Probiotics and composition for promoting ocular surface injury repair and preparation method

Abstract

The invention discloses probiotics and a composition for promoting ocular surface injury repair and a preparation method, and belongs to the field of microorganisms, and the probiotics comprise at least one of bifidobacterium animalis subsp. Lactis and lactobacillus reuteri. In the application process, the probiotics can be prepared into a liquid or fluid composition, the composition can be coated on the ocular surface, the composition containing the probiotics can repair ocular surface injury, the repairing effect is good, symptoms such as pain and inflammation are avoided, and the life quality of a patient is improved.

Description

Technical Field

[0001] The invention belongs to the field of microorganisms, and particularly relates to a probiotic and a composition for promoting the repair of ocular surface damage and a preparation method thereof. Background Art

[0002] The cornea is one of the important refractive media, and its transparency depends on the integrity of the epithelial, endothelial and stromal structures and the normality of their functions. The corneal epithelium is one of the most important biological barriers of the eye. The large number of tight junctions between epithelial cells can form a protective barrier, which can resist the invasion of chemicals and microorganisms into the cornea to a certain extent. Therefore, the integrity of the corneal epithelium is particularly important for eye health. Mild defects of the corneal epithelium can often heal themselves in a short period of time. Severe damage will cause symptoms such as pain and inflammation. If not treated in time, it will also cause serious complications such as infection, ulcers and even perforation. Patients often need amniotic membrane transplantation or even corneal transplantation. A large number of clinical studies have shown that neurotrophic keratopathy, corneal surface trauma, abnormal epithelial cell adhesion function, severe dry eye, and lack of limbal stem cells can cause corneal epithelial damage and repair disorders, which seriously threaten eye health and affect the quality of life of patients. The treatment of corneal damage is mainly based on local medications such as eye gels and eye drops, but the existing drugs are not effective in treating ocular surface damage. Summary of the invention

[0003] The purpose of the embodiments of the present invention is to provide a probiotic and a composition and a preparation method for promoting the repair of ocular surface damage, so as to solve the problem of poor treatment effect of ocular surface damage.

[0004] In a first aspect, an embodiment of the present invention provides a probiotic for promoting the repair of ocular surface damage, comprising:

[0005] Probiotics, wherein the probiotics include at least one of Bifidobacterium animalis subspecies lactis and Lactobacillus reuteri.

[0006] Optionally, the Bifidobacterium animalis subsp. lactis includes Bifidobacterium animalis subsp. lactis CP-9 strain, which is deposited in China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC NO: M2014588; and / or

[0007] The Lactobacillus reuteri includes the Lactobacillus reuteri GL-104 strain, which is preserved in the China Center for Type Culture Collection (Wuhan) with a preservation number of CCTCC NO: M209138.

[0008] In a second aspect, an embodiment of the present invention provides a probiotic composition for promoting the repair of ocular surface damage, the composition comprising:

[0009] The probiotics described in the above embodiments.

[0010] Optionally, the composition further comprises:

[0011] Carboxymethyl cellulose, sodium N-hydroxysulfosuccinimide.

[0012] Optionally, the composition further comprises:

[0013] gelatin.

[0014] Optionally, the mass ratio of the probiotics to the gelatin is (0.75-3):1.

[0015] Optionally, the mass concentration of gelatin in the composition is 5-15%; and / or

[0016] The content of the probiotics is 1.5-45 billion CFU / ml.

[0017] Optionally, the composition is fluid or semi-fluid; and / or

[0018] The probiotics are dead bacteria.

[0019] In a third aspect, an embodiment of the present invention provides a method for preparing a probiotic composition, comprising:

[0020] mixing the probiotics with the gelatin solution to obtain a mixed solution;

[0021] Adding carboxymethyl cellulose and sodium salt of N-hydroxysulfosuccinimide to the mixed solution, and mixing to prepare a composition;

[0022] Wherein, the probiotics include at least one of Bifidobacterium animalis subsp. lactis and Lactobacillus reuteri.

[0023] Optionally, the Bifidobacterium animalis subsp. lactis includes Bifidobacterium animalis subsp. lactis CP-9 strain, which is deposited in China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC NO: M2014588; and / or

[0024] The Lactobacillus reuteri includes the Lactobacillus reuteri GL-104 strain, which is deposited in China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC NO: M209138; and / or

[0025] Optionally, the mass ratio of the probiotics to the gelatin is (0.75-3):1; and / or

[0026] The mass concentration of gelatin in the composition is 5-15%.

[0027] The probiotics of the embodiment of the present invention include at least one of Bifidobacterium animalis subsp. animalis, Bifidobacterium animalis subsp. lactis, and Lactobacillus reuteri. During application, the probiotics can be configured into a liquid or fluid composition, and the composition containing the probiotics can be applied to the ocular surface. The probiotic composition can repair ocular surface damage with good repair effect, avoid causing symptoms such as pain and inflammation, and improve the quality of life of patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The results of the corneal epithelial cell scratch test with different concentrations of live bacteria of CP-9 strain at different times, the control group used phosphate buffer solution (PBS);

[0029] Figure 2 The results of the corneal epithelial cell scratch test with different concentrations of dead CP-9 strains at different times, the control group used a phosphate buffer solution;

[0030] Figure 3 The corneal repair effect after eye drops of different probiotic solutions, the control group used phosphate buffer solution;

[0031] Figure 4 To establish the corneal injury model, the mouse eyeballs were grossly photographed 24 hours after oral administration of CP-9 strain, and the control group used phosphate buffer solution;

[0032] Figure 5 After 2 weeks of intragastric administration of CP-9 strain, the corneal injury model was established, and the mouse eyeballs were photographed 24 hours later. The control group used phosphate buffer solution;

[0033] Figure 6 The scanning electron micrographs of the probiotic hydrogel prepared in Experimental Example 8 at different concentrations, the control is a phosphate buffer solution, and the control group uses a phosphate buffer solution;

[0034] Figure 7 To establish the corneal injury model, the mouse eyeballs were grossly photographed 24 hours after the CP-9 live bacteria hydrogel in Experimental Example 7 and the dead bacteria hydrogel in Experimental Example 8 were dripped into the eyes. The control group used a phosphate buffer solution.

[0035] Figure 8These are photos of the mouse cornea OCT (Optical Coherence Tomography) after the dead bacteria hydrogel and PBS were respectively dripped into the eyes in Experimental Example 9. DETAILED DESCRIPTION

[0036] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] The terms "first", "second", etc. in the specification and claims of the present invention are used to distinguish similar objects, and are not used to describe a specific order or sequence. It should be understood that the terms used in this way are interchangeable under appropriate circumstances, so that the embodiments of the present invention can be implemented in an order other than those illustrated or described herein. In addition, the term "and / or" in the specification and claims means at least one of the connected objects, and the character " / " generally indicates that the objects associated with each other are in an "or" relationship.

[0038] The probiotics for promoting the repair of ocular surface damage according to the embodiment of the present invention include:

[0039] Probiotics, wherein the probiotics include at least one of Bifidobacterium animalis subspecies lactis and Lactobacillus reuteri.

[0040] The probiotics for promoting the repair of ocular surface damage in the embodiment of the present invention include at least one of Bifidobacterium animalis subsp. lactis and Lactobacillus reuteri. During application, the probiotics can be configured into a liquid or fluid composition, and the composition containing the probiotics can be applied to the ocular surface. The probiotic composition can repair ocular surface damage with good repair effect, avoid causing symptoms such as pain and inflammation, and improve the quality of life of patients.

[0041] In some embodiments, the Bifidobacterium animalis subsp. lactis may include Bifidobacterium animalis subsp. lactis CP-9 strain, which is deposited in China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC NO: M2014588 and a deposit date of 2014.11.30.

[0042] Optionally, the Lactobacillus reuteri may include the Lactobacillus reuteri GL-104 strain, which is deposited in China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC NO: M209138 and a deposit date of 2009.08.07.

[0043] Probiotics may include Bifidobacterium animalis subsp. lactis CP-9 strain, Lactobacillus reuteri GL-104 strain.

[0044] An embodiment of the present invention provides a probiotic composition for promoting the repair of ocular surface damage, the composition comprising:

[0045] The probiotics described in the above embodiments. During the application process, the composition can be applied to the ocular surface, and the probiotic composition can repair ocular surface damage with good repair effect, avoiding symptoms such as pain and inflammation. In some embodiments, the composition may also include:

[0046] Carboxymethyl cellulose, sodium N-hydroxysulfosuccinimide.

[0047] Optionally, the composition may further include:

[0048] Gelatin. By adding gelatin, the composition can be made into a colloidal state, which is convenient for coating, conducive to contact with the ocular surface, and can increase the contact time between the composition and the ocular surface.

[0049] In some embodiments, the mass ratio of the probiotics to the gelatin may be (0.75-3): 1. For example, the mass ratio of the probiotics to the gelatin may be 0.75: 1, 1.5: 1, 2: 1, 2.5: 1 or 3: 1, and the specific mass ratio may be selected according to actual conditions.

[0050] Optionally, the mass concentration of gelatin in the composition may be 5-15%. For example, the mass concentration of gelatin in the composition may be 5%, 10%, 12%, or 15%, which may be selected according to actual conditions.

[0051] Optionally, the content of the probiotics can be 15-45 billion CFU / ml. For example, the content of the probiotics can be 15CFU / ml, 100CFU / ml, 200CFU / ml, 300CFU / ml, 400CFU / ml or 45 billion CFU / ml. 1mg of probiotics can contain 200-400 million CFU, for example, 1mg of probiotics can contain 300 million CFU. In actual application, the CFU content of probiotics in 1ml can be formulated according to the CFU content of probiotics in 1mg, and the specific probiotic content unit can be converted. The content of probiotics can be 5-150mg / ml, 1mg of probiotics can contain 200-400 million CFU, for example, 1mg of probiotics can contain 300 million CFU, for example, the content of the probiotics can be 5mg / ml, 50mg / ml, 100mg / ml or 150mg / ml, and the specific content can be adjusted as needed.

[0052] Optionally, the composition can be fluid or semi-fluid, and the composition can be hydrogel-like, which is easy to apply and is conducive to contact with the ocular surface. The composition can be used as eye drops, eye washes, etc., which have a good promoting and repairing effect on ocular surface damage. The composition can be used as eye drops, and the dosage of probiotics in the composition can be 15-45 billion CFU / ml. For example, the content of probiotics can be 15CFU / ml, 100CFU / ml, 200CFU / ml, 300CFU / ml, 400CFU / ml or 45 billion CFU / ml. 1mg of probiotics can contain 200-400 million CFU, and the CFU content of probiotics in 1ml can be formulated according to the CFU content of probiotics in 1mg. The specific content of probiotics in the composition can be selected according to actual conditions, and can also be diluted as needed.

[0053] Optionally, the probiotics are dead bacteria, which are more effective than live bacteria and can better repair ocular surface damage.

[0054] The method for preparing a probiotic composition for promoting the repair of ocular surface damage according to an embodiment of the present invention comprises:

[0055] mixing the probiotics with the gelatin solution to obtain a mixed solution;

[0056] Adding carboxymethyl cellulose and sodium salt of N-hydroxysulfosuccinimide to the mixed solution, and mixing to prepare a composition;

[0057] Wherein, the probiotics may include at least one of Bifidobacterium animalis subsp. lactis and Lactobacillus reuteri. For example, the probiotics may be Bifidobacterium animalis subsp. lactis or Lactobacillus reuteri, and the probiotics may be Bifidobacterium animalis subsp. lactis and Lactobacillus reuteri.

[0058] The probiotic composition for promoting the repair of ocular surface damage prepared by the preparation method in the embodiment of the present invention can be configured into a liquid or fluid during application, and the composition can be applied to the ocular surface. The probiotic composition can repair ocular surface damage with good repair effect, avoid causing symptoms such as pain and inflammation, and improve the quality of life of patients.

[0059] In some embodiments, the Bifidobacterium animalis subsp. lactis may include Bifidobacterium animalis subsp. lactis CP-9 strain, which is deposited in China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC NO: M2014588.

[0060] Optionally, the Lactobacillus reuteri may include the Lactobacillus reuteri GL-104 strain, which is deposited in China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC NO: M209138.

[0061] The probiotics may include Bifidobacterium animalis subsp. lactis CP-9 strain, Lactobacillus reuteri GL-104 strain, and the probiotics may include Lactobacillus reuteri GL-104 strain.

[0062] In some embodiments, the mass ratio of the probiotics to the gelatin may be (0.75-3): 1. For example, the mass ratio of the probiotics to the gelatin may be 0.75: 1, 1.5: 1, 2: 1, 2.5: 1 or 3: 1, and the specific mass ratio may be selected according to actual conditions.

[0063] Optionally, the mass concentration of gelatin in the composition may be 5-15%. For example, the mass concentration of gelatin in the composition may be 5%, 10%, 12%, or 15%, which may be selected according to actual conditions.

[0064] Optionally, the content of the probiotics is 1.5-45 billion CFU / ml. For example, the content of the probiotics can be 15 CFU / ml, 100 CFU / ml, 200 CFU / ml, 300 CFU / ml, 400 CFU / ml or 45 billion CFU / ml. 1 mg of probiotics can contain 200-400 million CFU, for example, 1 mg of probiotics can contain 300 million CFU. In actual application, the CFU content of probiotics in 1 ml can be formulated according to the CFU content of probiotics in 1 mg, and the specific content can be adjusted as needed.

[0065] Optionally, the composition can be in a fluid or semi-fluid state, and the composition can be in a hydrogel state, which is easy to apply and is conducive to contact with the ocular surface. The composition can be used as eye drops, eye washes, etc., which have a good promoting and repairing effect on ocular surface damage. The composition can be used as eye drops, and the dosage of probiotics in the composition can be 15-45 billion CFU / ml. For example, the content of probiotics can be 15CFU / ml, 150CFU / ml, 260CFU / ml, 350CFU / ml or 45 billion CFU / ml. 1mg of probiotics can contain 200-400 million CFUs. For example, 1mg of probiotics can contain 300 million CFUs. In actual application, the CFU content of probiotics in 1ml can be formulated according to the CFU content of probiotics in 1mg. The specific content of probiotics in the composition can be selected according to actual conditions, and can also be diluted as needed.

[0066] The specific preparation process of the composition can be as follows:

[0067] 1. Add 75-300 mg of probiotics to the centrifuge tube;

[0068] 2. Add 0.8-1.2 mL of water to the centrifuge tube in step 1 and shake until evenly mixed;

[0069] 3. Add 0.8-1.2 mL of gelatin solution to the centrifuge tube in step 1 and shake until mixed;

[0070] 4. Add carboxymethyl cellulose (CMC) and sodium N-hydroxysulfosuccinimide (NHS) solution to the centrifuge tube in step 3, with the mass ratio of CMC / NHS being (4.5-72.0):(1.1-18.0), and shake until well mixed;

[0071] The mixture in step 4 was allowed to react at room temperature for 5-30 minutes.

[0072] Wherein, the probiotics in step 2 may include at least one of Bifidobacterium animalis subsp. lactis CP-9 strain and Lactobacillus reuteri GL-104 strain. The probiotics may include live bacteria and dead bacteria after high-pressure sterilization, and the dosage form of the probiotics may be fluid or semi-fluid.

[0073] The technical solution of the present invention is further described below through some specific embodiments.

[0074] The strain Bifidobacterium animalis subsp. animalis BB115 is deposited in China General Microbiological Culture Collection Center with the deposit number CGMCC NO: 21840.

[0075] Experimental Example 1

[0076] 1. Add 150 mg of CP-9 strain to the centrifuge tube; 1 mg of probiotics contains 300 million CFU;

[0077] 2. Add 1.0 mL of water to the centrifuge tube in step 1 and shake until mixed to obtain a suspension solution;

[0078] 3. Apply the suspension solution prepared in step 2 to evaluate the repair of ocular surface damage.

[0079] Experimental Example 2

[0080] 1. Add 150 mg of CP-9 strain to the centrifuge tube; 1 mg of probiotics contains 300 million CFU;

[0081] 2. Sterilize the CP-9 strain in step 1 by high pressure;

[0082] 3. Add 1.0 mL of water to the centrifuge tube in step 2 and shake until mixed to obtain a suspension solution;

[0083] 4. Apply the suspension solution prepared in step 3 to evaluate the repair of ocular surface damage.

[0084] Experimental Example 3

[0085] 1. Add 150 mg of BB-115 strain to the centrifuge tube; 1 mg of probiotics contains 300 million CFU;

[0086] 2. Add 1.0 mL of water to the centrifuge tube in step 1 and shake until mixed to obtain a suspension solution;

[0087] 3. Apply the suspension solution prepared in step 2 to evaluate the repair of ocular surface damage.

[0088] Experimental Example 4

[0089] 1. Add 150 mg of GL-104 strain to the centrifuge tube; 1 mg of probiotics contains 300 million CFU;

[0090] 2. Add 1.0 mL of water to the centrifuge tube in step 1 and shake until mixed to obtain a suspension solution;

[0091] 3. Apply the suspension solution prepared in step 2 to evaluate the repair of ocular surface damage.

[0092] Experimental Example 5

[0093] 1. Add 150 mg of CP-9 strain and 150 mg of GL-104 strain to the centrifuge tube; 1 mg of probiotics contains 300 million CFU;

[0094] 2. Add 1.0 mL of water to the centrifuge tube in step 1 and shake until mixed to obtain a suspension solution;

[0095] 3. Apply the suspension solution prepared in step 2 to evaluate the repair of ocular surface damage.

[0096] Experimental Example 6

[0097] 1. Add 150 mg of GL-104 strain and 150 mg of BB-115 strain to the centrifuge tube; 1 mg of probiotics contains 300 million CFU;

[0098] 2. Add 1.0 mL of water to the centrifuge tube in step 1 and shake until mixed to obtain a suspension solution;

[0099] 3. Apply the suspension solution prepared in step 2 to evaluate the repair of ocular surface damage.

[0100] Experimental Example 7

[0101] 1. Add 150 mg of CP-9 live bacteria strain to the centrifuge tube; 1 mg of probiotics contains 300 million CFU;

[0102] 2. Add 1.0 mL of 10% gelatin solution to the centrifuge tube in step 1 and shake until mixed;

[0103] 3. Add a solution containing 18.0 mg CMC and 4.5 mg NHS to the centrifuge tube in step 2 and shake until mixed;

[0104] 4. The mixed solution in step 3 was allowed to stand and react at room temperature for 10 minutes to obtain a hydrogel;

[0105] 5. The obtained hydrogel is applied to the evaluation of ocular surface damage repair.

[0106] Experimental Example 8

[0107] 1. Add 150 mg of CP-9 dead bacteria strain to the centrifuge tube; 1 mg of probiotics contains 300 million CFU;

[0108] 2. Sterilize the CP-9 strain in step 1 by high pressure;

[0109] 3. Add 1.0 mL of 10% gelatin solution to the centrifuge tube in step 2 and shake until mixed;

[0110] 4. Add a solution containing 18.0 mg CMC and 4.5 mg NHS to the centrifuge tube in step 3 and shake until mixed;

[0111] 5. The mixed solution in step 4 was allowed to stand at room temperature for 10 minutes to obtain a hydrogel;

[0112] 6. The obtained hydrogel is applied to the evaluation of ocular surface damage repair.

[0113] Experimental Example 9

[0114] 1. Add 150 mg of CP-9 dead bacteria strain to the centrifuge tube; 1 mg of probiotics contains 300 million CFU;

[0115] 2. Sterilize the CP-9 strain in step 1 by high pressure;

[0116] 3. Add 1.0 mL of 10% gelatin solution to the centrifuge tube in step 2 and shake until mixed;

[0117] 4. Add a solution containing 18.0 mg CMC and 4.5 mg NHS to the centrifuge tube in step 3 and shake until mixed;

[0118] 5. The mixed solution in step 4 was allowed to stand at room temperature for 10 minutes to obtain a hydrogel;

[0119] 6. Add 1.0 mL of PBS solution to the centrifuge tube in step 2 and shake until mixed;

[0120] 7. Take 5 μL of the obtained hydrogel and drop it onto the surface of the anesthetized mouse cornea (the solution in step 6 serves as the control group).

[0121] 8. OCT photographs of the corneas of mice in the hydrogel group and PBS group were recorded at different times (0, 15min, 30min, 60min, and 120min).

[0122] Experiments were carried out using the materials prepared above.

[0123] The suspension solution in Experimental Example 1 was prepared into solutions of different concentrations and evaluated by cell scratch test. The specific results are as follows: Figure 1 As shown in , from the experimental results, it can be seen that when the concentration of the live bacteria solution is 0.2 mg / mL, the corneal epithelial cell damage repair effect is better.

[0124] The suspension solution in Experimental Example 2 was prepared into solutions of different concentrations and evaluated by cell scratch test. The specific results are as follows: Figure 2 As shown in , from the experimental results, it can be seen that when the concentration of the dead bacteria solution is 0.2 mg / mL, the corneal epithelial cell damage repair effect is better.

[0125] The solutions prepared in Experimental Example 1 and Experimental Examples 3-6 were evaluated for ocular surface damage repair. Figure 3After establishing the corneal injury model, the mouse eyeballs were photographed 24 hours after eye drops of BB-115, CP-9, and GL-104 strain solutions. The experimental results show that CP-9 and GL-104 strains have better effects on promoting corneal repair, while BB-115 strain is not as effective as CP-9 and GL-104 strains in promoting corneal repair. The combination of BB-115 and GL-104 strains and the combination of GL-104 and CP-9 strains; the combination of BB-115 and GL-104 strains and the combination of GL-104 and CP-9 strains all have the effect of promoting corneal repair, but the combination of BB-115 and GL-104 strains is not as effective as the combination of GL-104 and CP-9 strains.

[0126] Figure 4 To establish a corneal injury model, the mouse eyeballs were photographed 24 hours after oral gavage with the CP-9 strain solution. The CP-9 strain had little effect on promoting corneal injury repair by oral gavage; the CP-9 strain content in the CP-9 strain solution was 0.75 mg, and 1 mg of probiotics contained 300 million CFU.

[0127] Figure 5 After 2 weeks of oral administration of CP-9 solution, a corneal injury model was established, and the mouse eyeballs were taken for gross examination 24 hours later. It was found that the effect of promoting corneal injury repair by oral administration of CP-9 strain for 2 weeks and then establishing the injury model was small. The CP-9 strain content in the CP-9 strain solution was 0.75 mg, and 1 mg of probiotics contained 300 million CFU.

[0128] The hydrogel prepared in Experimental Example 8 was prepared into different concentrations. Figure 6 The scanning electron micrographs of the probiotic hydrogels are shown in FIG. 1 . The scanning electron micrographs of the probiotic hydrogels with different CP-9 strain contents (A) 75 mg / mL, (B) 150 mg / mL, and (C) 300 mg / mL) can be shown as follows: Figure 6 ,from Figure 6 It can be seen that the hydrogel is evenly dispersed.

[0129] The CP-9 live bacteria hydrogel prepared in Experimental Example 7 and the dead bacteria hydrogel prepared in Experimental Example 8 were respectively dripped into the eyes of mice. Figure 7 To establish a corneal injury model, the CP-9 strain live bacteria hydrogel prepared in Experimental Example 7 and the dead bacteria hydrogel prepared in Experimental Example 8 were respectively instilled into the eyes for 24 hours and then the eyeballs of mice were photographed. It was found that the dead bacteria of the CP-9 strain had a better effect on promoting corneal injury repair than the live bacteria.

[0130] In Experimental Example 9, the PBS solution of CP-9 strain almost completely disappeared after 30 minutes, while the hydrogel eye drops of CP-9 strain still had an obvious hydrogel layer after 120 minutes, which shows that the hydrogel can more significantly increase the residence time of probiotics on the ocular surface.

[0131] The embodiments of the present invention are described above in conjunction with the accompanying drawings, but the present invention is not limited to the above-mentioned specific implementation modes, which are merely illustrative rather than restrictive. Under the guidance of the present invention, ordinary technicians in this field can also make many forms without departing from the scope of protection of the present invention and the claims, all of which are within the protection of the present invention.

Claims

1. A probiotic for promoting the repair of ocular surface damage, characterized in that: include: Probiotics, wherein the probiotics include at least one of Bifidobacterium animalis subspecies lactis and Lactobacillus reuteri.

2. The probiotic according to claim 1, characterized in that The Bifidobacterium animalis subsp. lactis includes the Bifidobacterium animalis subsp. lactis CP-9 strain, which is deposited in the China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC NO: M2014588; and / or The Lactobacillus reuteri includes the Lactobacillus reuteri GL-104 strain, which is preserved in the China Center for Type Culture Collection (Wuhan) with a preservation number of CCTCC NO: M209138.

3. A probiotic composition for promoting the repair of ocular surface damage, characterized in that: The composition comprises: The probiotic according to any one of claims 1 to 2.

4. The composition according to claim 3, characterized in that The composition further comprises: carboxymethyl cellulose and sodium salt of N-hydroxysulfosuccinimide.

5. The composition according to claim 3, characterized in that The composition further comprises: gelatin.

6. The composition according to claim 5, characterized in that The mass ratio of the probiotics to the gelatin is (0.75-3):

1.

7. The composition according to claim 5, characterized in that The mass concentration of gelatin in the composition is 5-15%; and / or The content of the probiotics is 1.5-45 billion CFU / ml.

8. The composition according to claim 3, characterized in that The composition is fluid or semi-fluid; and / or The probiotics are dead bacteria.

9. A method for preparing a probiotic composition for promoting the repair of ocular surface damage, characterized in that: include: mixing the probiotics with the gelatin solution to obtain a mixed solution; Adding carboxymethyl cellulose and sodium salt of N-hydroxysulfosuccinimide to the mixed solution, and mixing to prepare a composition; Wherein, the probiotics include at least one of Bifidobacterium animalis subsp. lactis and Lactobacillus reuteri.

10. The preparation method according to claim 9, characterized in that The Bifidobacterium animalis subsp. lactis includes the Bifidobacterium animalis subsp. lactis CP-9 strain, which is deposited in the China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC NO: M2014588; and / or The Lactobacillus reuteri includes the Lactobacillus reuteri GL-104 strain, which is deposited in China Center for Type Culture Collection (Wuhan) with a deposit number of CCTCC NO: M209138; and / or The mass ratio of the probiotics to the gelatin is (0.75-3):1; and / or The mass concentration of gelatin in the composition is 5-15%.

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