Mesenchymal stem cell engineering transformation method based on lipoic acid polymerization reaction and application of mesenchymal stem cell engineering transformation method

Abstract

The invention discloses a mesenchymal stem cell engineering transformation method based on lipoic acid polymerization reaction and application thereof. The mesenchymal stem cell engineering modification method based on the lipoic acid polymerization reaction comprises the following steps: (1) preparing an alpha-lipoic acid modified hyaluronic acid solution and a polymerization reaction solution; 2) dispersing the mesenchymal stem cells in an alpha-lipoic acid modified hyaluronic acid solution for reaction to obtain the mesenchymal stem cells with the surfaces modified by alpha-lipoic acid; and (3) dispersing the mesenchymal stem cells of which the surfaces are modified by alpha-lipoic acid into a polymerization reaction solution, and performing ring-opening polymerization reaction by ultraviolet irradiation to obtain the engineered mesenchymal stem cells. The mesenchymal stem cell engineering modification method based on the lipoic acid polymerization reaction has the advantages of being easy to operate, mild in condition, short in consumed time, high in modification efficiency and the like, and the obtained engineering mesenchymal stem cells have lasting and stable oxidation resistance and can effectively resist oxidative stress damage in a transplantation environment.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cell engineering, in particular to a mesenchymal stem cell engineering method based on a lipoic acid polymerization reaction and application thereof. BACKGROUND

[0002] Mesenchymal stem cells have multiple differentiation, anti-inflammatory, immune regulation and other functions, and show good clinical application prospects in the treatment of ischemic diseases, nervous system injuries, autoimmune diseases and other diseases. Studies have shown that the therapeutic effect of mesenchymal stem cells after in vivo transplantation depends largely on their survival rate in the lesion tissue. However, a large number of studies have shown that the oxidative stress state and the high level of active oxygen accompanying it in the lesion tissue will seriously affect the survival of transplanted cells, and the survival rate of transplanted cells in the body will rapidly decrease within 1 day after transplantation, and the survival rate is usually less than 1% after 4-6 weeks, which has become one of the bottlenecks restricting the clinical application of mesenchymal stem cells.

[0003] At present, in order to improve the survival ability of mesenchymal stem cells in oxidative stress environment, small molecule antioxidants are often used for pretreatment. Commonly used small molecule antioxidants include glutathione, melatonin, vitamin E and alpha-lipoic acid, etc. This kind of small molecule antioxidant can scavenge active oxygen and can regulate the activity of endogenous antioxidant enzymes, so as to reduce the damage of transplanted cells and ultimately improve the state of transplanted cells. However, the above-mentioned small molecule antioxidants usually have problems such as easy degradation, unstable function, short duration of action, etc., and it is difficult to achieve long-term effective protection of transplanted mesenchymal stem cells, which cannot meet the needs of clinical stability and treatment persistence of mesenchymal stem cells.

[0004] Therefore, it is of great significance to develop an engineering method that can endow mesenchymal stem cells with persistent antioxidant properties to improve the survival rate and therapeutic effect of transplanted mesenchymal stem cells in vivo. SUMMARY

[0005] One of the purposes of the present application is to provide a mesenchymal stem cell engineering method based on a lipoic acid polymerization reaction.

[0006] The second purpose of the present application is to provide an application of the above-mentioned mesenchymal stem cell engineering method based on a lipoic acid polymerization reaction in the preparation of a drug for treating ischemic diseases, nervous system injuries or autoimmune diseases.

[0007] The third purpose of the present application is to provide an engineered mesenchymal stem cell prepared by the above-mentioned mesenchymal stem cell engineering method based on a lipoic acid polymerization reaction.

[0008] The fourth object of the present application is to provide a medicine comprising the engineered mesenchymal stem cell.

[0009] The technical solution adopted by the present application is:

[0010] A mesenchymal stem cell engineering method based on the polymerization reaction of alpha-lipoic acid comprises the following steps:

[0011] 1) Preparation of alpha-lipoic acid modified hyaluronic acid solution and polymerization reaction solution:

[0012] Preparation of alpha-lipoic acid modified hyaluronic acid solution: dissolve alpha-lipoic acid modified hyaluronic acid in PBS buffer to obtain alpha-lipoic acid modified hyaluronic acid solution;

[0013] Preparation of polymerization reaction solution: dissolve alpha-lipoic acid, N-(2-hydroxypropyl) methacrylamide and photoinitiator in PBS buffer to obtain polymerization reaction solution;

[0014] 2) Disperse the mesenchymal stem cells in the alpha-lipoic acid modified hyaluronic acid solution for reaction to obtain mesenchymal stem cells modified with alpha-lipoic acid on the surface;

[0015] 3) Disperse the mesenchymal stem cells modified with alpha-lipoic acid on the surface in the polymerization reaction solution, and then irradiate with ultraviolet light for ring-opening polymerization reaction to obtain engineered mesenchymal stem cells.

[0016] Preferably, the alpha-lipoic acid modified hyaluronic acid in step 1) is prepared by a preparation method comprising the following steps: dissolving hyaluronic acid tetrabutylammonium salt, alpha-lipoic acid, catalyst and carboxylic acid activator in a solvent, and then performing esterification reaction in a protective atmosphere to obtain alpha-lipoic acid modified hyaluronic acid.

[0017] Further preferably, the alpha-lipoic acid modified hyaluronic acid in step 1) is prepared by a preparation method comprising the following steps: dissolving hyaluronic acid tetrabutylammonium salt, alpha-lipoic acid, catalyst and carboxylic acid activator in a solvent, and then performing esterification reaction in a protective atmosphere, adding ice water to terminate the reaction, loading the reaction solution into a dialysis bag and placing it in water for dialysis, and then performing freeze-drying to obtain alpha-lipoic acid modified hyaluronic acid.

[0018] Preferably, the mass ratio of the hyaluronic acid tetrabutylammonium salt and alpha-lipoic acid is 1:1-2.

[0019] Preferably, the hyaluronic acid tetrabutylammonium salt is prepared by a preparation method comprising the following steps: dissolving hyaluronic acid in water, adding cation exchange resin for ion exchange, suction filtration, adjusting the pH value of the filtrate to 6.8-7.2 with tetrabutylammonium hydroxide, and then performing freeze-drying to obtain hyaluronic acid tetrabutylammonium salt.

[0020] Preferably, the catalyst is at least one of 4-dimethylaminopyridine, triethylamine, 4-pyrrolidinopyridine.

[0021] Preferably, the carboxylic acid activator is at least one of di-tert-butyl dicarbonate, N,N'-dicyclohexyl carbodiimide, N,N'-diisopropyl carbodiimide.

[0022] Preferably, the solvent is at least one of dimethyl sulfoxide, N,N-dimethylformamide, methanol.

[0023] Preferably, the protective atmosphere is an argon atmosphere or a nitrogen atmosphere.

[0024] Preferably, the esterification reaction is carried out at a temperature of 40-50°C, and the reaction time is 12-36h.

[0025] Preferably, the dialysis bag has a molecular weight cut-off of 3-14kD.

[0026] Preferably, the dialysis time is 2-4 days.

[0027] Preferably, the molar ratio of the alpha-lipoic acid to N-(2-hydroxypropyl) methacrylamide in step 1) is 1-9:1.

[0028] Preferably, the photoinitiator in step 1) is at least one of 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone, phenyl (2,4,6-trimethylbenzoyl) lithium phosphate, eosin Y.

[0029] Preferably, the reaction in step 2) is carried out at a temperature of 4-37°C, and the reaction time is 1-3h.

[0030] Preferably, the ring-opening polymerization reaction in step 3) is carried out at a UV light intensity of 5-10mW / cm 2 2 , and the reaction time is 2-30min.

[0031] Use of a mesenchymal stem cell engineering method based on lipoic acid polymerization as described above in the preparation of a drug for treating ischemic diseases, nervous system injuries, or autoimmune diseases.

[0032] An engineered mesenchymal stem cell prepared by the mesenchymal stem cell engineering method based on lipoic acid polymerization as described above.

[0033] A drug comprising the engineered mesenchymal stem cell as described above.

[0034] ​Principle of the present application: the present application adopts cell engineering method to construct degradable lipoic acid polymer on the surface of mesenchymal stem cells in situ, and the obtained engineered mesenchymal stem cells can realize sustained antioxidant protection of mesenchymal stem cells through slow release of alpha-lipoic acid after in vivo transplantation, thereby significantly improving the survival rate and treatment effect of mesenchymal stem cells in pathological microenvironment.

[0035] The present application has the advantages that the mesenchymal stem cell engineering method based on lipoic acid polymerization reaction of the present application has the advantages of simple operation, mild conditions, short time consumption, high modification efficiency, etc., and the obtained engineered mesenchymal stem cells have persistent and stable antioxidant properties, which can effectively resist oxidative stress damage in the transplantation environment.

[0036] Specifically:

[0037] 1) The present application adopts cell engineering method, and utilizes the reversible dynamic ring-opening polymerization-depolymerization characteristics of endogenous small molecule coenzyme alpha-lipoic acid itself to construct lipoic acid polymer on the surface of mesenchymal stem cells in situ, realizes controllable slow release of alpha-lipoic acid in pathological microenvironment, and slow release of alpha-lipoic acid can realize sustained antioxidant protection of mesenchymal stem cells, which can effectively resist oxidative stress damage in the transplantation environment, thereby significantly improving the survival rate and treatment effect of mesenchymal stem cells in pathological microenvironment;

[0038] 2) The mesenchymal stem cell engineering method based on lipoic acid polymerization reaction of the present application has the advantages of simple operation, mild conditions, short time consumption, high modification efficiency, good biocompatibility of the constructed lipoic acid polymer, etc., and has good clinical transformation application prospect. BRIEF DESCRIPTION OF DRAWINGS

[0039] Figure 1 The flow chart of the mesenchymal stem cell engineering method based on lipoic acid polymerization reaction of the present application.

[0040] Figure 2 The slow release schematic diagram of alpha-lipoic acid in the engineered mesenchymal stem cells of the present application.

[0041] Figure 3 The ultraviolet absorption spectrum diagram of alpha-lipoic acid solution before and after light irradiation.

[0042] Figure 4 The laser confocal diagram of the engineered mesenchymal stem cells in Example 1.

[0043] Figure 5 The modification effect test result diagram of the engineered mesenchymal stem cells in Example 1.

[0044] Figure 6Figure for cell viability test results of mesenchymal stem cells, pretreated mesenchymal stem cells and engineered mesenchymal stem cells in Example 1.

[0045] Figure 7 Figure for cell viability test results of mesenchymal stem cells, pretreated mesenchymal stem cells and engineered mesenchymal stem cells in Example 1.

[0046] Figure 8 Figure for intracellular reactive oxygen species level test results of mesenchymal stem cells, pretreated mesenchymal stem cells and engineered mesenchymal stem cells in Example 2. DETAILED DESCRIPTION

[0047] The application will be further explained and described with reference to specific examples.

[0048] Example 1:

[0049] A mesenchymal stem cell engineering method based on lipoic acid polymerization reaction, the steps are as follows (flow chart as shown in Figure 1

[0050] 1) Preparation of α-lipoic acid modified hyaluronic acid solution and polymerization reaction solution:

[0051] Preparation of α-lipoic acid modified hyaluronic acid solution: 10 mg of α-lipoic acid modified hyaluronic acid was dissolved in 1 mL of 1x PBS buffer, and then filtered with a needle filter with a pore size of 0.22 μm to obtain an α-lipoic acid modified hyaluronic acid solution (concentration of 1 wt%);

[0052] Preparation of polymerization reaction solution: 10.3 mg of α-lipoic acid, 1.4 mg of N-(2-hydroxypropyl) methyl acrylamide and 4.5 mg of 2-hydroxy-4'-(2-hydroxyethoxy)-2-methyl propiophenone were dissolved in 10 mL of 1x PBS buffer to obtain a polymerization reaction solution (concentrations of α-lipoic acid, N-(2-hydroxypropyl) methyl acrylamide and 2-hydroxy-4'-(2-hydroxyethoxy)-2-methyl propiophenone were 5 mmol / L, 1 mmol / L and 2 mmol / L, respectively);

[0053] 2) 1x 10 6 ​The mouse bone marrow-derived mesenchymal stem cells are dispersed in 200 μL of α-lipoic acid-modified hyaluronic acid solution to prepare a cell suspension, and then placed in a shaking metal bath under the condition of a temperature of 37°C and a shaking speed of 300 rpm. During the shaking process, the cell suspension is mixed by blowing with a pipette every 15 min, and then diluted with 200 μL of 1×PBS buffer. After centrifugation at a centrifugal force of 300 g for 4 min, the supernatant is removed, and then 400 μL of 1×PBS buffer is added to resuspend the cells. After centrifugation at a centrifugal force of 300 g for 4 min, the supernatant is removed to obtain the mesenchymal stem cells modified with α-lipoic acid on the surface;

[0054] 3) The mesenchymal stem cells modified with α-lipoic acid on the surface are resuspended in 400 μL of a polymerization reaction solution, and then the obtained cell suspension is placed under a UV LED light source with a wavelength of 365 nm. The irradiation intensity of the UV light is controlled to be 5 mW / cm 2 for 10 min. Then, 400 μL of complete culture medium (90% DMEM + 10% fetal bovine serum) is added to quench the reaction. After centrifugation at a centrifugal force of 300 g for 4 min, the supernatant is removed, and then 400 μL of 1×PBS buffer is added to resuspend the cells. After centrifugation at a centrifugal force of 300 g for 4 min, the supernatant is removed. The resuspension and centrifugation are repeated twice, and then the supernatant is removed to obtain the engineered mesenchymal stem cells (a schematic diagram of the slow release of α-lipoic acid is shown in Figure 2 ).

[0055] Note:

[0056] The preparation method of the α-lipoic acid-modified hyaluronic acid is as follows:

[0057] a) 2 g of hyaluronic acid (Macklin, product number H909935) is dissolved in 100 mL of deionized water under stirring, and then 6 g of Dowex 50WX8 ion exchange resin (Macklin, product number D766555) is added. The mixture is stirred at a temperature of 25°C and a stirring speed of 600 rpm for 2 h. After filtration, the filtrate is adjusted to a pH value of 7.03 with a 5% tetrabutylammonium hydroxide solution, and then freeze-dried to obtain hyaluronic acid tetrabutylammonium salt;

[0058] b) Dissolve 0.5 g of tetrabutylammonium hyaluronic acid, 0.817 g of α-lipoic acid, and 0.242 g of 4-dimethylaminopyridine in 25 mL of anhydrous dimethyl sulfoxide. Then add 121 μL of ditert-butyl dicarbonate. After the addition is complete, stir the mixture under an argon atmosphere at 45 °C and a stirring rate of 600 rpm for 20 h. Then add 25 mL of ice water to terminate the reaction. Then put the reaction solution into a dialysis bag with a molecular weight cutoff of 14 kD and dialyze it in deionized water for 3 days. Then freeze-dry to obtain α-lipoic acid modified hyaluronic acid.

[0059] The structural formula of α-lipoic acid-modified hyaluronic acid is as follows:

[0060]

[0061] Performance testing:

[0062] 1) Feasibility verification of photoinduced ring-opening polymerization of α-lipoic acid in solution: 0.064 g of α-lipoic acid was dissolved in 1 mL of 10% sodium hydroxide solution. The pH was adjusted to 7.4 with 1 mol / L hydrochloric acid, and then diluted with 1×PBS buffer to prepare a 100 mmol / L α-lipoic acid solution. 20 μL of the α-lipoic acid solution was added to a 48-well plate and diluted with 1×PBS buffer to a concentration of 5 mmol / L. The 48-well plate was then placed under a 365 nm UV LED light source, and the UV irradiation intensity was controlled at 0 °C at 5 mW / cm². 2 After irradiation for 10 minutes, the ultraviolet absorption spectra of the α-lipoic acid solution before and after irradiation, measured using an ultra-micro spectrophotometer, are shown below. Figure 3 As shown.

[0063] Depend on Figure 3 It can be seen that before light treatment, the α-lipoic acid solution has a significant absorption peak at 332 nm, which corresponds to the characteristic absorption of the dithiopentane structure in the α-lipoic acid molecule. After light treatment, the intensity of the characteristic peak at 332 nm of the α-lipoic acid solution decreased significantly, and the absorption signal of the dithiopentane weakened, indicating that the dithiopentane structure in the α-lipoic acid molecule broke under ultraviolet light and underwent a ring-opening polymerization reaction. This result provides experimental basis for subsequent in-situ polymerization modification on the surface of living cells.

[0064] 2) 5×10 5 The engineered mesenchymal stem cells in this embodiment were resuspended in 200 μL of 1×PBS buffer and then imaged under a laser confocal microscope. The resulting laser confocal image is shown below. Figure 4 As shown.

[0065] Depend onFigure 4 The uniform distribution of fluorescence signals on the cell surface indicates that the cell membrane is uniformly covered by continuous α-lipoic acid polymers, suggesting that the photo-induced in situ polymerization reaction successfully occurred on the surface of mesenchymal stem cells.

[0066] 3) Flow cytometry was used to characterize mesenchymal stem cells (before modification, i.e., original mouse bone marrow-derived mesenchymal stem cells) and engineered mesenchymal stem cells (after modification) in this embodiment. The cell modification effect test results are as follows: Figure 5 As shown.

[0067] Depend on Figure 5 It can be seen that the vast majority of engineered mesenchymal stem cells exhibited obvious positive fluorescence signals, and statistical results showed that the cell modification efficiency reached 99.8%.

[0068] 4) 5×10 5 Mesenchymal stem cells (before modification, i.e., original mouse bone marrow-derived mesenchymal stem cells) and engineered mesenchymal stem cells (after modification) in this example were resuspended in 200 μL of Calcein AM / PI assay working solution (Calcein / PI Cell Viability and Cytotoxicity Assay Kit, Beyotime, catalog number C2015M), and then placed in a shaking metal bath at 37°C and 300 rpm for 30 min. The cells were then characterized by flow cytometry, and the cell viability test results are shown below. Figure 6 As shown.

[0069] Depend on Figure 6 It can be seen that the viability of engineered mesenchymal stem cells is >95%, and the cell viability has not changed significantly compared with that of the unengineered mesenchymal stem cells, indicating that the mesenchymal stem cell engineering method of the present invention does not affect cell viability.

[0070] Example 2:

[0071] A method for engineering mesenchymal stem cells based on lipoic acid polymerization, comprising the following steps:

[0072] 1) Preparation of α-lipoic acid modified hyaluronic acid solution and polymerization reaction solution:

[0073] Preparation of α-lipoic acid modified hyaluronic acid solution: Dissolve 10 mg of α-lipoic acid modified hyaluronic acid in 1 mL of 1×PBS buffer, and then filter it through a 0.22 μm needle filter to obtain α-lipoic acid modified hyaluronic acid solution (concentration of 1 wt%).

[0074] Preparation of polymerization reaction solution: Dissolve 10.3 mg of α-lipoic acid, 2.8 mg of N-(2-hydroxypropyl)methacrylamide and 4.5 mg of 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylphenylacetone in 10 mL of 1×PBS buffer to obtain the polymerization reaction solution (the concentrations of α-lipoic acid, N-(2-hydroxypropyl)methacrylamide and 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylphenylacetone are 5 mmol / L, 2 mmol / L and 2 mmol / L respectively).

[0075] 2) Place 1×10 6 Mesenchymal stem cells derived from mouse bone marrow were dispersed in 200 μL of α-lipoic acid-modified hyaluronic acid solution to prepare a cell suspension. The suspension was then placed in a shaking metal bath and shaken at 25°C and 300 rpm for 1 h. During shaking, the cell suspension was mixed by pipetting every 15 min. 200 μL of 1×PBS buffer was added to dilute the cell suspension. The suspension was then centrifuged at 300 g for 4 min. After removing the supernatant, the cells were resuspended in 400 μL of 1×PBS buffer and centrifuged at 300 g for 4 min. The supernatant was then removed to obtain α-lipoic acid-modified mesenchymal stem cells.

[0076] 3) Mesenchymal stem cells modified with α-lipoic acid were resuspended in 400 μL of polymerization reaction solution, and the resulting cell suspension was placed under a 365 nm ultraviolet LED light source, with the ultraviolet light intensity controlled at 5 mW / cm². 2 Irradiate for 10 min, then add 400 μL of complete culture medium (90% DMEM + 10% fetal bovine serum) to quench the reaction, then centrifuge at 300 g for 4 min, remove the supernatant, add 400 μL of 1×PBS buffer to resuspend the cells, centrifuge at 300 g for 4 min, remove the supernatant, repeat the resuspension and centrifugation twice, remove the supernatant again, and obtain engineered mesenchymal stem cells.

[0077] Performance testing:

[0078] 1) Mesenchymal stem cells (referred to as Native MSCs; control group), pretreated mesenchymal stem cells (treated with complete medium (90% DMEM + 10% FBS) containing 10 μmol / L α-lipoic acid for 24 h, digested with trypsin and resuspended in 1×PBS buffer; referred to as LA-pretreated MSCs; control group), and engineered mesenchymal stem cells (referred to as pLA-MSCs) in this example were resuspended in 600 μmol / L hydrogen peroxide solution to prepare a cell concentration of 5×10⁻⁶ cells / year. 4Cell suspension was prepared at 100 μL / mL and seeded into 96-well plates. The plates were then incubated at 37°C with 5% CO2 for 4 hours. 10 μL of CCK-8 solution was added to each well, and incubation continued for another hour. Absorbance was measured at 450 nm using a multi-functional microplate reader to calculate cell viability. The cell viability results after hydrogen peroxide treatment are shown below. Figure 7 (In the figure, "ns" represents no significant difference, and "***" represents p<0.001).

[0079] Depend on Figure 7 It can be seen that after treatment with 600 μmol / L hydrogen peroxide solution for 4 hours, the cell viability of the two control groups was only 10% to 30%, while the cell viability of engineered mesenchymal stem cells (pLA-MSCs) remained at >70%. This indicates that under high concentration and continuous oxidative stress conditions, pretreatment with small molecule antioxidants is difficult to provide effective protection for cells, while engineering modification can still significantly improve cell survival.

[0080] 2) Mesenchymal stem cells were divided into three groups: a blank control group (denoted as Native MSCs), a control group + hydrogen peroxide treatment group (denoted as Native MSCs + H2O2), and the engineered mesenchymal stem cells + hydrogen peroxide treatment group in this embodiment (denoted as pLA-MSCs + H2O2). 1×10⁻⁶ cells were then taken from each group. 6 Cells were resuspended in 500 μL of 1×PBS buffer. Then, 30% hydrogen peroxide solution was added to the groups requiring treatment to bring the final concentration to 1 mmol / L. The mixture was thoroughly mixed by pipetting and incubated at room temperature for 1 h. The cells were then centrifuged at 300 g for 4 min, the supernatant was removed, and the cells were washed twice with 1×PBS buffer. The cells were then resuspended in 1 mL of 1×PBS buffer, and 1 μL of the DCFH-DA probe (Beyotime, catalog number S0033S-1) was added. The cells were incubated at 37 °C for 20 min, washed twice with 1×PBS buffer, and finally resuspended in 1 mL of PBS buffer. Flow cytometry was used to characterize each group. The intracellular reactive oxygen species (ROS) levels were measured as follows: Figure 8 (The “****” in the figure represents p<0.0001).

[0081] Depend on Figure 8 It can be seen that the fluorescence intensity of cells in the engineered mesenchymal stem cell + hydrogen peroxide treatment group was significantly lower than that in the control group + hydrogen peroxide treatment group, indicating that the level of reactive oxygen species in the former cells was much lower than that in the latter, suggesting that the engineered mesenchymal stem cells have a significantly enhanced ability to scavenge reactive oxygen species.

[0082] The above embodiments are the preferred embodiments of the present application, but the embodiments of the present application are not limited to the above embodiments, and any changes, modifications, substitutions, combinations, simplifications, etc. made without departing from the spirit and principles of the present application should be equivalent replacement manners and should be included in the protection scope of the present application.

Claims

1. A mesenchymal stem cell engineering method based on the polymeric reaction of thioctic acid, characterized by, The method comprises the following steps: 1) Preparation of alpha-lipoic acid modified hyaluronic acid solution and polymerization reaction solution: Preparation of alpha-lipoic acid modified hyaluronic acid solution: dissolve alpha-lipoic acid modified hyaluronic acid in PBS buffer to obtain alpha-lipoic acid modified hyaluronic acid solution; Preparation of polymerization reaction solution: dissolve alpha-lipoic acid, N-(2-hydroxypropyl) methacrylamide and photoinitiator in PBS buffer to obtain polymerization reaction solution; 2) Disperse mesenchymal stem cells in alpha-lipoic acid modified hyaluronic acid solution for reaction to obtain alpha-lipoic acid surface modified mesenchymal stem cells; 3) Disperse the alpha-lipoic acid surface modified mesenchymal stem cells in the polymerization reaction solution, and then irradiate with ultraviolet light for ring-opening polymerization reaction to obtain engineered mesenchymal stem cells.

2. The method of engineering mesenchymal stem cells based on the polymeric reaction of thioctic acid according to claim 1, characterized by the fact that: The alpha-lipoic acid modified hyaluronic acid in step 1) is prepared by a preparation method comprising the following steps: dissolve hyaluronic acid tetrabutylammonium salt, alpha-lipoic acid, catalyst and carboxylic acid activator in a solvent, and then place it in a protective atmosphere for esterification reaction to obtain alpha-lipoic acid modified hyaluronic acid.

3. The method of engineering mesenchymal stem cells based on the polyreaction of thioctic acid according to claim 2, characterized by the fact that: The mass ratio of the hyaluronic acid tetrabutylammonium salt to alpha-lipoic acid is 1:1-2.

4. The method of engineering mesenchymal stem cells based on the polyreaction of thioctic acid according to claim 2, characterized by the fact that: The esterification reaction is carried out at a temperature of 40-50°C, and the reaction time is 12-36h.

5. The method for engineering mesenchymal stem cells based on the lipoic acid polymerization reaction according to any one of claims 1 to 4, characterized in that: The molar ratio of alpha-lipoic acid to N-(2-hydroxypropyl) methacrylamide in step 1) is 1-9:

1.

6. The lipoic acid polymerization reaction-based mesenchymal stem cell engineering method according to claim 1, characterized by: The reaction in step 2) is carried out at a temperature of 4-37°C, and the reaction time is 1-3h.

7. The method of engineering mesenchymal stem cells based on the lipoic acid polymerization reaction according to claim 1, characterized in that: Step 3) said ring-opening polymerization reaction is carried out under the condition of UV light intensity of 5 mW / cm 2 ~ 10 mW / cm 2 and the reaction time is 2 min ~ 30 min.

8. Use of the mesenchymal stem cell engineering method based on lipoic acid polymerization reaction according to any one of claims 1-7 in the preparation of a drug for treating ischemic diseases, nervous system injuries or autoimmune diseases.

9. An engineered mesenchymal stem cell, characterized in that, The mesenchymal stem cell engineering method based on lipoic acid polymerization reaction according to any one of claims 1-7.

10. A medicament, characterized by comprising: The engineered mesenchymal stem cells according to claim 9. The engineered mesenchymal stem cells according to claim 9.

Images