Use of sclerostin in the preparation of a preparation for preventing and treating hair loss
By using periosteal proteins to regulate hair follicle cell activity, the problem of poor efficacy in existing hair loss treatments has been solved, and hair follicle cell proliferation and hair growth have been significantly promoted, providing a new hair loss treatment option.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- SOUTHERN MEDICAL UNIVERSITY
- Filing Date
- 2026-02-06
- Publication Date
- 2026-06-05
AI Technical Summary
Existing hair loss treatments such as hair transplantation and drug therapy have side effects and are not very effective. Furthermore, there is a lack of effective means to regulate hair follicle cell activity, resulting in varying treatment outcomes from person to person.
Osteomodulin (OMD) is used to regulate the activity of hair follicle papilla cells, hair matrix cells, and outer root sheath cells, promoting their proliferation and hair follicle cycle transition, and to prepare topical, patch, or injectable formulations for the treatment of hair loss.
Periostoma protein can significantly promote hair follicle cell proliferation, enhance the hair growth induction ability of dermal papilla cells, promote hair growth and hair follicle cycle transformation, and achieve the effectiveness and consistency of hair loss treatment.
Smart Images
Figure CN121648263B_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the fields of biomedicine and cosmetics, and specifically relates to the application of periosteal protein in the preparation of formulations for promoting hair follicle cell activity or for preventing and treating hair loss. Background Technology
[0002] Currently, hair loss treatment options mainly include hair transplantation, oral medications, and emerging PRP injections. Drug treatments have certain side effects, and their effectiveness often falls short of patient expectations. The effectiveness of hair transplantation is closely related to the condition of the hair follicles in the donor area, and the results of newer treatments vary from person to person (ZHOU Y, et al. Advances in microneedles research based on promoting hair regrowth [J]. Journal of Controlled Release, 2023, 353: 965-74.). Therefore, finding and developing new ingredients with preventative and therapeutic effects on hair loss is of great significance.
[0003] In mature individuals, hair follicles generally do not regenerate, but rather exhibit a cyclical pattern, including the anagen (growth) phase, catagen (regression) phase, and telogen (resting) phase. Hair grows rapidly during the anagen phase, stops growing during the catagen phase, and falls out during the telogen phase. New hair then grows back after entering the next anagen phase. Hair consists of two parts: the hair shaft and the hair root. The part exposed on the skin surface is the hair shaft, and the part embedded in the skin is the hair root. The tissue structure surrounding the hair root is the hair follicle (HF). The hair follicle is an accessory organ of the skin, distributed in most parts of the human skin. It is a complex microorganism with a tissue structure consisting of two layers: an inner epithelial sheath and an outer connective tissue sheath (HAMIDA OB, et al. Hair Regeneration Methods Using Cells Derived from Human Hair Follicles and Challenges to Overcome [J]. Cells, 2025, 14(1): 7.). The upper end of the epithelial sheath is continuous with the skin epidermis, and the lower end wraps around the hair root and fuses with the hair root, swelling to form a hair bulb. The epithelial cells in the hair bulb are hair matrix cells, which can differentiate into hair root and epithelial sheath cells. Therefore, hair matrix cells are the hair growth point and the only source cells responsible for the actual synthesis and extension of hair. The activity and differentiation direction of hair matrix cells directly determine the thickness, length and growth rate of hair. Almost all types of hair loss are ultimately directly or indirectly manifested as impaired function and weakened activity of hair matrix cells (MA S, et al. Transcriptomic Analysis Reveals Candidate Ligand-Receptor Pairs and SignalingNetworks Mediating Intercellular Communication between Hair Matrix Cells and Dermal Papilla Cells from Cashmere Goats [J]. Cells, 2023, 12(12): 1645.).
[0004] Connective tissue protrudes from the base of the hair bulb to form the dermal papilla, which contains abundant capillaries, nerve endings, and dermal papilla cells. The dermal papilla cells are the signal command center of the hair follicle cycle. During the transition of the hair follicle from the resting phase to the anagen phase, the dermal papilla cells induce the hair follicle to enter the anagen phase. At this time, the activity of alkaline phosphatase (ALP) is enhanced, and a series of growth factors and signaling molecules (such as Wnt, FGF, HGF, VEGF, etc.) are secreted, activating hair follicle stem cells and hair matrix cells, and starting a new growth cycle (MEHTA A, et al. Revolutionary Approaches to Hair Regrowth: Follicle Neogenesis, Wnt / β-Catenin Signaling, and Emerging Therapies [J]. Cells, 2025,14(11): 779.). Studies have found that human dermal papilla cells in the "bald area" of androgenetic alopecia patients show an earlier aging phenotype and lose their hair follicle induction function (PAPPALARDO A, et al. Restoration of hair follicle inductive properties by depletion). (of senescent cells [J]. Aging Cell, 2025, 24(1):e14353.), therefore, regulating the vitality and secretory capacity of dermal papilla cells can promote the hair cycle and hair growth.
[0005] The outer root sheath cells of hair follicles are the main reservoir of hair follicle stem cells. They receive signals from dermal papilla cells and translate them into instructions for proliferation and differentiation, providing a cellular source for the periodic regeneration of hair follicles (AGRAMUNT J, et al. Mechanical stimulation of human hair follicle outer root sheath culturesactivates adjacent sensory neurons [J]. Science Advances, 2023, 9(43): eadh3273.). Therefore, regulating and maintaining the vitality of dermal papilla cells, hair matrix cells, and outer root sheath cells is crucial for the treatment of hair loss.
[0006] Osteomodulin (OMD), also known as osteomodulatory protein or bone adhesion protein, is a distinctive member of the extracellular matrix (ECM). Its structural features include leucine-rich repeat sequences, which enable it to interact with ECM components such as type I collagen and participate in ECM tissue organization and mineralization processes. It is primarily active in mineralized tissues like bones and teeth, but studies have also found its association with skin condition. OMD exerts its anti-fibrotic effect by inhibiting the TGF-β / Smad pathway and regulating ECM deposition in dermatofibrotic diseases such as keloids (LI Q, et al. Osteomodulin contributes to keloid development by regulating p38 MAPKsignaling [J]. Journal of Dermatology, 2023, 50(7): 895-905.). OMD expression is significantly downregulated in various epithelial cancers (including bladder cancer), suggesting that it may act as a factor in maintaining epithelial tissue homeostasis, but this has not yet been directly verified in skin cancer. Based on the commonalities of SLRP family proteins, OMD may be involved in skin ECM homeostasis (such as the aging process). However, there are currently no reports in the literature on the regulatory effects of periosteal proteins on hair follicle-associated cells, dermal papilla cells, hair follicle matrix cells, and outer root sheath of hair follicles. Summary of the Invention
[0007] The purpose of this invention is to overcome the shortcomings and deficiencies of the prior art and to provide the application of periosteal protein in the preparation of formulations for promoting hair follicle cell activity or for preventing and treating hair loss.
[0008] The objective of this invention is achieved through the following technical solution: the application of periosteal protein in the preparation of formulations for promoting hair follicle cell activity or for preventing and treating hair loss. This invention is the first to discover that periosteal protein can regulate the activity of dermal papilla cells, hair matrix cells, and outer root sheath cells in hair follicles, promote the proliferation of these cells, enhance the hair induction ability of dermal papilla cells, and promote hair follicle cycle transition, hair growth, and thickening. Therefore, it can be applied to the preparation of formulations for treating hair loss.
[0009] The aforementioned osteomodulin (OMD), also known as osteomodulatory protein or bone adhesion protein, with the gene symbol OMD, is located on human chromosome 9 (9q22.31). Its core protein consists of 421 amino acids with a molecular weight of approximately 90 kDa. Its primary structure contains characteristic leucine-rich repetitive sequences. These LRR domains are the molecular basis for its interaction with other components in the extracellular matrix, such as collagen. After secretion, the protein is glycosylated and linked with keratin sulfate chains, which transforms it into a proteoglycan and enhances its hydrophilicity and structural stability.
[0010] The promotion of hair follicle cell activity refers to promoting the proliferation of at least one of the dermal papilla cells, hair matrix cells, and outer root sheath cells, and / or enhancing the ability of dermal papilla cells to induce hair growth.
[0011] The preferred type of preparation is a topical liniment, topical patch, or injectable reagent.
[0012] The preferred dosage form of the topical liniment is a liquid, cream, or ointment.
[0013] The preferred topical patch is a transdermal patch.
[0014] The injectable formulation is preferably a skin injection formulation; more preferably, it is at least one of a microneedle injection formulation and a dot injection formulation.
[0015] The concentration of periosteal protein in the preparation is preferably 1 to 10,000 ng / mL; more preferably 10 to 10,000 ng / mL.
[0016] The present invention has the following advantages and effects compared with the prior art:
[0017] (1) The present invention uses CCK8 experiments to clarify that periosteal protein has the effect of promoting the proliferation of hair follicle dermal papilla cells, hair matrix cells and outer root sheath cells, and determines the optimal concentration.
[0018] (2) The present invention has demonstrated by alkaline phosphatase staining that periosteal proteins have the ability to induce hair growth in hair follicles and dermal papillae.
[0019] (3) The present invention found through mouse experiments that periosteal protein has the effect of promoting hair follicle cycle transition and hair growth thickening.
[0020] In summary, this invention is the first to discover a protein—periosteal protein—that regulates the growth of dermal papilla cells, hair matrix cells, and outer root sheath cells in hair follicles. The compound enhances the cell viability of these cells, promotes cell proliferation, and strengthens the ability of dermal papilla cells to induce hair growth. Furthermore, this invention utilizes hair cycle experiments in C57BL / 6 mice to demonstrate that periosteal protein promotes hair growth, thickening, and hair follicle cycle transition on the back of mice. The effect of periosteal protein on the hair follicle cycle was determined by HE staining of back skin sections, and the influence of periosteal protein on hair growth was detected by measuring the diameter and length of back hair. Therefore, periosteal protein can be used in the preparation of formulations for treating hair loss and has significant application value. Attached Figure Description
[0021] Figure 1 This is an image showing the effect of periosteal protein promoting the proliferation of dermal papilla cells.
[0022] Figure 2 This is an image showing the effect of periosteum protein promoting the proliferation of hair matrix cells.
[0023] Figure 3 This is an image showing the effect of periosteal protein promoting the proliferation of outer root sheath cells in hair follicles.
[0024] Figure 4 This is a graph showing the detection results of cellular alkaline phosphatase in dermal papilla cells after treatment with periosteal protein; where A is the activity staining graph and B is the statistical graph of A.
[0025] Figure 5 The images show the effect of periosteal protein on promoting hair growth and thickening on the back of mice; where A is a photograph, B is a statistical result of hair diameter, and C is a statistical result of hair length.
[0026] Figure 6 This is a photograph of H&E staining of hair follicles on the back of mice after treatment with periosteal protein.
[0027] In the figure, * indicates a comparison with the control group. P <0.05, ** indicates P <0.01, *** indicates P <0.001, **** indicates P <0.0001. Detailed Implementation
[0028] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto. Example 1
[0029] (1) Periosteal protein was dissolved in serum-free medium (DMEM high glucose medium, Gibco) to obtain periosteal protein solutions with concentrations of 1 ng / mL, 10 ng / mL and 100 ng / mL, respectively.
[0030] (2) Human hair follicle papilla cells (purchased from NEWGAINBIO, IH1006, Wuxi Xinrun Biotechnology Co., Ltd.) were seeded in 96-well plates at a density of 2000 cells per well. When the cells reached 50%–60% confluence, they were starved in serum-free medium (DMEM high glucose medium, Gibco) for 8 hours. The cells were then divided into a control group and a periosteal protein treatment group. The original medium was discarded. Periosteal protein solutions at concentrations of 1 ng / mL, 10 ng / mL, and 100 ng / mL were added to the periosteal protein treatment group, 100 μL / well, with three replicates for each concentration. The control group received only serum-free medium. The 96-well plates were incubated at 37°C in a 5% CO2 incubator for 24 hours for CCK8 assay. Absorption at 450 nm was measured using a microplate reader (BLJ BIO-RECH, China, DLJ-200). Data analysis was performed using SPSS 25 statistical software. For the measurement data, paired measurements were used for comparison at different time points. t Test. Data showing statistically significant differences are indicated by different asterisks (*). P <0.05,** P <0.01, *** P <0.001, **** P <0.0001. Experimental results are shown in [link to results]. Figure 1 The results showed that the addition of periosteal protein could promote the proliferation of hair follicle dermal papilla cells; when the periosteal protein concentration was 10 ng / mL, it significantly promoted the proliferation of hair follicle dermal papilla cells. Example 2
[0031] CCK8 assay was used to detect the effect of different concentrations of periosteal protein on the proliferation of human hair follicle matrix cells:
[0032] Human hair follicle matrix cells (purchased from Qingqi (Shanghai) Biotechnology Development Co., Ltd., BLUEFBIO) TM Cells (BFN60803943) were seeded at a density of 2000 cells per well in 96-well plates. When the cells reached 50%–60% confluence, they were starved for 8 hours in serum-free medium (DMEM high-glucose medium, Gibco). The cells were then divided into a control group and a periosteum protein treatment group. The original medium was discarded. Periosteum protein solutions at concentrations of 1 ng / mL, 10 ng / mL, and 100 ng / mL were added to each well (100 μL / well), with three replicates for each concentration. The control group received only serum-free medium. The 96-well plates were incubated at 37°C in a 5% CO2 incubator for 24 hours before CCK8 assay. Absorbance at 450 nm was measured using a microplate reader (BLJ BIO-RECH, China, DLJ-200). Data analysis was performed using SPSS 25 statistical software. For comparisons of measurements at different time points, paired measurements were used.t Test. Data showing statistically significant differences are indicated by different asterisks (*). P <0.05,** P <0.01, *** P <0.001. Experimental results are shown in [link to results]. Figure 2 . Figure 2 The results showed that the addition of periosteal protein could promote the proliferation of hair follicle matrix cells; when the periosteal protein concentration was 10 ng / mL, it significantly promoted the proliferation of hair follicle matrix cells. Example 3
[0033] CCK8 assay was used to detect the effect of different concentrations of periosteal protein on the proliferation of human hair follicle outer root sheath cells:
[0034] Human hair follicle outer root sheath cells (purchased from Applied Biological Materials (abm), T9233) were seeded at a density of 2000 cells per well in 96-well plates. When the cells reached 50%–60% confluence, they were starved for 8 hours in serum-free medium (DMEM high-glucose medium, Gibco). The cells were then divided into a control group and a periosteum protein treatment group. The original medium was discarded. Periosteum protein solutions at concentrations of 1 ng / mL, 10 ng / mL, and 100 ng / mL were added to each well (100 μL / well), with three replicates for each concentration. The control group received only serum-free medium. The 96-well plates were incubated at 37°C in a 5% CO2 incubator for 24 hours, and CCK8 assays were performed. Absorbance at 450 nm was measured using a microplate reader (BLJ BIO-RECH, China, DLJ-200). Data analysis was performed using SPSS 25 statistical software. For comparisons of measurements at different time points, paired measurements were used. t Test. Data showing statistically significant differences are indicated by different asterisks (*). P <0.05,** P <0.01, *** P <0.001, **** P <0.0001. Experimental results are shown in [link to results]. Figure 3 . Figure 3 The results showed that the addition of periosteal protein could promote the proliferation of outer root sheath cells of hair follicles; when the periosteal protein concentration was 10 ng / mL, it significantly promoted the proliferation of outer root sheath cells of hair follicles. Example 4
[0035] An alkaline phosphatase (ALP) activity assay was conducted to detect the effect of periosteal proteins on the ability of human hair follicle dermal papilla cells to induce hair growth.
[0036] Human dermal papilla cells (purchased from Wuxi Xinrun Biotechnology Co., Ltd., NEWGAINBIO, IH1006) were seeded into a 6-well plate at a density of 5×10 4 cells per well. When the cells grew to 50% - 60% confluence, they were starved in serum-free medium (DMEM high-glucose medium, Gibco) for 8 h, and then divided into a control group and a periostin treatment group. The original medium was discarded, and 1.5 mL of serum-free medium containing 10 ng / mL periostin was added to the periostin treatment group for treatment. After 24 h of culture, the original medium was discarded, and the cells were washed 3 times with washing solution, and then fixed with 4% paraformaldehyde solution (PFA) for 10 minutes. BCIP / NBT staining working solution was added successively according to the ratio, and incubated at room temperature in the dark for 45 minutes. The BCIP / NBT staining working solution was removed, and the cells were washed 2 times with distilled water to terminate the color reaction. The BCIP / NBT staining working solution can stain dermal papilla cells with high alkaline phosphatase activity dark blue-violet. The results are as Figure 4 shown: The alkaline phosphatase staining results showed that periostin treatment could increase the number of dermal papilla cells with positive alkaline phosphatase staining. Example 5
[0037] Experimental study on the effect of periostin on the hair length and diameter of the back of C57BL / 6 mice using a hair follicle cycle model:
[0038] C57BL / 6 mice (purchased from the Experimental Animal Center of Southern Medical University, SCXK (Beijing) 2021-0059) were anesthetized by intraperitoneal injection of 2% pentobarbital (Sigma, St. Louis, MO, USA; w / v; 0.01 mL / g body mass). The back hair shafts of the mice were removed using depilatory cream without damaging the hair roots, and the hair follicles in the back of the mice were identified as being in the telogen phase by the pink skin color. After depilation, the mice were randomly divided into two groups, with five mice in each group. Each mouse in the treatment group was injected with 50 μL of periostin solution at a concentration of 10 μg / mL (the solvent was PBS), and the control group was injected with 50 μL / mouse of PBS. The injection was performed once a day for 24 days. On the 24th day, 30 hairs (6 hairs per mouse) were randomly plucked from the two treatment areas on the back of the mice, photographed under a stereomicroscope, and the length and diameter of the hairs were measured using LitoDigital software. For the measured data, paired t tests were used to compare the measured values at different time points. For data with significant differences after statistics, * indicates P <0.05, *** P <0.01, **** P <0.001. The experimental results are shown in Figure 5 . Figure 5Figures A, B, and C show that treatment of mouse dorsal skin with a 10 µg / mL periosteal protein solution significantly promoted hair growth and increased hair diameter, and this effect was statistically significant. Example 6
[0039] Experimental study of the effect of periosteal proteins on the hair follicle cycle using the C57BL / 6 mouse hair follicle cycle model:
[0040] C57BL / 6 mice were anesthetized by intraperitoneal injection of 2% pentobarbital (Sigma, St. Louis, MO, USA; w / v; 0.01 mL / g body mass). Hair removal cream was used to remove the hair shafts on the back of the mice without damaging the hair roots. The pink skin color was used to identify whether the hair follicles on the back of the mice were in the resting phase. After hair removal, the mice were randomly divided into two groups of five mice each. Each mouse in the treatment group was injected with 50 μL of 10 µg / mL periosteal protein solution (solvent: PBS), while the control group received 50 μL / mouse of PBS. Injections were given once daily for 5 days. On day 5, the mice were euthanized by cervical dislocation. Skin from each treatment group on the back of the mice was collected, fixed with 4% paraformaldehyde, embedded in paraffin, and sectioned into 5 μm sections. The paraffin sections were dewaxed with xylene, rehydrated with ethanol of progressively higher concentrations, and then stained with H&E to observe the morphology of the hair follicle tissue. The experimental results are shown in […]. Figure 6 . Figure 6 The results showed that hair follicles on the back of mice treated with a periosteal protein solution at a concentration of 10 µg / mL were in the growth phase, while those in the control group were in the resting phase.
[0041] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.
Claims
1. The application of osteomodulin in the preparation of formulations for the prevention and treatment of hair loss, characterized in that: The osteomodulatory protein Osteomodulin is derived from humans.
2. The application according to claim 1, characterized in that: The aforementioned prevention and treatment of hair loss is achieved by promoting the activity of hair follicle cells; The promotion of hair follicle cell activity is to promote the proliferation of at least one of the dermal papilla cells, hair matrix cells, and outer root sheath cells, and / or enhance the ability of dermal papilla cells to induce hair growth.
3. The application according to claim 1, characterized in that: The preparation is of the type of topical liniment, topical patch, or injectable preparation.
4. The application according to claim 3, characterized in that: The dosage form of the topical liniment is liquid, cream, or ointment.
5. The application according to claim 3, characterized in that: The aforementioned external patch is a transdermal patch.
6. The application according to claim 3, characterized in that: The injectable preparation is a skin injection preparation.
7. The application according to claim 6, characterized in that: The skin injection preparation is at least one of microneedle injection preparation and dot injection preparation.
8. The application according to any one of claims 1 to 7, characterized in that: The concentration of osteomodulin in the formulation is 1–10,000 ng / mL.
9. The application according to claim 8, characterized in that: The concentration of osteomodulin in the formulation is 10–10,000 ng / mL.