Products to help improve oral mucosa-associated fibrosis

Abstract

Provided is a product containing lysozyme that helps improve oral mucosa-related fibrosis. Lysozyme is found for the first time to have a significant improvement effect on oral mucosa-related fibrosis. Compared with anti-inflammatory drugs such as glucocorticoids commonly used at present, the product has a better improvement effect. Meanwhile, lysozyme is highly safe and can be used as both a drug and food, and has a good application prospect in improving oral mucosa-related fibrosis.

Description

Technical Field

[0001] This application relates to a product that helps improve oral mucosa-associated fibrosis. Background Technology

[0002] The oral mucosa is composed of oral epithelial cells and subepithelial soft tissue, forming a barrier on the surface of the oral cavity that protects the oral tissues from foreign substances.

[0003] Normal oral mucosa is soft and elastic, but it can change under the stimulation of certain factors. For example, the keratin layer may thicken, the number of epithelial cells may decrease, the softness may decrease, blood vessels may atrophy, and fibrosis may occur, affecting normal life (such as causing pain, dry mouth, and decreased mouth opening). Histological examination may reveal extensive fibrous deposition in the mucosal soft tissue.

[0004] Currently, there are few products available for improving oral mucosa-associated fibrosis, with corticosteroids and tanshinone being the most commonly recommended. However, there is still a need to develop more products with proven efficacy, low irritation, and few adverse reactions. Summary of the Invention

[0005] One of the purposes of this application is to provide an application. To achieve this purpose, the technical solution adopted by this application is as follows:

[0006] Application of lysozyme in the preparation of products that help improve oral mucosa-associated fibrosis.

[0007] In some embodiments, the oral mucosa-associated fibrosis is fibrosis of the oral mucosa and / or submucosal fibrosis.

[0008] In some implementations, the product may be a pharmaceutical or a non-pharmaceutical.

[0009] In some implementations, the product reduces the level of fibrosis in oral mucosal tissue.

[0010] The second objective of this application is to provide a product. To achieve this objective, the technical solution adopted in this application is:

[0011] A product that helps improve oral mucosa-associated fibrosis, the product comprising lysozyme.

[0012] A product for improving oral mucosa-associated fibrosis, the product comprising lysozyme.

[0013] In some implementations, the product may be a pharmaceutical or a non-pharmaceutical.

[0014] In some implementations, the product may also include acceptable excipients or applicable additives.

[0015] A medicine for the prevention or treatment of oral mucosa-associated fibrosis, the medicine comprising lysozyme.

[0016] In some implementations, the oral mucosa-associated fibrosis is not oral ulceration or oral mucositis.

[0017] In some embodiments, the oral mucosa-associated fibrosis is fibrosis of the oral mucosa and / or submucosal fibrosis.

[0018] In some implementations, the oral mucosa-associated fibrosis is not caused by bacterial or viral infection.

[0019] In some implementations, the oral mucosa-associated fibrosis is caused by areca nut, areca nut products, or chemical components in areca nut.

[0020] In some implementations, the oral mucosa-associated fibrosis is caused by the use of areca nut or areca nut products.

[0021] The third objective of this application is to provide a method. To achieve this objective, the technical solution adopted in this application is:

[0022] A method for improving oral mucosa-associated fibrosis includes applying a product containing an effective amount of lysozyme to the desired subject.

[0023] In some implementations, the product may be a pharmaceutical or a non-pharmaceutical.

[0024] In some implementations, the product may also include acceptable excipients or applicable additives.

[0025] In some embodiments, the daily dosage of the lysozyme may be 0.001g-100g.

[0026] In some implementations, the subject is a user of areca nut or areca nut products.

[0027] In some embodiments, the oral mucosa-associated fibrosis is fibrosis of the oral mucosa and / or submucosal fibrosis.

[0028] In some implementations, the improvement of oral mucosa-associated fibrosis includes reducing the level of fibrosis in the oral mucosa.

[0029] The beneficial effects of this application are:

[0030] 1. This application is the first to discover that lysozyme has a significant ameliorative effect on oral mucosa-associated fibrosis, and can dose-dependently reduce the collagen level and Masson staining optical density value of the oral mucosa, and significantly reverse the fibrosis process.

[0031] 2. The lysozyme of this application has a better inhibitory effect on oral mucosal fibrosis than commonly used glucocorticoids (such as triamcinolone acetonide), and has a better effect on improving oral mucosal fibrosis.

[0032] 3. The lysozyme of this application has high safety and can be used as both medicine and food, and has a good application prospect in improving oral mucosa-associated fibrosis. Attached Figure Description

[0033] Figure 1 These are Masson-stained light micrographs (400x magnification) of oral mucosal sections from various groups in the golden hamster oral mucosa-associated fibrosis model. A is the normal control group, B is the model control group, C is the triamcinolone group, D is the low-dose lysozyme group, E is the medium-dose lysozyme group, and F is the high-dose lysozyme group. Detailed Implementation

[0034] The present application will now be described in detail with reference to specific embodiments. It should be understood that the content of the specific embodiments section is illustrative and not restrictive, that is, it does not limit the content of the present application in any way.

[0035] definition:

[0036] "Improvement": In this application, "improvement" means mitigating an adverse condition. For example, in the case of mucosa-associated fibrosis, this includes reducing the degree of mucosa-associated fibrosis. Improvement includes both therapeutic and non-therapeutic improvements.

[0037] "Oral mucosa-associated fibrosis": The oral mucosa-associated fibrosis in this application includes oral submucous fibrosis (OSF). Oral mucosa-associated fibrosis differs from oral ulcers or oral mucositis. Oral ulcers or oral mucositis are mainly characterized by destruction of oral mucosal epithelial cells, hyperemia, and edema, and are related to inflammatory reactions, bacterial infections, and viral infections; while the main characteristic of oral mucosa-associated fibrosis is the fibrous proliferation and deposition of soft tissues in the oral submucosa, with betel nut chewing being one of the important risk factors.

[0038] "Lysozyme": The lysozyme in this application may be a lysozyme derived from animals, plants, or microorganisms, or a recombinant of natural lysozyme. For example, it may be egg white lysozyme, human lysozyme, recombinant human lysozyme, bacteriophage lysozyme, etc. The lysozyme in this application includes its pharmaceutical salts, such as hydrochloride, sulfate, or amino acid salts.

[0039] Lysozyme, first discovered by Fleming, is an endogenous enzyme widely found in living organisms. It has been approved worldwide for use as a food or pharmaceutical substance. In the United States, it is recognized as a Generally Recognized As Safe (GRAS) substance. The WHO, several European countries, Japan, and China permit its use as a food additive. It has also been approved for medicinal use in China, Japan, Singapore, and other countries.

[0040] "Product": The product in this application includes pharmaceuticals or non-pharmaceuticals. The product in this application includes topical products, oral products, or injectable products, etc. The product in this application includes liquid products, solid products, or semi-solid products. Non-pharmaceuticals may include food.

[0041] Example 1: Study on the effect of lysozyme on oral mucosa-associated fibrosis

[0042] This experiment used an animal model to investigate the effect of lysozyme on oral mucosal fibrosis.

[0043] Experimental animals: SPF-grade golden hamsters, weighing 80g-120g, half male and half female.

[0044] Test products: Lysozyme: Lysozyme lozenges (commercially available, Shōhoku Wellman Pharmaceutical Co., Ltd.) were ground into powder and set aside. Blank lozenges (from Shōhoku Wellman Pharmaceutical Co., Ltd., which have the same components as the lysozyme lozenges except that they do not contain lysozyme) were ground into powder and set aside. Triamcinolone: ​​0.1% triamcinolone oral ointment (commercially available, Aomei Pharmaceutical Co., Ltd.)

[0045] Experimental Methods: Animals in all groups except the normal control group underwent modeling to induce oral submucosal fibrosis. A bristle brush was used to apply concentrated areca nut extract (commercially available areca nut boiled in 10 times its volume of water, with the extract concentrated to a concentration of 1g areca nut / mL) to the oral mucosa of the animals 10 times, once every 2 days for 16 weeks. Starting in week 5, 0.2mL of concentrated areca nut extract was injected submucosally into the animals' oral mucosa once daily until week 8. After 16 weeks, successfully modeled animals were randomly divided into a model control group, a triamcinolone group, a low-dose lysozyme group, a medium-dose lysozyme group, and a high-dose lysozyme group, with 10 animals in each group. Ten normal animals that did not develop the model at the same time were used as the normal control group. The triamcinolone group received 0.2g of 0.1% triamcinolone oral ointment. The low-dose lysozyme group received 2mg / kg of lysozyme. The medium-dose lysozyme group received 10mg / kg of lysozyme. The high-dose lysozyme group was given 50 mg / kg of lysozyme. The model control group and the normal control group were given blank tablets at the same dosage as the high-dose lysozyme group. All animals were given the medication once daily for 6 consecutive weeks.

[0046] Testing: The following day after the last administration of the test product, various indicators were tested:

[0047] 1. Take oral mucosal tissue from the modeling site, wash it, add it to an EP tube, add 9 times the amount of physiological saline, homogenize, centrifuge, take the supernatant, and use an ELISA kit to detect the level of type I collagen in the supernatant.

[0048] 2. Oral mucosal tissue from the modeling site was collected, fixed in 10% formalin solution, embedded in paraffin, sectioned, and stained with Masson's stain. The optical density per unit area of ​​the Masson-stained sections was measured. Five fields of view were randomly selected for each animal, and the average optical density of the Masson-positive (blue) areas in each field of view was measured using Image-pro plus software. A higher Masson staining optical density value indicates a higher degree of fibrosis in the sample.

[0049] Experimental results: The levels of type I collagen in oral mucosa tissue are shown in Table 1. Masson staining optical density values ​​of oral mucosa are shown in Table 2. Observation of oral mucosa sections (Masson staining, 400x magnification) are shown in... Figure 1 In the figure, A is the normal control group, B is the model control group, C is the triamcinolone group, D is the low-dose lysozyme group, E is the medium-dose lysozyme group, and F is the high-dose lysozyme group; blue in the figure represents oral mucosal fibrosis signal.

[0050] Table 1: Type I collagen levels in oral mucosa

[0051]

[0052] Note: Comparison between the model control group and the normal control group. ++ P≤0.01; Comparison between each experimental product group and the model control group, ** P≤0.01,

[0053] * P≤0.05.

[0054] Table 2: Masson staining optical density values ​​of oral mucosa

[0055]

[0056] Note: Comparison between the model control group and the normal control group. ++ P≤0.01; Comparison between each experimental product group and the model control group, ** P≤0.01.

[0057] Experimental conclusion:

[0058] As shown in Table 1, compared with the normal control group, the type I collagen level in the model control group was significantly increased (P≤0.01). Compared with the model control group, the type I collagen levels in the triamcinolone group and each dose group of lysozyme were significantly decreased (P≤0.05), indicating that all tested products can reduce the collagen level of the oral mucosa.

[0059] Figure 1 The blue color represents the fiberization signal, from Figure 1 It can be seen that the blue areas in each lysozyme dosage group (D, E, F) are significantly less than those in the model control group (B) and also significantly less than those in the triamcinolone group (C), indicating that lysozyme can significantly improve oral mucosal fibrosis and is superior to triamcinolone.

[0060] Table 2 shows that Masson staining optical density values ​​can quantitatively reflect the degree of mucosal fibrosis. Compared with the normal control group, the optical density value of the model control group was significantly increased (P≤0.01), indicating that the oral mucosa-associated fibrosis model was successfully constructed. The optical density value of the triamcinolone group showed no significant change compared with the model control group, suggesting that triamcinolone was essentially ineffective in improving fibrosis; while the optical density values ​​of all lysozyme dose groups were significantly decreased compared with the model control group (P≤0.01), indicating that lysozyme has a significant effect on improving oral mucosal fibrosis.

[0061] This study demonstrates that while anti-inflammatory products such as triamcinolone acetonide can reduce oral mucosal collagen levels, they are largely ineffective in improving oral mucosal fibrosis. In contrast, lysozyme can significantly reduce the degree of oral mucosal fibrosis and reverse the fibrotic process. Furthermore, lysozyme can be used both as a medicine and food, and its safety profile is far superior to that of glucocorticoids. Therefore, developing lysozyme products to improve oral mucosal fibrosis shows great promise.

[0062] The present application has been described in detail above with general descriptions and specific embodiments. Based on this, those skilled in the art can make some changes or improvements. All such changes or improvements made without departing from the scope of protection of the present application shall fall within the scope of protection claimed in the present application.

Claims

1. The application of lysozyme in the preparation of drugs that help improve oral mucosa-associated fibrosis.

2. The application according to claim 1, characterized in that: The oral mucosa-associated fibrosis is oral submucosal fibrosis.

3. A method for improving oral mucosa-associated fibrosis, characterized in that: The method involves administering a drug containing an effective amount of lysozyme to the desired object.

4. The method according to claim 3, characterized in that: The medicine also includes acceptable excipients or applicable additives.

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