Stem cell-containing eye care solution and method of making same

Abstract

The application relates to the technical field of eye drops, in particular to an eye nursing liquid containing stem cells and a preparation method thereof, which comprises physiological saline, stem cells, punicalin, porous adsorption material and traditional Chinese medicine extract according to mass parts, the traditional Chinese medicine extract comprises psyllium, atractylodes, radix falcaliae, cistanche, stone lotus, radix paeoniae and poria cocos, and the porous adsorption material is obtained by swelling and drying treatment of solid endosperm of a coconut. The application repairs eye injuries through stem cells, reduces cell apoptosis and inflammatory reactions, promotes blood vessel regeneration and improves the repair ability of eye tissues; the natural porous adsorption material is used as a carrier to adsorb the effective components such as traditional Chinese medicine extract and stem cells on the eyes, prolongs the drug residence time, has a moisturizing effect at the same time, and relieves the dry eye symptoms; the eye nursing liquid does not contain preservatives and has a longer sustained effect after use.

Description

Technical Field

[0001] This invention relates to the field of eye drop technology, specifically to an eye care solution containing stem cells and its preparation method. Background Technology

[0002] Eye care solutions are used to relieve eye fatigue and for the prevention and treatment of some eye diseases. These solutions typically contain multiple active ingredients such as antibacterial and antiviral components, along with moisturizing ingredients like hyaluronic acid and glycerin to alleviate dryness and discomfort. However, existing eye care solutions have some limitations. For example, their effectiveness is relatively short-lived, requiring frequent use and increasing the burden on patients. Furthermore, long-term, frequent use of eye care solutions may lead to dry eyes because the medicinal components can irritate eye cells. Moreover, to extend shelf life, commercially available eye care solutions often contain preservatives, most of which can harm the eyes. Summary of the Invention

[0003] (1) Technical problems to be solved

[0004] The purpose of this invention is to provide an eye care solution containing stem cells and its preparation method, so as to solve the problems of short action time and short shelf life of eye care solutions without the addition of preservatives.

[0005] (2) Technical solution

[0006] To achieve the above objectives, in one aspect, the present invention provides an eye care solution containing stem cells, wherein the eye care solution comprises, by weight, the following components:

[0007] 60–90 parts physiological saline, 0.7–2.3 parts stem cells, 1–4 parts pungent glycoside, 0.5–1.4 parts porous adsorption material, and 3–8 parts traditional Chinese medicine extract;

[0008] The herbal extract comprises, by weight, the following:

[0009] 10-20 parts Plantago seed, 13-30 parts Atractylodes lancea, 8-16 parts Polygonum multiflorum, 17-24 parts Cistanche deserticola, 16-30 parts Nelumbo nucifera, 10-25 parts Paeonia suffruticosa, and 16-30 parts Poria cocos;

[0010] The porous adsorbent material is obtained by swelling and drying the solid endosperm of coconut, and the particle size of the porous adsorbent material is 10-20 μm.

[0011] Furthermore, the eye care solution further includes 0.3 to 1.2 parts of modified sodium hyaluronate and 1.5 to 4.5 parts of ε-aminocaproic acid by weight.

[0012] Furthermore, the preparation method of the modified sodium hyaluronate includes:

[0013] S11. Dissolve sodium hyaluronate in an acetic acid / sodium acetate buffer solution with a pH of 4-6 to form a solution with a concentration of 1-3%.

[0014] S12, continue adding 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide, wherein the molar ratio of sodium hyaluronate, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide is 5:

[0015] (1~2): (3~5);

[0016] S13. After reacting at 25-40℃ for 2-6 hours, add an equal volume of anhydrous ethanol to precipitate and filter.

[0017] S14. The obtained product is vacuum dried at 40-60℃ for 12-24 hours to obtain modified sodium hyaluronate.

[0018] Furthermore, the preparation method of the porous adsorbent material includes:

[0019] S21. Soak 5-10 month old animals in deionized water to swell until constant weight is achieved, then remove them to obtain swollen material.

[0020] S22. The swollen material is dried at 40-80°C to obtain a natural porous adsorbent material;

[0021] The outer layer of the porous adsorbent material has pores ranging from 500 nm to 3 μm.

[0022] Furthermore, the preparation method of the traditional Chinese medicine extract includes:

[0023] S31. Take Plantago seed, Atractylodes lancea, Polygonum multiflorum, Cistanche deserticola, Nelumbo nucifera, Paeonia suffruticosa and Poria cocos, wash and dry them, crush and sieve them to prepare coarse powder, add them to ethanol for pre-soaking, then heat and filter to obtain the first alcohol extract and the first filter residue.

[0024] S32. Add the first filter residue to ethanol and heat, then filter to obtain the second alcohol extract and the second filter residue.

[0025] S33. Mix the first and second alcohol extracts, filter thoroughly, and evaporate and concentrate to obtain a paste or slurry-like extract stock solution to obtain the Chinese herbal extract.

[0026] Furthermore, the ethanol pre-soaking time is 0.5 to 1 hour, the amount of ethanol used in the extraction process is 8 to 12 times the weight of the object to be extracted, and the concentration of ethanol is 50 to 80%.

[0027] Furthermore, the heating time during the alcohol extraction process is 1 to 2 hours, and the heating temperature is 60 to 80°C.

[0028] Based on the same inventive concept, in another aspect, the present invention also provides a method for preparing an eye care solution containing stem cells, applicable to the preparation of the aforementioned eye care solution containing stem cells, the preparation method comprising:

[0029] S1. Place the Chinese herbal extract and pomegranate glycoside into physiological saline and stir to mix evenly to obtain a water-soluble substance;

[0030] S2. Place the porous adsorbent material into the aqueous solution and stir to mix evenly to obtain a dispersion.

[0031] S3. Add modified sodium hyaluronate and ε-aminocaproic acid to the dispersion, then add stem cells and stir to mix evenly to obtain the eye care solution.

[0032] (3) Beneficial effects

[0033] Compared with the prior art, the beneficial effects of the present invention are:

[0034] 1. Through the use of traditional Chinese medicine components such as Plantago asiatica, Atractylodes lancea, Polygonum multiflorum, Cistanche deserticola, Nelumbo nucifera, Paeonia suffruticosa, and Poria cocos, it has anti-inflammatory and anti-infective effects, promotes blood circulation, and helps improve eye health.

[0035] 2. Stem cells can repair eye damage, reduce cell apoptosis and inflammatory response, promote angiogenesis, and improve the repair capacity of eye tissues.

[0036] 3. Using natural porous adsorption materials as carriers, the active ingredients such as Chinese herbal extracts and stem cells are effectively adsorbed onto the eye, prolonging the drug's residence time and simultaneously providing moisturizing effects to relieve dry eye symptoms.

[0037] 4. Modified sodium hyaluronate prolongs the pre-keratotic residence time of other components. The modified sodium hyaluronate binds to stem cell surface receptors and promotes stem cell proliferation and differentiation. Attached Figure Description

[0038] Figure 1 This is a block diagram illustrating a method for preparing a stem cell-containing eye care solution according to Embodiment 1 of the present invention.

[0039] Figure 2 This is a SEM image of the porous adsorbent material prepared in Example 1 of the present invention; Detailed Implementation

[0040] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0041] Example 1: This example discloses an eye care solution containing stem cells, wherein the eye care solution comprises, by weight, the following:

[0042] 60 parts physiological saline, 0.7 parts stem cells, 1 part pungent glycoside, 0.5 parts porous adsorption material and 3 parts traditional Chinese medicine extract;

[0043] The herbal extract comprises, by weight, the following:

[0044] 10 parts Plantago seed, 13 parts Atractylodes lancea, 8 parts Polygonum multiflorum, 17 parts Cistanche deserticola, 16 parts Nelumbo nucifera, 10 parts Paeonia suffruticosa, and 16 parts Poria cocos;

[0045] The porous adsorbent material is obtained by swelling and drying the solid endosperm of coconut, and the particle size of the porous adsorbent material is 10-20 μm.

[0046] The eye care solution also includes 0.3 parts modified sodium hyaluronate and 1.5 parts ε-aminocaproic acid by weight.

[0047] The preparation method of the modified sodium hyaluronate includes:

[0048] S11. Dissolve sodium hyaluronate in an acetic acid / sodium acetate buffer solution with a pH of 4-6 to form a solution with a concentration of 1-3%.

[0049] S12, continue to add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide, wherein the molar ratio of sodium hyaluronate, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide is 5:(1~2):(3~5);

[0050] S13. After reacting at 25-40℃ for 2-6 hours, add an equal volume of anhydrous ethanol to precipitate and filter.

[0051] S14. The obtained product is vacuum dried at 40-60℃ for 12-24 hours to obtain modified sodium hyaluronate.

[0052] The method for preparing the porous adsorption material includes:

[0053] S21. Soak 5-10 month old animals in deionized water to swell until constant weight is achieved, then remove them to obtain swollen material.

[0054] S22. The swollen material is dried at 40-80°C to obtain a natural porous adsorbent material;

[0055] The outer layer of the porous adsorbent material has pores ranging from 500 nm to 3 μm.

[0056] like Figure 1 As shown, the preparation method of the traditional Chinese medicine extract includes:

[0057] S31. Take Plantago seed, Atractylodes lancea, Polygonum multiflorum, Cistanche deserticola, Nelumbo nucifera, Paeonia suffruticosa and Poria cocos, wash and dry them, crush and sieve them to prepare coarse powder, add them to ethanol for pre-soaking, then heat and filter to obtain the first alcohol extract and the first filter residue.

[0058] S32. Add the first filter residue to ethanol and heat, then filter to obtain the second alcohol extract and the second filter residue.

[0059] S33. Mix the first and second alcohol extracts, filter thoroughly, and evaporate and concentrate to obtain a paste or slurry-like extract stock solution to obtain the Chinese herbal extract.

[0060] The ethanol pre-soaking time is 0.5 to 1 hour, the amount of ethanol used in the extraction process is 8 to 12 times the weight of the material to be extracted, and the concentration of ethanol is 50 to 80%. The heating time in the extraction process is 1 to 2 hours, and the heating temperature is 60 to 80°C.

[0061] The method for preparing the stem cell-containing eye care solution includes:

[0062] S1. Place the Chinese herbal extract and pomegranate glycoside into physiological saline and stir to mix evenly to obtain a water-soluble substance;

[0063] S2. Place the porous adsorbent material into the aqueous solution and stir to mix evenly to obtain a dispersion.

[0064] S3. Add modified sodium hyaluronate and ε-aminocaproic acid to the dispersion, then add stem cells and stir to mix evenly to obtain the eye care solution.

[0065] The polysaccharides and flavonoids in Plantago asiatica have the effects of clearing heat and detoxifying, promoting diuresis and reducing swelling, and can scavenge free radicals and enhance ocular immune function; the volatile oil and atractylone in Atractylodes lancea have the effects of drying dampness, resolving turbidity, and relieving pain, and can inhibit the release of inflammatory mediators and prevent eye infections; the stilbene glycosides and lecithin in Polygonum multiflorum have the effects of nourishing essence and blood, detoxifying and stopping malaria, and can inhibit inflammatory signaling pathways and reduce ocular inflammatory reactions; the phenylethyl glycosides and echinacosides in Cistanche deserticola have the characteristics of tonifying yang without drying, moistening the intestines and promoting bowel movements, and can promote blood circulation and enhance... It strengthens the nutritional supply to the eye tissues; the alkaloids and flavonoids in the lotus have the effects of clearing heat and detoxifying, stopping bleeding and dysentery, and can inhibit the growth of pathogenic microorganisms and prevent eye infections; the paeonol and salvianolic acid in the peony bark have the effects of clearing heat and cooling blood, promoting blood circulation and removing blood stasis, and can inhibit the vascular endothelial growth factor (VEGF) signaling pathway and reduce pathological angiogenesis; the poria polysaccharides and triterpenoids in the poria have the effects of promoting diuresis and removing dampness, strengthening the spleen and calming the mind, and can enhance the immune function of the eye tissues, reduce inflammatory response, and promote eye health.

[0066] Simultaneously, stem cells are added to repair damaged areas of the eye, promote the reduction of cell apoptosis, alleviate inflammatory response, promote angiogenesis, inhibit fibrosis, enhance tissue repair potential, and promote new blood vessels in the eye, thus achieving the goal of combining "treatment" and "nourishment".

[0067] However, eye care solutions tend to slip off the corners of the eyes during use, and due to the presence of the tear film (which has a relatively smooth surface), it is difficult for the active ingredients to adhere to the eye and achieve the aforementioned therapeutic effects. Therefore, natural porous absorbent materials are added to the eye care solution. These materials have excellent skin affinity, preventing any foreign body sensation in the eyes, and possess a strong adsorption capacity. They adsorb the active ingredients from traditional Chinese medicine extracts and stem cells, allowing them to be quickly absorbed onto the tear film upon contact with the eyes, and then released, enabling the stem cells and traditional Chinese medicine extracts to exert their corresponding effects. Furthermore, these natural porous absorbent materials have good moisturizing properties, preventing dryness and effectively relieving and preventing dry eye symptoms. Figure 2 As shown, the microscopic image of the natural porous adsorbent material obtained by viewing it under a zoom microscope reveals that the surface of the natural porous adsorbent material has many pores that can accommodate and absorb effective ingredients, and even large molecular clusters of traditional Chinese medicine ingredients.

[0068] However, natural porous adsorbent materials often contain allergens, which can easily cause allergic symptoms in the eyes. Eye allergies prompt the eyes to secrete mucus and tears in an attempt to flush out the allergens. This secretion of mucus can inhibit stem cell differentiation. This application adds pungent glycoside to the eye care solution. Pungent glycoside promotes stem cell differentiation, enhancing the eye care effect; simultaneously, it synergistically works with the stem cells to achieve receptor blocking, thus achieving an anti-allergic effect. Furthermore, pungent glycoside has strong antibacterial and antioxidant properties, so its addition eliminates the need for preservatives in the eye care solution, improving its safety. After entering the eye, pungent glycoside is decomposed into ellagic acid under the action of human enzymes. This ellagic acid, in synergy with traditional Chinese medicine extracts, exerts a powerful anti-infective effect, effectively enhancing the efficacy against ophthalmic diseases.

[0069] During use, after the patient instills this eye care solution, the saline solution rapidly dissolves and disperses the active ingredients, ensuring full contact with the ocular surface tissue. Modified sodium hyaluronate forms a uniform hydrogel film on the ocular surface, delaying tear evaporation and solution loss, thereby prolonging the corneal residence time of other components. Modified sodium hyaluronate, after its carboxyl groups are activated by EDC / NHS, cross-links with amino groups, increasing intermolecular chemical bonds, improving molecular weight and viscoelasticity, thus enhancing the stability and bioadhesion of the hydrogel film. Simultaneously, modified sodium hyaluronate can specifically bind to receptors such as CD44 and CD168 on the surface of stem cells, activating signaling pathways such as Src and ERK, promoting stem cell proliferation; and activating the STAT3 pathway, maintaining the multipotent differentiation potential of stem cells. The HA-CD44 binding-induced signaling pathway synergistically enhances stem cell proliferation and differentiation with ε-aminocaproic acid (EACA). EACA, as a histone deacetylase inhibitor, enhances HA-induced stem cell pluripotency gene expression by weakening histone deacetylation and relaxing chromatin structure.

[0070] While modified sodium hyaluronate and EACA synergistically promote stem cell proliferation and differentiation, porous adsorbent materials, in conjunction with pungent glycosides, construct a microenvironment conducive to tissue repair. The porous adsorbent material possesses numerous nano- to micron-sized pores, with pores ranging from 500 nm to 3 μm distributed on the outer layer, capable of accommodating and sustaining the release of macromolecular proteins such as EGF and bFGF growth factors. This porous structure makes the porous adsorbent material an ideal drug delivery system, creating a microenvironment of high concentrations of cytokines and active substances on the ocular surface, which is beneficial for stem cell proliferation, differentiation, and tissue repair. Simultaneously, polyphenolic substances (catechols, proanthocyanidins, etc.) in pungent glycosides can be adsorbed onto the surface of the porous material through hydrophobic interactions and hydrogen bonding, making the porous adsorbent material a sustained-release carrier for pungent glycosides and prolonging its efficacy. The sustained-release pungent glycosides and modified sodium hyaluronate complement each other, jointly maintaining the stability of the stem cell microenvironment and promoting tissue repair.

[0071] In addition, porous adsorbent materials can adsorb inflammatory mediators and metabolic waste from tears, reducing ocular surface inflammation and improving the microenvironment. Their surface can also be modified with antimicrobial peptides, endowing the material with antimicrobial properties and preventing secondary infections.

[0072] Traditional Chinese medicine extracts exhibit synergistic effects with their main components through multi-pathway and multi-target mechanisms. For example, astaxanthin in Plantago asiatica extract scavenge reactive oxygen species (ROS), reduce oxidative stress, protect stem cells and corneal epithelium from oxidative damage, and synergistically maintain cell viability with modified sodium hyaluronate and EACA. Stilbene glycosides in Polygonum multiflorum extract inhibit PAR2, block inflammatory responses, and synergistically exert anti-inflammatory effects with Poria cocos polysaccharides. Danshensu B in Paeonia suffruticosa extract inhibits the EGFR signaling pathway, reduces pathological angiogenesis, and synergistically inhibits ocular scar formation with oleanolic acid in Nelumbo nucifera extract.

[0073] Modified sodium hyaluronate, upon binding to receptors such as CD44, triggers a cascade amplification of signaling pathways including Src, ERK, and STAT3. EACA, by inhibiting histone deacetylation, synergistically enhances these pathways, jointly regulating the expression of genes such as cyclin D1, Sox2, and Oct4, thus promoting stem cell proliferation and differentiation. Porous adsorption materials can physically adsorb and enrich growth factors such as EGF and bFGF, activating receptor tyrosine kinases such as EGFR and FGFR, triggering signaling pathways such as RAS-MAPK and PI3K-AKT, and synergistically promoting stem cell proliferation, differentiation, and migration through the HA-CD44-induced signaling pathway. Active components in traditional Chinese medicine extracts exert anti-inflammatory effects at the gene expression level by inhibiting key receptors such as EGFR and PAR2, downregulating the activity of transcription factors such as NF-κB and STAT3, and negatively regulating the transcription of inflammatory factor genes such as IL-6 and TNF-α, thereby improving stem cell survival rates. Meanwhile, Chinese herbal extracts can also activate antioxidant transcription factors such as Nrf2, upregulate the expression of antioxidant enzymes such as HO-1 and NQO1, clear ROS, reduce oxidative stress, maintain the redox homeostasis of stem cells and corneal epithelial cells, and provide a stable environment for tissue repair.

[0074] Example 2: This example discloses an eye care solution containing stem cells, wherein the eye care solution comprises, by weight, the following:

[0075] 90 parts physiological saline, 2.3 parts stem cells, 4 parts pungent glycoside, 1.4 parts porous adsorption material and 8 parts traditional Chinese medicine extract;

[0076] The herbal extract comprises, by weight, the following:

[0077] 10 parts Plantago seed, 13 parts Atractylodes lancea, 8 parts Polygonum multiflorum, 17 parts Cistanche deserticola, 16 parts Nelumbo nucifera, 10 parts Paeonia suffruticosa, and 16 parts Poria cocos;

[0078] The porous adsorbent material is obtained by swelling and drying the solid endosperm of coconut, and the particle size of the porous adsorbent material is 10-20 μm.

[0079] The eye care solution also includes 0.3 parts modified sodium hyaluronate and 1.5 parts ε-aminocaproic acid by weight.

[0080] The preparation methods for the porous adsorption material, the traditional Chinese medicine extract, the modified sodium hyaluronate, and the stem cell-containing eye care solution are the same as in Example 1.

[0081] Example 3: This example discloses an eye care solution containing stem cells, wherein the eye care solution comprises, by weight, the following:

[0082] 60 parts physiological saline, 0.7 parts stem cells, 1 part pungent glycoside, 0.5 parts porous adsorption material and 3 parts traditional Chinese medicine extract;

[0083] The herbal extract comprises, by weight, the following:

[0084] 20 parts Plantago seed, 30 parts Atractylodes lancea, 16 parts Polygonum multiflorum, 24 parts Cistanche deserticola, 30 parts Nelumbo nucifera, 25 parts Paeonia suffruticosa, and 30 parts Poria cocos;

[0085] The porous adsorbent material is obtained by swelling and drying the solid endosperm of coconut, and the particle size of the porous adsorbent material is 10-20 μm.

[0086] The eye care solution also includes 0.3 parts modified sodium hyaluronate and 1.5 parts ε-aminocaproic acid by weight.

[0087] The preparation methods for the porous adsorption material, the traditional Chinese medicine extract, the modified sodium hyaluronate, and the stem cell-containing eye care solution are the same as in Example 1.

[0088] Example 4: This example discloses an eye care solution containing stem cells, wherein the eye care solution comprises, by weight, the following:

[0089] 60 parts physiological saline, 0.7 parts stem cells, 1 part pungent glycoside, 0.5 parts porous adsorption material and 3 parts traditional Chinese medicine extract;

[0090] The herbal extract comprises, by weight, the following:

[0091] 10 parts Plantago seed, 13 parts Atractylodes lancea, 8 parts Polygonum multiflorum, 17 parts Cistanche deserticola, 16 parts Nelumbo nucifera, 10 parts Paeonia suffruticosa, and 16 parts Poria cocos;

[0092] The porous adsorbent material is obtained by swelling and drying the solid endosperm of coconut, and the particle size of the porous adsorbent material is 10-20 μm.

[0093] The eye care solution also includes 1.2 parts modified sodium hyaluronate and 4.5 parts ε-aminocaproic acid by weight.

[0094] The preparation methods for the porous adsorption material, the traditional Chinese medicine extract, the modified sodium hyaluronate, and the stem cell-containing eye care solution are the same as in Example 1.

[0095] Control Group 1: The difference between this control group and Example 1 is that it does not contain porous adsorbent materials, and the prepared eye care solution comprises, by weight:

[0096] 60 parts physiological saline, 0.7 parts stem cells, 1 part pomegranate glycoside and 3 parts traditional Chinese medicine extract;

[0097] The herbal extract comprises, by weight, the following:

[0098] 10 parts Plantago seed, 13 parts Atractylodes lancea, 8 parts Polygonum multiflorum, 17 parts Cistanche deserticola, 16 parts Nelumbo nucifera, 10 parts Paeonia suffruticosa, and 16 parts Poria cocos;

[0099] The eye care solution also includes 0.3 parts modified sodium hyaluronate and 1.5 parts ε-aminocaproic acid by weight.

[0100] The traditional Chinese medicine extract and modified sodium hyaluronate are the same as in Example 1, but porous adsorption materials are not added in the preparation method of the stem cell-containing eye care solution.

[0101] Control Group 2: This control group differs from Example 1 in that it does not contain modified sodium hyaluronate and ε-aminocaproic acid. The eye care solution in this control group, by weight, comprises:

[0102] 60 parts physiological saline, 0.7 parts stem cells, 1 part pungent glycoside, 0.5 parts porous adsorption material and 3 parts traditional Chinese medicine extract;

[0103] The herbal extract comprises, by weight, the following:

[0104] 10 parts Plantago seed, 13 parts Atractylodes lancea, 8 parts Polygonum multiflorum, 17 parts Cistanche deserticola, 16 parts Nelumbo nucifera, 10 parts Paeonia suffruticosa, and 16 parts Poria cocos;

[0105] The porous adsorbent material is obtained by swelling and drying the solid endosperm of coconut, and the particle size of the porous adsorbent material is 10-20 μm.

[0106] The porous adsorption material and the traditional Chinese medicine extract are the same as in Example 1. Modified sodium hyaluronate and ε-aminocaproic acid are not added in the preparation method of the stem cell-containing eye care solution.

[0107] Control Group 3: The difference between this control group and Example 1 is that it does not contain pungent glycosides, and the prepared eye care solution comprises, by weight:

[0108] 60 parts physiological saline, 0.7 parts stem cells, 0.5 parts porous adsorption material, and 3 parts traditional Chinese medicine extract;

[0109] The herbal extract comprises, by weight, the following:

[0110] 10 parts Plantago seed, 13 parts Atractylodes lancea, 8 parts Polygonum multiflorum, 17 parts Cistanche deserticola, 16 parts Nelumbo nucifera, 10 parts Paeonia suffruticosa, and 16 parts Poria cocos;

[0111] The porous adsorbent material is obtained by swelling and drying the solid endosperm of coconut, and the particle size of the porous adsorbent material is 10-20 μm.

[0112] The eye care solution also includes 0.3 to 1.2 parts of modified sodium hyaluronate and 1.5 to 4.5 parts of ε-aminocaproic acid by weight.

[0113] The preparation methods for porous adsorption materials, traditional Chinese medicine extracts, and modified sodium hyaluronate are the same as in Example 1. However, pungent glycosides are not added in the preparation method of the stem cell-containing eye care solution.

[0114] Experiment 1: 200 ml of the ophthalmic care solution prepared in Example 1 and Control Group 1 was used as a sample. Twenty healthy adult rabbits were selected and divided into two groups, and both groups were general anesthetized with anesthetic drugs. Using an electronic balance, equal amounts of the sample were instilled into the eyes of each rabbit in both Example 1 and Control Group 1. A constant temperature and humidity chamber was used to set the experimental environment to high dryness (relative humidity <20%) to simulate environmental dehydration. Liquid samples were collected from the corners of the rabbits' eyes at regular intervals using a liquid sampler, and the drug concentration was measured using high-performance liquid chromatography (HPLC). The sampling time points were designed according to the experiment. Simultaneously, standard medical methods for measuring dry eye syndrome were used, and a tear film breakup time meter was used to assess the moisturizing effect on the ocular surface. The experimental equipment included an electronic balance (Mettler Toledo, model XS204), a temperature and humidity chamber (ThermoFisher Scientific, model Heratherm), animal subjects (rabbits, obtained in accordance with animal experimentation ethics), anesthetic drugs (Zoetis, model Isoflurane), a liquid sampler (Eppendorf, model Research plus), a high-performance liquid chromatograph (Agilent, model 1260 Infinity II), and a tear film breakup time meter (Topcon, model KR-1W). The experimental results are shown in Tables 1 and 2.

[0115] Table 1: Drug Concentration Measurement Data

[0116] Time / d Example 1: Drug concentration (mg / L) Drug concentration in control group 1 (mg / L) 0 31.2±0.5 50.6±0.5 1 49.5±0.5 43.3±0.5 2 50.8±0.5 37.9±0.5 3 45.6±0.5 31.5±0.5 24 32.3±0.5 14.2±0.5 48 29.1±0.5 6.7±0.5 72 26.4±0.5 2.5±0.5

[0117] The above drug concentrations were inferred by measuring the components of two or three drugs in the total ratio. Additionally, due to the precision of the measuring instruments, there was measurement error in the drug concentration measurements. The data for each group were obtained by averaging data from multiple experimental rabbits. Table 1 shows that the eye care solution of Example 1 has a sustained-release effect, initially releasing slowly until reaching a peak at approximately 2 hours, and then continuing to release slowly thereafter.

[0118] Table 2: Tear film breakup time

[0119] Time (d) Example 1: Tear film breakup time (s) Tear film breakup time (s) in control group 1 0 10.8±0.2 11.1±0.2 3 10.0±0.2 7.5±0.2 6 9.2±0.2 5.2±0.2 12 8.6±0.2 3.6±0.2

[0120] Because under high dryness conditions, the tear film breakup time in Example 1 of Example 1 was relatively normal with good moisturizing effect, while in Control Group 1, under dry environment and without moisturizing eye care solution, the tear film breakup time was greatly shortened, indicating poor moisturizing of the ocular surface.

[0121] Experiment 2: 150 ml of the ophthalmic care solution prepared in Example 1 and Control Group 3 was used as a sample. Twenty healthy adult rabbits were randomly divided into two groups of 10 each. Administration began on day 1, with 0.5 ml of the corresponding ophthalmic care solution added according to the grouping, four times daily for seven consecutive days. Before administration and on days 1, 3, 5, and 7 after administration, corneal and conjunctival congestion and edema were observed and scored using a slit-lamp microscope, along with corneal epithelial damage. On day 7 after administration, the animals were sacrificed, and rabbit eye tissue was collected for pathological anatomical examination to observe pathological changes in the cornea and conjunctiva. The experimental equipment included a slit-lamp microscope (Topcon SL-D701), an ophthalmic surgical microscope (Leica M844 F40), an optical microscope (Zeiss Primo Star), a pathological microtome (Leica RM2235), and a pathological slide warmer (Leica HI1210). The experimental results are shown in Table 3.

[0122] Table 3: Observation Record of Corneal Conjunctiva

[0123] Time / d Example 1 Control group 3 Day 1 There was no congestion or edema in the cornea or conjunctiva. Slightly bloodshot cornea Day 3 There was no congestion or edema in the cornea or conjunctiva. Slightly bloodshot cornea Day 5 There was no congestion or edema in the cornea or conjunctiva. Slight bloodshot eyes on cornea and conjunctiva Day 7 There was no congestion or edema in the cornea or conjunctiva. Slight bloodshot eyes on cornea and conjunctiva After dissection No obvious pathological changes were observed. Slight congestion of the cornea and conjunctiva

[0124] As can be seen from Table 3, the eye care solution in control group 3, which does not contain pungent glycoside, caused slight irritation to the eyes. After adding pungent glycoside, the irritation to the eyes was reduced to the point that no adverse reactions occurred.

[0125] Experiment 3: 200 ml of the ophthalmic care solution prepared in Example 1 and Control Group 2 was used as a sample. Twenty healthy adult rabbits were randomly divided into two groups of 10 each. The rabbit eyes were chemically burned using a 0.1% benzalkonium chloride solution to induce an acute corneal inflammation model. On days 1, 2, 3, 5, and 7, the ophthalmic care solutions from Example 1 and Control Group 2 were instilled, 3 drops per eye, 4 times daily. Before administration and on days 1, 3, 5, and 7 after administration, corneal opacity, neovascularization, epithelial defects, conjunctival hyperemia and edema were observed using a slit-lamp microscope. Photographs were taken and recorded. The degree of corneal opacity was scored using a grayscale value of 0-5. The length of neovascularization was expressed as the distance from the limbus to the anterior end of the neovascularization. The area of ​​epithelial defects was stained with sodium fluorescein and measured under a slit lamp with a steel ruler. The degree of conjunctival hyperemia and edema was scored using a severity value of 0-4. The rabbits were sacrificed on day 7, and the eyes were enucleated. Corneal and conjunctival tissues were fixed in 4% paraformaldehyde and embedded in paraffin. Routine sections were prepared. Histopathological changes were observed under a light microscope after HE staining. Corneal tissue homogenate was collected. The levels of inflammatory factors TNF-α, IL-6, and IL-1β were detected by ELISA. Experimental equipment included benzalkonium chloride solution (Sigma, 0.1%), a slit-lamp microscope (Topcon, SL-D701), sodium fluorescein eye drops (Alcon, 0.5%), an optical microscope (Olympus, BX53), and TNF-α, IL-6, and IL-1β ELISA kits (R&D Systems). The experimental results after drug administration are shown in Tables 4 and 5.

[0126] Table 4: Records of Simulated Eye Injury Experiments

[0127]

[0128] Table 5: Experimental Records for Detecting Inflammatory Factors

[0129] Group TNF-α IL-6 IL-1β Example 1 85.2 62.5 45.8 Control group 2 102.6 88.3 68.6

[0130] The data for each group were obtained by averaging the data from multiple experimental rabbits. As shown in Tables 4 and 5, the modified sodium hyaluronate and ε-aminocaproic acid in Example 1 significantly improved the effect of the eye care solution.

[0131] Experiment 4: 200ml of the eye care solutions prepared in Examples 1-4 were used as samples. Twenty volunteers with easily irritated, red, and swollen eyes and mild dry eye syndrome were recruited at a hospital. An eye symptom rating scale and a visual acuity chart were used to assess the volunteers' eye symptoms and visual acuity, and initial symptom scores and visual acuity values ​​were recorded. The volunteers were divided into four groups of five, each using one of the eye care solutions from Examples 1-4. The stem cell-containing eye care solution was used twice daily, morning and evening, for two weeks. Before and two weeks after use, the volunteers' eye symptoms were subjectively assessed using an eye symptom rating scale (scores 1-5: 1 for red, swollen eyes with excessive tearing and impaired vision; 2 for mild redness and dryness with impaired vision; 3 for redness without swelling, mild dryness, and slight impaired vision; 4 for bloodshot eyes and dryness; 5 for mild or no bloodshot eyes and no other discomfort). The appearance of the eyes of each group of volunteers was observed, and the improvement rate of eye appearance was obtained by objectively scoring the appearance of the eyes. The score was also based on the similarity to the appearance of normal eyes. The results are shown in Table 6.

[0132] Table 6: Comparison of the effects of eye care solutions in Examples 1-4

[0133]

[0134] By comparing the data from Examples 1 to 4, it can be seen that the stem cell-containing eye care solution has a significant therapeutic effect on eye damage. Among them, the treatment effect of Example 1 group was the best, while the efficacy varied slightly among Examples 2 to 4. This indicates that the formulation of the eye care solution affects the therapeutic effect.

[0135] Finally, it should be noted that although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An eye care solution containing stem cells, characterized in that, The eye care solution comprises, by weight, the following: 60-90 parts physiological saline, 0.7-2.3 parts stem cells, 1-4 parts pungent glycoside, 0.5-1.4 parts porous adsorption material and 3-8 parts traditional Chinese medicine extract; The herbal extract is made from the following components by weight: 10-20 parts Plantago asiatica, 13-30 parts Atractylodes lancea, 8-16 parts Polygonum multiflorum, 17-24 parts Cistanche deserticola, 16-30 parts Nelumbo nucifera, 10-25 parts Paeonia suffruticosa, and 16-30 parts Poria cocos. The porous adsorbent material is obtained by swelling and drying the solid endosperm of coconut, and the particle size of the porous adsorbent material is 10~20μm.

2. The eye care solution containing stem cells according to claim 1, characterized in that, The eye care solution also includes 0.3 to 1.2 parts of modified sodium hyaluronate and 1.5 to 4.5 parts of ε-aminocaproic acid by weight.

3. The eye care solution containing stem cells according to claim 2, characterized in that, The preparation method of the modified sodium hyaluronate includes: S11. Dissolve sodium hyaluronate in an acetic acid / sodium acetate buffer solution with a pH of 4-6 to form a solution with a concentration of 1-3%. S12, continue to add 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide, wherein the molar ratio of sodium hyaluronate, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and N-hydroxysuccinimide is 5:(1~2):(3~5); S13. After reacting at 25~40℃ for 2~6 hours, add an equal volume of anhydrous ethanol to precipitate and filter. S14. The obtained product is vacuum dried at 40~60℃ for 12~24 hours to obtain modified sodium hyaluronate.

4. The eye care solution containing stem cells according to claim 1, characterized in that, The method for preparing the porous adsorbent material includes: S21. The solid endosperm of coconuts aged 5-10 months is soaked in deionized water to swell until constant weight is reached and then removed to obtain swollen material; the swollen material is sheared, sand-milled and dispersed to the micron level. S22. The swollen material is dried at 40~80℃ to obtain a natural porous adsorbent material; The outer layer of the porous adsorbent material has pores ranging from 500 nm to 3 μm.

5. The eye care solution containing stem cells according to claim 1, characterized in that, The preparation method of the traditional Chinese medicine extract includes: S31. Take Plantago seed, Atractylodes lancea, Polygonum multiflorum, Cistanche deserticola, Nelumbo nucifera, Paeonia suffruticosa and Poria cocos, wash and dry them, crush and sieve them to prepare coarse powder, add them to ethanol for pre-soaking, then heat and filter to obtain the first alcohol extract and the first filter residue. S32. Add the first filter residue to ethanol and heat, then filter to obtain the second alcohol extract and the second filter residue. S33. Mix the first and second alcohol extracts, filter thoroughly, and evaporate and concentrate to obtain a paste or slurry-like extract stock solution to obtain the Chinese herbal extract.

6. The eye care solution containing stem cells as described in claim 5, characterized in that, The ethanol pre-soaking time is 0.5 to 1 hour, the amount of ethanol used in the extraction process is 8 to 12 times the weight of the object to be extracted, and the concentration of ethanol is 50 to 80%.

7. The eye care solution containing stem cells as described in claim 5, characterized in that, The heating time during the alcohol extraction process is 1-2 hours, and the heating temperature is 60-80℃.

8. A method for preparing an eye care solution containing stem cells, applied to the preparation of the eye care solution containing stem cells as described in any one of claims 2-3, characterized in that, The preparation method includes: S1. Place the Chinese herbal extract and pomegranate glycoside into physiological saline and stir to mix evenly to obtain a water-soluble substance; S2. Place the porous adsorbent material into the aqueous solution and stir to mix evenly to obtain a dispersion. S3. Add modified sodium hyaluronate and ε-aminocaproic acid to the dispersion, then add stem cells and stir to mix evenly to obtain the eye care solution.

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