A hair growth white tea nanoparticle / Ca 2+ Method for preparing and using a composite system

Abstract

The present application relates to the fields of medicine and cosmetics, and particularly relates to a white tea nanoparticle / Ca 2+ Composite system and preparation method and application thereof. The present application discloses a white tea nanoparticle / Ca 2+ Composite system and preparation method and application thereof. The present application discloses a white tea nanoparticle / Ca 2+ Composite system and preparation method and application thereof. The present application discloses a white tea nanoparticle / Ca 2+ Composite system and preparation method and application thereof. The present application discloses a white tea nanoparticle / Ca

Description

Technical Field

[0001] This invention relates to the fields of medicine and cosmetics, and more particularly to a white tea nanoparticle / Ca nanoparticle with hair growth function. 2+ Composite systems, their preparation methods, and applications. Background Technology

[0002] The scalp ecosystem is characterized by dense hair, abundant glands, and a complex structure. Hair follicles, located at the hair root and responsible for anchoring hair, are crucial appendages of the scalp. A mature hair follicle is a complex micro-organ whose growth cycle is strictly regulated. During postnatal development, the hair follicle undergoes a continuous cycle of anagen (growth), catagen (transitional), and telogen (resting) phases. The proper functioning of this process determines whether hair can successfully undergo cyclical growth. Therefore, research on anchoring the microstructure of hair follicles to promote hair growth is key in the field of hair care. However, currently, there is a lack of products on the market that anchor deeply into the hair follicle, and most hair care products contain certain toxic chemicals, which can have negative health effects with long-term use. Therefore, developing safe and effective shampoos and conditioners that can penetrate deep into the hair follicle, based on the close relationship between hair follicles and hair, is of paramount importance.

[0003] With the development of nanotechnology, the application of nanomaterials has gradually attracted attention. Encapsulating substances into nanoparticles holds promise for improving the bioavailability of locally applied products. While nanoparticles cannot penetrate the intact skin barrier, they can reach hair follicles and release active substances within them. Studies have shown that medium-sized particles (around 640 nm) have the strongest ability to enter hair follicles, and by changing the particle size, selective targeting to different parts of the hair follicle can be achieved. Therefore, nanotechnology holds promise for providing new strategies for the development of hair growth products that anchor to the hair follicle structure.

[0004] In recent years, with the deepening research on the health benefits of tea, white tea has gradually entered the public eye due to its excellent nutritional components and active functions. White tea is a type of tea made through withering and drying processes. During processing, a series of physicochemical changes occur, resulting in its unique quality and style. Numerous studies have shown that white tea contains more polyphenols, caffeine, amino acids, and other functional substances than other tea varieties. It has functions such as anti-oxidation, antibacterial and anti-inflammatory effects, repairing the skin barrier, and protecting damaged cells, and is of great significance for the regulation of scalp microecology. Existing research has proven that effective components in white tea, such as EGCG, can reduce the concentration of cellular inflammatory factors and can play a certain role in alleviating inflammatory responses in blood vessels, intestines, and nerves in animals. However, the therapeutic effect of a single white tea component is not ideal, and a complex system needs to be constructed to enhance the therapeutic effect. Existing research has shown that Ca... 2+It is a key factor in regulating cell proliferation, maturation, and promoting epidermal lipid barrier function, playing an important regulatory role in enhancing wound healing, epidermal regeneration, and dermal reconstruction; meanwhile, Ca... 2+ It can also enhance EGCG's ability to scavenge free radicals, thereby improving its antioxidant and anti-inflammatory effects. Summary of the Invention

[0005] The technical problem to be solved by this invention is to provide white tea nanoparticles / Ca with hair growth function. 2+ Composite systems, their preparation methods, and applications.

[0006] To solve the above technical problems, the present invention provides a hair growth white tea nanoparticle / Ca 2+ The preparation method of the composite system includes the following steps:

[0007] (1) Grind the white tea (put it into a high-speed grinder for grinding and then sieve it) to obtain tea powder;

[0008] (2) Mix the tea powder obtained in step (1) with water at a material-to-liquid ratio of 1g:25-35mL, heat the mixture at 90-100℃ for 35-40min, and then filter to obtain the extract.

[0009] Preferred ratio: 1g:30mL, extraction at 100℃ for 40min;

[0010] (3) Gradient centrifugation: The extract obtained in step (2) is centrifuged for the first time at 4000-5000 rpm for 10±2 min. The supernatant obtained from the first centrifugation is then centrifuged for the second time at 6000-7000 rpm for 10±2 min. The precipitate obtained from the second centrifugation is ultrasonically cleaned (pure water that does not cover the precipitate is added for ultrasonic cleaning), and then freeze-dried to obtain white tea nanoparticle dry powder (stored at -80℃).

[0011] Gradient centrifugation preferred: After centrifuging the extract obtained in step (2) for the first time at 4500 rpm for 10 min, take the supernatant and centrifuge the supernatant a second time at 6000 rpm for 10 min;

[0012] (4) The white tea nanoparticle powder obtained in step (3) is resuspended in pure water to prepare a (5±0.5)mg / ml white tea nanoparticle suspension.

[0013] Take 6 ml of white tea nanoparticle suspension, stir magnetically (for about 3 ± 1 min); add (dropwise) 1–2 ml of CaCl2 solution with a concentration of (5 ± 0.5) mg / ml and stir for 30 ± 5 min; then centrifuge at 6000–7000 rpm for 10 ± 2 min, take the precipitate and ultrasonically clean it (add pure water to cover the precipitate for ultrasonic cleaning), freeze-dry to obtain white tea nanoparticles / CaCl2 with hair growth function. 2+ Composite system.

[0014] As the hair-growth white tea nanoparticles / Ca of the present invention 2+ Improvement of the preparation method of the composite system: In step (1), the particle size of the tea powder is 30-50 mesh.

[0015] As the hair-growth white tea nanoparticles / Ca of the present invention 2+ Improved preparation method of composite system: Ultrasonic cleaning is performed by adding pure water to the precipitate (to resuspend the nanoparticles), repeatedly blowing while sonicating for 5±1 min, centrifuging at 6000-7000 rpm for 10±2 min; the resulting precipitate is then subjected to the above ultrasonic cleaning operation once more.

[0016] The ultrasonic cleaning in steps (3) and (4) is as described above.

[0017] This invention also provides hair-growth white tea nanoparticles / Ca prepared by the above method. 2+ Composite system.

[0018] This invention also provides hair growth white tea nanoparticles / Ca 2+ Application of composite systems in the preparation of drugs or reagents that promote hair growth.

[0019] As an improvement to the application of the present invention: drugs or reagents that promote hair growth include pharmaceutical preparations, cosmetics, care products, and beauty products.

[0020] The cosmetics are selected from any one of the following: shampoo, conditioner, hair serum, hair oil, scalp and hair nourishing products, hair gel, hair mask, and eyelash / eyebrow care serum.

[0021] The white tea nanoparticles / Ca with hair growth function of the present invention 2+ When used in scalp care, the compound system can be prepared as a 2.5–10 mg / ml aqueous solution and applied to the scalp 1–3 times daily.

[0022] This invention first considers extracting nanoparticles from white tea and constructing white tea nanoparticle / Ca 2+ The composite system utilizes white tea nanoparticles and Ca 2+It has a synergistic effect in penetrating deep into the hair follicle, repairing the skin barrier, and promoting hair growth, thus promoting hair follicle development and hair growth, providing a new method for solving hair growth problems, and providing a theoretical basis for related research on regulating the scalp microecology using nanoparticles.

[0023] The beneficial effects of this invention are: it successfully prepares composite nanomaterials based on white tea nanoparticles, expanding the application fields of white tea nanoparticles. The nanoscale composite system prepared by this invention can effectively penetrate deep into the hair follicle, promote hair follicle cell division, and effectively accelerate hair growth. It can be developed into a natural hair growth promoter, replacing artificially synthesized hair care and hair growth products. This invention is scientifically and rationally designed and has great practical application value.

[0024] In summary, this invention extracts white tea nanoparticles from white tea infusion and combines them with Ca... 2+ After centrifugation and washing, white tea nanoparticles / Ca were obtained. 2+ Composite system. White tea nanoparticles / Ca 2+ The composite system can promote hair follicle reconstruction and accelerate hair growth. White tea nanoparticles / Ca 2+ The compound system has a significant effect on promoting hair growth. Attached Figure Description

[0025] The specific embodiments of the present invention will be further described in detail below with reference to the accompanying drawings.

[0026] Figure 1 For characterization of experimental diagrams;

[0027] Figure 1 In the diagram, a is the UV-Vis spectrum experimental diagram; b is the Zeta potential analysis experimental diagram; and c is the nanoparticle size analysis experimental diagram.

[0028] Figure 2 The back views of mice in different drug administration groups on days 2, 4, 6, 8, 10 and 12 after hair removal.

[0029] Figure 3 Images showing hair growth in mice in different drug administration groups on days 2, 4, 6, 8, 10, and 12 after hair removal;

[0030] Figure 3 In the image, a is a rating chart for hair growth; b is a chart showing the percentage of hair coverage.

[0031] Figure 4 Microscopic images of hair follicles in mice from different drug administration groups on day 12 after hair removal;

[0032] Figure 4In the image, a is a micrograph of hair follicles in the model group (M) mice on day 12 after hair removal; b is a micrograph of hair follicles in the white tea nanoparticle group (WT) mice on day 12 after hair removal; c is a micrograph of hair follicles in the white tea nanoparticle and CaCl2 mixed group (WT+CaCl2) mice on day 12 after hair removal; and d is a micrograph of hair follicles in the white tea nanoparticle / CaCl2 mixed group. 2+ Composite system group (WT / Ca) 2+ Microscopic image of hair follicles in mice on day 12 after hair removal;

[0033] Figure 5 H&E stained sections of skin from mice in different drug administration groups on day 12 after hair removal;

[0034] Figure 5 In the image, a is a H&E stained section of skin from mice in the model group (M) on day 12 after hair removal; b is a H&E stained section of skin from mice in the white tea nanoparticle group (WT) on day 12 after hair removal; c is a H&E stained section of skin from mice in the white tea nanoparticle and CaCl2 mixed group (WT+CaCl2) on day 12 after hair removal; and d is a section of skin from mice in the white tea nanoparticle / CaCl2 mixed group. 2+ Composite system group (WT / Ca) 2+ H&E stained skin section of a mouse on day 12 after hair removal;

[0035] Figure 6 Quantitative graphs of hair follicle length on day 12 after hair removal in mice of different drug administration groups. Detailed Implementation

[0036] The present invention will be further described below with reference to specific embodiments, but the scope of protection of the present invention is not limited thereto:

[0037] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; the reagents and materials described are commercially available unless otherwise specified.

[0038] The high-speed pulverizer is set to a speed of 15,000 to 18,000 rpm.

[0039] Example 1: A white tea nanoparticle / Ca 2+ The preparation method of the composite system comprises the following steps:

[0040] (1) Weigh out white tea, put it into a high-speed grinder for grinding, and pass it through a 40-mesh sieve to obtain tea powder for later use;

[0041] (2) The tea powder obtained in step (1) is mixed with pure water at a ratio of 1g:30mL to obtain a mixture; the mixture is heated and extracted at 100℃ for 40min, and then filtered to obtain a clear extract.

[0042] (3) Gradient centrifugation: After centrifuging the extract obtained in step (2) at 4500 rpm for 10 min, take the supernatant and centrifuge the supernatant a second time at 6000 rpm for 10 min.

[0043] Add pure water (enough to cover the precipitate) to the precipitate obtained from the second centrifugation to resuspend the nanoparticles. While sonicating, repeatedly blow and agitate for 5 minutes. Centrifuge at 6000 rpm for 10 minutes to obtain the precipitate, and repeat the operation once more. Finally, freeze-dry the precipitate obtained from centrifugation to obtain white tea nanoparticle powder, and store it at -80℃ for later use.

[0044] (4) Resuspend the white tea nanoparticle powder obtained in (3) in pure water to prepare a 5 mg / ml white tea nanoparticle suspension; take 6 ml of the white tea nanoparticle suspension, stir magnetically for 3 min, add 1 ml of 5 mg / ml CaCl2 solution and stir for 30 min, centrifuge at 6000 rpm for 10 min; add pure water to the precipitate to resuspend the nanoparticles, repeatedly blow and blow while sonicating for 5 min, centrifuge at 6000 rpm for 10 min to obtain the precipitate, and repeat the operation once more to obtain white tea nanoparticles with anti-hair loss and hair growth function / CaCl2. 2+ The composite system, after freeze-drying, is stored at -80℃ for later use.

[0045] Experiment 1: White Tea Nanoparticles / Ca 2+ Characterization of composite systems

[0046] The white tea nanoparticles / Ca prepared in Example 1 were used... 2+ The composite system was characterized by ultraviolet absorption spectroscopy, zeta potential analysis, and nanoparticle size analysis.

[0047] Ultraviolet (UV) absorption spectroscopy was used to characterize the optical properties of nanoparticles. A UV-Vis spectrophotometer was used to measure the UV absorption spectrum (200–1000 nm) of the white tea nanoparticles / Ca nanoparticles prepared in Example 1 through a quartz microporous plate. 2+ The composite system and the white tea nanoparticles were measured three times each.

[0048] Nanoparticle size analysis and Zeta potential analysis were used to characterize the particle size distribution and Zeta potential of nanoparticles. Using nanoparticle size and Zeta potential analyzers, the particle size distribution and Zeta potential of well-dispersed white tea nanoparticles / Ca were analyzed. 2+ The composite system resuspension and the white tea nanoparticle resuspension were measured three times respectively.

[0049] according to Figure 1 (a) Figure 1 (b) and Figure 1 (c) It can be seen that Ca 2+The addition of [a substance] altered the ultraviolet spectrophotometer and potential value of the white tea nanoparticles, proving that the white tea nanoparticles / Ca [a substance] [is a potential precursor]. 2+ Successful preparation of the composite system; white tea nanoparticles / Ca 2+ The composite particles have a similar particle size to the white tea nanoparticles, around 500 nm.

[0050] Experiment 2: White Tea Nanoparticles / Ca 2+ Effects of the composite system on hair growth in hair-removed mice

[0051] Eight-week-old male C57BL / 6 mice were used in the experiment. After routine acclimatization culture (7 days), the mice were randomly divided into the model group (M), white tea nanoparticle group (WT, 5 mg / ml white tea nanoparticle suspension), white tea nanoparticle and CaCl2 mixed group (WT + CaCl2, 5 mg / ml white tea nanoparticle suspension and 5 mg / ml CaCl2 solution mixed at a volume ratio of 6:1), and the white tea nanoparticle / CaCl2 mixture obtained in Example 1. 2+ Composite system group (WT / Ca) 2+ 5mg / ml white tea nanoparticles / Ca 2+ (Composite suspension), 9 mice in each group. Before the experiment, the back hair of each group was shaved with a small animal razor, and depilatory cream was applied for thorough hair removal to establish a hair removal model; white tea nanoparticle group, white tea nanoparticle and CaCl2 mixture group, and white tea nanoparticle / CaCl2 mixture group. 2+ The composite system group was treated by applying absorbent cotton soaked in the corresponding solution to the hairless back of mice for 2 minutes; the model group was treated with distilled water in the same way. The experiment lasted for 12 days. During the experiment, the hair growth on the back of the mice was recorded daily by taking photos. The mice had free access to food and water during the experiment.

[0052] according to Figure 2 It can be seen that, compared with the model group, the white tea nanoparticle group, the white tea nanoparticle and CaCl2 mixture group, and the white tea nanoparticle / CaCl2 mixture group are significantly different. 2+ All composite systems promoted hair growth in mice, but the white tea nanoparticles / Ca... 2+ The composite system group showed the best results, indicating that the white tea nanoparticles / Ca 2+ The complex system has the best effect on promoting hair growth.

[0053] Experiment 3: White Tea Nanoparticles / Ca 2+ The effect of composite systems on hair coverage

[0054] Model group (I), white tea nanoparticle group (II), white tea nanoparticle and CaCl2 mixture group (III), white tea nanoparticle / CaCl2 mixture group 2+The setup of the composite system group (IV) was the same as in Experiment 2; during the experiment, the hair growth of mice was recorded by taking photos every day, the grade was evaluated, the hair growth area was measured and the percentage was calculated.

[0055] according to Figure 3 (a) and Figure 3 (b) It can be seen that, compared with the model group, the white tea nanoparticle group, the white tea nanoparticle and CaCl2 mixture group, and the white tea nanoparticle / CaCl2 mixture group are significantly different. 2+ All components of the composite system promoted hair growth in mice, gradually progressing through four stages with increasing treatment time: resting phase, pigment deposition, hair shaft growth, and full coverage, with the percentage of hair coverage gradually increasing. Furthermore, the white tea nanoparticles / Ca... 2+ The composite system group showed the best results, exhibiting the fastest hair regrowth speed and ultimately achieving 100% hair coverage, indicating that the white tea nanoparticles / Ca 2+ The complex system has the best effect on promoting hair growth.

[0056] Experiment 4: White Tea Nanoparticles / Ca 2+ Effects of the composite system on hair follicle growth in hair-removed mice

[0057] Model group (M), white tea nanoparticle group (WT), white tea nanoparticle and CaCl2 mixture group (WT+CaCl2), white tea nanoparticle / Ca 2+ Composite system group (WT / Ca) 2+ The setup method was the same as in Experiment 2; after 12 days of treatment, skin tissue from each group of mice was taken and observed and photographed under an optical microscope.

[0058] according to Figure 4 (a) Figure 4 (b) Figure 4 (c) and Figure 4 (d) It can be seen that in the model group, after 12 days of hair removal treatment, the mouse hair follicles were still in the resting phase, the hair follicles atrophied, and the hair was short or no hair grew; in the white tea nanoparticle group and the white tea nanoparticle and CaCl2 mixture group, the hair follicles were in the early growth stage, the hair bulbs became larger, and hair began to grow but was fine; however, the white tea nanoparticle / CaCl2 mixture group... 2+ In the composite system group, the hair follicles are in the late growth stage, with rapid cell division, enlarged and plumped hair bulbs, and longer hair, indicating that the white tea nanoparticles / Ca 2+ The composite system has an optimal synergistic effect in promoting hair follicle growth.

[0059] Experiment 5: White Tea Nanoparticles / Ca 2+ Effects of the composite system on the skin tissue of hair-removed mice

[0060] Model group (M), white tea nanoparticle group (WT), white tea nanoparticle and CaCl2 mixture group (WT+CaCl2), white tea nanoparticle / Ca 2+ Composite system group (WT / Ca) 2+ The setup method was the same as in Experiment 2; 12 days after treatment, H&E staining was performed on the skin tissue of mice in each group for observation.

[0061] according to Figure 5 (a) Figure 5 (b) Figure 5 (c) Figure 5 (d) and Figure 6 It can be seen that in the model group, 12 days after hair removal treatment, the mouse hair follicles were still in the resting phase, the hair bulbs were atrophied and small in diameter, and most hair follicles had not yet differentiated to grow hair; the white tea nanoparticle group, the white tea nanoparticle and CaCl2 mixed group, and the white tea nanoparticle / CaCl2 group... 2+ After treatment with the composite system, hair follicle cells gradually divided, increased in diameter, and increased in length; among them, white tea nanoparticles / Ca 2+ The longest hair follicles in the composite system group indicate that the white tea nanoparticles / Ca 2+ The composite system has the best synergistic effect in promoting hair follicle growth.

[0062] Comparative Example 1: The amount of "5 mg / ml CaCl2 solution" in Example 1 was changed from 1 ml to 2 ml, 3 ml, and 0.5 ml, respectively; the rest remained the same as in Example 1. The resulting white tea nanoparticles / Ca... 2+ The composite system was then subjected to the corresponding tests as described above, and the performance results obtained were compared with those of Example 1 as shown in Table 1 below.

[0063] Table 1

[0064]

[0065] Comparative Example 2: In Example 1, CaCl2 was replaced with CaSO4 and Ca(NO3)2, while the calcium solution concentration and amount remained unchanged; everything else was the same as in Example 1. The resulting white tea nanoparticles / Ca 2+ The composite system was then subjected to the corresponding tests as described above, and the performance results obtained were compared with those of Example 1 as shown in Table 2 below.

[0066] Table 2

[0067]

[0068] Comparative Example 3: The "stirring for 30 min" in Example 1 was changed to 15 min, 60 min, and 120 min (as shown in Table 3 below). The calcium solution concentration and amount remained unchanged, and the rest were the same as in Example 1. The resulting white tea nanoparticles / Ca2+ The composite system was then tested according to the above experiments, and the performance results obtained were compared with those of Example 1 as shown in Table 3 below.

[0069] Table 3

[0070]

[0071] Comparative Example 4:

[0072] During the invention process, it was discovered that: changing the gradient centrifugation in step (3) to "centrifugation at 4500 rpm for 20 min" or "centrifugation at 6000 rpm for 20 min", while maintaining the same procedure as step (3) in Example 1, resulted in a significant difference in the particle size of the obtained white tea nanoparticles, meaning that the relatively uniform nanoparticles described in Example 1 could not be obtained. The obtained white tea nanoparticles / Ca 2+ The composite system was tested according to the above experiments, and the hair coverage percentage was approximately 85% on day 12.

[0073] Finally, it should be noted that the above examples are merely some specific embodiments of the present invention. Obviously, the present invention is not limited to the above embodiments and many variations are possible. All variations that can be directly derived or conceived by those skilled in the art from the disclosure of this invention should be considered within the scope of protection of this invention.

Claims

1. A type of hair-growth white tea nanoparticles / Ca 2+ The method for preparing the composite system is characterized by... Includes the following steps: (1) Grind the white tea to obtain tea powder; (2) Mix the tea powder obtained in step (1) with water at a material-to-liquid ratio of 1g:25~35mL, heat the mixture at 90~100℃ for 35~40min, and then filter to obtain the extract. (3) The extract obtained in step (2) is centrifuged for the first time at 4000~5000 rpm for 10±2 min, and then the supernatant obtained from the first centrifugation is centrifuged for the second time at 6000~7000 rpm for 10±2 min; the precipitate obtained from the second centrifugation is ultrasonically cleaned and then freeze-dried to obtain white tea nanoparticle dry powder. (4) The white tea nanoparticle powder obtained in step (3) is resuspended in pure water to prepare a white tea nanoparticle suspension of 5±0.5mg / ml. Take 6 ml of white tea nanoparticle suspension, stir magnetically for 3 ± 1 min; add 1-2 ml of CaCl2 solution with a concentration of 5 ± 0.5 mg / ml and stir for 30 ± 5 min; then centrifuge at 6000-7000 rpm for 10 ± 2 min, take the precipitate, ultrasonically wash, and freeze-dry to obtain white tea nanoparticles / CaCl2 solution. 2+ Composite system.

2. The hair-growth white tea nanoparticles / Ca according to claim 1 2+ The method for preparing the composite system is characterized by: In step (1), the particle size of the tea powder is 30-50 mesh.

3. The hair-growth white tea nanoparticles / Ca according to claim 2 2+ The method for preparing the composite system is characterized by: Ultrasonic cleaning is performed as follows: Add pure water to the precipitate, cover it completely, and repeatedly blow it while sonicating for 5±1 min. Centrifuge at 6000~7000 rpm for 10±2 min. Repeat the ultrasonic cleaning operation once more.

4. The hair-growth white tea nanoparticles / Ca prepared by the method described in any one of claims 1 to 3 2+ Composite system.

5. The hair-growth white tea nanoparticles / Ca prepared by the method described in any one of claims 1 to 3 2+ Application of composite systems in the preparation of drugs or cosmetics that promote hair growth.

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