A composite nutrient microneedle hair growth patch and its application

Abstract

The present invention belongs to the field of biomedicine and relates to the treatment of androgenic alopecia (hair loss). A composite nutrient microneedle hair growth patch and its application are disclosed. The composite nutrient microneedle hair growth patch of the present application comprises a non-woven fabric layer, a sodium hyalate base and a conical microneedle, wherein the conical microneedle is evenly distributed on one side of the sodium hyalate base, and the other side of the sodium hyalate base is covered with a non-woven fabric layer; the conical microneedle contains wheat germ albumin, anti-hypoxia peptide, vitamin B7, ferrous lactate, vitamin C and zinc sulfate. The composite nutrient of the present application can directly act on the tissue around the hair follicles, promote the growth and repair of the hair follicles, and improve the treatment effect of male baldness. The puncture effect of the microneedle hair growth patch designed by 3D printing can increase the permeability of the skin, improve the absorption efficiency of the drug, and reduce the dosage and side effects of the drug.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine and relates to the treatment of androgenic alopecia (hair loss). Background Art

[0002] The age of people suffering from hair loss is getting younger. Androgenic alopecia is a common hair loss disease, which is mainly manifested by the gradual thinning and loss of hair, seriously affecting the patient's appearance and mental health. Hair loss: People born in the 1980s and 1990s have become the main force of hair loss, with hair loss accounting for 38.5% for those born in the 1980s, 36.1% for those born in the 1990s, and 0.4% for those born in the 2000s. Currently, the treatments for hair loss mainly include drug therapy and hair transplantation, but these methods have certain limitations. Drug therapy may have side effects and limited effects; hair transplantation surgery is costly and risky, and is not suitable for all patients, and hair loss will occur again after three years. While there is still hope for transdermal hair follicles, the advantages and disadvantages of drug therapy are shown in Table 1 below:

[0003] Table 1 Current situation of medication for hair loss in China

[0004]

[0005] Therefore, finding a safe, effective and convenient method for treating hair loss has important practical significance. Summary of the invention

[0006] In order to solve the above technical problems, the present invention proposes a composite nutrient microneedle hair growth patch and its application.

[0007] The technical solution of the present invention is achieved in this way:

[0008] A composite nutrient microneedle hair growth patch, comprising a non-woven fabric layer, a sodium hyalate base and conical microneedles, wherein the conical microneedles are evenly distributed on one side of the sodium hyalate base, and the other side of the sodium hyalate base is covered with a non-woven fabric layer;

[0009] The conical microneedles contain wheat germ albumin, anti-hypoxia peptide, vitamin B7, ferrous lactate, vitamin C and zinc sulfate.

[0010] Regarding the raw materials in the conical microneedles of this application, from the perspective of Western medicine, hair loss is caused by male hormones. Abnormal androgen metabolism is closely related to the onset of androgenic alopecia. Under the action of type II 5α-reductase, testosterone becomes dihydrotestosterone, which is more compatible with AR, breaking the normal operation of the internal signaling pathway, leading to vascular atrophy and accelerating hair loss. In traditional Chinese medicine theory, "hair is the rest of blood". Blood plays a role in nourishing the whole body, including hair. Qi and blood fullness is an important basis for hair growth. With age and other reasons leading to insufficient qi and blood, the blood cannot fully nourish the hair follicles on the head, and the hair follicle cells atrophy, which will cause the hair to lose nourishment and fall off. Just like the water in the land has dried up and crops cannot grow, nutrients such as protein cannot be transported to the roots of the hair, and the hair falls off naturally, forming a bald head over a long period of time.

[0011] Wheat germ protein: The main component of hair is keratin. When protein is lacking, the raw materials for hair follicle cells to synthesize keratin are insufficient. Without enough amino acids to build keratin, hair will gradually become soft, fragile, and eventually fall off. Because keratin is like the "building material" of hair, lack of it will damage the structure of hair. Therefore, albumin (also called wheat germ albumin) extracted from wheat germ, which is similar to the composition of human protein, is preferred in the nutrient formula. Albumin is an important nutrient in human plasma, which can maintain plasma osmotic pressure and can be combined with a variety of nutrients, hormones and drugs. It can reflect the nutritional status of the body and cells. The usage in the present invention refers to the previous paper published by this research group (Li Xiaoxiao. Preparation and anti-exercise fatigue effect of wheat germ albumin and its peptides [D]. Henan University of Technology, 2022.), 50 mg / kg for mice, 25g for mice in this experiment, and the daily dose is calculated to be 1.25 mg.

[0012] Vitamin B family: Biotin (vitamin B7) is essential for hair health. It is involved in cell growth, fat synthesis and amino acid metabolism. Lack of biotin can lead to decreased hair follicle function and cause hair loss. Adults take 100-300μg of vitamin B7 a day, which is equivalent to a daily dose of 1.138μg for mice.

[0013] The calculation formula is as follows:

[0014] Note: M1: adult dose based on body weight; M2: mouse dose; W1: adult body weight (calculated at 60kg); W2: mouse body weight (25g in this experiment); K is the conversion factor of 9.1 between humans and mice.

[0015] Vitamin C: It is an essential nutrient for collagen synthesis. The connective tissue around the hair follicles needs collagen to maintain its structure and function. Lack of vitamin C will affect the synthesis of collagen, weaken the supporting tissue around the hair follicles, and the hair follicles will not be well supported, which will affect the growth of hair and lead to hair loss.

[0016] An adult consumes 100 mg of vitamin C per day, which, according to the above formula, translates to a daily dose of 0.3792 mg (379.2 μg) for mice.

[0017] Iron (ferrous lactate): Iron is a key raw material for making hemoglobin. Hemoglobin is responsible for transporting oxygen to various tissues in the body, including hair follicle cells. When iron is deficient, hair follicle cells will be in a state of hypoxia, metabolic activities such as aerobic respiration will be affected, energy production will be insufficient, and normal cell function cannot be maintained, resulting in hair follicle atrophy and hair loss. This situation is more common in women with excessive menstrual flow or long-term vegetarians. Adult males need to consume 15-20 mg of iron per day; according to the above formula, it is converted into a daily iron dose of 75.88 μg for mice, which is converted into a daily dose of ferrous lactate of 316.9 μg.

[0018] Anti-hypoxia peptide: solves the problem of hair follicle cells being in a state of hypoxia, insufficient energy production, and inability to maintain normal cell function, which leads to hair follicle atrophy. Anti-hypoxia peptide comes from the preparation method of the invention patent of our research group "A rice anti-hypoxia peptide and its application (ZL 202411406713.8)". Calculated at 10μg / kg, the daily dose in this experiment is 0.25μg.

[0019] Zinc (zinc sulfate): Zinc participates in the synthesis and metabolism of many enzymes in the body and plays an important role in the growth, differentiation and repair of cells. For hair follicle cells, zinc can regulate their growth cycle. Zinc deficiency will hinder the renewal and differentiation of hair follicle cells, leading to sparse hair and hair loss. The daily zinc intake of adult men is about 15 mg. According to the above formula, it is converted into a daily zinc dose of 56.88140 μg for mice, which is 140 μg for mice.

[0020] According to the conventional dosage ratio of the above nutrients: 1.25 mg of wheat germ albumin, 0.25 μg of anti-hypoxia peptide, 1.138 μg of vitamin B7, 205.7 μg of ferrous lactate, 379.2 μg of vitamin C, 140 μg of zinc sulfate in 1 ml of pure water, a complex nutrient is prepared and then added to 30 ml of hyaluronic acid solution (35%) to form a gel for 3D printing.

[0021] The steps of 3D printing are: select suitable biodegradable materials and hyaluronic acid that is beneficial to hair follicle absorption as the matrix material of microneedles. The purpose is to improve the puncture performance of microneedles and drug release efficiency without causing pain. Design and optimize the shape (cone), size and structure of microneedles (such as Figure 1 ), using 3D printing technology, the biodegradable materials containing complex nutrients are printed into microneedle hair growth patches. The printed microneedle hair growth patches are post-processed, dried, affixed with non-woven adhesive, sterilized, etc. to ensure the quality and safety of the microneedle hair growth patches.

[0022] The present invention has the following beneficial effects:

[0023] 1. The composite nutrients of the present application can directly act on the tissues around the hair follicles and be absorbed by the cells in the tissues. Wheat germ protein provides the raw materials for synthesizing keratin, vitamin B7 participates in the growth of cells, vitamin C synthesizes collagen in the connective tissue around the hair follicles, ferrous lactate produces hemoglobin, and hemoglobin is responsible for transporting oxygen to the hair follicle cells. Anti-hypoxia peptides solve the problem of insufficient energy production when the hair follicle cells are in an oxygen-deficient state and maintain normal cell function. Zinc sulfate regulates the growth cycle and differentiation of hair follicle cells. The puncture effect of the microneedle hair growth patch designed by 3D printing can increase the permeability of the skin, and the nutrients are fully absorbed without being metabolized in the body, reducing the dosage and side effects of the drug.

[0024] 2. 3D printed composite nutrient microneedles are easy to use, and patients can use them at home without the need for professional operation and guidance. Moreover, the microneedle nutrition hair growth of this application is made of biodegradable, orally available, absorbable nutrients, which is safe to use and will not cause harm to the skin. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0026] Figure 1 Schematic diagram of transdermal microneedle design.

[0027] Figure 2 This is an electron microscope picture of the nutrient microneedle patch.

[0028] Figure 3 Sample pictures of nutritional hair growth stickers taken from different shooting angles.

[0029] Figure 4 This is the electron microscope image of Example 2.

[0030] Figure 5 This is the electron microscope image of Example 3.

[0031] Figure 6 This is the electron microscope image of Example 4.

[0032] Figure 7 This is the electron microscope image of Example 5.

[0033] Figure 8 Representative images of skin tissues of different groups on the 14th day after hair removal; a is a representative image of 4-HNE in skin tissues of different groups on the 14th day after hair removal (red: 4-HNE; blue: DAPI), scale bar = 100 μm; b is a representative image of Ki67 in skin tissues of different groups on the 14th day after hair removal (green: Ki67; blue: DAPI). Scale bar = 100 μm.

[0034] Fig. 9 Representative images and vascular expressions of CD31 in skin tissues of different groups on the 14th day after hair removal; a is a representative image of CD31 in skin tissues of different groups on the 14th day after hair removal (red: CD31; blue: DAPI staining), scale bar = 100 μm; b is the expression of blood vessels in skin tissues of different groups, scale bar = 50 μm.

[0035] Fig.10 Shows the hair growth conditions of different groups; Group A is the model group, Group B is the blank group, Group C is the positive control group, Group D1 is the microneedle hair growth patch without wheat germ albumin, and Group D2 is the microneedle hair growth patch containing wheat germ albumin. DETAILED DESCRIPTION

[0036] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] Unless otherwise specified, the experimental methods used in the following experimental examples are all conventional methods; the materials and reagents used are reagents and materials that can be obtained from commercial channels unless otherwise specified.

[0038] The wheat germ albumin used in this application is prepared with reference to the paper previously published by this research group (Li Xiaoxiao. Study on the preparation and anti-exercise fatigue effect of wheat germ albumin and its peptides [D]. Henan University of Technology, 2022.); the dosage is: 50 mg / kg for mice, the mice in this experiment are 25g, and the daily dose is calculated to be 1.25 mg.

[0039] The anti-hypoxia peptide used in this application is prepared with reference to patent ZL 202411406713.8; the dosage is: calculated at 10 μg / kg, the experimental mouse is 25g, and the daily dose is calculated to be 0.25 μg.

[0040] Hyaluronic acid, purity ≥95% (HPLC) Molecular weight: 1000Da, 30ml hyaluronic acid solution (35%wt);

[0041] Example 1

[0042] The preparation steps of the composite nutrient microneedle hair growth patch of this embodiment are as follows:

[0043] (1) Preparation of compound nutrient solution: dissolve 1.25 mg of wheat germ albumin, 0.25 μg of anti-hypoxia peptide, 1.138 μg of vitamin B7, 205.7 μg of ferrous lactate, 379.2 μg of vitamin C and 140 μg of zinc sulfate in 1 ml of pure water to prepare a compound nutrient solution;

[0044] (2) 3D printing raw materials: Add the composite nutrient solution to 30 ml of hyaluronic acid solution to form a gel-like composite nutrient for 3D printing;

[0045] (3) 3D printing: Using hyaluronic acid as the matrix material of microneedles, design and optimize the shape (conical), size and structure of microneedles (such as Figure 1 ), print the gel-like composite nutrients into a microneedle hair growth patch, post-process the printed microneedle hair growth patch, dry, paste the non-woven fabric adhesive, sterilize, etc., to ensure the quality and safety of the microneedle hair growth patch; the 3D printer parameter settings used in this embodiment are as follows: the printing temperature is 200°C, the printing speed is 10mm / s, and the layer height is 50μm. Inject the mixture of hyaluronic acid and composite nutrients into the feed port of the printer and automatically print it into the designed microneedle shape. The shape of the microneedle is conical, with a height of 500μm, a bottom diameter of 300μm, and a needle tip of 5μm;

[0046] (4) Post-processing: Wash the printed microneedles with deionized water three times to remove the remaining uncured material. Dry the washed microneedles in a windless environment at 35°C for 48 hours to remove moisture. Observe the patch and, after drying and forming, demold, cut, disinfect, and package.

[0047] Note: The dried microneedles are sterilized using ethylene oxide sterilization for 12 hours.

[0048] The electron microscope image of the nutrient microneedle patch prepared in this application is as follows Figure 2As shown in the figure, the arrangement is very neat, presenting a regular cone shape. The shape of the patch is very complete, the surface is smooth and uniform, and there are no obvious defects. In addition, the height of these microstructures is maintained at 600 microns, reflecting the high manufacturing precision and consistency. This morphological design is conducive to the effective penetration and nutrient release of the hair growth patch, thereby achieving better improvement effects, and it does not contact the pain nerves, and there is no tingling feeling (the transdermal schematic diagram is shown in the figure). Figure 1 as shown).

[0049] Figure 3 Sample pictures of nutritional hair growth stickers from different shooting angles, by Figure 3 It can be seen that the nutritional hair growth patch is transparent as a whole, the needle-shaped structure is complete, and the backing is uniform and has no cracks.

[0050] Example 2

[0051] The preparation steps of the composite nutrient microneedle hair growth patch of this embodiment are as follows:

[0052] (1) Preparation of compound nutrient solution: Dissolve 1.27 mg of wheat germ albumin, 0.30 μg of anti-hypoxia peptide, 1.5 μg of vitamin B7, 206 μg of ferrous lactate, 380 μg of vitamin C and 143 μg of zinc sulfate in 1 ml of pure water to prepare a compound nutrient solution;

[0053] (2) 3D printing raw materials: Add the composite nutrient solution to 30 ml of hyaluronic acid solution to form a gel-like composite nutrient for 3D printing;

[0054] (3) 3D printing: Using hyaluronic acid as the matrix material of microneedles, design and optimize the shape (conical), size and structure of microneedles (such as Figure 1 ), print the gel-like composite nutrients into a microneedle hair growth patch, post-process the printed microneedle hair growth patch, dry, paste the non-woven fabric adhesive, sterilize, etc., to ensure the quality and safety of the microneedle hair growth patch; the 3D printer parameter settings used in this embodiment are as follows: the printing temperature is 200°C, the printing speed is 10mm / s, and the layer height is 50μm. Inject the mixture of hyaluronic acid and composite nutrients into the feed port of the printer and automatically print it into the designed microneedle shape. The shape of the microneedle is conical, with a height of 500μm, a bottom diameter of 300μm, and a needle tip of 5μm;

[0055] (4) Post-processing: The printed microneedles (such as Figure 4 ) Wash with deionized water three times to remove the remaining uncured material. Dry the cleaned microneedles in a windless environment at 35°C for 48 hours to remove moisture. Observe the patch and after drying and forming, demold, cut, disinfect and package.

[0056] Note: The dried microneedles are sterilized using ethylene oxide sterilization for 12 hours.

[0057] Example 3

[0058] The preparation steps of the composite nutrient microneedle hair growth patch of this embodiment are as follows:

[0059] (1) Preparation of compound nutrient solution: dissolve 1.29 mg of wheat germ albumin, 0.35 μg of anti-hypoxia peptide, 2.0 μg of vitamin B7, 206 μg of ferrous lactate, 380 μg of vitamin C and 143 μg of zinc sulfate in 1 ml of pure water to prepare a compound nutrient solution;

[0060] (2) 3D printing raw materials: Add the composite nutrient solution to 30 ml of hyaluronic acid solution to form a gel-like composite nutrient for 3D printing;

[0061] (3) 3D printing: Using hyaluronic acid as the matrix material of microneedles, design and optimize the shape (conical), size and structure of microneedles (such as Figure 1 ), print the gel-like composite nutrients into a microneedle hair growth patch, post-process the printed microneedle hair growth patch, dry, paste the non-woven fabric adhesive, sterilize, etc., to ensure the quality and safety of the microneedle hair growth patch; the 3D printer parameter settings used in this embodiment are as follows: the printing temperature is 200°C, the printing speed is 10mm / s, and the layer height is 50μm. Inject the mixture of hyaluronic acid and composite nutrients into the feed port of the printer and automatically print it into the designed microneedle shape. The shape of the microneedle is conical, with a height of 500μm, a bottom diameter of 300μm, and a needle tip of 5μm;

[0062] (4) Post-processing: The printed microneedles (such as Figure 5 ) Wash with deionized water three times to remove the remaining uncured material. Dry the cleaned microneedles in a windless environment at 35°C for 48 hours to remove moisture. Observe the patch and after drying and forming, demold, cut, disinfect and package.

[0063] Note: The dried microneedles are sterilized using ethylene oxide sterilization for 12 hours.

[0064] Example 4

[0065] The preparation steps of the composite nutrient microneedle hair growth patch of this embodiment are as follows:

[0066] (1) Preparation of compound nutrient solution: dissolve 1.3 mg of wheat germ albumin, 0.4 μg of anti-hypoxia peptide, 2.0 μg of vitamin B7, 208 μg of ferrous lactate, 387 μg of vitamin C and 148 μg of zinc sulfate in 1 ml of pure water to prepare a compound nutrient solution;

[0067] (2) 3D printing raw materials: Add the composite nutrient solution to 30 ml of hyaluronic acid solution to form a gel-like composite nutrient for 3D printing;

[0068] (3) 3D printing: Using hyaluronic acid as the matrix material of microneedles, design and optimize the shape (conical), size and structure of microneedles (such as Figure 1 ), print the gel-like composite nutrients into a microneedle hair growth patch, post-process the printed microneedle hair growth patch, dry, paste the non-woven fabric adhesive, sterilize, etc., to ensure the quality and safety of the microneedle hair growth patch; the 3D printer parameter settings used in this embodiment are as follows: the printing temperature is 200°C, the printing speed is 10mm / s, and the layer height is 50μm. Inject the mixture of hyaluronic acid and composite nutrients into the feed port of the printer and automatically print it into the designed microneedle shape. The shape of the microneedle is conical, with a height of 500μm, a bottom diameter of 300μm, and a needle tip of 5μm;

[0069] (4) Post-processing: The printed microneedles (such as Figure 6 ) Wash with deionized water three times to remove the remaining uncured material. Dry the cleaned microneedles in a windless environment at 35°C for 48 hours to remove moisture. Observe the patch and after drying and forming, demold, cut, disinfect and package.

[0070] Note: The dried microneedles are sterilized using ethylene oxide sterilization for 12 hours.

[0071] Example 5

[0072] The preparation steps of the composite nutrient microneedle hair growth patch of this embodiment are as follows:

[0073] (1) Preparation of compound nutrient solution: dissolve 1.31 mg of wheat germ albumin, 0.38 μg of anti-hypoxia peptide, 2.8 μg of vitamin B7, 209 μg of ferrous lactate, 388 μg of vitamin C and 141 μg of zinc sulfate in 1 ml of pure water to prepare a compound nutrient solution;

[0074] (2) 3D printing raw materials: Add the composite nutrient solution to 30 ml of hyaluronic acid solution to form a gel-like composite nutrient for 3D printing;

[0075] (3) 3D printing: Using hyaluronic acid as the matrix material of microneedles, design and optimize the shape (conical), size and structure of microneedles (such as Figure 7), print the gel-like composite nutrients into a microneedle hair growth patch, post-process the printed microneedle hair growth patch, dry, paste the non-woven fabric adhesive, sterilize, etc., to ensure the quality and safety of the microneedle hair growth patch; the 3D printer parameter settings used in this embodiment are as follows: the printing temperature is 200°C, the printing speed is 10mm / s, and the layer height is 50μm. Inject the mixture of hyaluronic acid and composite nutrients into the feed port of the printer and automatically print it into the designed microneedle shape. The shape of the microneedle is conical, with a height of 500μm, a bottom diameter of 300μm, and a needle tip of 5μm;

[0076] (4) Post-processing: Wash the printed microneedles with deionized water three times to remove the remaining uncured material. Dry the washed microneedles in a windless environment at 35°C for 48 hours to remove moisture. Observe the patch and, after drying and forming, demold, cut, disinfect, and package.

[0077] Note: The dried microneedles are sterilized using ethylene oxide sterilization for 12 hours.

[0078] Comparative Example 1

[0079] The preparation steps of the composite nutrient microneedle hair growth patch of this embodiment are as follows:

[0080] (1) Preparation of composite nutrient solution: dissolve 0.25 μg of anti-hypoxia peptide, 1.138 μg of vitamin B7, 205.7 μg of ferrous lactate, 379.2 μg of vitamin C and 140 μg of zinc sulfate in 1 ml of pure water to prepare composite nutrient solution;

[0081] (2) 3D printing raw materials: Add the composite nutrient solution to 30 ml of hyaluronic acid solution to form a gel-like composite nutrient for 3D printing;

[0082] (3) 3D printing: Using hyaluronic acid as the matrix material of microneedles, design and optimize the shape (conical), size and structure of microneedles (such as Figure 1 ), print the gel-like composite nutrients into a microneedle hair growth patch, post-process the printed microneedle hair growth patch, dry, paste the non-woven fabric adhesive, sterilize, etc., to ensure the quality and safety of the microneedle hair growth patch. The 3D printer parameter settings used in this embodiment are as follows: the printing temperature is 200°C, the printing speed is 10mm / s, and the layer height is 50μm. Inject the mixture of hyaluronic acid and composite nutrients into the feed port of the printer and automatically print it into the designed microneedle shape. The shape of the microneedle is conical, with a height of 500μm, a bottom diameter of 300μm, and a needle tip of 5μm;

[0083] (4) Post-processing: Wash the printed microneedles with deionized water three times to remove the remaining uncured material. Dry the washed microneedles in a windless environment at 35°C for 48 hours to remove moisture. Observe the patch and, after drying and forming, demold, cut, disinfect, and package.

[0084] Note: The dried microneedles are sterilized using ethylene oxide sterilization for 12 hours.

[0085] Application Examples

[0086] The animal experiment plan for the treatment of hair loss with microneedle nutrition hair growth patch is designed. The main experimental purpose is to verify the effectiveness and safety of microneedle nutrition hair growth patch in the treatment of androgenic alopecia (hair loss).

[0087] Experimental animals: Male c57bl / 6, SPF grade, initial body weight: 20g±2g, 25 mice (modeling 1-4 weeks, average body weight 25g, drug dosage calculated based on 25g), purchased from Beijing Huafukang Biotechnology Co., Ltd., No.: 110322241101082543, License No.: SCK (Beijing) 2019-0008, Experimental Unit Use License No.: SYXK (Beijing) 2021-0012. This strain of mice is often used in hair-related research.

[0088] Detection reagents and instruments: such as skin tissue sectioning equipment and microscopes.

[0089] Animal Grouping:

[0090] The mice were randomly divided into the following 5 groups:

[0091] Group A (model group, male baldness model was established but no treatment was given), Group B (blank group, i.e., negative control group), Group C (positive control group, treated with minoxidil solution), Group D1 (5 nutrients hair growth patch prepared in Comparative Example 1, excluding wheat germ albumin), Group D2 (6 nutrients prepared in Example 1, nutrient hair growth patch containing wheat germ albumin).

[0092] Establishment of male baldness model: First, part of the hair on the back of each group of mice was shaved, and the male hormone testosterone was applied to the hair removal area once a day for two weeks. It can miniaturize the hair follicles of animals, simulate the pathological and physiological process of male baldness in humans, and induce the male baldness model.

[0093] Treatment process:

[0094] All groups (except group B or the blank group) were given an application of the male hormone testosterone once a day.

[0095] Group C: Minoxidil solution (50 mg minoxidil per 1 ml) was applied to the hair removal area according to the regular dose.

[0096] Group D1: The microneedle nutrition hair growth patch prepared with compound nutrients 1 was applied to the hair removal area of ​​the mice and kept for more than 4 hours before being removed. Once a day (apply after 9:00 pm and remove before 6:00 am).

[0097] Group D2: The microneedle nutrition hair growth patch prepared with compound nutrients 2 was applied to the hair removal area of ​​the mice; it was kept for more than 4 hours and then removed. Once a day (apply after 9:00 pm and remove before 6:00 am).

[0098] Group B: blank microneedle patch was given and the operation was the same as that of the microneedle nutrition hair growth patch group.

[0099] Histopathological examination: After the experiment, the skin tissue of the mouse hair removal site was sliced ​​and DAPI staining was used to observe the changes in the morphology, structure and number of hair follicles.

[0100] Experimental schedule: 1-4 weeks after the establishment of the male pattern baldness model: treatment, weekly observation and record of hair appearance. 4th week: end of the experiment, collect samples for various index tests.

[0101] The fixation and processing methods of tissue samples for observing cytokines such as 4-HNE and CD31 are as follows:

[0102] Cryofixation: The tissue sample is quickly frozen in liquid nitrogen and then transferred to a -80℃ refrigerator for storage. This method can better preserve the antigenicity and ultrastructure of the tissue, but special processing methods are required in the subsequent sectioning and staining process, such as using a freezing microtome for sectioning. Dehydration: The fixed tissue needs to be dehydrated with alcohol to remove the water in the tissue. Transparency: After dehydration, the tissue sample will become opaque, and a transparent agent (such as xylene) is needed to remove the alcohol and other solvents in it. Embedding: The transparent tissue can be embedded in paraffin to form a tissue block that is easy to section. Put the tissue sample in the melted paraffin, let the paraffin fully penetrate into the tissue sample, and then pour it into the mold. When the paraffin cools and solidifies, the tissue sample is embedded in the paraffin. Sectioning: Use a microtome to cut the embedded tissue into 3-5 micron thick slices, and attach the slices to a glass slide for subsequent staining, analysis and observation.

[0103] 4-HNE, or 4-hydroxynonenal, is a highly active lipid peroxidation product. 4-HNE has strong cytotoxicity and can react with biomacromolecules such as proteins and nucleic acids in cells, thereby affecting the normal function of cells. Although 4-HNE has many important functions in the body, excessive production or continued presence may cause damage to cells and tissues. Representative images of 4-HNE in skin tissues of different groups on the 14th day after hair removal, such as Figure 8As shown, on the 14th day, the positive expression of 4-HEN in group A was evenly distributed, while that in group B was mainly concentrated around the hair follicles. The three intervention groups (C, D1, and D2) had weaker positive expression of 4-HEN around the hair follicles, indicating that these treatments reduced the damage of ROS to the hair follicles, and the Ki67 expression in these three groups was significantly higher than that in the Model group, indicating that the proliferation of hair follicle cells was promoted. Ki67 is a nuclear protein that is closely related to cell proliferation. It is expressed in all stages of the cell cycle, but not in the cell resting phase (G0 phase). The expression level of Ki67 reflects the proliferation activity of the cells. The higher its expression, the faster the cell proliferation.

[0104] On day 14, the positive expression of 4-HEN in group A was evenly distributed, while that in group B was mainly concentrated around the hair follicles. The positive expression of 4-HEN around the hair follicles in the three intervention groups (C, D1, and D2) was weaker, indicating that these treatments reduced the damage of ROS to the hair follicles; the expression of Ki67 in the three intervention groups (C, D1, and D2) was significantly higher than that in group A (model), indicating that the proliferation of hair follicle cells was promoted.

[0105] CD31 is a special marker for vascular endothelial cells and can indicate the distribution of blood vessels in tissues. Fig. 9 As shown, there is only a small amount of CD31 positive expression around the hair follicles in group B, and the CD31 positive expression around the hair follicles in the three intervention groups (C, D1, and D2) is strongly positive. At the same time, the number of blood vessels around the hair follicles in these three groups increased (black arrows), indicating that angiogenesis around the hair follicles was induced. CD31, also known as platelet endothelial cell adhesion molecule-1 (PECAM-1), is a transmembrane glycoprotein that is mainly expressed on the surface of endothelial cells, platelets, some white blood cells, and some hematopoietic stem cells.

[0106] In hair research, CD31 is mainly used as a marker for vascular endothelial cells. Its role in normal hair growth: (1) Nutritional supply: Normal hair growth requires adequate nutrients and oxygen supply, which are mainly provided by blood vessels around the hair follicles. CD31-positive vascular endothelial cells form a rich capillary network around the hair follicles to provide the necessary nutritional support for hair growth. During the hair growth period, the blood vessel density around the hair follicles is usually high, and the expression of CD31 is relatively strong to ensure that hair can grow rapidly. (2) Growth factor regulation: The vascular endothelial cells around the hair follicles can also secrete a variety of growth factors and cytokines, which have an important regulatory effect on hair growth. For example, vascular endothelial growth factor (VEGF) can promote angiogenesis, increase blood supply around the hair follicles, and thus stimulate hair growth. CD31-positive endothelial cells may indirectly affect hair growth by regulating the secretion of growth factors such as VEGF.

[0107] By intervening in the androgen mouse model, Fig.10 It can be seen that:

[0108] (1) Group C (minoxidil patch), group D1 (nutritional hair growth patch), and group D2 (nutritional hair growth patch containing wheat germ protein) had a stronger stimulating effect on hair growth than the model group and the blank patch group.

[0109] (2) The three groups of mice treated with patches, namely group C (minoxidil patch), group D1 (nutritional hair growth patch), and group D2 (nutritional hair growth patch containing wheat germ protein), began to develop dark gray skin on the 13th day, accompanied by the regeneration of a small amount of hair.

[0110] (3) On the 16th day, hair regeneration was observed in all three groups.

[0111] (4) By the 27th day, no obvious hair growth occurred in the model group, and some areas of the blank patch-treated area still did not show hair regeneration (the reason why the blank group grew hair is that hyaluronic acid itself has an anti-inflammatory effect, which can relieve the inflammatory response. Secondly, the micro-trauma caused by the microneedle may also have a promoting effect, and the mechanical tension during the patch operation also has a promoting effect.).

[0112] (5) The hair regeneration area in group C (minoxidil patch), group D1 (nutritional hair growth patch), and group D2 (nutritional hair growth patch containing wheat germ protein) was greater than that in the blank patch group.

[0113] (6) Compared with the D1 group (nutritional hair growth patch) and the D2 group (nutritional hair growth patch, containing wheat germ protein), on the 16th day, the D2 group (nutritional hair growth patch, containing wheat germ protein) was already fully covered with hair, while the D1 group (nutritional hair growth patch) had no growth; on the 27th day, the D2 group (nutritional hair growth patch, containing wheat germ protein) was completely covered with hair, while the D1 group (nutritional hair growth patch) was not completely covered.

[0114] (7) The effects of group C (minoxidil patch) and group D2 (nutritional hair growth patch containing wheat germ protein) were basically equivalent, indicating that the nutritional hair growth patch achieved the effects of drugs without the side effects of drugs.

[0115] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A composite nutrient microneedle hair growth patch, characterized in that: It includes a non-woven fabric layer, a sodium hyalate base and conical microneedles, wherein the conical microneedles are evenly distributed on one side of the sodium hyalate base, and the other side of the sodium hyalate base is covered with a non-woven fabric layer; The conical microneedles contain wheat germ albumin, anti-hypoxia peptide, vitamin B7, ferrous lactate, vitamin C and zinc sulfate; the mass ratio of wheat germ albumin, anti-hypoxia peptide, vitamin B7, ferrous lactate, vitamin C and zinc sulfate in the composite nutrient solution is: 1250-1350: 0.25-0.50: 1-3: 205-210: 375-390: 140-150; The preparation method of the tapered microneedle is: Wheat germ albumin, anti-hypoxia peptide, vitamin B7, ferrous lactate, vitamin C and zinc sulfate are dissolved in water to prepare a composite nutrient solution, and then hyaluronic acid solution is added to obtain a gel-like composite nutrient, which is then produced by 3D printing to obtain a composite nutrient microneedle hair growth patch.

2. The composite nutrient microneedle hair growth patch according to claim 1, characterized in that: The concentration of wheat germ albumin in the composite nutrient solution is 1.25-1.35 mg / mL.

3. The composite nutrient microneedle hair growth patch according to claim 2, characterized in that: The mass percentage of the hyaluronic acid solution is 25%-35%.

4. The composite nutrient microneedle hair growth patch according to claim 3, characterized in that: The volume ratio of the composite nutrient solution to the hyaluronic acid solution is 1:30-50.

5. The composite nutrient microneedle hair growth patch according to any one of claims 2 to 4, characterized in that: The cone-shaped microneedle has a height of 500 μm, a bottom diameter of 300 μm, and a needle tip of 5 μm.

6. The composite nutrient microneedle hair growth patch according to claim 5, characterized in that: The parameters of the 3D printing are: printing temperature 200° C., printing speed 10 mm / s, and layer height 50 μm.

7. Use of the composite nutrient microneedle hair growth patch according to claim 1 in the preparation of a drug or medical device for treating male pattern baldness.

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