application Example 1 in terms of the number of hair follicles and the hair growth rate.

[0089] Six male volunteers with hair loss were selected. They sprayed 1ml of the hair loss prevention and hair growth lotion prepared in this application once daily before bedtime. Before and after use, photos were taken to compare hair growth progress. Figures 1-6It is evident that hair volume increased significantly after using the anti-hair loss and hair growth essence.

[0090] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.

Claims

1. A microneedle for preventing hair loss and promoting hair growth, characterized in that, It includes a microneedle matrix and hair loss prevention and hair growth ingredients loaded in the microneedle matrix. The raw material of the microneedle matrix is ​​selected from at least one of chitosan, sodium hyaluronate, type I and type II collagen complex, silk fibroin, Ganoderma lucidum polysaccharide, sodium alginate and PLGA. The ingredients for preventing hair loss and promoting hair growth include walnut shell vinegar, *Ulmus pumila*, zinc-loaded eggshell membrane powder, and lactic acid bacteria fermentation liquid in a mass ratio of 1:0.7-1.2:0.3-0.6:0.1-0.

5.

2. The anti-hair loss and hair growth microneedle according to claim 1, characterized in that: The zinc-loaded eggshell membrane powder is prepared by the following method: The eggshell membrane was soaked in alkaline solution, washed with water, and then soaked in a mixture containing zinc gluconate, ε-polylysine and water. The mixture was then freeze-dried to obtain the carrier material. The carrier material was immersed in a treatment solution containing adipose-derived mesenchymal stem cell exosomes and a film-forming agent, then filtered, dried, and pulverized to obtain zinc-loaded eggshell membrane powder.

3. The anti-hair loss and hair growth microneedle according to claim 2, characterized in that: In the zinc-loaded eggshell membrane, the mass ratio of eggshell membrane, zinc gluconate, and ε-polylysine is 1:0.01-0.05:0.02-0.1; The mass ratio of the carrier material to adipose-derived mesenchymal stem cell exosomes and film-forming agent is 1:0.1-0.3:0.05-0.

1.

4. The anti-hair loss and hair growth microneedle according to claim 2, characterized in that: The mass ratio of zinc gluconate to ε-polylysine is 1:1-2.

5. The anti-hair loss and hair growth microneedle according to claim 1, characterized in that: The raw materials for the microneedle matrix include chitosan and sodium hyaluronate in a mass ratio of 6:4 to 7:

3.

6. The method for preparing the anti-hair loss and hair growth microneedles according to any one of claims 1-5, characterized in that: Includes the following steps: Mix the black vine and bamboo shoots in a 1:1 mass ratio, extract the juice, filter to remove impurities, add compound bacteria at an inoculation rate of 0.05-0.08%, ferment at 35-38℃ for 46-48 hours, filter, and obtain mixed enzyme liquid. Mix the mixed enzyme solution with walnut shell vinegar solution, zinc-loaded eggshell membrane powder and lactic acid bacteria fermentation broth evenly, and sonicate for 20-30 minutes to obtain the active ingredient solution. The raw materials for the microneedle matrix are dissolved to prepare a matrix solution; The active ingredient solution and the matrix solution are mixed evenly at a mass ratio of 0.5-1:3 to obtain the precursor solution; The precursor solution was added to the microneedle mold, defoamed under negative pressure, dried at 30-35℃ for 24-28 hours, and then demolded to obtain anti-hair loss and hair growth microneedles.

7. The method for preparing the anti-hair loss and hair growth microneedles according to claim 6, characterized in that: The compound bacteria include Lactobacillus plantarum, Lactobacillus paracasei, and Lactobacillus rhamnosus in a mass ratio of 4-5:2-4:0.5-1. The viable bacteria count in the mixed enzyme solution was (3-5) × 10⁻⁶. 11 cfu / ml.

8. A hair loss prevention and hair growth essence, characterized in that, The hair loss prevention and hair growth essence contains the hair loss prevention and hair growth microneedles as described in any one of claims 1-5 or the hair loss prevention and hair growth microneedles prepared by the preparation method described in any one of claims 6-7.

9. The anti-hair loss and hair growth essence according to claim 8, characterized in that: The hair loss prevention and hair growth essence contains the following ingredients by weight percentage: 10-20% hair loss prevention and hair growth microneedles, 3-6% glycerin, 2-4% fucoidan, 1-3% niacinamide, 1.5-2% eggshell membrane polypeptide, 0.1-3% betaine, 0.1-1% PEG-40 hydrogenated castor oil, 0.2-1% preservative, 0.05-0.2% citric acid, 0.1-0.6% fragrance, and the balance being water.

10. The anti-hair loss and hair growth essence according to claim 9, characterized in that: The method for preparing the hair loss prevention and hair growth essence includes the following steps: Glycerin, fucoidan, nicotinamide, betaine, and water are heated to 70-80℃ and stirred evenly. PEG-40 hydrogenated castor oil, glycerin, preservatives, and citric acid are added. After homogenization at 60-65℃ for 1-2 minutes, anti-hair loss and hair growth microneedles, eggshell membrane peptides, and fragrance are added. After homogenization at 60-65℃ for 1-2 minutes, the mixture is cooled to room temperature to obtain the anti-hair loss and hair growth essence.

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