CN116392588A - Photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair and preparation method and application thereof - Google Patents
Photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair, which comprises a hydrogel carrier and AIE vesicles loaded in the hydrogel carrier; the AIE vesicle is an AIE molecule with an outer surface wrapped with an artificial stem cell vesicle; the mass ratio of the AIE molecules to the artificial stem cell vesicles is in the range of 1: 3-50, wherein the maximum final concentration of AIE molecules in the AIE vesicle is not more than 400 mu mol/L. The hydrogel not only has excellent functions of resisting infection, resisting bacteria and promoting repair and regulation, but also has good biosafety; and has good stability and long-acting property. In addition, the hydrogel is simple to prepare, low in cost, good in universality and good in practical popularization significance in the field of treatment of various infections.
Description
Technical Field
The invention belongs to the technical field of hydrogel materials, and in particular relates to photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair, and a preparation method and application thereof.
Background
Serious skin injury such as scald, burn and the like is caused by dermis destruction, concurrent repeated infection and the like, and the double problems of repeated infection and difficult healing are faced after healing, so that the epidermis structure is difficult to recover, contracture deformity and the like can be generated, and the physiological and mental burden of a patient is aggravated. The traditional wound repair means mainly comprise drug treatment, dressing application, pressure therapy, skin transplantation and the like, the complete repair is still difficult to realize, and large-area deep infection is easy to cause sepsis and other concurrent diseases so as to prolong the treatment process.
The current novel repair-promoting biological treatment such as platelet-rich plasma therapy, extracellular matrix therapy, stem cell extracellular vesicle therapy and the like has the advantage of difficult replacement and obvious treatment effect because the novel repair-promoting biological treatment can provide autologous comprehensive healing-promoting biological treatment components for the wound surface. Wherein, the stem cells can paracrine extracellular vesicles containing signal molecules such as protein, RNA and the like, and the extracellular vesicles are reported to have low immune response and homing targeting, activating a certain signal path, promoting up-regulation of expression of various cell growth factors and the like, thereby realizing synchronous proliferation and repair of various cells; because of the smaller volume, the tissue penetration performance is better, the stability and the safety are better than those of cells, and the delivery of small molecular drugs, proteins, nucleic acids and the like is facilitated. But still has the problems of high price, lack of anti-infection function, limited acting time and the like. Tang Benzhong et al reported targeting and photodynamic therapy of tumor cells after modification of tumor extracellular vesicles with AIE material (Angew.chem.int.ed.2020, 59, 13836-13843), but no photodynamic therapy vesicles suitable for wound surface antibacterial and repair promotion were reported.
The extracellular vesicles used for the wound surface are usually injected clinically, but have short acting time and are easy to lose efficacy. The hydrogel is a polymer sustained-release material with a three-dimensional network structure, and is widely applied to the fields of medicines and tissue engineering such as cell scaffolds, drug delivery systems, soft tissue substitutes, wound dressings and the like. The hydrogel has good biocompatibility and degradability, and can form a drug reservoir at the lesion site to realize the local controllable release of the drug, thereby improving the effective accumulation of the drug at the lesion site and achieving the treatment purpose. However, in order to load the vesicle into the gel, it is necessary to ensure that the components of the vesicle are not damaged in the preparation process of the gel, and usually, temperature-sensitive hydrogels such as PF-127 and its complex (nanoscales, 2021,13,14866-14878) have low selectivity and cannot realize targeted controllable slow release according to the wound microenvironment, while other controllable hydrogels mostly require light irradiation, heating or catalyst to assist in curing, so that the effective components in the vesicle may be damaged. In addition, although most laboratories have developed various separation and extraction techniques such as volume exclusion chromatography, microfluidic and immunocapture, the cost of obtaining stem extracellular vesicles is still high and the flow is complex. Therefore, how to provide a hydrogel material with excellent functions for combining extracellular vesicles is a technical problem to be solved urgently.
Disclosure of Invention
Aiming at the prior art problems, the primary aim of the invention is to provide a photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair, which has excellent functions of anti-infection, antibacterial, repair promotion and regulation, good biological safety and stability; and the preparation process is mild and rapid, and the stability and long-acting property of the carried vesicle are ensured.
The second aim of the invention is to provide a preparation method of the photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair.
The third object of the invention is to provide the application of the photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair in preparing wound repair materials.
A photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair comprises a hydrogel carrier and AIE vesicles loaded in the hydrogel carrier; the AIE vesicle is an AIE molecule with an outer surface wrapped with an artificial stem cell vesicle; the mass ratio of the AIE molecules to the artificial stem cell vesicles is in the range of 1: 3-50, wherein the maximum final concentration of AIE molecules in the AIE vesicle is not more than 400 mu mol/L.
The invention provides an artificial stem cell vesicle hydrogel with a wound repair promoting function, which is prepared by adopting an artificial extrusion preparation technology, subjecting stem cells to subculture, then performing hypotonic liquid pyrolysis, and then extruding separated cell components and a phospholipid membrane through a liposome extruder again to obtain an artificial stem cell vesicle, wherein the operation is simple, the yield is high, the large-scale preparation is convenient while the multiple healing promotion components of the original stem cells are maintained, and meanwhile, the problems of insufficient safety and controllability of the traditional stem cell treatment are overcome. Further, the artificial stem cell vesicles were combined with a specific AIE molecule (Aggregation-Induced Emission) to prepare the AIE vesicles. In this process, the inventors found that the maximum final concentration of AIE in AIE vesicles needs not to exceed 400. Mu. Mol/L, which is the limit of loading, and that further increases in concentration result in AIE not being well dispersed, thereby precipitating solid precipitates, affecting AIE vesicles and subsequent complex preparation with hydrogels. Finally, the AIE vesicle is entrapped by a long-acting slow-release hydrogel carrier, so that the problems of insufficient anti-infection capability, limited stability, short half-life and the like when the vesicle directly acts on a body surface are solved.
Preferably, the AIE molecule has the structural formula (I):
wherein R is - To balance the charge of anions selected from Cl - 、Br - 、I - 、PF6 - One of them. The AIE molecule has outstanding anti-infective ability and good biocompatibility; in addition, the water-based vesicle material has excellent water medium dispersing capability and can be better combined with vesicles.
Preferably, the hydrogel carrier is prepared from natural water-soluble high molecular compounds containing amino or sulfhydryl groups and water-soluble compounds containing ester alkynyl according to the functional group ratio of 1-10: 10-1 is prepared by click chemistry reaction. The hydrogel carrier is obtained by utilizing the click reaction between the activated ester alkynyl and the high molecular compound containing amino or sulfhydryl, the condition is milder, the process is simpler, no heating, illumination or catalysis of an additional catalyst is needed, the extra step and the residue of a toxic catalyst are avoided, and the formed amino propylene ester bond has acid responsiveness, so that the controllable release characteristic of the hydrogel is endowed.
Further preferably, the natural water-soluble polymer compound containing amino or mercapto is selected from one or more of chitosan and derivatives thereof, and sodium carboxymethyl cellulose.
Preferably, the water-soluble compound containing an ester alkynyl group has one or more of the structures shown in any one of the following formulas (II):
wherein n is an integer of 1 or more.
Preferably, the artificial stem cell vesicles are derived from one or more of adipose stem cells, fibroblasts, epidermal stem cells.
Preferably, the mass ratio of AIE vesicles to hydrogel carrier is 0.67-10: 1.
furthermore, the application also claims a preparation method of the photodynamic anti-infective artificial stem cell vesicle hydrogel for promoting wound repair, which comprises the following preparation steps:
s1, splitting the artificial stem cells to obtain membrane suspension, and extruding the membrane suspension through a liposome extruder to obtain artificial stem cell vesicles;
s2, after the artificial stem cell vesicle is incubated with the AIE molecules, the artificial stem cell vesicle is extruded by a liposome extruder to realize the coating of the artificial stem cell vesicle on the AIE molecules, so as to obtain the AIE vesicle;
s3, adding an aqueous solution of AIE vesicles or a PBS (phosphate buffered saline) solution of the AIE vesicles into a precursor solution of the hydrogel carrier, standing and curing to obtain the photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair.
Preferably, the concentration of the AIE vesicles in the aqueous solution of the AIE vesicles or the PBS solution of the AIE vesicles is 1-300 mug/mL (the mass percentage concentration is 0.1% -30%).
Preferably, the precursor solution of the hydrogel carrier is 3-15% of the mass percentage concentration of the hydrogel carrier.
Preferably, the precursor solution of the hydrogel carrier is obtained by mixing a natural water-soluble high molecular compound containing amino or sulfhydryl and a water-soluble compound containing ester alkynyl.
Preferably, in the step S3, the standing time is 4 to 24 hours.
Preferably, in the step S1 and the step S2, the liposome extruder is a 100nm to 900nm liposome extruder.
Further preferably, in the step S1, the liposome extruder is a 600-800 nm liposome extruder; in the step S2, the liposome extruder is a 100-200 nm liposome extruder.
In addition, the application of the photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair in preparing a wound repair material is also claimed. The hydrogel plays a role in wound repair by regulating and releasing AIE vesicle components for sterilization, bacteriostasis and promoting tissue self-repair at a wound site, and isolating exposure of the wound in air and reducing pathogen infection.
Compared with the prior art, the invention has the following beneficial effects: the invention provides a photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair; the hydrogel not only has excellent functions of resisting infection, resisting bacteria and promoting repair and regulation, but also has good biosafety; and has good stability and long-acting property. In addition, the hydrogel is simple to prepare, low in cost, good in universality and good in practical popularization significance in the field of treatment of various infections.
Drawings
FIG. 1 is a nuclear magnetic resonance hydrogen spectrum of AIE molecules in example 1.
FIG. 2 is a representation of the vesicles in example 1, wherein FIG. 2 (a) is a transmission electron microscope image of the vesicles; FIG. 2 (b) shows the results of the vesicle NTA test.
FIG. 3 is a nuclear magnetic resonance hydrogen spectrum of PEG4000-DA of example 1.
Fig. 4 is a transmission electron microscope topography of an artificial stem cell vesicle hydrogel with a function of promoting wound repair in example 1.
FIG. 5 is a graph showing the antibacterial ability of THB vesicles to Staphylococcus aureus and Escherichia coli in example 1.
FIG. 6 is a graph showing the antibacterial ability of the TCB vesicle of comparative example 2 against Staphylococcus aureus and Escherichia coli.
FIG. 7 is a statistical result of promotion of cell proliferation by AIE vesicles in example 1 and vesicles in comparative example 1.
FIG. 8 is a graph showing scratch test of AIE vesicles in example 1 and vesicles in comparative example 1.
FIG. 9 is a graph showing the sustained release capacity of the artificial stem cell vesicle hydrogel with the function of promoting wound repair in example 1.
Fig. 10 is a graph showing the repairing effect of each treatment group after the treatment of the scalded wound surface of the mice.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Example 1 preparation of Artificial Stem cell vesicle hydrogel with wound repair promoting function
(1) Synthesis of AIE molecules (THB): 4-methylpyridine (5 mmol, 487. Mu.L, commercially available) and 2-bromoethanol (5 mmol, 354. Mu.L, commercially available) were heated in acetonitrile at 50℃for 24h, most of the solvent was removed by spinning, 5- (4- (diphenylamino) phenyl) thiophene-2-carbaldehyde (142 mg,0.4mmol, commercially available) was added, redissolved with ethanol, and a catalytic amount of piperidine was added, and the mixture was heated to 80℃and stirred under nitrogen for 16 h. After removal of the solvent by rotary evaporator, the product was purified by neutral alumina column using dichloromethane/methanol (v/v=4/1) as eluent to give a red solid, THB (i.e. AIE molecule). The AIE molecule is prepared as follows:
(2) Preparation of AIE vesicles: the method comprises the following steps of obtaining membrane suspension by splitting artificial stem cells, extruding the membrane suspension by a liposome extruder, and obtaining artificial stem cell vesicles, wherein the preparation steps are as follows: extracting primary adipose-derived stem cells from rat adipose tissue, subculturing in vitro, collecting cell precipitate, and adding hypotonic cell lysis buffer (every 5×10) 6 About 100ul of lysate is added to each cell), after the cells are fully lysed overnight at 4 ℃, the cell suspension is subjected to 20 regrind using a tissue mill pestle, centrifuged at 3500g for 5min at 4 ℃, the precipitate removed, and the supernatant retained; the artificial stem cell vesicles are obtained by passing a Millipore Polycarbonate (PCTE) membrane with a pore diameter of 800nm through a liposome extruder, repeatedly extruding and sieving to complete the preliminary reconstruction of cell membranes and remove the solution obtained by impurity collection;
adding a small amount of the DMSO solution of the AIE molecules obtained in the step (1) into the artificial stem cell vesicles, wherein the mass ratio of the AIE molecules to the artificial stem cell vesicles is 1:5, after 30 minutes incubation, the final reconstitution of the vesicles was accomplished by passing through a Millipore Polycarbonate (PCTE) membrane with a pore size of 200nm using a liposome extruder, reciprocating extrusion sieving. Purifying the obtained vesicles by using a gradient centrifugation method or a 20kDa ultrafiltration membrane, removing components which are not wrapped in vesicle structures, and obtaining a pure AIE functionalized adipose stem cell vesicle solution (namely AIE vesicles or THB vesicles); wherein the final concentration of AIE in AIE vesicles was found to be about 125. Mu. Mol/L.
(3) Preparation of artificial stem cell vesicle hydrogel with function of promoting wound repair: PEG-4000 (10 g,2.5mmol, commercially available) and excess propiolic acid (1.75 g,25mmol, commercially available) and catalytic amounts of p-toluenesulfonic acid (0.29 g,1.5mmol, commercially available) were dissolved in toluene, the mixed solution was stirred and refluxed for 48h, and the water produced was continuously removed. And concentrating the solution under vacuum, removing most of the solvent, precipitating the product with diethyl ether, recrystallizing with isopropanol, and drying in vacuum to obtain the target product PEG4000-DA (water-soluble compound containing ester alkynyl). The preparation equation for PEG4000-DA is shown below:
mixing water-soluble chitosan (CMC) with equal functional group ratio (namely functional group ratio is 1:1) and a cross-linking agent (PEG 4000-DA) to obtain a hydrogel carrier, adding the hydrogel carrier into PBS suspension of AIE vesicles (wherein the concentration of the AIE vesicles is 40 mug/mL and the mass percentage concentration of the hydrogel carrier is 4%), dissolving, standing and curing for 8 hours, and obtaining the artificial stem cell vesicle hydrogel with the function of promoting wound repair. Wherein, the equations of CMC and PEG4000-DA for amine-alkyne click reaction are as follows:
example 2
The difference between this embodiment and embodiment 1 is that: in the step (2), the mass ratio of AIE molecules to artificial stem cell vesicles is 1:50.
example 3
The difference between this embodiment and embodiment 1 is that: in the step (2), the mass ratio of AIE molecules to artificial stem cell vesicles is 1:3.
example 4
The difference between this embodiment and embodiment 1 is that: in step (2), the final AIE concentration in AIE vesicles was found to be about 400. Mu. Mol/L.
Example 5
The difference between this embodiment and embodiment 1 is that: in the step (2), the mass ratio of AIE molecules to artificial stem cell vesicles is 1:30.
comparative example 1
The difference between this comparative example and example 1 is that: and (3) adding no AIE molecule in the step (2), and directly mixing the AIE molecule with water-soluble chitosan (CMC) and a cross-linking agent (PEG 4000-DA) with equal functional group ratio in redundant PBS suspension after the artificial stem cell vesicle is obtained.
Comparative example 2
The difference between this comparative example and example 1 is that: in this comparative example, TCB molecules (TCB molecules belonging to one of AIE molecules having the structural formula shown below) were used instead of THB molecules in example 1 to prepare TCB vesicles.
Comparative example 3
The difference between this comparative example and example 1 is that: the final AIE concentration in AIE vesicles was found to be about 500. Mu. Mol/L. It can be observed from the preparation process that the final concentration of AIE is too high, exceeding the load limit, AIE cannot be well dispersed, and a solid precipitate is precipitated.
Test example 1
The detection of THB (AIE molecule) in step (1) of example 1 using nmr hydrogen spectroscopy, as shown in the nmr hydrogen spectroscopy of fig. 1, shows that THB was successfully prepared.
Characterization of vesicles in example 1 step (2) Using Transmission Electron microscopyFIG. 2 is a representation of the vesicles of example 1, as shown in FIG. 2 (a), showing the formation of a 100-200 nm sized teacup support, demonstrating successful preparation of vesicles; from the NTA test of FIG. 2 (b), it was calculated that the vesicle yield was 1.99X10 10 /10 6 And (3) cells. The protein concentration of vesicles detectable by BCA assay was about 201 μg/10 10 。
PEG4000-DA in step (3) of example 1 was detected using nuclear magnetic resonance hydrogen spectroscopy. FIG. 3 is a graph of PEG4000-DA of example 1, as shown in the hydrogen nuclear magnetic resonance spectrum of FIG. 3, illustrating successful preparation of PEG 4000-DA.
The hydrogel prepared in example 1 was characterized by transmission electron microscopy, as shown in fig. 4, in which uniformly distributed porosity was observed, demonstrating the successful preparation of the hydrogel crosslinked network.
Test example 2 antibacterial test of Artificial Stem cell vesicle hydrogel with wound repair promoting function
The antibacterial properties of the THB vesicles in step (2) of example 1 and the TCB vesicles in comparative example 2 were tested as follows: preparation of AIE vesicles, TCB vesicles and 10 at different concentrations in 48 well plates 6 Incubating the mixed solution of CFU/mL bacterial liquid for 10min, and then using 100mW/cm 2 Is irradiated by white light for 10min, and then the bacterial liquid is diluted to 10 4 CFU/mL, 100. Mu.L of the mixture was applied to LB plates to observe colony growth.
FIG. 5 is a graph showing the antimicrobial ability of THB vesicles against Staphylococcus aureus and Escherichia coli, and as can be seen from FIG. 5, AIE vesicles exhibit significant dark toxicity against gram-positive Staphylococcus aureus, and have enhanced antimicrobial ability after illumination; meanwhile, the antibacterial agent also has good antibacterial capability on gram-negative escherichia coli under the illumination condition.
Fig. 6 is a graph showing the antibacterial ability of TCB vesicles against staphylococcus aureus and escherichia coli, and as can be seen from fig. 6, TCB vesicles show significant dark toxicity against gram-positive staphylococcus aureus, and the antibacterial ability after illumination is enhanced, however, the antibacterial effect is significantly worse than THB vesicles in example 1. While TCB vesicles do not exhibit significant killing capacity against gram-negative escherichia coli. It can be seen that the THB vesicles in example 1 are killing against both gram negative and gram positive bacteria, whereas the TCB vesicles in comparative example 2 are killing against only gram positive bacteria.
The same antibacterial experiments are carried out by adopting the artificial stem cell vesicle hydrogel with the function of promoting wound repair, which is prepared in the examples 2-5, and the same effect is achieved.
Test example 3 cell proliferation and scratch test of Artificial Stem cell vesicle hydrogel with wound repair promoting function
The AIE vesicles of step (2) of example 1 and the vesicles of comparative example 1 were subjected to a test for promoting cell proliferation, specifically as follows: haCaT cells were grown at 2.0X10 4 Cell density/mL was inoculated in 96-well plates, treated with AIE vesicles from step (2) of example 1 and vesicles from comparative example 1 after incubation for 24 hours, CCK-8 reagent was added at 24 hours, and absorbance was measured at 450nm after incubation for 1 hour at 37 ℃.
The AIE vesicles in step (2) of example 1 and the vesicles in comparative example 1 were subjected to migration performance test, specifically as follows: the wound healing effect of the AIE vesicles in step (2) of example 1 and the vesicles in comparative example 1 was evaluated in vitro using a scratch test. HaCaT cells were grown at 1.0X10 5 Cell density was seeded in 6-well plates, cultured to 90%, parallel scratches were formed in each well with a 200 μl pipette tip, washed once with PBS, and then treated with AIE vesicles in step (2) of example 1, vesicles in comparative example 1, or PBS, respectively. Photographs were taken using an optical microscope after 12 hours, 24 hours. A blank group and an AIE molecular control group were simultaneously set.
FIG. 7 is a statistical result of promotion of cell proliferation by AIE vesicles in step (2) of example 1 and vesicles in comparative example 1. As can be seen from fig. 7, the cell number significantly increased in a dose-dependent manner, and the AIE vesicles in step (2) of example 1 and the vesicles in comparative example 1 had a significantly cell proliferation promoting function.
FIG. 8 shows scratch tests of AIE vesicles in step (2) of example 1 and vesicles in comparative example 1. As can be seen from fig. 8, the AIE vesicles in example 1, step (2) and the vesicles in comparative example 1 have a significant promoting effect on cell migration compared to the blank group and the AIE molecule control group, and the addition of the AIE molecule does not adversely affect cell proliferation and migration.
Test example 4 acid response sustained release test of Artificial Stem cell vesicle hydrogel with wound repair promoting function
The same volume of the photodynamic anti-infective artificial stem cell vesicle hydrogel of example 1 was immersed in PBS buffer solutions with pH values of 6, 7 and 8, respectively, sampled at different intervals for absorption measurement, and converted into the concentration of the released AIE molecules, thereby calculating the percentage of the gel to the slow release of the AIE vesicles.
FIG. 9 is a graph depicting the sustained release capacity of the photodynamic anti-infective artificial stem cell vesicle hydrogel prepared in example 1 in terms of acid response. As can be seen from fig. 9, the proportion of hydrogel to AIE vesicle sustained release was significantly increased in the acidic environment at pH 6 relative to pH 7 and pH 8, demonstrating that the targeted release of photodynamic anti-infective artificial stem cell vesicle hydrogel at the wound site was facilitated in the acidic environment.
Test example 5 wound treatment test of Artificial Stem cell vesicle hydrogel with wound repair promoting function
The specific operation steps of the construction of the wound surface of the rat scalds are as follows: the deep II degree standard scald wound surface molding of the rat is carried out by using a scald instrument on the 0 th day, the setting condition is 500g,95 ℃ and 15s, the scald epidermis is yellow and hardened, meanwhile, the tissue slice can be fused into a piece by the collagen fiber, most sebaceous glands, sweat glands and hair follicles except the deep dermis are destroyed, and the molding is proved to be successful. Treating scalded wound on day 1, cutting off the scalded necrotic skin, dividing the wound into 4 groups, and respectively applying a blank hydrogel with the same area as the wound and the thickness of 2mm, the vesicle-containing hydrogel prepared in comparative example 1 or the AIE vesicle-containing hydrogel prepared in example 1 to the other 3 groups except that the control group is treated with normal saline only; AIE vesicle hydrogel group was incubated for 12h on wound surface with 100mW/cm 2 The white light irradiation is carried out for 10min for photodynamic sterilization. Then, gel is replaced again and photodynamic therapy is carried out on the 3 rd, 7 th, 14 th and 21 th days of treatment respectively, and wound recovery conditions on the 7 th and 14 th days are recorded.
Fig. 10 is a graph showing the healing promotion effect of the scalded wound surface of each treatment group, and compared with the control group and the blank hydrogel group, the encapsulated vesicle or THB vesicle can obviously improve the healing speed of the wound surface, the area of the wound surface of the hydrogel group containing the vesicle or THB vesicle can be observed to be obviously smaller than that of the other two groups on the 7 th day of the healing of the wound surface, and the healing promotion capability of the vesicle components encapsulated by the hydrogel on the living body layer is proved; meanwhile, the thickness and distribution of wound infection bacteria of the THB vesicle group are obviously improved, and the THB can be proved to have killing and inhibiting effects on bacteria on the wound by photodynamic therapy. The results show that the repair promoting material has potential application value in the field of scald wound treatment.
The artificial stem cell vesicle hydrogel with the function of promoting wound repair prepared in examples 2-5 is used for carrying out the same wound treatment test, and has the same effect.
The foregoing examples are illustrative only and serve to explain some features of the method of the invention. The claims that follow are intended to claim the broadest possible scope as conceivable and the embodiments presented herein are demonstrated for the applicant's true test results. It is, therefore, not the intention of the applicant that the appended claims be limited by the choice of examples illustrating the features of the invention. Some numerical ranges used in the claims also include sub-ranges within which variations in these ranges should also be construed as being covered by the appended claims where possible.
Claims (10)
1. A photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair, which is characterized by comprising a hydrogel carrier and AIE vesicles loaded in the hydrogel carrier; the AIE vesicle is an AIE molecule with an outer surface wrapped with an artificial stem cell vesicle;
the mass ratio of the AIE molecules to the artificial stem cell vesicles is in the range of 1: 3-50, wherein the maximum final concentration of AIE molecules in the AIE vesicle is not more than 400 mu mol/L.
2. The hydrogel of claim 1, wherein the AIE molecule has the formula (I):
wherein R is - To balance the charge of anions selected from Cl - 、Br - 、I - 、PF6 - One of them.
3. The hydrogel according to claim 1, wherein the hydrogel carrier is prepared from natural water-soluble polymer compounds containing amine groups or sulfhydryl groups and water-soluble compounds containing ester alkynyl groups according to a functional group ratio of 1-10: 10-1 is prepared by click chemistry reaction.
4. The hydrogel according to claim 3, wherein the natural water-soluble polymer compound containing amine groups or thiol groups is selected from one or more of chitosan and its derivatives, sodium carboxymethyl cellulose.
5. The hydrogel of claim 3, wherein the ester-alkynyl containing water-soluble compound has one or more of the structures of any of the following formulas (II):
wherein n is an integer of 1 or more.
6. The hydrogel of claim 1, wherein the artificial stem cell vesicles are derived from one or more of adipose stem cells, fibroblasts, epidermal stem cells.
7. The hydrogel of claim 1, wherein the mass ratio of AIE vesicles to hydrogel carrier is 0.67-10: 1.
8. the method for preparing the photodynamic anti-infective artificial stem cell vesicle hydrogel for promoting wound repair according to any one of claims 1-7, which is characterized by comprising the following steps:
s1, splitting the artificial stem cells to obtain membrane suspension, and extruding the membrane suspension through a liposome extruder to obtain artificial stem cell vesicles;
s2, after the artificial stem cell vesicle is incubated with the AIE molecules, the artificial stem cell vesicle is extruded by a liposome extruder to realize the coating of the artificial stem cell vesicle on the AIE molecules, so as to obtain the AIE vesicle;
s3, adding an aqueous solution of AIE vesicles or a PBS (phosphate buffered saline) solution of the AIE vesicles into a precursor solution of the hydrogel carrier, standing and curing to obtain the photodynamic anti-infection artificial stem cell vesicle hydrogel for promoting wound repair.
9. The method according to claim 8, wherein in the step S3, the AIE vesicle is contained in an aqueous solution or a PBS solution of AIE vesicles, and the concentration of AIE vesicles is in the range of 1 to 300. Mu.g/mL; the precursor solution of the hydrogel carrier is prepared from 3-15% of the hydrogel carrier by mass percent.
10. Use of the photodynamic anti-infective artificial stem cell vesicle hydrogel for promoting wound repair according to any one of claims 1-7 in preparing wound repair material.
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