Eyeball refraction system model based on two-aqueous-phase droplet fusion and preparation method

Abstract

The invention discloses an eyeball refraction system model based on two-aqueous-phase droplet fusion and a preparation method, and belongs to the technical field of medical models. The preparation method comprises the following steps: firstly preparing a shell solution, a core solution, a receiving solution and a crystalline lens / cornea solution in sequence, then injecting the core solution into liquid drops of the shell solution for cross-linking in the receiving solution, then preparing a crystalline lens and a cornea in sequence through photocuring cross-linking, wrapping a hyaluronic acid solution between the crystalline lens and a cornea layer as aqueous humor, and preparing. The process is simple, operation is easy, special large-scale equipment is not needed, large-scale production is supported, and the prepared eyeball refraction system model is stable in size, high in sphericity degree and stable in material, has the refractive index, transmittance and transparency similar to those of human cornea and human crystalline lenses, has good water retention and biocompatibility, and is suitable for being used as an eyeball refraction system model. The structure is highly similar to that of a human eyeball, and the method can be applied to the aspects of organoid construction, drug delivery, cell culture, tissue repair and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of medical models, and particularly relates to an eyeball refractive system model based on aqueous two-phase droplet fusion and a preparation method thereof. Background Art

[0002] More than 90% of the information obtained by humans from the outside world comes from vision. Therefore, the human eye, which functions with appropriate light stimulation, is an important sensory organ for humans to obtain the most external information. From an optical perspective, the human eye is a complex optical system, and its main components from the outside to the inside are: the cornea, aqueous humor, lens, and vitreous body. Each interface from the cornea to the retina in the fundus is a part of this composite optical system.

[0003] The research on optical eye models has been ongoing. Existing models include animal models, 3D printing models, organoid models, and partial structure models of the eyeball constructed from various materials. Among them, although animal models can study the eyeball within the body system, there are problems such as interspecies differences; 3D printing models have certain requirements for materials, and the materials required need a certain strength to be printed by a 3D printer, but the strength of the eyeball materials is not high, so some properties of existing models do not meet the requirements of the eyeball model; organoid models are models similar to organs constructed with cells. Since the models are in vitro and the cells lacking external support are difficult to truly form the shape of the eyeball; in recent years, the research and development of hydrogels have greatly increased the possibility of using hydrogels to simulate the eyeball refractive system model. Many scientists have used various materials of hydrogels to simulate various parts of the eyeball structure, including corneas / artificial lenses / aqueous humor, etc. made of various materials. However, there is still a lack of a model that can simulate the various parts of the eyeball refractive system model in a layered and zoned manner and whose physical properties are close to those of a real eyeball. Summary of the Invention

[0004] In order to solve the above technical problems, the purpose of the present invention is to provide an eyeball refractive system model based on aqueous two-phase droplet fusion and a preparation method thereof, so as to solve problems such as the complex preparation process of existing optical eye models and the lack of a model that can simulate the various parts of the eyeball refractive system model in a layered and zoned manner and whose physical properties are closer to those of a real eyeball.

[0005] The technical solution of the present invention for solving the above technical problems is as follows:

[0006] A preparation method of an eyeball refractive system model based on aqueous two-phase droplet fusion includes the following steps:

[0007] (1) Mix two groups of polyethylene glycol, dextran, and water evenly, and let them stand for phase separation to obtain a polyethylene glycol solution and dextran solutions I and II with different concentrations.

[0008] (2) Mix the alginic acid powder and the dextran solution I obtained in step (1) evenly to prepare the outer shell solution of the large sphere core-shell structure;

[0009] (3) Mix the hyaluronic acid hydrogel, alginic acid and the dextran solution I obtained in step (1) evenly to prepare the core solution of the large sphere core-shell structure;

[0010] (4) Mix anhydrous calcium chloride and the polyethylene glycol solution obtained in step (1) evenly to prepare the receiving solution;

[0011] (5) Mix the mixed solution containing the photoinitiator and methacrylated gelatin with the dextran solution II obtained in step (1) evenly to prepare the lens / cornea solution;

[0012] (6) First, inject the core solution of the large sphere core-shell structure obtained in step (3) into the droplets of the outer shell solution of the large sphere core-shell structure obtained in step (2), then drop it into the receiving solution obtained in step (4) for crosslinking to obtain the core-shell microspheres. Then, make an incision on the core-shell microspheres, inject the lens / cornea solution obtained in step (5) onto the incision, and perform photocuring crosslinking to obtain the gel. Finally, inject the hyaluronic acid solution onto the gel, and inject and wrap the hyaluronic acid solution with the lens / cornea solution obtained in step (5), and perform photocuring crosslinking to prepare it.

[0013] The beneficial effects of the present invention are as follows: The present invention uses calcium alginate gel with a porous structure as the outer shell, and uses alginic acid, hyaluronic acid and dextran as the core to prepare double-layer gel core-shell microspheres. The corneal structure of the prepared eye refractive system model has a contact angle similar to that of the human cornea. The corneal structure and the lens have refractive indices, transmittances and transparencies similar to those of the human cornea and the human lens. At the same time, the preparation materials have good water retention and biocompatibility, and have a high degree of similarity to the human eye structure in terms of structure, and can be applied to organoid construction, cell culture, drug delivery and tissue repair, etc.

[0014] Further, in step (1), the mass concentration of the polyethylene glycol solution is 12%-18%, the molecular weight of the polyethylene glycol is 5-10 kDa; the mass concentration of the dextran solution I is 7%-9%, and the mass concentration of the dextran solution II is 12%-18%.

[0015] Preferably, in step (1), the mass concentration of the polyethylene glycol solution is 15%, the molecular weight of the polyethylene glycol is 8 kDa; the mass concentration of the dextran solution I is 8%, the mass concentration of the dextran solution II is 15%, and the molecular weight of the dextran is 400-600 kDa.

[0016] Further, in step (2), the mass concentration of alginic acid in the outer shell solution of the large sphere core-shell structure is 2.5%-3.5%.

[0017] Preferably, the mass concentration of alginic acid in the outer shell solution of the large sphere core-shell structure in step (2) is 3%.

[0018] Further, in step (3), the mass fraction of the hyaluronic acid hydrogel is 1%-2%, the molecular weight of hyaluronic acid is 800,000-1.5 million; the mass ratio of the hyaluronic acid hydrogel to the dextran solution I is 1:1.5-2.5; the mass concentration of alginic acid in the core solution of the large sphere core-shell structure is 1%-3%.

[0019] Preferably, in step (3), the mass fraction of the hyaluronic acid hydrogel is 1.5%, the molecular weight of hyaluronic acid is 1.2 million; the mass ratio of the hyaluronic acid hydrogel to the dextran solution I is 1:2; the mass concentration of alginic acid in the core solution of the large sphere core-shell structure is 2%.

[0020] Further, in step (4), the mass concentration of anhydrous calcium chloride in the receiving solution is 7%-15%.

[0021] Preferably, in step (4), the mass concentration of anhydrous calcium chloride in the receiving solution is 10%.

[0022] Further, in step (5), the mass concentration of methacrylated gelatin in the lens / cornea solution is 8%-12%, the mass concentration of dextran is 6.5%-8.5%, and the concentration of the photoinitiator is 0.1%-0.15%.

[0023] Preferably, in step (5), the mass concentration of methacrylated gelatin in the lens / cornea solution is 10%, the mass concentration of dextran is 7.5%, and the concentration of the photoinitiator is 0.125%.

[0024] The beneficial effects of adopting the above further technical solutions are as follows: By regulating the concentrations of methacrylated gelatin and dextran in the lens / cornea solution, the present invention can change the transparency, transmittance, contact angle, etc. of the cornea and lens in the eye refractive system model to a certain extent, so as to meet different application requirements. When the concentration of dextran is too large, the dripping model is likely to fail, which is not conducive to the construction of the eye refractive system model.

[0025] Further, the mixed solution containing the photoinitiator and methacrylated gelatin in step (5) is prepared by the following method: adding methacrylated gelatin to the photoinitiator solution, heating and stirring at 40-50°C for 30-90 min to obtain it.

[0026] Preferably, the mixed solution containing the photoinitiator and methacrylated gelatin in step (5) is prepared by the following method: adding methacrylated gelatin to the photoinitiator solution, heating and stirring at 45°C for 60 min to obtain it.

[0027] Further, in step (6), the concentration of the hyaluronic acid solution is 1.2% - 1.8%.

[0028] Preferably, in step (6), the concentration of the hyaluronic acid solution is 1.5%.

[0029] The beneficial effect of adopting the above further technical solution is that: in the present invention, hyaluronic acid is used as the aqueous humor part of the eye refractive system model. This part of the gel material does not crosslink during preparation. After the rest of the model crosslinks and forms, the aqueous humor material flows out of the model through pores, enabling the exchange of substances with external culture media and other substances, and very well simulating the role of aqueous humor as the energy supply material for the lens in a real eye.

[0030] An eye refractive system model based on the fusion of aqueous two-phase droplets is prepared by the above preparation method.

[0031] The above eye refractive system model based on the fusion of aqueous two-phase droplets is applied in organoid construction, cell culture, drug delivery carriers, and tissue repair medical devices.

[0032] The present invention has the following beneficial effects:

[0033] (1) The eye refractive system model prepared by the present invention has stable dimensions, high sphericity, stable materials, a simple preparation process, is easy to operate, does not require special large-scale equipment, and supports mass production.

[0034] (2) The corneal structure of the eye refractive system model prepared by the present invention has a contact angle (hydrophilicity) similar to that of the human cornea, and the corneal structure and lens structure have refractive index, transmittance, transparency, etc. similar to those of the human cornea and human lens.

[0035] (3) The materials used in the eye refractive system model prepared by the present invention have good water retention, and there is an obvious layered structure between each part of the structure, which is very suitable for subsequent research on the eye refractive system.

[0036] (4) The eye refractive system model prepared by the present invention has very good biocompatibility. Its outer shell is made of calcium alginate gel with pores, which can carry out the exchange and transmission of nutrients, is very suitable for encapsulating cells in the model, and can culture different cells in different regions, enabling it to simulate the interaction of different cells in the eye.

[0037] (5) The eye refractive system model prepared by the present invention can be adjusted in size within a certain range, further expanding the application scope. It has a high degree of similarity to the human eye structure in terms of structure and can be used in applications such as organoid construction, drug delivery, cell culture, and tissue repair. Description of the Drawings

[0038] Figure 1 The silicone flat plate, the curved-edge knife with a curvature, and the microsphere clamp used for preparing the eye refractive system model in the embodiment, where A is the curved-edge knife, B is the silicone flat plate, and C is the microsphere clamp;

[0039] Figure 2 Schematic diagram of the eye refractive system model prepared in Example 1;

[0040] Figure 3 Actual picture of the eye refractive system model prepared in Example 1;

[0041] Figure 4 Scanning electron microscope characterization results of the eye refractive system model prepared in Example 1, where A is the internal section structure of the microsphere, B is the complete external structure of the microsphere, and C is the boundary between the outer shell and the core of the core-shell microsphere;

[0042] Figure 5 Actual diameter diagram and statistical chart of the core-shell microspheres of the eye refractive system model prepared in Example 1, where A is the actual diagram and B is the statistical chart;

[0043] Figure 6 Actual thickness diagram and statistical chart of the outer shell thickness of the eye refractive system model prepared in Example 1, where A is the actual diagram and B is the statistical chart;

[0044] Figure 7 Statistical chart of the contact angle of the GelMA gel of the corneal structure of the eye refractive system model prepared in Example 1. Detailed implementation mode

[0045] The principles and features of the present invention are described below with reference to the accompanying drawings. The examples given are only used to explain the present invention and are not intended to limit the scope of the present invention. For those conditions not specified in the examples, they are carried out according to conventional conditions or the conditions recommended by the manufacturer. For the reagents or instruments whose manufacturers are not specified, they are all conventional products that can be obtained by purchasing in the market.

[0046] Example 1:

[0047] A preparation method of an eye refractive system model based on the fusion of aqueous two-phase droplets, comprising the following steps:

[0048] (1) Mix polyethylene glycol and dextran, add deionized water thereto, then place it on a magnetic stirrer. After the polyethylene glycol and dextran are mixed evenly, let it stand for 12 h for the aqueous two-phase separation to prepare an aqueous two-phase solution with a mass ratio of polyethylene glycol of 8% and a mass ratio of dextran of 8%. Among them, the molecular weight of polyethylene glycol is 8 kDa, and the molecular weight of dextran is 500 kDa. Separate the two phases to obtain dextran solution I with a mass fraction of 8%.

[0049] (2) Mix polyethylene glycol and dextran, add deionized water thereto, then place it on a magnetic stirrer. After the polyethylene glycol and dextran are mixed evenly, let it stand for 12 h until the two-phase aqueous system separates to form a two-phase aqueous solution with a mass ratio of polyethylene glycol of 15% and a mass ratio of dextran of 15%. Among them, the molecular weight of polyethylene glycol is 8 kDa, and the molecular weight of dextran is 500 kDa. Separate the two phases to obtain a dextran solution II with a mass fraction of 15% and a polyethylene glycol solution with a mass fraction of 15%.

[0050] (3) Add alginic acid powder to a part of the dextran solution I obtained in step (1) to prepare a solution with an alginic acid mass ratio of 3%, which is used as the outer shell solution of the large spherical core-shell structure.

[0051] (4) Add hyaluronic acid powder to deionized water to prepare a hyaluronic acid gel with a mass ratio of 1.5%. Among them, the molecular weight of hyaluronic acid is 1.2 million. Then mix the hyaluronic acid gel with a part of the dextran solution I obtained in step (1) at a mass ratio of 1:2 to obtain a mixed solution with an alginic acid mass fraction of 0.5% and a dextran mass fraction of 5.3%. Finally, add alginic acid to prepare a mixed solution containing 2% (mass fraction) alginic acid, which is used as the core solution of the large spherical core-shell structure.

[0052] (5) Add anhydrous calcium chloride to the polyethylene glycol solution obtained in step (2) to prepare a mixed solution with a calcium chloride mass ratio of 10%, which is used as the receiving liquid.

[0053] (6) Under light-shielded conditions, first weigh the photoinitiator powder (LAP) and add it to deionized water to prepare a photoinitiator solution with a mass ratio of 0.25%. Then weigh the methacrylated gelatin powder and add it to the photoinitiator solution. After heating and stirring at 45 °C for 60 min, it is dissolved to prepare a mixed solution with a methacrylated gelatin mass ratio of 20%. Then mix it with the dextran solution II obtained in step (2) at a mass ratio of 1:1 to obtain a mixed solution with a methacrylated gelatin mass ratio of 10%, a dextran mass ratio of 7.5%, and a photoinitiator mass ratio of 0.125%, which is used as the lens / cornea solution.

[0054] (7) Prepare a silica gel plate with small holes having a diameter slightly larger than the diameter of the core-shell microspheres. The silica gel plate is used to fix the microspheres during operation, and the depth of the small holes is sufficient to prevent the upper part of the microspheres from being covered, as shown in Figure B in Figure 1 as shown in Figure B.

[0055] (8) Inject the large spherical core-shell structure core solution obtained in step (4) into the droplets formed by the large spherical core-shell structure outer shell solution obtained in step (3) through a syringe, and then drop it into the receiving liquid obtained in step (5) for crosslinking, and take out the prepared core-shell microspheres.

[0056] (9) First, use a microsphere clamp (as shown in Figure C of Figure 1 ) to place the core-shell microspheres on the silica gel plate obtained in step (7), fix them through the small holes, expose the upper half, and use a tool knife with a curvature (as shown in Figure A of Figure 1 ) to cut small holes in the core-shell microspheres, with the incision size being 0.875 mm; then inject the lens / cornea solution prepared in step (6) onto the incision, and quickly transfer the silica gel plate to under the ultraviolet lamp for photocuring crosslinking to prepare the gel; then inject a hyaluronic acid solution with a mass fraction of 1.5% onto the gel; subsequently, inject another layer of the lens / cornea solution prepared in step (6) on the surface of the hyaluronic acid solution, and make the lens / cornea solution wrap the hyaluronic acid solution; finally, transfer the silica gel plate to under the ultraviolet lamp again for photocuring crosslinking to prepare the gel, obtaining a multi-layer framework and preparing an eyeball refractive system model. The schematic diagram of the model is as shown in Figure 2 , and the actual diagram of the model is as shown in Figure 3 .

[0057] Example 2:

[0058] A preparation method of an eyeball refractive system model based on the fusion of aqueous two-phase droplets, comprising the following steps:

[0059] (1) Mix polyethylene glycol and dextran, add deionized water thereto, then place it on a magnetic stirrer. After the polyethylene glycol and dextran are mixed evenly, let it stand for 12 h for the aqueous two-phase separation to prepare an aqueous two-phase solution with a mass ratio of polyethylene glycol being 7% and a mass ratio of dextran being 7%. Among them, the molecular weight of polyethylene glycol is 8 kDa, and the molecular weight of dextran is 500 kDa. Separate the two phases to obtain dextran solution I with a mass fraction of 7%.

[0060] (2) Mix polyethylene glycol and dextran, add deionized water thereto, then place it on a magnetic stirrer. After the polyethylene glycol and dextran are mixed evenly, let it stand for 12 h for the aqueous two-phase separation to prepare an aqueous two-phase solution with a mass ratio of polyethylene glycol being 13% and a mass ratio of dextran being 13%. Among them, the molecular weight of polyethylene glycol is 8 kDa, and the molecular weight of dextran is 500 kDa. Separate the two phases to obtain dextran solution II with a mass fraction of 13% and polyethylene glycol solution with a mass fraction of 13%.

[0061] (3) Add alginic acid powder to a part of the dextran solution I obtained in step (1) to prepare a solution with a mass ratio of alginic acid being 2.5% as the outer shell solution of the large sphere core-shell structure.

[0062] (4) Add hyaluronic acid powder to deionized water to prepare a hyaluronic acid gel with a mass ratio of 1.0%. Among them, the molecular weight of hyaluronic acid is 1.2 million. Then mix the hyaluronic acid gel with a part of the dextran solution I obtained in step (1) at a mass ratio of 1:1.5. Finally, add alginic acid to prepare a mixed solution containing 2% (mass fraction) of alginic acid as the core solution of the large spherical core-shell structure.

[0063] (5) Add anhydrous calcium chloride to the polyethylene glycol solution obtained in step (2) to prepare a mixed solution with a mass ratio of calcium chloride of 7% as the receiving solution.

[0064] (6) Under light-shielded conditions, first weigh the photoinitiator powder (LAP) and add it to deionized water to prepare a photoinitiator solution with a mass ratio of 0.2%. Then weigh the methacrylated gelatin powder and add it to the photoinitiator solution. After heating and stirring at 45 °C for 60 min, it is dissolved to prepare a mixed solution with a mass ratio of methacrylated gelatin of 16%. Then mix it with the dextran solution II obtained in step (2) at a mass ratio of 1:1 to obtain a mixed solution with a mass ratio of methacrylated gelatin of 8%, a mass ratio of dextran of 6.5%, and a mass ratio of photoinitiator of 0.10% as the lens / cornea solution.

[0065] (7) Prepare a silica gel plate with small holes having a diameter slightly larger than that of the core-shell microspheres. The silica gel plate is used to fix the microspheres during operation, and the depth of the small holes is sufficient to prevent the upper part of the microspheres from being covered.

[0066] (8) Inject the core solution of the large spherical core-shell structure obtained in step (4) into the droplets formed by the outer shell solution of the large spherical core-shell structure obtained in step (3) through a syringe, and then drop it into the receiving solution obtained in step (5) for crosslinking, and take out the prepared core-shell microspheres.

[0067] (9) First, use a microsphere clamp to place the core-shell microspheres on the silica gel plate obtained in step (7), fix them through the small holes, expose the upper part, and cut a small hole on the core-shell microspheres with a tool knife having a curvature. The size of the cut is 0.75 mm; then inject the lens / cornea solution prepared in step (6) onto the cut, and quickly transfer the silica gel plate to under the ultraviolet lamp for photocuring crosslinking to prepare the gel; then inject a hyaluronic acid solution with a mass fraction of 1.2% onto the gel; subsequently, inject another layer of the lens / cornea solution prepared in step (6) on the surface of the hyaluronic acid solution, and make the lens / cornea solution wrap the hyaluronic acid solution; finally, transfer the silica gel plate to under the ultraviolet lamp again for photocuring crosslinking to prepare the gel, obtain a multi-layer framework, and prepare an eyeball refractive system model.

[0068] Example 3:

[0069] A preparation method of an eye refractive system model based on the fusion of aqueous two-phase droplets, comprising the following steps:

[0070] (1) Mix polyethylene glycol and dextran, add deionized water thereto, then place it on a magnetic stirrer. After the polyethylene glycol and dextran are mixed evenly, let it stand for 12 h for the aqueous two-phase separation to prepare an aqueous two-phase solution with a mass ratio of polyethylene glycol of 9% and a mass ratio of dextran of 9%. Among them, the molecular weight of polyethylene glycol is 8 kDa, and the molecular weight of dextran is 500 kDa. Separate the two phases to obtain dextran solution I with a mass fraction of 9%.

[0071] (2) Mix polyethylene glycol and dextran, add deionized water thereto, then place it on a magnetic stirrer. After the polyethylene glycol and dextran are mixed evenly, let it stand for 12 h for the aqueous two-phase separation to prepare an aqueous two-phase solution with a mass ratio of polyethylene glycol of 17% and a mass ratio of dextran of 17%. Among them, the molecular weight of polyethylene glycol is 8 kDa, and the molecular weight of dextran is 500 kDa. Separate the two phases to obtain dextran solution II with a mass fraction of 17% and polyethylene glycol solution with a mass fraction of 17%.

[0072] (3) Add alginic acid powder to a part of the dextran solution I obtained in step (1) to prepare a solution with a mass ratio of alginic acid of 3% as the outer shell solution of the large sphere core-shell structure.

[0073] (4) Add hyaluronic acid powder to deionized water to prepare a hyaluronic acid gel with a mass ratio of 1.8%. Among them, the molecular weight of hyaluronic acid is 1.2 million. Then mix the hyaluronic acid gel with a part of the dextran solution I obtained in step (1) according to a mass ratio of 1:2.5, and finally add alginic acid to prepare a mixed solution containing 2% (mass fraction) of alginic acid as the core solution of the large sphere core-shell structure.

[0074] (5) Add anhydrous calcium chloride to the polyethylene glycol solution obtained in step (2) to prepare a mixed solution with a mass ratio of calcium chloride of 15% as the receiving liquid.

[0075] (6) Under light-shielded conditions, first weigh the photoinitiator powder (LAP) and add it to deionized water to prepare a photoinitiator solution with a mass ratio of 0.30%. Then weigh the methacrylated gelatin powder and add it to the photoinitiator solution. After heating and stirring at 45 °C for 60 min, it is dissolved to prepare a mixed solution with a mass ratio of methacrylated gelatin of 24%. Then mix it with the dextran solution II obtained in step (2) according to a mass ratio of 1:1 to obtain a mixed solution with a mass ratio of methacrylated gelatin of 12%, a mass ratio of dextran of 8.5% and a mass ratio of photoinitiator of 0.15% as the lens / cornea solution.

[0076] (7) Prepare a silica gel flat plate with small holes having a diameter slightly larger than the diameter of the core-shell microspheres. The silica gel flat plate is used to fix the microspheres during operation, and the depth of the small holes is sufficient to ensure that the upper half of the microspheres is not covered.

[0077] (8) Inject the core solution of the large sphere core-shell structure obtained in step (4) into the droplet formed by the shell solution of the large sphere core-shell structure obtained in step (3) through a syringe, and then drop it into the receiving solution obtained in step (5) for crosslinking, and take out the prepared core-shell microspheres.

[0078] (9) First, use a microsphere clamp to place the core-shell microspheres on the silica gel flat plate obtained in step (7), fix them through the small holes, expose the upper half, and cut a small hole on the core-shell microspheres with a tool knife having a curvature. The size of the cut is 1.0 mm; then inject the lens / cornea solution prepared in step (6) onto the cut, and quickly transfer the silica gel flat plate to under an ultraviolet lamp for photocuring crosslinking to prepare a gel; then inject a hyaluronic acid solution with a mass fraction of 1.8% onto the gel; subsequently, inject another layer of the lens / cornea solution prepared in step (6) on the surface of the hyaluronic acid solution, and make the lens / cornea solution wrap the hyaluronic acid solution; finally, transfer the silica gel flat plate to under the ultraviolet lamp again for photocuring crosslinking to prepare a gel, obtaining a multi-layer framework, and preparing an eye refractive system model.

[0079] Experimental example:

[0080] Take the eye refractive system model prepared in Example 1 for characterization. Characterize the model structure by scanning electron microscopy, and count the diameter of the core-shell structure microspheres and the thickness of the shell. Measure the refractive index, transmittance, and contact angle of the methacrylated gelatin (GelMA) gel in the same group of samples. The experimental results are shown in Table 1 - Table 2 and as Figures 4 - 7 shown.

[0081] Table 1 Statistical table of refractive index of GelMA gel

[0082]

[0083] Table 2 Statistical table of transmittance of GelMA gel

[0084]

[0085] In Table 2, Tt is the total light transmittance, representing all the light amounts passing through the sample; Td is the diffused light transmittance, representing the diffused light in the light passing through the sample; Tp is the parallel light transmittance, representing the parallel light in the light passing through the sample.

[0086] The natural light (400 - 700 nm) transmittance and refractive index of the human cornea are approximately between 1.390 - 1.400, the natural light (400 - 700 nm) transmittance is approximately 81% - 94%, and the contact angle is 55 - 65°. For the GelMA gel used for the cornea in the model prepared in Example 1 of the present invention, the natural light refractive index is approximately 1.331 - 1.367, the natural light transmittance is 93%, and the contact angle at 120 s is 70.3°. All the data are very close to those of the natural cornea. At the same time, according to the statistical results of the core-shell thickness and size, it can be seen that the model ratio prepared by the present invention has significant similarity with the ratio of the real eyeball sclera and vitreous body.

[0087] According to the scanning electron microscope images, it can be known that the eye refractive system model in Example 1 of the present invention has been successfully constructed. In the electron microscope images, since the gel used for the aqueous humor part is not cross-linked, after the rest of the model is cross-linked and formed, the aqueous humor material flows out of the model through the pores, which can achieve the function of exchanging substances with the external culture medium and other substances, and very well simulates the function of the aqueous humor as the energy supply material for the lens in the real eyeball.

[0088] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for preparing an eyeball refractive system model based on two-phase aqueous droplet fusion, characterized in that: The following steps are involved: (1) respectively mixing two groups of polyethylene glycol, dextran and water evenly, and allowing to stand for phase separation to prepare polyethylene glycol solution and dextran solution I and dextran solution II with different concentrations; (2) mixing the alginate powder and the dextran solution I obtained in step (1) uniformly to obtain a shell solution of a large spherical core-shell structure; (3) mixing the hyaluronic acid hydrogel, alginate and the dextran solution I obtained in step (1) to obtain a core solution of a macrospherical core-shell structure; (4) mixing anhydrous calcium chloride and the polyethylene glycol solution obtained in step (1) to obtain a receiving solution; (5) uniformly mixing the mixed solution containing the photoinitiator and the methacrylated gelatin with the dextran solution II obtained in step (1) to prepare a lens / cornea solution; (6) First, the core solution of the macrosphere core-shell structure obtained in step (3) is injected into the droplet of the shell solution of the macrosphere core-shell structure obtained in step (2), and then dropped into the receiving solution obtained in step (4) for cross-linking to obtain core-shell microspheres, and then an incision is made on the core-shell microspheres, and the lens / cornea solution obtained in step (5) is injected into the incision, and photocured and cross-linked to obtain a gel, and finally a hyaluronic acid solution is injected into the gel, and the hyaluronic acid solution is wrapped by the lens / cornea solution obtained in step (5) injection, and photocured and cross-linked to obtain.

2. The method for preparing an eyeball refractive system model based on two-phase aqueous droplet fusion according to claim 1, characterized in that: In the step (1), the mass concentration of the polyethylene glycol solution is 12%-18%, and the molecular weight of the polyethylene glycol is 5-10 kDa; the mass concentration of the dextran solution I is 7%-9%, the mass concentration of the dextran solution II is 12%-18%, and the molecular weight of the dextran is 400-600 kDa.

3. The method for preparing an eyeball refractive system model based on two-phase aqueous droplet fusion according to claim 1, characterized in that: In the step (2), the mass concentration of alginate in the shell solution of the large spherical core-shell structure is 2.5%-3.5%.

4. The method for preparing an eyeball refractive system model based on two-phase aqueous droplet fusion according to claim 1, characterized in that: In the step (3), the mass fraction of the hyaluronic acid hydrogel is 1%-2%, the molecular weight of the hyaluronic acid is 800,000-1.5 million; the mass ratio of the hyaluronic acid hydrogel to the dextran solution I is 1:1.5-2.5; the mass concentration of alginate in the core solution of the macrospherical core-shell structure is 1%-3%.

5. The method for preparing an eyeball refractive system model based on two-phase aqueous droplet fusion according to claim 1, characterized in that: The mass concentration of anhydrous calcium chloride in the receiving solution in step (4) is 7%-15%.

6. The method for preparing an eyeball refractive system model based on two-phase aqueous droplet fusion according to claim 1, characterized in that: In the step (5), the mass concentration of methacryloyl gelatin in the lens / cornea solution is 8%-12%, the mass concentration of dextran is 6.5%-8.5%, and the concentration of the photoinitiator is 0.1%-0.15%.

7. The method for preparing an eyeball refractive system model based on two-phase aqueous droplet fusion according to claim 1, characterized in that: The mixed solution containing the photoinitiator and methacrylic gelatin in step (5) is prepared by the following method: adding methacrylic gelatin to the photoinitiator solution, heating and stirring at 40-50° C. for 30-90 minutes.

8. The method for preparing an eyeball refractive system model based on two-phase aqueous droplet fusion according to claim 1, characterized in that: The concentration of the hyaluronic acid solution in step (6) is 1.2%-1.8%.

9. An eye refractive system model based on two-phase aqueous droplet fusion, characterized in that: The method is prepared by the preparation method according to any one of claims 1 to 7.

10. Application of the eye refractive system model based on two-phase aqueous droplet fusion as described in claim 9 in organoid construction, cell culture, drug delivery carrier and tissue repair medical device.

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