TWI899523B - Using growth factors to treat arthritis - Google Patents

Abstract

The present invention discloses that epidermal growth factor can be used to treat arthritis.

Description

Using growth factors to treat arthritis

The present invention relates to the use of epidermal growth factor (EGF) to treat arthritis, and more particularly to the use of a composition containing EGF, glycosaminoglycan, and hyaluronic acid (HA).

Arthritis is a common chronic disease that can occur in various joints (e.g., hands, feet, wrists, elbows, knees, hips, and ankles). Common types include degenerative arthritis (also known as osteoarthritis (OA)), rheumatoid arthritis (RA), and gouty arthritis (GA). These conditions primarily cause joint inflammation, swelling, deformity, pain, atrophy, and stiffness, and may even lead to loss of mobility.

Current clinical treatments for arthritis generally involve the use of steroids to reduce inflammation and hyaluronic acid (HA) to provide lubrication, combined with physical rehabilitation to control cartilage damage. However, the effectiveness of these treatments remains unsatisfactory, and severe cases may even require surgical replacement of artificial joints.

Platelet-rich plasma (PRP) is a concentrated fluid obtained by removing red blood cells from blood. It contains a large number of platelets and various growth factors, with insulin-like growth factor I (IGF-I) being the highest, followed by transforming growth factor-β (TGF-β). Epidermal growth factor (EGF) is extremely low, approximately 1/450 of the level of IGF-I (Ha CW et al . (2019), Arthroscopy , 35:2878-2884). Although PRP has been used to improve arthritis, its effectiveness remains to be fully verified in detailed studies. In a recent study published by Gazendam A. et al . (2021), Br. J. Sports Med. , 55:256-261, Gazendam A. et al. screened 1,782 studies on hip arthritis using the PRISMA methodology, conducted a systematic review, and conducted a network meta-analysis (NMA). The results of this analysis found no statistical difference between PRP injections and placebo (i.e., saline injections) in improving joint function or pain. In particular, the combination of PRP and hyaluronic acid was less effective than PRP alone.

On the other hand, studies have shown that EGF is expressed in large quantities in patients with RA and is believed to be involved in the pathogenesis of arthritis, especially the inflammatory process of RA (Nah SS et al . (2010), Rheumatol. Int. , 30:443-449).

In Swanson CD et al . (2012), J. Immunol. , 188:3513-3521, they found that epidermal growth factor receptor (EGFR) is highly expressed in mice with collagen-induced arthritis and in patients with RA, suggesting that EGFR may be involved in the pathogenesis of arthritis. Swanson CD et al. further attempted to treat the disease by inhibiting EGFR and found that the EGFR inhibitor erlotinib could alleviate collagen-induced arthritis, suggesting that EGFR could be a novel therapeutic target.

On the other hand, Sun H. et al . (2018), EBioMedicine , 32:223-233, found that EGFR is highly activated in OA patients and suggested that EGFR activation may lead to joint damage. They also attempted to treat OA by inhibiting EGFR and found that the EGFR inhibitor gefitinib could improve OA in mice, suggesting that it could be used as an effective treatment strategy.

Summary of the Invention

Despite the aforementioned prior research on epidermal growth factor (EGF) and its receptors, the applicants unexpectedly discovered in the present invention that direct use of EGF can effectively treat arthritis, and that further combining it with glycosaminoglycans and hyaluronic acid (HA) can significantly enhance its therapeutic efficacy.

Therefore, the present invention provides an epidermal growth factor for use in preparing a composition for treating arthritis. Preferably, the composition further comprises glycosaminoglycan and hyaluronic acid.

The present invention also provides a method for treating arthritis, comprising administering a composition as described above to a subject in need thereof.

Preferably, the arthritis is degenerative arthritis or rheumatoid arthritis.

Detailed description of the invention

It is to be understood that if any prior publication is cited herein, that publication does not constitute an admission that the publication forms part of the common general knowledge in the art in Taiwan or any other country.

For the purposes of this specification, it will be expressly understood that the word "comprising" means "including but not limited to," and that the word "comprises" has a corresponding meaning.

Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art to which this invention belongs. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein that can be used to practice the present invention. However, the present invention is in no way limited to the methods and materials described.

The present invention provides an epidermal growth factor (EGF) for use in preparing a composition for treating arthritis.

According to the present invention, the arthritis can be selected from the group consisting of degenerative arthritis (also known as osteoarthritis (OA)), rheumatoid arthritis (RA), gouty arthritis (GA), psoriatic arthritis (PA), infectious arthritis (IA), and combinations thereof.

In a preferred embodiment of the present invention, the arthritis is degenerative arthritis. In another preferred embodiment of the present invention, the arthritis is rheumatoid arthritis.

As used herein, "treating" or "treatment" means preventing, reducing, alleviating, ameliorating, relieving, or controlling one or more clinical signs of a disease or disorder, as well as lowering, stopping, or reversing the progression of the severity of a condition or symptom being treated.

According to the present invention, the EGF can be derived from humans or various animals, plants, and microorganisms and can be a commercially available product or a product produced by techniques well known and commonly used by those skilled in the art, such as a natural product isolated from biological materials or a recombinant protein obtained through genetic engineering. In a preferred embodiment of the present invention, the EGF is recombinant human EGF (rhEGF).

According to the present invention, the composition may further comprise glycosaminoglycan and hyaluronic acid (HA).

Preferably, the glycosaminoglycan is selected from the group consisting of chondroitin sulfate, keratan sulfate, heparan sulfate, glucosamine sulfate, and combinations thereof. In a preferred embodiment of the present invention, the glycosaminoglycan is chondroitin sulfate.

According to the present invention, the weight ratio of the epidermal growth factor, the glycosaminoglycan, and the hyaluronic acid can be in a range of 1:10:100 to 1:1000:5000. In a preferred embodiment of the present invention, the weight ratio of the epidermal growth factor, the glycosaminoglycan, and the hyaluronic acid is 1:100:1000.

According to the present invention, the composition does not contain platelet-rich plasma (PRP).

According to the present invention, the composition can be a food composition, for example, in the form of a food additive, which can be added to an edible material to prepare a food product for human or animal consumption. According to the present invention, the types of food products can include, but are not limited to: milk powder, fermented milk, yogurt, butter, beverages (e.g., tea, coffee), functional beverages, flour products, baked foods, confectionery, candies, fermented foods, animal feeds, health foods, infant food, and dietary supplements.

According to the present invention, the composition can be a pharmaceutical composition.

According to the present invention, the pharmaceutical composition may be in a dosage form suitable for parenteral administration, oral administration, or topical administration.

According to the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier widely used in drug manufacturing technology. For example, the pharmaceutically acceptable carrier may comprise one or more agents selected from the group consisting of a solvent, a buffer, an emulsifier, a suspending agent, a decomposer, a disintegrating agent, a dispersing agent, a binding agent, an excipient, a stabilizing agent, a chelating agent, a diluent, a gelling agent, a preservative, a wetting agent, a lubricant, an absorption delaying agent, a liposome, and the like. The selection and amount of these reagents are within the professional and routine skills of those skilled in the art.

According to the present invention, the pharmaceutical composition can be prepared into a dosage form suitable for parenteral administration [including injection, for example, sterile aqueous solution or dispersion] by a technique well known to those skilled in the art, and administered by a route selected from the group consisting of intraperitoneal injection, intrapleural injection, intramuscular injection, intravenous injection, intraarterial injection, intraarticular injection, intrasynovial injection, intrathecal injection, intracranial injection, intraepidermal injection, subcutaneous injection, intradermal injection, Preferably, the pharmaceutical composition is formulated for administration by intraperitoneal injection or intraarticular injection.

According to the present invention, the pharmaceutical composition can be prepared into a dosage form suitable for oral administration using techniques well known to those skilled in the art, including, but not limited to, sterile powders, tablets, troches, lozenges, pellets, capsules, dispersible powders or granules, solutions, suspensions, emulsions, syrups, elixirs, slurries, and the like.

According to the present invention, the pharmaceutical composition can also be formulated into an external preparation suitable for topical application to the skin using techniques well known to those skilled in the art, including, but not limited to, emulsions, gels, ointments, creams, patches, liniments, powders, aerosols, sprays, lotions, serums, pastes, foams, drops, suspensions, salve, and bandages.

The present invention also provides a method for treating arthritis, comprising administering a composition containing epidermal growth factor as described above to a subject in need thereof.

As used herein, the terms "administering" and "administration" are used interchangeably and mean introducing, providing, or delivering a predetermined active ingredient to a subject by any appropriate route to perform its intended effect.

As used herein, the term "subject" means any mammal of interest, such as humans, monkeys, cows, sheep, horses, pigs, goats, dogs, cats, mice, rats, rabbits, elephants, bears, deer, whales, and dolphins; reptiles, such as turtles and lizards; amphibians, such as frogs and salamanders; fish; and birds.

The dosage and frequency of administration of the composition according to the present invention will vary depending on the severity of the disease to be treated, the route of administration, and the age, physical condition, and response of the individual to be treated . In general, the composition according to the present invention can be in the form of a single dose or divided into several doses and can be administered orally, parenterally, or topically.

The present invention will be further described with reference to the following examples. However, it should be understood that these examples are for illustration only and should not be construed as limiting the implementation of the present invention .

The following ingredients were purchased from Leye Biochemical Technology Co., Ltd.: epidermal growth factor (EGF) (Cat. No. 02-108); chondroitin sulfate (CS) (Cat. No. 05-103); and hyaluronic acid (HA) (Cat. No. 07-113). General experimental methods: 1. Statistical analysis:

In the following examples, experimental data were statistically analyzed using GraphPad Prism version 8.4 (GraphPad Software, San Diego, CA) and presented as mean ± standard error of the mean (SEM). All data were analyzed by one-way analysis of variance (ANOVA) followed by Tukey's test to assess differences between groups. A p < 0.05 result was considered statistically significant. Example 1. Evaluation of the efficacy of epidermal growth factor in the treatment of degenerative arthritis ( also known as osteoarthritis , OA) Experimental Materials: 1. Composition containing EGF , CS , and HA :

The composition containing EGF, CS, and HA used in this example was prepared by mixing the EGF, CS, and HA from the "General Experimental Materials" above at a weight ratio of 1:100:1000 and adding the mixture to sterile water. The mixture contained 20 μg/g of EGF. 2. Experimental Animals:

Male Sprague Dawley rats (6 weeks old, weighing approximately 200 to 300 g) used in this example were purchased from the National Laboratory Animal Center (ROC). All experimental animals were housed individually in an animal room with a 12-hour light and dark cycle, a room temperature of 21-24°C, and a relative humidity of 45-70%, and were fed water and feed ad libitum . All experimental procedures involving laboratory animals were approved by the Institutional Animal Care and Use Committee of National Chung Hsing University and conducted in accordance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health (NIH). Experimental Methods: A. Induction of OA and Administration of Epidermal Growth Factor:

First, male Sprague-Dawley rats were randomly divided into one normal control group, one pathological control group, one comparison group, and two experimental groups (i.e., Experimental Groups 1 and 2) (n = 5 per group). Next, 30 μL of a 50 g/L monosodium iodoacetate (MIA) solution (dissolved in 0.9% saline) was injected into the right knee joint of the rats in the pathological control group, the comparison group, and each experimental group via intraarticular injection. The normal control group received an equal volume of 0.9% saline.

On days 3 and 14 after the MIA solution injection, the control group rats were injected with 30 μL of HA into the right knee joint. The same site was injected with 30 μL of 20 μg/g EGF and 30 μL of a combination of EGF, CS, and HA (containing 20 μg/g EGF) into the same area in experimental groups 1 and 2, respectively. The pathological control group rats were injected with an equal volume of saline. The normal control group rats received no treatment. B. Evaluation of Clinical Signs :

At the end of the 28th day after the injection of MIA solution, the diameter of the right knee joint of each group of rats was measured with a caliper. The experimental data were then analyzed according to the method described in the first item "Statistical Analysis" of "General Experimental Methods" above. C. Preparation of biological samples:

At the end of the 28th day after the injection of the MIA solution, after completing the clinical symptom assessment in Section B above, the rats in each group were sacrificed by carbon dioxide inhalation. Blood was then collected from the heart of each group of rats by puncture with a 25G needle and centrifuged at 1,500 × g for 10 minutes at 25°C to obtain serum samples.

Finally, the right knee joint tissues of rats in each group were removed and washed with 0.9% saline. D. Histopathological evaluation:

First, the right knee joint tissues of the rats in each group obtained in Item C above were fixed with 10% neutral buffered formalin (BiOTnA Biotech, Cat No. TABS06-4000) at room temperature for 24 hours. The fixed tissue samples were then embedded in paraffin and sectioned to obtain 5 μm thick tissue sections.

The tissue sections were then stained with hematoxylin-eosin (Sigma Aldrich, Cat. No. 1.09249) and safranin O-fast green (Sigma Aldrich, S8884, Cat. No. F7258) according to the techniques known and commonly used by those skilled in the art. The sections were then observed under an Olympus CKX41 microscope at a magnification of 200 times. Two areas were randomly selected for photographing and the modified Mankin scoring system described in Pauli C. et al. (2012), Osteoarthritis Cartilage, 20:476-85 was used for evaluation. The OA pathological features of cartilage structure (including cartilage structure damage, chondrocyte loss, and proteoglycan loss) are scored using a 3D skeletal muscle assessment system to assess the progression of MIA-induced OA. E. Determination of serum OA biomarkers and inflammatory factors:

First, the serum samples from each group of rats obtained in section C above were diluted 5-fold with RPMI 1640 medium. The OA biomarker COMP in the serum of each group of rats was measured using a rat cartilage oligomeric matrix protein (COMP) ELISA kit (Cusabio, Cat. No. CSB-E13833r) according to the manufacturer's instructions. The levels of proinflammatory cytokines in these sera were measured using TNF-α, IL-1β, IL-6, and IL-17A ELISA kits (Thermo Fisher Scientific, Invitrogen, Cat. No. 88-7340-88, 88-6010-22, 88-50625-88, and 88-7170-22). The levels of PGE2 were also measured using a PGE2 ELISA kit (Cusabio, Cat. No. CSB-E07967r) was used to measure the level of proinflammatory mediators prostaglandin E2 (PGE2) in serum.

In addition, the level of nitric oxide (NO), a pro-inflammatory mediator, was measured using the Griess assay. Briefly, 100 μL of serum sample was collected from each group of rats and mixed with equal volumes of Griess reagent (containing 0.2% N- (1-naphthyl)ethylenediamine, 2 % sulfanilamide, and 5% phosphoric acid). The mixture was reacted at room temperature for 10 minutes, and the absorbance of each well was read at a wavelength of 540 nm using a spectrophotometer (OD ₅₄₀ ). The obtained _OD540 values were converted into nitrite concentrations based on a pre-made correlation curve of sodium nitrate relative to their own _OD540 values.

Afterwards, the experimental data were analyzed according to the method described in the first item "Statistical Analysis" of "General Experimental Methods" above. Results: A. Evaluation of clinical symptoms:

Figure 1 shows the right knee joint diameters measured for each group of rats. As shown in Figure 1, the knee joints of the rats in the pathological control group were significantly swollen compared to the normal control group, indicating that MIA has successfully induced OA in the rats. Compared to the pathological control group, the knee joint swelling of the rats in the comparison group and experimental group 1 was partially improved, while the knee joint swelling of experimental group 2 was significantly improved, even approaching that of the normal control group. This experimental result shows that EGF can treat OA and improve joint swelling, and its efficacy is similar to that of drugs known to be used to treat arthritis. In addition, EGF can show a synergistic therapeutic effect when used in combination with CS and HA. B. Histopathological evaluation:

Staining of knee joint tissue samples from rats in each group revealed that, compared to the normal control group, the cartilage in the pathological control group exhibited multiple distinct cracks and an irregular surface, along with severe loss of chondrocytes and proteoglycans. This indicates that MIA had successfully induced OA and associated tissue pathology. In contrast, the cartilage in the control group and experimental groups 1 and 2 exhibited fewer cracks and a smoother surface, with less pronounced loss of chondrocytes and proteoglycans. The cartilage in experimental group 2 exhibited the best condition, with a smooth surface and a substantial retention of chondrocytes and proteoglycans (data not shown).

Figure 2 shows the results of cartilage structure assessment of knee joint tissue samples from rats in each group using the modified Mankin scoring system. As shown in Figure 2, the cartilage tissue of rats in the normal control group showed no pathological signs. In contrast, the cartilage tissue of rats in the pathological control group had the most severe pathological signs. On the other hand, the pathological signs of cartilage tissue in rats in the comparison group and experimental group 1 showed a slight reduction in symptoms, while the cartilage tissue of rats in experimental group 2 had the mildest pathological signs, with a score of approximately 1 point.

These experimental results show that EGF can treat OA and improve related cartilage tissue lesions, and its efficacy is similar to that of drugs commonly used to treat arthritis. Furthermore, EGF, CS, and HA can demonstrate synergistic therapeutic effects when used in combination. C. Determination of OA biomarkers and inflammatory factors in serum :

Figure 3 shows the serum COMP levels measured in rats from each group. As shown, serum COMP levels in the pathological control group were significantly elevated compared to the normal control group, indicating that MIA had successfully induced OA and led to the onset of cartilage pathology and degeneration. Compared to the pathological control group, serum COMP levels in both the control group and experimental group 1 decreased slightly, with experimental group 2 experiencing the greatest decrease and returning to levels similar to those in the normal control group.

Figure 4 shows the serum levels of TNF-α, IL-1β, IL-6, and IL-17A measured in rats from each group. As shown in Figure 4, serum levels of inflammatory cytokines in the pathological control group were significantly elevated compared to those in the normal control group, indicating that MIA successfully induced OA and produced an inflammatory response in the knee joint. Compared to the pathological control group, serum levels of inflammatory cytokines in both the comparison group and experimental group 1 showed a downward trend, with the greatest decrease in serum inflammatory cytokine levels in experimental group 2.

Figure 5 shows the serum levels of pro-inflammatory mediators PGE2 and NO measured in each group of rats. As shown in Figure 5, compared to the normal control group, the pathological control group showed a significant increase in serum PGE2 and NO levels. However, compared to the pathological control group, the serum PGE2 and NO levels in the comparison group and experimental group 1 showed a downward trend, with the greatest decrease in serum PGE2 and NO levels in experimental group 2.

These experimental results demonstrate that EGF can effectively reduce OA and improve associated inflammatory responses, with efficacy comparable to that of conventional drugs used to treat arthritis. Furthermore, EGF exhibits synergistic therapeutic effects when used in combination with CS and HA. Example 2. Evaluation of the efficacy of epidermal growth factor in the treatment of rheumatoid arthritis (RA) Experimental animals:

Female DBA/1 mice (8 weeks old, weighing approximately 20 g) used in this example were purchased from the National Laboratory Animal Center. All experimental animals were individually housed in an animal room with a 12-hour light and dark cycle, a room temperature maintained at 21-23°C, and a relative humidity maintained at 45-60%, and were fed water and feed ad libitum. All experimental procedures involving experimental animals were approved by the Laboratory Animal Care and Use Committee of National Chung Hsing University and were conducted in accordance with the Standards for the Care and Use of Laboratory Animals of the National Institutes of Health of the United States. Experimental methods: A. Induction of RA and administration of epidermal growth factor:

First, a 2 mg/mL chicken type II collagen (CII) solution (Chondrex, Cat. No. 20012) in 10 mM acetic acid was mixed with complete Freund's adjuvant (Sigma Aldrich, Cat. No. F5881-10ML) at a 1:1 (v/v) ratio to produce a CII first emulsion. A 2 mg/mL CII solution was then mixed with incomplete Freund's adjuvant (ChemCruzx, Cat. No. sc-24648) at a 1:1 (v/v) ratio to produce a CII second emulsion.

Female DBA/1 mice were randomly divided into one normal control group, one pathological control group, one comparison group, and two experimental groups (i.e., Experimental Groups 1 and 2) (n=6 per group). 200 μL of the first CII emulsion was then subcutaneously injected into the tails of the pathological control group, the comparison group, and each experimental group for the first immunization. Twenty-one days after the initial immunization, mice were given a booster immunization with an equal volume of the second CII emulsion to induce RA. Mice in the normal control group received essentially the same injections, except that saline was used instead of CII solution in the emulsion preparation.

On day 21 after the initial immunization, after completing the booster immunization, mice in the control group received an intraperitoneal injection of 100 μL of HA. Mice in experimental groups 1 and 2 received intraperitoneal injections of 100 μL of 20 μg/g EGF and 100 μL of a composition containing EGF, CS, and HA (containing 20 μg/g EGF), respectively. Each group of mice received injections once daily for a total of 20 days. B. Assessment of Clinical Symptoms:

At the end of day 21 after the initial immunization, the visual arthritis scoring system described in Brand DD et al . (2007), Nat. Protoc. , 2:1269-1275 and Milici AJ et al . (2008), Arthritis Res. Ther. , 10:R14 was used to score the erythema and edema of the foot joints and ankle-to-knee joints of each group of mice. Thereafter, visual arthritis scoring was performed every three days until day 42 after the initial immunization, for a total of eight scoring sessions. C. Histopathological Assessment:

After completing the clinical symptom assessment in Section B above, mice in each group were sacrificed by carbon dioxide inhalation, and hind knee joint tissue was removed. Tissue sections were prepared according to the method described in Section D of Example 1 above, followed by hematoxylin-eosin staining and microscopic observation. Subsequently, the degree of RA pathological features (including inflammatory cell infiltration, synovial hyperplasia, and cartilage surface erosion) was scored according to the arthritis histological scoring method described in Deng GM et al. (2005), Nat Med., 11:1066-1072, to assess the disease progression of CII-induced RA. D. Determination of pro-inflammatory cytokines in the soles of the feet:

After sacrificing each group of mice, 100 mg of paw tissue was removed, frozen in liquid nitrogen, and then ground. 1 mL of tissue lysis buffer (Gold Biotechnology, Inc., Cat. No. GB-181-100) (containing a protease inhibitor cocktail) was added and mixed thoroughly. Next, TNF-α, IL-1β, IL-6, IL-17A, and IFN-γ ELISA kits (BioLegend, Cat No. 430904, 432604, 431304, 432504, 430807) and IL-8 ELISA kit ( MyBioSource , Cat No. MBS261967) were used to measure the levels of these pro-inflammatory cytokines in the paw tissues of each group of mice.

Figure 6 shows the results of visual arthritis scoring in each group of mice from days 21 to 42 after the initial immunization. As shown in Figure 6, the normal control group mice showed no signs of arthritis. In contrast, the arthritis scores of the pathological control group mice gradually increased over time, indicating that CII had successfully induced RA in the mice. Compared to the pathological control group, the arthritis scores of the comparison group and experimental group 1 increased more slowly over time. The progression of arthritis in experimental group 2 was the slowest, and the arthritis score curve between days 33 and 42 after the initial immunization was flat and significantly lower than that of the pathological control group.

The results of this experiment show that EGF can treat and alleviate RA and improve various clinical symptoms caused by RA. Its efficacy is similar to that of drugs commonly used to treat arthritis. In addition, EGF can show a synergistic therapeutic effect when used in combination with CS and HA. B. Histopathological evaluation:

Staining of cartilage tissue samples from each group of mice revealed that, compared to the normal control group, the cartilage structure of the pathological control group was fragmented, with no smooth cartilage surface observed. This indicates that CII had successfully induced RA in the mice and caused related tissue lesions. In contrast, the cartilage structure of the comparison group and experimental group 1 showed only partial damage, with a partially intact cartilage surface observed. The cartilage structure of experimental group 2 was the most intact, with a smooth surface, similar to that of the normal control group (data not shown).

Figure 7 shows the results of histological evaluation of knee joint cartilage tissue samples from each group of mice using arthritis scoring. As shown in Figure 7, the cartilage tissue of the normal control group exhibited no pathological signs. In contrast, the scores for inflammatory cell infiltration, synovial hyperplasia, and cartilage erosion in the pathological control group were significantly elevated. Furthermore, compared to the pathological control group, the scores for various pathological features in the control group and experimental group 1 decreased significantly, with the most significant decreases in the scores for synovial hyperplasia and cartilage surface erosion in experimental group 2.

These experimental results show that EGF can treat RA and improve related cartilage lesions, and its efficacy is similar to that of drugs known to treat arthritis. In addition, EGF, when used in combination with CS and HA, can show synergistic effects in treating RA cartilage lesions. C. Determination of pro-inflammatory cytokines in the soles of the feet:

Figures 8A and 8B show the levels of pro-inflammatory cytokines TNF-α, IL-6, and IL-1β, as well as IL-8, IL-17A, and IFN-γ, measured in the paw tissues of each group of mice. As shown in Figure 8 , compared to the normal control group, the pathological control group showed a significant increase in pro-inflammatory cytokine levels, indicating that CII successfully induced RA and produced an inflammatory response. Compared to the pathological control group, the pro-inflammatory cytokine levels in the control group and experimental groups 1 and 2 all showed a downward trend, with the most significant decrease in pro-inflammatory cytokine levels in experimental group 2.

This experimental result shows that EGF can effectively reduce the inflammatory response caused by RA, and its efficacy is similar to that of drugs commonly used to treat arthritis. In addition, EGF can show synergistic therapeutic effects when used in combination with CS and HA.

Based on the above experimental results, the applicant believes that epidermal growth factor has high potential for treating arthritis (especially OA and RA), and its efficacy can be further enhanced by combining it with glycosaminoglycans (especially chondroitin sulfate) and hyaluronic acid.

All patents and references cited in this specification are incorporated herein by reference in their entirety. In the event of any conflict, the detailed description in this specification (including definitions) will prevail.

Although the present invention has been described with reference to the specific embodiments above, it is apparent that many modifications and variations can be made without departing from the scope and spirit of the invention. It is therefore intended that the present invention be limited only as indicated by the appended patent claims.

The above and other objects, features and advantages of the present invention will become apparent after referring to the following detailed description and preferred embodiments and the accompanying drawings, in which: Figure 1 shows the right knee joint diameters of rats in each group measured on the 28th day after injection of monosodium iodoacetate (MIA), wherein "*", "**" and "***" respectively indicate that when compared with the pathological control group, p <0.05, p <0.01 and p <0.001; Figure 2 shows the results of the evaluation of cartilage structure of knee joint tissue samples of rats in each group on the 28th day after injection of MIA using the modified Mankin scoring system, wherein "*" and "***" respectively indicate that when compared with the pathological control group, p <0.05 and p <0.001. ＜0.001; Figure 3 shows the serum cartilage oligomeric matrix protein (COMP) levels of the rats in each group measured on the 28th day after MIA injection, where "**" indicates p ＜0.01 when compared with the pathological control group; Figure 4 shows the serum TNF-α, IL-1β, IL-6, and IL-17A levels of the rats in each group measured on the 28th day after MIA injection, where "*", "**", and "***" indicate p ＜0.05, p ＜0.01, and p ＜0.001, respectively, when compared with the pathological control group; Figure 5 shows the serum PGE2 and NO levels of the rats in each group measured on the 28th day after MIA injection, where "*", "**", and "***" indicate p ＜0.05, p ＜0.01, and p ＜ 0.001, respectively, when compared with the pathological control group. ＜0.01 and p ＜0.001; Figure 6 shows the results of the visual arthritis scoring system for each group of mice from 21 to 42 days after the initial immunization with chicken type II collagen (CII) emulsion; Figure 7 shows the results of the arthritis histological scoring system for the cartilage tissue samples of the knee joint of each group of mice on the 42nd day after the initial immunization with CII emulsion, where "*" and "***" respectively indicate p ＜0.05 and p ＜0.001; and Figures 8A and 8B respectively show the levels of TNF-α, IL-6, IL-1β, IL-8, IL-17A, and IFN-γ measured in the paw tissues of each group of mice on the 42nd day after the initial immunization with CII emulsion. "*", "**", and "***" indicate p ＜0.05, p ＜0.01, and p ＜0.001, respectively, when compared with the pathological control group.

Claims (8)

An epidermal growth factor is provided for use in preparing a composition for treating arthritis. The use of claim 1, wherein the arthritis is degenerative arthritis. The use according to claim 1, wherein the arthritis is rheumatoid arthritis. The use of claim 1, wherein the composition is composed of epidermal growth factor. An epidermal growth factor is provided for use in preparing a composition for treating arthritis, wherein the composition further comprises chondroitin sulfate and hyaluronic acid, wherein the weight ratio of epidermal growth factor, chondroitin sulfate, and hyaluronic acid is 1:100:1000, and the content of epidermal growth factor is 20 μg/g. The use of claim 5, wherein the composition is composed of epidermal growth factor, chondroitin sulfate and hyaluronic acid. The use according to claim 1 or 5, wherein the composition is a food composition or a pharmaceutical composition. The use of claim 7, wherein the pharmaceutical composition is in a dosage form for parenteral administration, oral administration or topical administration.