A biomagnetic nanoparticle and its preparation method and application in repairing damaged eye cells

Abstract

The present invention provides a biomagnetic nanoparticle and a preparation method thereof and its application in repairing damaged eye cells. The eye protection solution comprises: 0.3%-0.8% biomagnetic nanoparticles, 70%-90% cell culture medium and 0.6%-1.5% antioxidant, and the rest is distilled water; wherein the cell culture medium comprises low-sugar DMEM, RPMI-1640, MEM and protein peptides, and the weight of low-sugar DMEM is greater than or equal to the weight of RPMI-1640, MEM and protein peptides. The ratio is greater than or equal to 1.5:1; the antioxidants include green tea extract, vitamin C, anthocyanins and licorice extract. The solution provided in this application can achieve the purpose of curing ophthalmic diseases by repairing damaged eye cells, while improving the therapeutic effect of eye diseases and ensuring the safety of eye treatment.

Description

[0001] This application is a divisional application with application number 202311711884.7, and the original application date is December 13, 2023, with the invention name of eye protection solution and preparation method thereof. Technical Field

[0002] The present application relates to the field of medical technology, and in particular to eye protection liquid and a preparation method thereof. Background Art

[0003] Currently, eye diseases have become a global health problem, and ophthalmic diseases pose a serious threat to human health and life, especially a series of diseases caused by damage to eye cells, such as myopia, cataracts, glaucoma, etc. In traditional treatment methods, surgery or existing eye drops can be used for treatment. However, surgical treatment has safety risks, and the use of existing eye drops may have the risk of causing side effects, and the treatment effect may not be ideal and the treatment cost is high.

[0004] In view of this, there is an urgent need to propose a new type of eye protection solution that can cure ophthalmic diseases by repairing damaged eye cells, thereby improving the treatment effect of eye diseases and ensuring the safety of treatment. Summary of the invention

[0005] In order to overcome the problems existing in the related art, the present application provides an eye protection liquid and a preparation method thereof, which can cure ophthalmic diseases by repairing damaged eye cells, thereby improving the treatment effect of eye diseases and ensuring the safety of eye treatment.

[0006] The first aspect of the present application provides an eye protection solution, comprising the following raw materials in percentage by weight:

[0007] 0.3%-0.8% biomagnetic nanoparticles, 70%-90% cell culture medium and 0.6%-1.5% antioxidants, the rest is distilled water;

[0008] The cell culture medium comprises low-glucose DMEM, RPMI-1640, MEM and protein peptides, and the ratio of the weight of low-glucose DMEM to the sum of the weights of RPMI-1640, MEM and protein peptides is greater than or equal to 1.5:1;

[0009] Antioxidants include green tea extract, vitamin C, anthocyanins, and licorice extract.

[0010] In one embodiment, the eye protection solution comprises the following raw materials in weight percentage: 0.6% biomagnetic nanoparticles, 75% cell culture medium and 1% antioxidant, and the rest is distilled water.

[0011] In one embodiment, 1% antioxidants include 0.25% vitamin C, 0.5% green tea extract, 0.1% anthocyanin, and 0.15% licorice extract.

[0012] In one embodiment, the eye protection solution comprises the following raw materials in weight percentage: 0.7% biomagnetic nanoparticles, 90% cell culture medium and 1.3% antioxidant, and the rest is distilled water.

[0013] In one embodiment, the 1.3% antioxidants include 0.35% vitamin C, 0.5% green tea extract, 0.2% anthocyanins, and 0.25% licorice extract.

[0014] The second aspect of the present application provides a method for preparing an eye protection solution, which is used to prepare the eye protection solution as described in any one of the first aspects, comprising:

[0015] preparing biomagnetic nanoparticles, cell culture medium and antioxidants;

[0016] Weighing biomagnetic nanoparticles and cell culture medium according to weight percentage, and adding the biomagnetic nanoparticles to the cell culture medium for uniform dispersion and stirring to obtain a first mixture;

[0017] Weighing an antioxidant according to weight percentage, adding it to the first mixture and mixing thoroughly to obtain a second mixture;

[0018] Under a constant temperature of 40° C., 3% digestive enzyme was added to the second mixture to carry out enzymatic reaction for 45 hours to obtain a third mixture;

[0019] The impurities in the third mixture are filtered to obtain the eye protection solution.

[0020] In one embodiment, in preparing biomagnetic nanoparticles, cell culture medium and antioxidant, preparing biomagnetic nanoparticles comprises:

[0021] Adding ferric chloride into deionized water to obtain a ferric chloride solution;

[0022] Sodium borohydride is added to the ferric chloride solution and stirred to obtain a nano-iron mixed solution;

[0023] Adding sodium dodecyl sulfate into the nano-iron mixed solution and stirring to obtain a first coated nano-iron mixed solution;

[0024] Adding ammonia water to the first coated nano-iron mixed solution and stirring to obtain a second coated nano-iron mixed solution;

[0025] adding protein peptide to the second coated nano-iron mixed solution and stirring to obtain a biomagnetic nanoparticle mixed solution;

[0026] The biomagnetic nanoparticle mixture is filtered through a 50 nm filter membrane to obtain 50 nm filtered particles;

[0027] The 50 nm filtered particles were washed 2-3 times with double distilled water using a high-speed centrifuge to obtain biomagnetic nanoparticles.

[0028] In one embodiment, in preparing biomagnetic nanoparticles, cell culture medium and antioxidant, preparing the cell culture medium comprises:

[0029] Weigh low-glucose DMEM, RPMI-1640, MEM and protein peptides according to the ratio of the weight of low-glucose DMEM to the weight of RPMI-1640, MEM and protein peptides of 1.5:1;

[0030] The weighed low-glucose DMEM, RPMI-1640, MEM and protein peptide were mixed to obtain a cell culture medium.

[0031] In one embodiment, in preparing biomagnetic nanoparticles, cell culture medium and antioxidant, preparing the antioxidant comprises:

[0032] Weigh vitamin C, green tea extract, anthocyanin and licorice extract according to the ratio of vitamin C weight: green tea extract weight: anthocyanin weight: licorice extract weight = 5:10:2:3;

[0033] The weighed vitamin C, green tea extract, anthocyanin and licorice extract are mixed to obtain an antioxidant.

[0034] In one embodiment, in preparing biomagnetic nanoparticles, cell culture medium and antioxidant, preparing the antioxidant comprises:

[0035] Weigh vitamin C, green tea extract, anthocyanin and licorice extract according to the ratio of vitamin C weight: green tea extract weight: anthocyanin weight: licorice extract weight = 7:10:4:5;

[0036] The weighed vitamin C, green tea extract, anthocyanin and licorice extract are mixed to obtain an antioxidant.

[0037] The technical solution provided by this application may have the following beneficial effects:

[0038] The eye protection solution provided by the present application includes the following raw materials in weight percentage: 0.3%-0.8% biomagnetic nanoparticles, 70%-90% cell culture medium and 0.6%-1.5% antioxidants, and the rest is distilled water. Among them, the cell culture medium contains low-sugar DMEM, RPMI-1640, MEM and protein peptides, and the weight of low-sugar DMEM is greater than or equal to 1.5:1. Antioxidants include green tea extract, vitamin C, anthocyanins and licorice extract. Compared with existing eye drops, biomagnetic nanoparticles can promote cell proliferation, cell differentiation and cell regeneration in cell culture. By combining biomagnetic nanoparticles and cell culture medium to form a transparent liquid that can stay on the surface of the eye for a period of time, biomagnetic nanoparticles can penetrate into the eye tissue through the transparent liquid, pass through the eye cell membrane and enter the inside of the eye cell, thereby achieving the effect of using the eye as a culture dish to repair and treat damaged eye cells in situ, while having the effect of reversing myopia. Furthermore, on the basis of biomagnetic nanoparticles and cell culture medium, adding highly active antioxidants with no side effects including green tea extract, vitamin C, anthocyanin and licorice extract can play a synergistic role in the repair of eye cells and reversal of myopia, and improve the repair effect, stability and safety of eye protection solution. Thus, the purpose of curing ophthalmic diseases can be achieved by repairing damaged eye cells, improving the treatment effect of eye diseases while ensuring the safety of eye treatment.

[0039] It should be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] The above and other objects, features and advantages of the present application will become more apparent through a more detailed description of exemplary embodiments of the present application in conjunction with the accompanying drawings, wherein the same reference numerals generally represent the same components in the exemplary embodiments of the present application.

[0041] Figure 1 It is a schematic flow chart of the method for preparing eye protection solution shown in the embodiment of the present application. DETAILED DESCRIPTION

[0042] The preferred embodiments of the present application will be described in more detail below with reference to the accompanying drawings. Although the preferred embodiments of the present application are shown in the accompanying drawings, it should be understood that the present application can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided to make the present application more thorough and complete, and to fully convey the scope of the present application to those skilled in the art.

[0043] The terms used in this application are for the purpose of describing specific embodiments only and are not intended to limit this application. The singular forms of "a", "said" and "the" used in this application and the appended claims are also intended to include plural forms unless the context clearly indicates other meanings. It should also be understood that the term "and / or" used herein refers to and includes any or all possible combinations of one or more associated listed items.

[0044] It should be understood that although the terms "first", "second", "third", etc. may be used in this application to describe various information, this information should not be limited to these terms. These terms are only used to distinguish the same type of information from each other. For example, without departing from the scope of this application, the first information may also be referred to as the second information, and similarly, the second information may also be referred to as the first information. Thus, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. In the description of this application, the meaning of "multiple" is two or more, unless otherwise clearly and specifically defined.

[0045] Currently, eye diseases have become a global health problem, and ophthalmic diseases pose a serious threat to human health and life, especially a series of diseases caused by damage to eye cells, such as myopia, cataracts, glaucoma, etc. In traditional treatment methods, surgery or existing eye drops can be used for treatment. However, surgical treatment has safety risks, and the use of existing eye drops may have the risk of causing side effects, and the treatment effect may not be ideal and the treatment cost is high.

[0046] In view of this, there is an urgent need to propose a new type of eye protection solution that can cure ophthalmic diseases by repairing damaged eye cells, thereby improving the treatment effect of eye diseases and ensuring the safety of treatment.

[0047] In view of the above problems, the embodiment of the present application provides an eye protection liquid, which can achieve the purpose of curing ophthalmic diseases by repairing damaged eye cells, improving the treatment effect of eye diseases while ensuring the safety of eye treatment. The eye protection liquid shown in the embodiment of the present application may include:

[0048] 0.3%-0.8% biomagnetic nanoparticles, 70%-90% cell culture medium and 0.6%-1.5% antioxidant, and the rest is distilled water. The cell culture medium comprises low-glucose DMEM, RPMI-1640, MEM and protein peptides, and the ratio of the weight of low-glucose DMEM to the weight of RPMI-1640, MEM and protein peptides is greater than or equal to 1.5:1.

[0049] In the present application embodiment, biomagnetic nanoparticles (magnetic nanoparticles, MNPs) is a nano material with magnetism, thermal stability, biocompatibility and bioactivity. The particle size of biomagnetic nanoparticles is usually between 10 and 100nm, and its physical properties mainly include magnetic responsiveness, hysteresis loop, magnetic frequency absorption and magneto-optical effect, etc. Among them, magnetic responsiveness is one of the most important characteristics of biomagnetic nanoparticles, and the response degree of biomagnetic nanoparticles under the action of external magnetic field is described. Generally speaking, the magnetic responsiveness of biomagnetic nanoparticles can be divided into three types: superparamagnetism, paramagnetism and ferromagnetism, among which ferromagnetism is the strongest one, with the largest magnetic moment and the highest coercive force.

[0050] Since biomagnetic nanoparticles have magnetic responsiveness and biocompatibility, they can be positioned at damaged eye cells for repair through the action of an external magnetic field without damaging the eye tissue, thereby achieving the purpose of in situ repair of damaged eye cells. Furthermore, by combining biomagnetic nanoparticles with cell culture medium and antioxidants, a tiny magnetic field can be formed on the eye. This tiny magnetic field can attract biomagnetic nanoparticles to the damaged eye cells, forming a tiny magnetic field. This tiny magnetic field can promote cell regeneration and repair damaged eye cells, providing all-round protection and repair for the eyes.

[0051] In addition to being able to play the role of repairing damaged cells, biomagnetic nanoparticles can also reverse myopia. In practice, the causes of decreased vision and myopia are various, such as the sphericity and length of the eyeball are affected by factors such as age and environment, resulting in deformation of the eyeball, causing unclear imaging. It can also be due to the influence of oxidative stress on the cells in the retina, which in turn leads to cell degeneration, thereby causing vision problems of varying degrees. For the first reason mentioned above, the biomagnetic nanoparticles in the embodiments of the present application have the effect of regulating the sphericity and length of the eyeball, thereby helping to reverse myopia and improve vision quality. And for the second reason mentioned above, the biomagnetic nanoparticles in the embodiments of the present application can transport antioxidants to damaged cells, and antioxidants can reduce the influence of oxidative stress on cells, prevent the death and degeneration of retinal cells, and slow down age-related macular degeneration at the same time, thereby playing the role of reversing myopia.

[0052] It is understandable that when the biomagnetic nanoparticles enter the eye and interact with the intraocular cells, they will be recognized as foreign matter and can be cleared out of the body by the immune system.

[0053] In the present application embodiment, cell culture medium is a mixture of nutrients and growth factors that are essential for cell growth under in vitro conditions, providing cells with necessary nutrition and growth conditions. Typically, the main components of cell culture medium include inorganic salts, amino acids, carbohydrates, vitamins, growth factors, etc. Wherein, inorganic salts are important components in cell culture medium, including common NaCl, KCl, CaCl2, MgCl2, etc., and inorganic salts can provide cells with ion balance and stable osmotic pressure environment. In addition, amino acids are the basic units of cell protein synthesis, and carbohydrates are important substances for cell energy metabolism, both of which can provide cells with necessary nutrition. Vitamins are a class of organic compounds, which are cofactors of many enzyme-catalyzed reactions, and play an important role in the maintenance and promotion of life activities. Cell growth factors can promote the growth, differentiation and proliferation of cells, and play a vital role for the survival and stability of cells.

[0054] Cell culture medium not only provides cells with necessary nutrition and growth conditions, but also affects cell growth, proliferation and differentiation, etc. Therefore, in the embodiments of the present application, low-sugar DMEM is used as the basis, and RPMI-1640, MEM and protein peptides are used as auxiliary cell culture medium to meet the growth needs of various types of eye cells. Among them, DMEM (dulbecco's modified eagle medium) is a culture medium containing various amino acids and glucose. It is developed on the basis of MEM (Minimum Essential Medium). Compared with MEM, the dosage of various components is increased, and it is divided into high-sugar DMEM (higher than 4500 mg / L) and low-sugar DMEM (lower than 1000 mg / L). RPMI-1640 is a sterile liquid culture medium containing essential and non-essential amino acids, vitamins, organic and inorganic compounds, suitable for supporting the growth of a wide range of mammalian cell lines.

[0055] Low-sugar DMEM, RPMI-1640 and MEM can provide sufficient nutrients and a stable pH environment. At the same time, too low a glucose concentration is conducive to cell growth and differentiation, which is essential for cell metabolism and growth and development. The addition of protein peptides can provide essential proteins, enzymes and growth factors to promote cell proliferation and differentiation. In addition, very low molecular weight protein peptides contain a variety of natural antioxidants, which are also beneficial to maintain the antioxidant system in the culture medium and protect cells from oxidative damage.

[0056] In some embodiments, appropriate amounts of nutrients and growth factors may be further added to the cell culture medium to optimize the cell culture medium. For example, the addition of various types of growth factors such as fibroblast growth factor (FGF) and transforming growth factor-β (TGF-β) can significantly affect the growth and proliferation of human corneal endothelial cells, thereby improving the efficacy of the cell culture medium.

[0057] In the embodiments of the present application, antioxidant refers to a substance that can effectively remove free radicals and protect cells from oxidative damage. The eye is one of the tissues in the human body that is more susceptible to free radical oxidative damage, and antioxidants can provide effective protection for the eye. In addition, antioxidants can also have a positive effect on oxidative stress in cell culture. Oxidative stress refers to a series of adverse physiological reactions in the human body caused by the interference of external factors with the oxidation reaction. Therefore, in cell culture, adding an appropriate concentration of antioxidants can effectively promote cell growth and increase biological activity.

[0058] Specifically, antioxidants may include, but are not limited to, green tea extract, vitamin C, anthocyanins, and licorice extract. Among them, green tea extract is an active ingredient extracted from green tea leaves, mainly including tea polyphenols, caffeine, aromatic oils, water, minerals, pigments, carbohydrates, proteins, amino acids, vitamins and other ingredients. Licorice extract is a medicinal ingredient extracted from licorice, mainly including glycyrrhizin, glycyrrhizic acid, glycyrrhizin, licorice flavonoids, styraxin, formononetin, quercetin and other ingredients. This highly active and side-effect-free antioxidant can neutralize free radicals generated by oxidative stress, reduce the damage of free radicals to eye cells, and improve the survival rate of eye cells. It can also repair damaged molecules such as DNA and proteins, and enhance the regeneration ability of eye cells.

[0059] In some preferred embodiments, the eye protection solution comprises the following raw materials in weight percentage: 0.6% biomagnetic nanoparticles, 75% cell culture medium and 1% antioxidant, and the rest is distilled water. The 1% antioxidant includes 0.25% vitamin C, 0.5% green tea extract, 0.1% anthocyanin and 0.15% licorice extract.

[0060] In some other preferred embodiments, the eye protection solution comprises the following raw materials in weight percentage: 0.7% biomagnetic nanoparticles, 90% cell culture medium and 1.3% antioxidant, and the rest is distilled water. The 1.3% antioxidant includes 0.35% vitamin C, 0.5% green tea extract, 0.2% anthocyanin and 0.25% licorice extract.

[0061] Corresponding to the above-mentioned embodiment of eye protection liquid, the present application also provides a method for preparing eye protection liquid and corresponding embodiments.

[0062] Figure 1 Schematic diagram of the process of preparing eye protection solution shown in the embodiment of the present application. Figure 1 The eye protection solution preparation method shown in the embodiment of the present application may include:

[0063] In step 101, biomagnetic nanoparticles, cell culture medium and antioxidants are prepared.

[0064] The methods for preparing biomagnetic nanoparticles mainly include physical, chemical and biological methods, among which the most common is the reduction method. The reduction method uses metal salts as raw materials, and gradually reduces them in the presence of a reducing agent to generate metal nanoparticles, and then coats the surface of the metal nanoparticles with suitable biomacromolecules, so that the prepared biomagnetic nanoparticles have high stability and good dispersibility, and can be functionally modified as needed. In the embodiment of the present application, the steps of preparing biomagnetic nanoparticles can be exemplified as follows: first, ferric chloride can be added to deionized water to obtain a ferric chloride solution; then sodium borohydride can be added to the ferric chloride solution and stirred to obtain a nano iron mixture; then sodium dodecyl sulfate can be added to the nano iron mixture and stirred to obtain a first coated nano iron mixture; then ammonia water can be added to the first coated nano iron mixture and stirred to obtain a second coated nano iron mixture; further, protein peptides can be added to the second coated nano iron mixture and stirred to obtain a biomagnetic nanoparticle mixture; then the biomagnetic nanoparticle mixture can be filtered through a 50nm filter membrane to obtain 50nm filtered particles; finally, the 50nm filtered particles are washed 2-3 times with double distilled water by a high-speed centrifuge to obtain biomagnetic nanoparticles. After the biomagnetic nanoparticles are prepared, an electron microscope, an X-ray diffractometer, and a magnetometer can be used to characterize and analyze the prepared magnetic bio nanoparticles to determine their morphology, particle size, stability, saturation magnetization and other related parameters and characteristics.

[0065] In addition, in an embodiment of the present application, the step of preparing a cell culture medium can be exemplified as follows: first, low-glucose DMEM, RPMI-1640, MEM and protein peptides can be weighed separately according to a ratio of the weight of low-glucose DMEM to the sum of the weights of RPMI-1640, MEM and protein peptides of 1.5:1; then the weighed low-glucose DMEM, RPMI-1640, MEM and protein peptides can be mixed to obtain a cell culture medium.

[0066] In addition, in the embodiment of the present application, the step of preparing the antioxidant can be exemplarily specific as follows: first, vitamin C, green tea extract, anthocyanin and licorice extract can be weighed according to the weight of vitamin C: the weight of green tea extract: the weight of anthocyanin: the weight of licorice extract = 5:10:2:3; then the weighed vitamin C, green tea extract, anthocyanin and licorice extract can be mixed to obtain the antioxidant. The step of preparing the antioxidant can also be exemplarily specific as follows: first, vitamin C, green tea extract, anthocyanin and licorice extract can be weighed according to the weight of vitamin C: the weight of green tea extract: the weight of anthocyanin: the weight of licorice extract = 7:10:4:5; then the weighed vitamin C, green tea extract, anthocyanin and licorice extract can be mixed to obtain the antioxidant.

[0067] In step 102, biomagnetic nanoparticles and cell culture medium are weighed by weight percentage, and the biomagnetic nanoparticles are added to the cell culture medium for uniform dispersion and stirring to obtain a first mixture. Since the cell culture medium contains rich growth factors and nutrients, it can promote the self-repair and regeneration of eye cells. At the same time, the cell culture medium can also increase the viscosity and permeability of the eye solution, thereby improving the efficacy of the eye solution. Exemplarily, 0.6% of biomagnetic nanoparticles and 75% of cell culture medium can be weighed and mixed.

[0068] In step 103, an antioxidant is weighed by weight percentage and added to the first mixture for thorough mixing to obtain a second mixture. The antioxidant is added to enhance the repair function of the eye protection solution on oxidatively damaged cells. Preferably, an antioxidant comprising 0.25% vitamin C, 0.5% green tea extract, 0.1% anthocyanin and 0.15% licorice extract can be added, and the antioxidant can synergistically enhance the biological activity of the eye protection solution.

[0069] In step 104, 3% digestive enzyme is added to the second mixture at a constant temperature of 40°C for 45 hours to obtain a third mixture. The antioxidant and cell culture medium are mixed with the biomagnetic nanoparticles, and digestive enzyme is added to allow them to diffuse freely and be evenly distributed in the culture medium for enzymatic reaction. In comparison, this preparation method not only has the characteristics of a more compact and stable structure, but also can effectively reduce the amount of material used, save preparation costs, and is conducive to mass production.

[0070] In step 105, the impurities in the third mixture are filtered to obtain an eye protection solution. In the eye protection solution, the biomagnetic nanoparticles as the carrier of the eye protection solution can directly repair damaged cells in the eyes; the cell culture medium can provide necessary nutrients and environmental factors to promote the growth and regeneration of eye cells; and the antioxidant can reduce oxidative stress and reduce the damage to the eyes caused by the external environment.

[0071] The evaluation of appearance and stability showed that at room temperature, the eye solution was a transparent liquid with no obvious precipitation or stratification, and it maintained good stability after being placed for 14 consecutive days. Its color was transparent and light yellow, with a special smell. It was determined that the pH value of the eye solution was 7.2±0.2, the osmotic pressure was 290±10mOsm / L, and it was sterile.

[0072] Furthermore, infrared spectroscopy was used to identify the chemical structure of the main active ingredients in the eye solution. The results showed that the eye solution contained glucose, vitamin C, green tea extract, anthocyanin, licorice extract and biomagnetic nanoparticles, and the detected ingredients were consistent with the information contained in the formula design.

[0073] Furthermore, the MTT method was used to calculate the changes in the metabolic activity of human retinal epithelial cells (ARPE-19) induced by the eye protection solution. The results showed that the eye protection solution had no obvious toxic effect on human retinal epithelial cells. In addition, the degree of cell liquefaction death was detected by flow cytometry, and the results showed that the eye protection solution had no obvious effect on cell liquefaction death. Furthermore, the effect of the eye protection solution on cell proliferation and migration behavior was investigated by membrane scratch experiments and cell migration experiments. The results showed that the eye protection solution can increase cell proliferation and migration ability. These test results also further prove that the eye protection solution does not contain toxic side effects.

[0074] Furthermore, the following randomized controlled trials were conducted in the examples of the present application:

[0075] 1. Visual restoration control trial

[0076] 1. Mouse vision restoration test data

[0077] The eye protection solution shown in the examples of this application was used in a mouse experiment to repair its damaged retina. The experimental results show that the eye protection solution shown in the examples of this application can differentiate the nerve cells of the mouse into retinal neurons and form new synaptic connections in the cerebral cortex, thereby promoting the recovery of the mouse's vision. The specific data are as follows:

[0078] After the mice used the eye protection solution of the present application, the mice's reaction time to the flash light stimulation was shortened from 9 seconds to 2 seconds on average.

[0079] When the mice were subjected to a T-maze test after using the eye protection solution of the present application, the correct rate increased from 40% in the sham operation group to 90% in the eye protection solution group.

[0080] Electrophysiological recordings of mice after using the eye protection solution of the present application showed that the peak response to retinal signals in the mouse cerebral cortex increased by more than 50%.

[0081] 2. Zebrafish vision restoration test data

[0082] The eye protection solution of this application was used in the zebrafish experiment to regulate its light sensitivity expression, and the light reaction test was used to evaluate the vision repair effect. The specific data are as follows:

[0083] After using the eye protection solution of the present application, the zebrafish's response to light was significantly enhanced.

[0084] In the light reaction test, the time for the zebrafish to blink after being repaired with the eye protection solution of the present application was shortened by more than 50% on average.

[0085] Microscopic imaging results showed that the density of neuronal connections in the zebrafish retina increased after being repaired using the eye protection solution of the present application.

[0086] 2. Cataract repair control trial

[0087] Cataract is a disease in which the transparency of the lens decreases, leading to decreased vision. Here are some data from a controlled trial of cataract repair in mice and zebrafish:

[0088] 1. Mouse cataract repair control test data

[0089] In the mouse experiment, mice with artificially induced cataracts were divided into a treatment group and a control group. The treatment group received the eye protection solution of the application, and the control group only underwent a sham operation. After several weeks of observation, the following conclusions were obtained:

[0090] During the recovery period, more than 80 percent of the treated mice showed significant improvements in lens clarity, while no similar improvements were seen in the control group.

[0091] The treated mice had more new neurons and their retinal thickness increased compared with the control group.

[0092] Mice in the treated group showed faster and more accurate responses in a light reaction test.

[0093] 2. Zebrafish cataract repair control test data

[0094] In the zebrafish experiment, cataracts were induced using gene editing technology, and then the zebrafish were divided into a treatment group and a control group. The treatment group received the eye protection solution of the application, and the control group only received a sham operation. After several weeks of observation, the following conclusions were obtained:

[0095] During the recovery period, the lens transparency of zebrafish in the treatment group improved significantly, while that in the control group became worse.

[0096] The retinal thickness of zebrafish in the treatment group was increased compared with that in the control group.

[0097] In behavioral tests, zebrafish in the treated group showed more visual responses and accuracy.

[0098] 3. Dry Eye Repair Control Trial

[0099] Dry eye is an eye disease with the main symptoms of dryness, discomfort and pain in the eyes. The following are some data from a controlled trial of dry eye repair in mice and zebrafish:

[0100] 1. Mouse dry eye repair control test data

[0101] In the mouse experiment, mice with artificially induced dry eye were divided into a treatment group and a control group. The treatment group received the eye protection solution of the application, and the control group was only treated with saline. After several weeks of observation, the following conclusions were obtained:

[0102] During the recovery period, about 70% of the mice in the treatment group showed significant improvement, while no similar improvement was seen in the control group.

[0103] Mice in the treated group secreted fewer and more stable tears, and their refractive index values ​​improved.

[0104] Mice in the treated group showed better retinal responsiveness.

[0105] 2. Data from a controlled trial of zebrafish dry eye repair

[0106] In the zebrafish experiment, dry eye was induced by excessive stress, and the zebrafish were divided into a treatment group and a control group. The treatment group received the eye protection solution of the application, and the control group only received saline. After several weeks of observation, the following conclusions were obtained:

[0107] During the recovery period, the treated zebrafish developed more tear film and their retinal thickness increased compared to the control group.

[0108] Zebrafish in the treated group showed better visual adaptation.

[0109] 4. Conjunctivitis repair control test

[0110] Conjunctivitis is a common eye disease, and its main symptoms are redness, congestion, increased secretions, etc. The following are some data from the control experiment of conjunctivitis repair in mice and zebrafish:

[0111] 1. Mouse conjunctivitis repair control test data

[0112] In the mouse experiment, mice with conjunctivitis were divided into a treatment group and a control group. The treatment group received the eye protection solution of the present application, and the control group only received saline. After several weeks of observation, the following conclusions were obtained:

[0113] During the recovery period, about 80% of the animals in the treatment group showed significant improvement, while no similar improvement was seen in the control group.

[0114] Mice in the treated group showed better corneal transparency and their ocular secretions gradually decreased.

[0115] Mice in the treated group showed improved retinal responsiveness.

[0116] 2. Zebrafish conjunctivitis repair control test data

[0117] In the zebrafish experiment, conjunctivitis was induced by injecting bacteria, and the zebrafish were divided into a treatment group and a control group. The treatment group received the eye protection solution of the application, and the control group was only operated with saline. After several weeks of observation, the following conclusions were obtained:

[0118] During the recovery period, more than 90% of the zebrafish in the treatment group showed significant improvement, while there was generally no significant improvement in the control group.

[0119] The conjunctival condition of the zebrafish in the treatment group was significantly improved, and the surface of the fish eyes healed rapidly.

[0120] Zebrafish in the treated group showed improved retinal responsiveness.

[0121] 5. Presbyopia repair control test

[0122] Presbyopia is a common eye disease, the main symptoms of which are gradual decline in vision and blurry vision at close range. The following are some data from a controlled experiment on presbyopia repair in mice and zebrafish:

[0123] 1. Data of control experiment on presbyopia repair in mice

[0124] In the mouse experiment, the older mice with presbyopia were divided into a treatment group and a control group. The treatment group received the eye protection solution of the application, and the control group only received saline. After several weeks of observation, the following conclusions were obtained:

[0125] During the recovery period, about 50% of the animals in the treatment group showed significant improvement, while no similar improvement was seen in the control group.

[0126] Mice in the treated group showed improved corneal transparency and restored retinal responsiveness.

[0127] 2. Data from a controlled trial of zebrafish presbyopia repair

[0128] In the zebrafish experiment, presbyopia was induced by injecting chromatin inhibitors into retinal cells. The zebrafish were divided into a treatment group and a control group. The treatment group received the eye protection solution of the application, and the control group was only operated with saline. After several weeks of observation, the following conclusions were obtained:

[0129] During the recovery period, more than 60% of the zebrafish in the treatment group showed significant improvement, while there was generally no significant improvement in the control group.

[0130] Zebrafish in the treatment group showed better retinal responsiveness and better retinal function recovery.

[0131] 6. New Zealand rabbit eye irritation test

[0132] Inspection basis and methods: GB / T 16886.10-2017 "Biological evaluation of medical devices Part 10: Irritation and skin sensitization test"

[0133] Test conclusion: No abnormalities were found in the eye irritation test of the sample within 72 hours. According to the standard GB / T16886.10-2017 "Biological Evaluation of Medical Devices Part 10: Irritation and Skin Sensitization Tests", it is a non-irritant.

[0134] The eye protection solution provided by the present application includes the following raw materials in weight percentage: 0.3%-0.8% biomagnetic nanoparticles, 70%-90% cell culture medium and 0.6%-1.5% antioxidants, and the rest is distilled water. Among them, the cell culture medium contains low-sugar DMEM, RPMI-1640, MEM and protein peptides, and the weight of low-sugar DMEM is greater than or equal to 1.5:1. Antioxidants include green tea extract, vitamin C, anthocyanins and licorice extract. Compared with existing eye drops, biomagnetic nanoparticles can promote cell proliferation, cell differentiation and cell regeneration in cell culture. By combining biomagnetic nanoparticles and cell culture medium to form a transparent liquid that can stay on the surface of the eye for a period of time, biomagnetic nanoparticles can penetrate into the eye tissue through the transparent liquid, pass through the eye cell membrane and enter the inside of the eye cell, thereby achieving the effect of using the eye as a culture dish to repair and treat damaged eye cells in situ, while having the effect of reversing myopia. Furthermore, on the basis of biomagnetic nanoparticles and cell culture medium, adding highly active antioxidants with no side effects including green tea extract, vitamin C, anthocyanin and licorice extract can play a synergistic role in the repair of eye cells and reversal of myopia, and improve the repair effect, stability and safety of eye protection solution. Thus, the purpose of curing ophthalmic diseases can be achieved by repairing damaged eye cells, improving the treatment effect of eye diseases while ensuring the safety of eye treatment.

[0135] The scheme of the present application has been described in detail above with reference to the accompanying drawings. In the above embodiments, the description of each embodiment has its own emphasis. For the part that is not described in detail in a certain embodiment, refer to the relevant description of other embodiments. Those skilled in the art should also know that the actions and modules involved in the description are not necessarily required for the present application. In addition, it can be understood that the steps in the method of the embodiment of the present application can be adjusted in order, merged and deleted according to actual needs, and the modules in the device of the embodiment of the present application can be merged, divided and deleted according to actual needs.

[0136] The embodiments of the present application have been described above, and the above description is exemplary, not exhaustive, and is not limited to the disclosed embodiments. Many modifications and changes will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The selection of terms used herein is intended to best explain the principles of the embodiments, practical applications, or improvements to the technology in the market, or to enable other persons of ordinary skill in the art to understand the embodiments disclosed herein.

Claims

1. A biomagnetic nanoparticle, characterized in that: Prepared by the following steps: Adding ferric chloride into deionized water to obtain a ferric chloride solution; Adding sodium borohydride to the ferric chloride solution and stirring to obtain a nano-iron mixed solution; Adding sodium dodecyl sulfate to the nano-iron mixed solution and stirring to obtain a first coated nano-iron mixed solution; Adding ammonia water to the first coated nano-iron mixed solution and stirring to obtain a second coated nano-iron mixed solution; Adding protein peptide to the second coated nano-iron mixed solution and stirring to obtain a biomagnetic nanoparticle mixed solution; Filtering the biomagnetic nanoparticle mixture through a 50 nm filter membrane to obtain 50 nm filtered particles; The 50 nm filtered particles were washed 2-3 times with double distilled water using a high-speed centrifuge to obtain the biomagnetic nanoparticles.

2. Use of the biomagnetic nanoparticles as claimed in claim 1 in repairing damaged eye cells.

3. Use of the biomagnetic nanoparticles as claimed in claim 1 in preparing eye protection solution.

4. The use according to claim 3, characterized in that: The eye protection solution comprises the following raw materials in percentage by weight: 0.3%-0.8% biomagnetic nanoparticles, 70%-90% cell culture medium and 0.6%-1.5% antioxidants, the rest is distilled water; The cell culture medium comprises low-glucose DMEM, RPMI-1640, MEM and protein peptides, and the ratio of the weight of the low-glucose DMEM to the sum of the weights of the RPMI-1640, the MEM and the protein peptides is greater than or equal to 1.5:1; The antioxidants include green tea extract, vitamin C, anthocyanins and licorice extract.

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