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Optic nerve repair composition as well as preparation method and application thereof

A composition and optic nerve technology, which can be used in drug combinations, pharmaceutical formulations, medical raw materials derived from mammals, etc., can solve problems such as the inability to effectively repair the optic nerve.

Pending Publication Date: 2021-03-09
GUANGZHOU ZHUNYOU BIOLOGICAL TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing treatment methods cannot effectively repair the damaged optic nerve, so it is urgent to find a method to effectively repair the damage and reconstruct the function of the optic nerve, which is of great significance

Method used

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  • Optic nerve repair composition as well as preparation method and application thereof
  • Optic nerve repair composition as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0020] The preparation method of the optic nerve repair composition according to one embodiment of the present invention comprises the following steps: mixing Rho kinase inhibitor, activated platelet-rich plasma and stem cells and culturing at 2°C to 37°C for 8 to 72 hours to obtain optic nerve repair combination. The aforementioned stem cells have the ability to differentiate into epithelial cells.

[0021] Endothelial progenitor cells (EPCs) are a type of precursor stem cells between stem cells and vascular endothelial cells. They can differentiate into mature vascular endothelial cells, form blood vessel-like structures, promote vascular endothelial proliferation, and participate in endogenous repair of blood vessels. It can also promote angiogenesis by providing cytokines and trophic factors.

[0022] Platelet-rich plasma (platelet-rich plasma, PRP) is a platelet concentrate separated from autologous blood by centrifugation. It contains a variety of growth factors and pr...

Embodiment 1

[0040] This embodiment provides a preparation method of an optic nerve repair composition, which specifically includes the following steps:

[0041] (1) Before drawing peripheral blood, the volunteers signed the informed consent;

[0042] (2) Take 100 mL of autologous peripheral blood and place it in 12 15 mL tubes, centrifuge at 200 g / min for 15 minutes;

[0043] (3) In 8 tubes, carefully absorb the plasma layer and buffy coat layer, and discard the red blood cell layer;

[0044] (4) Place the plasma buffy coat liquid in two 15mL centrifuge tubes, centrifuge at 200g / min for 10 minutes;

[0045] (5) Take the supernatant, pass it through a 30 μm filter to obtain peripheral blood stem cells, and store it at 4°C for later use;

[0046] (6) For the other 4 tubes, absorb the supernatant, place them in two 15ml test tubes on average, centrifuge at 2500g / min for 10 minutes;

[0047] (7) Discard the supernatant after centrifugation, leaving only the bottom sediment and 3mL serum; ...

Embodiment 2～4

[0091] Examples 2-4 are basically the same as Example 1, except that when the stem cells are cultured at low temperature, the concentrations of the Rho kinase inhibitor AR-12286 are 0.05wt%, 0.15wt%, and 0.35wt%, respectively.

[0092] The cells in the final products of Examples 1-4, Comparative Example 2 and Comparative Example 5 were immunostained with CD34-PE and CD133-FITC respectively, and the percentage of double positive cells was analyzed by flow cytometry. The results are shown in Table 1 below.

[0093] Table 1

[0094] Percentage of double positive cells Comparative example 2 stem cell 4.5％ Comparative example 5 stem cells+PRP 7.2％ Example 2 Stem cells + PRP + 0.05% Rho inhibitor 7.9％ Example 3 Stem cells + PRP + 0.15% Rho inhibitor 10.6％ Example 1 Stem cells + PRP + 0.25% Rho inhibitor 15.3％ Example 4 Stem cells + PRP + 0.35% Rho inhibitor 14.7％

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Abstract

The invention relates to an optic nerve repair composition as well as a preparation method and application thereof. The preparation method comprises the following steps of mixing an Rho kinase inhibitor, activated platelet-rich plasma and stem cells, and performing culture at 2-37 DEG C for 8-72 hours to obtain the optic nerve repair composition. Specifically for pathogenic factors of vascular dysfunction, intraocular pressure rise and optic nerve cell injury of ophthalmic disease patients, the stem cells, the PRP and the Rho kinase inhibitor are combined for the first time, and a set of new three-dimensional cascade interaction mechanism is established. The tight synergistic effect of the PRP, the stem cells and the Rho kinase inhibitor prompts that the PRP and the Rho kinase inhibitor can jointly secrete cytokines, promote vascular endothelium and optic nerve cell repair, reconstruct vascular functions, reduce intraocular pressure, reduce retinal damage and reduce retinal cell apoptosis by promoting the function of EPC.

Description

technical field [0001] The invention relates to the technical field of ophthalmology, in particular to an optic nerve restoration composition and its preparation method and application. Background technique [0002] Glaucoma is one of the three major irreversible blinding eye diseases, with a large number of patients worldwide. Glaucoma is an optic nerve disease, and pathological increased intraocular pressure and optic nerve insufficiency are the most important primary risk factors. Animal and human studies suggest that vascular factors, including ischemia and vascular dysfunction, play a role in the pathogenesis of glaucoma, and vascular dysfunction is closely related to endothelial progenitor cells (EPCs). Studies have shown that endothelial dysfunction and reduced number in glaucoma patients lead to abnormal blood vessel regulation and elevated intraocular pressure. Therefore, reducing intraocular pressure and promoting ocular blood flow are the keys to treatment. [0...

Claims

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Application Information

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IPC IPC(8): A61K45/06A61K31/4409A61K31/4725A61K35/16A61K35/28A61P27/02A61P27/06
CPCA61K45/06A61K35/16A61K35/28A61K31/4725A61K31/4409A61P27/06A61P27/02A61K2300/00
Inventor 吴道贫鲁坤康太郎陶然覃绍君赖小华肖泽鹏莫乾强邹洁宜谢秀凤
Owner GUANGZHOU ZHUNYOU BIOLOGICAL TECH CO LTD
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