CN115337255A - Self-electrogenesis enzyme-linked microneedle patch and preparation method and application thereof - Google Patents
Self-electrogenesis enzyme-linked microneedle patch and preparation method and application thereof Download PDFInfo
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- CN115337255A CN115337255A CN202211057304.2A CN202211057304A CN115337255A CN 115337255 A CN115337255 A CN 115337255A CN 202211057304 A CN202211057304 A CN 202211057304A CN 115337255 A CN115337255 A CN 115337255A
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- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
- A61K9/0021—Intradermal administration, e.g. through microneedle arrays, needleless injectors
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/44—Oxidoreductases (1)
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/44—Oxidoreductases (1)
- A61K38/443—Oxidoreductases (1) acting on CH-OH groups as donors, e.g. glucose oxidase, lactate dehydrogenase (1.1)
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
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- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
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- C—CHEMISTRY; METALLURGY
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Abstract
The invention relates to the field of biomedicine, and particularly discloses a self-electrogenesis enzyme cascade microneedle patch and a preparation method and application thereof, wherein the microneedle patch consists of a microneedle array consisting of a conductive substrate without a target medicament and at least one needle body containing the target medicament, the substrate is prepared from polypyrrole modified by polydopamine, hyaluronic acid and polyvinyl alcohol, the needle body is prepared from polypyrrole modified by polydopamine, hyaluronic acid and polyvinyl alcohol, glucose oxidase and horseradish peroxidase/catalase, an external power supply is not required to be provided for the self-electrogenesis enzyme cascade microneedle patch, physiological current can be generated through enzyme cascade reaction, migration and growth of various cells are promoted, and the self-electrogenesis enzyme cascade microneedle patch has the advantages of safety, effectiveness, portability, low cost, high efficiency, simplicity and convenience in operation and the like, has good biocompatibility, no in vivo toxicity and has a wide application prospect in the field of biomedicine.
Description
Technical Field
The invention relates to the technical field of biomedical materials, in particular to a self-generating enzyme-linked microneedle patch as well as a preparation method and application thereof.
Background
The micro-needle is a novel transdermal drug delivery system, when the drug is delivered, the micro-needle can pierce the stratum corneum of the skin, so that a micro-channel from the stratum corneum to the epidermis layer or the dermis shallow layer is opened for the delivery of the drug, and then the drug is absorbed by capillary vessels, and the drug active molecules reach the focus part of the human body along with the blood circulation to play a role in treatment. The micro-needle administration has the advantages of micro-wound, no pain, convenience and the like. In order to promote the release of drugs in the microneedles, the bioelectricity conductive material is added into the microneedles, and the prepared microneedles have good in-vitro stability, excellent electrochemical performance and biocompatibility, and are concerned in aspects of biomedical sensors, neural prostheses, wound dressings, drug controlled release devices and the like.
For example, the invention patent CN110201296a discloses a conductive polymer microneedle patch for controllable drug release, wherein the microneedle is a conductive polymer microneedle, and comprises a polymer solid microneedle, an inert metal layer and a drug-carrying conductive polymer film layer; controlled release of the drug is achieved by electrical stimulation. However, the micro-needle needs to provide an external power supply, and the outer layer of the micro-needle is provided with the inert metal layer and the drug-loaded conductive polymer film layer, so that the preparation method is complex, and the plating layer is easy to drop; in the earlier research of the subject group of the inventor, a double-layer conductive microneedle patch for treating subcutaneous tumors and a preparation method and application thereof are developed (CN 114848577A), and conductive polymer materials adopted at the substrate and the microneedle tips close to the substrate provide an external power supply, so that electroporation is facilitated, the cell membrane permeability is increased, and the biological agent or drug absorption rate is increased; however, the microneedle patch also requires an external power source, and is inconvenient to use. In addition, the conventional conductive microneedle patch found by the inventors during the research process has the following problems: the conductive polymer PPy most commonly used for preparing the conductive micro-needle has poor dispersibility in water, so that the conductive micro-needle is difficult to manufacture.
In order to solve the technical problems, the invention prepares an advanced self-electrogenic enzyme-linked microneedle patch system, the self-electrogenic enzyme-linked microneedle patch does not need to provide an external power supply, can be communicated with enzyme-linked reaction to generate physiological current, promotes the migration and growth of various cells, has the advantages of safety, effectiveness, portability, low cost, high efficiency, simple and convenient operation and the like, has good biocompatibility, no in-vivo toxicity and wide application prospect in the field of biomedicine.
Disclosure of Invention
In order to achieve the above object, a primary object of the present invention is to provide an electrogenic enzyme cascade microneedle patch, which comprises a microneedle array comprising a conductive substrate without a target drug and at least one needle body containing the target drug, wherein the substrate is made of poly-dopamine-modified polypyrrole (DA-PPy), hyaluronic Acid (HA) and polyvinyl alcohol (PVA), and the needle body is made of poly-dopamine-modified polypyrrole (DA-PPy), hyaluronic Acid (HA) and polyvinyl alcohol (PVA), glucose oxidase, horseradish peroxidase/catalase.
Preferably, the glucose oxidase and the horseradish peroxidase/catalase are fixed on the metal organic framework material ZIF-8.
Preferably, in the substrate, the mass ratio of the dopamine-modified polypyrrole (DA-PPy), the Hyaluronic Acid (HA) and the polyvinyl alcohol (PVA) is 0.01-1: 1 to 100: 0.5-50, wherein in the needle body, the mass ratio of dopamine-modified polypyrrole (DA-PPy), hyaluronic Acid (HA), polyvinyl alcohol (PVA), glucose oxidase, horseradish peroxidase/catalase and ZIF-8 is 0.01-1: 1 to 100:0.5 to 50:0.005 to 0.5: 0.005-0.5.
The second purpose of the invention is to provide a preparation method of the self-electrogenesis enzyme-linked microneedle patch, which comprises the following steps:
(1) Under the conditions of water bath and magnetic stirring, proportionally adding polydopamine-modified polypyrrole, hyaluronic acid and polyvinyl alcohol into deionized water to obtain microneedle patch substrate gel;
(2) Mixing the microneedle patch substrate gel prepared in the step (1) with glucose oxidase to obtain glucose oxidase needle body gel; mixing with horse radish peroxidase/catalase to obtain horse radish peroxidase/catalase needle gel;
(3) Sucking 5-20ul of glucose oxidase needle gel prepared in the step (2) by using a pipette, dripping the glucose oxidase needle gel on a half pinhole of a PDMS microneedle template, and putting the PDMS microneedle template into a vacuum drying oven for vacuum negative pressure injection molding at a pressure of-1 to-0.8 MPa: repeatedly vacuumizing for 2-3 times, each time for 3-5 min, and naturally drying at room temperature to obtain a microneedle template carrying glucose oxidase;
(4) Sucking 5-20ul of the prepared horseradish peroxidase/catalase needle body gel in the step (2) by using a pipette, dripping the horseradish peroxidase/catalase needle body gel on the other half of the pinholes of the microneedle template in the step (3), and putting the microneedle template into a vacuum drying oven for vacuum negative pressure injection molding at-1 to-0.8 MPa: repeatedly vacuumizing for 2-3 times, each time for 3-5 min, and naturally drying at room temperature to obtain a glucose oxidase-horse radish peroxidase/catalase-loaded conductive microneedle patch needle body;
(5) Placing the substrate gel prepared in the step (1) on the microneedle body template obtained in the step (4), placing the microneedle body template in a vacuum drying box again, setting the vacuum to be-1 to-0.8 MPa, performing negative pressure injection molding and vacuumizing for 10-30 min, and naturally drying at room temperature to obtain the self-electrogenesis enzyme cascade microneedle patch needle body;
(6) And (3) paving the conductive hydrogel prepared in the step (1) on the microneedle template of which the needle body is filled with the conductive polymer matrix gel in the step (5), and naturally drying to obtain the self-electrogenesis enzyme-linked microneedle patch.
Preferably, the temperature of the water bath in the step (1) is 0-90 ℃.
The third purpose of the invention is to provide the application of the self-generating enzyme cascade microneedle patch in preparing a medicine for treating local diseases.
The fourth purpose of the invention is to provide the application of the self-generating enzyme cascade microneedle patch in preparing a medicament for treating skin injury.
The fifth purpose of the invention is to provide the application of the self-generating enzyme cascade microneedle patch in preparing a medicament for treating nerve injury.
The invention has the beneficial effects that: (1) The invention provides a self-generating enzyme-linked microneedle patch, which can generate physiological current by enzyme-linked reaction without adding external electrical stimulation equipment; (2) According to the self-electrogenesis enzyme cascade microneedle patch, a base material is polydopamine modified (DA) polypyrrole (PPy) is added into Hyaluronic Acid (HA) and polyvinyl alcohol (PVA) gel to prepare a polymer (DPPH) with good conductivity, one half of a needle body is doped with ZG nano particles with ZIF-8 fixed glucose oxidase, the other half of the needle body is doped with ZH nano particles with ZIF-8 fixed horseradish peroxidase, ZIF-8 can protect the activity and stability of enzyme, and ZIF-8 solves the problem that free enzyme is easy to inactivate and denature under strong acid, strong base and high temperature to influence the catalytic effect of the free enzyme; the reuse rate of free enzyme is not high, the purification and separation process is complex, and the use cost is high. (3) The ZG released after absorbing body fluid and swelling after acting on a human body by the self-generating enzyme cascade microneedle patch can quickly generate oxidation-reduction reaction when meeting a substrate to generate hydrogen peroxide which can be reduced into water and oxygen by ZH released from the other half of the needle body, and electrons generated in the process can be transferred through DPPH hydrogel to form a complete current path so as to promote the migration and growth of various cells; (4) The prepared self-electricity-generating enzyme cascade microneedle patch is sensitive to the blood glucose concentration of a diabetic wound surface, can quickly respond, generates an enzyme cascade reaction, reduces local blood glucose, and can output physiological micro-current, so that the wound healing is better promoted; the ZGH-MN microneedle patch group realizes scar-free healing after 21 days, and other groups of skin surfaces still have small wounds or obvious scars on the skin surfaces after healing. (5) The self-generating enzyme-linked microneedle patch has good antibacterial performance; (6) The self-generating enzyme cascade microneedle patch has a good effect on repairing the sciatic nerve; (7) The microneedle patch has the advantages of safety, effectiveness, portability, low cost, high efficiency, simple and convenient operation and the like, and has wide application prospect in the biomedical field such as chronic wounds of diabetes and the like, nerve repair, myocardial repair, body fluid detection and diagnosis and detection of various physiological indexes.
Drawings
FIG. 1 is a diagram of a self-electrogenic enzyme-linked microneedle patch
FIG. 2 is a graph showing the difference between the dispersibility and solubility of PPy and DA-PPy in water
FIG. 3 scanning Electron microscopy of PPy and DAPPy
FIG. 4 self-electrogenic performance diagram of self-electrogenic enzyme-linked microneedle patch
FIG. 5 is a graph of the antibacterial results of ZG/ZH nanoparticles at different concentrations
FIG. 6 treatment of diabetic rat wound by electrogenic cascade microneedle patch
FIG. 7 repair of sciatic nerve by electrogenic enzyme cascade microneedle patch
FIG. 8 staining of HE section of gastrocnemius muscle by electrogenic cascade microneedle patch
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the following examples, glucose oxidase is immobilized on the metal organic framework material ZIF-8 and abbreviated as ZG nanoparticles, and horseradish peroxidase is immobilized on the metal organic framework material ZIF-8 and abbreviated as ZH nanoparticles.
Example 1 preparation of microneedle templates
(1) Preparing a microneedle mould by using a PDMS transfer process, wherein the solid-liquid mass ratio of PDMS is 10;
(2) And (5) placing the poured mould into a vacuum drying oven for drying for later use.
Example 2 preparation of ZIF-8, ZG and ZH nanoparticles
(1) Adding Zn (NO) 3 ) 2 ·6H 2 A mixture of O (40 mg) and 2-methylimidazole (0.77 g) was dissolved in 5 mlDeionized (DI) water. They were then stirred at room temperature for 5 minutes, centrifuged to obtain pale yellow nanocrystals, washed several times with deionized water, and dried to obtain ZIF-8 nanoparticles.
(2) Zn (NO) 3 ) 2 ·6H 2 A mixture of O (40 mg), 2-methylimidazole (0.77 g) and Glucose Oxidase (GOX) was dissolved in 5 ml of Deionized (DI) water. They were then stirred at room temperature for 5 minutes, centrifuged to obtain pale yellow nanocrystals, washed several times with deionized water, and dried to obtain ZG nanoparticles.
(3) Adding Zn (NO) 3 ) 2 ·6H 2 A mixture of O (40 mg), 2-methylimidazole (0.77 g) and horseradish peroxidase (HRP) was dissolved in 5 ml Deionized (DI) water. They were then stirred at room temperature for 5 minutes, centrifuged to obtain pale yellow nanocrystals, washed several times with deionized water, and dried to obtain ZH nanoparticles.
Example 3 preparation of self-electrogenic enzyme-linked microneedle Patch (ZGH-MN)
(1) The PDMS microneedle template prepared in example 1 was used;
(2) Weighing 0.01g of DA-PPy, 1g of Hyaluronic Acid (HA) and 0.5g of polyvinyl alcohol (PVA), dissolving in 10ml of deionized water, stirring for 24 hours under magnetic stirring to ensure sufficient swelling, and obtaining DPPH microneedle substrate gel;
(3) And mixing the hydrogel polymer matrix solution with ZG and ZH nanoparticles to obtain the drug-loaded needle body gel, wherein the mass of the added ZG and ZH is 5mg.
(4) And (3) sucking 5-20ul of the prepared ZG needle body gel in the step (3) by using a pipette, dripping the prepared ZG needle body gel on a half pinhole of a PDMS micro needle template, and putting the PDMS micro needle template into a vacuum drying oven for vacuum-1 MPa negative pressure injection molding: repeatedly vacuumizing for 3 times, 5min each time, and naturally drying at room temperature to obtain a microneedle template carrying ZG;
(5) Sucking 10ul of ZH needle body gel prepared in the step (3) by using a pipette, dripping the ZH needle body gel on the other half of needle holes of the microneedle template in the step (4), and putting the ZH needle body gel into a vacuum drying box for vacuum-1 MPa negative pressure injection molding: repeatedly vacuumizing for 3 times, each time for 3-5 min, and naturally drying at room temperature to obtain a needle tip ZG-ZH conductive microneedle patch (shown in figure 1);
(6) Placing the gel solution prepared in the step (2) on the microneedle template with the needle point loaded with the ZG-ZH nanoparticles in the step (5), placing the microneedle template into a vacuum drying box again, setting vacuum-1 MPa negative pressure for injection molding and vacuumizing for 30min, and naturally drying at room temperature to obtain the electrogenic enzyme cascade microneedle patch with the needle point filled with the ZG-ZH nanoparticles;
(7) Spreading the soluble polymer matrix solution prepared in the step (2) on the microneedle template with the needle point filled with the conductive polymer matrix gel in the step (6), and naturally drying to obtain the formed self-electrogenic enzyme-linked microneedle patch;
example 4 preparation of ZIF-8 nanoparticle-loaded conductive microneedle Patch (ZIF-MN)
(1) The PDMS microneedle template prepared in example 1 was used;
(2) Weighing 0.01g of DA-PPy, 1g of Hyaluronic Acid (HA) and 0.5g of polyvinyl alcohol (PVA), dissolving in 10ml of deionized water, stirring for 24 hours under magnetic stirring to ensure sufficient swelling, and obtaining DPPH microneedle substrate gel;
(3) And mixing the hydrogel polymer matrix solution with ZIF-8 nanoparticles to obtain drug-loaded needle gel, wherein the mass of ZIF-8 is 5mg.
(4) Sucking 10ul of the prepared ZIF-8 needle body gel in the step (3) by using a pipette, dripping the gel on micropores of a PDMS micro needle template, and putting the micro needle template into a vacuum drying oven for vacuum-1 MPa negative pressure injection molding: repeatedly vacuumizing for 3 times, 5min each time, and naturally drying at room temperature to obtain a ZIF-8-loaded microneedle template;
(5) Placing the gel solution prepared in the step (2) on the microneedle template with the needle point loaded with the ZIF-8 nanoparticles in the step (4), placing the microneedle template into a vacuum drying oven again, setting vacuum-1 MPa negative pressure for injection molding and vacuumizing for 30min, and naturally drying at room temperature to obtain a conductive microneedle patch with the needle point filled with the ZIF-8 nanoparticles;
(6) Paving the soluble polymer matrix solution prepared in the step (2) on a microneedle template with the conductive polymer matrix gel filled in the needle point in the operation of the step (5), and naturally drying to obtain a formed conductive microneedle patch loaded with ZIF-8 nano particles;
example 5 preparation of conductive microneedle Patch carrying ZG nanoparticles (ZG-MN)
(1) The PDMS microneedle template prepared in example 1 was used;
(2) Weighing 0.01g of DA-PPy, 1g of Hyaluronic Acid (HA) and 0.5g of polyvinyl alcohol (PVA), dissolving in 10ml of deionized water, stirring for 24 hours under magnetic stirring to ensure sufficient swelling, and obtaining DPPH microneedle substrate gel;
(3) And mixing the hydrogel polymer matrix solution with ZG nanoparticles to obtain the drug-loaded needle body gel, wherein the mass of ZG is 5mg.
(4) Sucking 10ul of the prepared ZG needle body gel in the step (3) by using a pipette, dripping the prepared ZG needle body gel on micropores of a PDMS micro needle template, and putting the micro needle template into a vacuum drying oven for vacuum-1 MPa negative pressure injection molding: repeatedly vacuumizing for 3 times, 5min each time, and naturally drying at room temperature to obtain a micro-needle template loaded with ZG;
(5) Placing the gel solution prepared in the step (2) on the microneedle template with the needle point loaded with the ZG nano-particles in the step (4), placing the microneedle template into a vacuum drying oven again, setting vacuum-1 MPa negative pressure for injection molding and vacuumizing for 30min, and naturally drying at room temperature to obtain a conductive microneedle patch with the needle point filled with the ZG nano-particles;
(6) Flatly paving the soluble polymer matrix solution prepared in the step (2) on a microneedle template with the needle point filled with the conductive polymer matrix gel in the operation of the step (5), and naturally drying to obtain a formed conductive microneedle patch loaded with ZG nano particles;
example 6 preparation of conductive microneedle Patch carrying ZH nanoparticles (ZH-MN)
(1) The first operation is implemented: the PDMS microneedle template prepared in example 1 was used;
(2) Weighing 0.01g of DA-PPy, 1g of Hyaluronic Acid (HA) and 0.5g of polyvinyl alcohol (PVA), dissolving in 10ml of deionized water, stirring for 24 hours under magnetic stirring to ensure sufficient swelling, and obtaining DPPH microneedle substrate gel;
(3) And mixing the hydrogel polymer matrix solution with ZH nanoparticles to obtain a drug-loaded needle body gel, wherein the mass of ZH is 5mg.
(4) Sucking 10ul of ZH needle body gel prepared in the step (3) by using a pipette, dripping the ZH needle body gel on micropores of a PDMS micro needle template, and putting the gel into a vacuum drying oven for vacuum-1 MPa negative pressure injection molding: repeatedly vacuumizing for 3 times, 5min each time, and naturally drying at room temperature to obtain a ZH-loaded microneedle template;
(5) Placing the gel polymer matrix solution prepared in the step (2) on the microneedle template with the needle point loaded with the ZH nanoparticles in the step (4), placing the microneedle template into a vacuum drying oven again, setting vacuum-1 MPa negative pressure for injection molding and vacuumizing for 30min, and naturally drying at room temperature to obtain a conductive microneedle patch with the needle point filled with the ZH nanoparticles;
(6) Flatly paving the hydrogel polymer matrix solution prepared in the step (2) on the microneedle template with the conductive polymer matrix gel filled in the needle point in the step (5), and naturally drying to obtain the formed conductive microneedle patch loaded with the ZH nanoparticles;
example 7 preparation of DPPH conductive microneedle Patch (DPPH-MN)
(1) The PDMS microneedle template prepared in example 1 was used;
(2) Weighing 0.01g of DA-PPy, 1g of Hyaluronic Acid (HA) and 0.5g of polyvinyl alcohol (PVA), dissolving in 10ml of deionized water, stirring for 24 hours under magnetic stirring to ensure sufficient swelling, and obtaining blank microneedle substrate gel;
(3) Sucking 10ul of the prepared needle body gel in the step (2) by using a pipette, dripping the prepared needle body gel on micropores of a PDMS (polydimethylsiloxane) microneedle template, and putting the prepared needle body gel into a vacuum drying oven for vacuum-1 MPa negative pressure injection molding: repeatedly vacuumizing for 3 times, each time for 5min, and naturally drying at room temperature to obtain a DPPH conductive microneedle template;
(4) Placing the gel polymer matrix solution prepared in the step (3) on the microneedle template with the needlepoint filled with DPPH in the step (3), placing the microneedle template into a vacuum drying oven again, setting vacuum-1 MPa negative pressure for injection molding and vacuumizing for 30min, and naturally drying at room temperature to obtain a conductive microneedle patch with the needlepoint filled with conductive polymer matrix solution;
(5) Spreading the polymer gel matrix solution prepared in the step (2) on the microneedle template with the needle point filled with the conductive polymer matrix gel in the step (4), and naturally drying to obtain a formed DPPH conductive microneedle patch, namely BLANK-MN in the following experimental example;
experimental example I comparison of dispersibility and solubility of PPy and DA-PPy in Water
Shaking and dispersing the prepared PPy and DA-PPy in deionized water at the same concentration (10 mg/mL), standing, and observing the difference immediately after standing, after standing for 1min and after standing for 5 min. And simultaneously, the PPy and DA-PPy are represented by a scanning electron microscope, and the change of the micro-morphology before and after the modification of the polydopamine is observed.
The experimental results show that: as shown in FIG. 2, PPy has poor solubility in aqueous solution, and dispersion is unstable and is not uniform soon; and the DAPPy has better solubility and stability in aqueous solution and can be uniformly dispersed for a long time. As shown in fig. 3, where (a) and (b) are PPy and (c) and (d) are DAPPy, the scale of (a, c) is 1 μm, and the scale of (b, d) is 100nm, it can be seen that DA modification can change the morphology of PPy from aggregated particles to a short fibrous structure, which will improve its hydrophilic dispersion and facilitate the fabrication of microneedles.
Experimental example II, test of current generation effect of self-electrogenic enzyme-linked microneedle patch
The prepared self-electrogenesis enzyme-linked microneedle patch ZGH-MN is placed in a culture dish, glucose solutions with different concentrations (0/18/20/24/26 mmol/L) are dripped every 1 minute, body fluid environments with different blood glucose concentrations are simulated, a high-precision desk type digital multimeter is used for measuring, and output current is recorded at the same time.
The experimental results are as follows: as shown in fig. 4, as the glucose concentration increases, the output current of the microneedles also increases. When the concentration of glucose is 0/18/20/24/26mmol/L, the output current generated by the microneedle patch through enzyme cascade reaction is divided into 1.01 muA, 1.74 muA, 2.01 muA, 2.32 muA, 2.64 muA and 3.53 muA, and the result shows that the homemade microneedle patch is sensitive to the blood glucose concentration of the diabetic wound and can quickly respond. The enzyme cascade reaction is generated, the physiological micro-current can be output while the local blood sugar is reduced, and therefore the wound healing is better promoted.
Experimental example III curative effect of self-electrogenic enzyme-linked microneedle patch on treatment of diabetic wounds
The micro-needle patches with different concentration ratios of ZIF-8, ZG and ZH nano-particles are required to be tested for antibacterial performance because bacterial infection is easy to occur to wounds after skin is damaged and the healing condition of the wounds is influenced, and the experimental method is to perform co-culture on the micro-needle patches, escherichia coli and staphylococcus aureus bacteria liquid and then evaluate the micro-needle patches by adopting a plate counting method.
In animal experiments, male Wister rats of 6 weeks old, weighing 180-220g, purchased from animal laboratories, inc. of Lanzhou university were selected, STZ was prepared as a 10mg/mL STZ solution in PBS and sterilized by filtration using a 0.22 μm filter. The preparation is carried out in dark, and the preparation is used immediately. Before the operation of 12h fasting, the model group rats are subjected to intraperitoneal injection according to the STZ with the dose of 60mg/kg, fasting blood glucose is measured 3 days and 7 days after the STZ injection, and if the rats are listened, slow in reaction, increased in appetite, remarkably increased in urine volume, reduced in weight and more than 13.5-25mmol/L in blood glucose concentration, the type I diabetes model is successfully established. The anesthetized diabetic rats are fixed on a rat frame, and two square full-thickness wounds with the same size of 1 multiplied by 1cm are prepared after the skin on the two sides of the spinal column of the back is shaved, prepared and disinfected.
Experimental grouping and treatment protocol: dividing the constructed diabetes wound animal model into 6 groups, wherein each group comprises 6 animals, the wounds of 5 groups are respectively treated by Blank-MN patch, ZIF-MN patch, ZG-MN patch, ZH-MN patch and ZGH-MN patch, and the rest group is Blank control group.
And (3) evaluating the curative effect: the wound was observed roughly on days 7, 14 and 21, respectively, and the size of the wound was measured and recorded.
The experimental results are as follows:
the result of the antibacterial experiment of FIG. 5 shows that the antibacterial rate almost reaches 100% when the concentration of the ZIF-8, ZG and ZH nanoparticles reaches 0.5mg/ml, and the antibacterial agent has good antibacterial performance.
From the results of fig. 6, it is understood that the diabetic wound became gradually smaller from the electrogenic enzyme-linked microneedle patch set. No scar healing is realized 21 days after the ZGH-MN microneedle patch group, but small wounds still exist on the skin surfaces of other groups or obvious scars exist on the skin surfaces after healing. Experiments show that the self-electrogenesis enzyme-linked microneedle patch provided by the invention has an obvious healing effect on diabetic wounds and realizes scar-free healing of the diabetic wounds.
Experimental example four, curative effect of self-electrogenic enzyme-linked microneedle patch for treating sciatic nerve
Male Wister rats at 6 weeks of age, weighing 180-220g, were purchased from the animal testing center, lanzhou university. Using a 10% chloral hydrate intraperitoneal injection (3 ml/kg), the skin was prepared after anesthetizing the rats, and the skin around the ischial tuberosities on the right side of each rat was exposed, with the right side being selected as the experimental side and the left side as the control side. The operation is performed according to the aseptic principle, the position of the ischial tuberosity is touched by fingers, the skin is incised backwards along the ischial tuberosity on the superficial layer of the femur, obvious intermuscular fascia white lines are visible, and the vessel forceps are used for blunt separation along the white line direction to expose the sciatic nerve trunk. Total sciatic nerve trunk at the lower end of the right sciatic tuberosity of the rat is dissociated and exposed, a 10mm nerve segment is cut in the middle of the sciatic nerve trunk, a prepared microneedle nerve tube is placed at the dissociated end of the nerve to replace the repaired and lost nerve, and the sciatic nerve trunk is layered and sutured.
Experimental grouping and treatment protocol: the established sciatic nerve rat model is divided into 7 groups, 3 nerves are arranged in each group, and the nerve broken ends of 5 groups are respectively treated by a Blank-MN patch, a ZIF-MNs patch, a ZG-MN patch, a ZH-MN patch and a ZGH-MN patch. The remaining one group was a blank control group, and the other group was an autograft group, in which nerves from normal rats were taken and transplanted to the free ends of sciatic nerve model rats.
And (3) evaluating the curative effect: the rats were sacrificed at week 6 and week 12, and the treated sciatic nerve and gastrocnemius muscle were observed in general, fixed with a gastrocnemius tissue fixing solution, and used for HE section staining to compare the muscle fiber condition.
The experimental results are as follows:
as can be seen from the results of fig. 7, from the gross photograph of the gastrocnemius muscle at week 6, the volume of the gastrocnemius muscle after treatment with the conductive microneedle patch was significantly larger than that of the blank control group, wherein the volume of the gastrocnemius muscle in the ZGH electrogenic cascade microneedle patch group was the largest; as can be seen from the staining result of the gastrocnemius HE section in fig. 8, gastrocnemius muscle fibers in the treatment group of the self-electrogenic enzyme-linked microneedle patch are significantly larger than those of the Blank control group of the ZG, ZH and Blank conductive microneedle patch groups at weeks 6 and 12, and especially, the gastrocnemius muscle fibers in the ZGH self-electrogenic enzyme-linked microneedle patch group are close to those in the autograft group in size and arrangement; the electrogenesis cascade conductive microneedle patch has a good effect on repairing the sciatic nerve. Animal experiments show that the self-generating enzyme-linked conductive microneedle patch provided by the invention has a good repairing effect on injury of peripheral nerves such as sciatic nerve and the like, and has an obvious curative effect.
In conclusion, the electrogenesis enzyme cascade microneedle patch prepared by the invention is sensitive to the blood glucose concentration of the diabetic wound surface and can quickly respond. The enzyme cascade reaction is generated, the physiological micro-current can be output while the local blood sugar is reduced, so that the wound healing is better promoted, and the antibacterial effect is good; the ZGH-MN microneedle patch group realizes scar-free healing after 21 days, and other groups of skin surfaces still have small wounds or obvious scars on the skin surfaces after healing. Experiments show that the self-electrogenesis enzyme-linked microneedle patch provided by the invention has an obvious healing effect on diabetic wounds and realizes scar-free healing of the diabetic wounds. From the 6 th week general picture of the gastrocnemius muscle, the volume of the gastrocnemius muscle treated by the conductive microneedle patch is obviously larger than that of a blank control group, wherein the volume of the gastrocnemius muscle of the ZGH self-electrogenic cascade microneedle patch group is the largest; the gastrocnemius muscle fibers of the self-electrogenesis enzyme cascade microneedle patch treatment group are obviously larger than those of a Blank control group of ZG, ZH and Blank conductive microneedle patch groups at the 6 th week and the 12 th week, and particularly the gastrocnemius muscle fibers of the ZGH self-electrogenesis enzyme cascade microneedle patch group are close to those of an autograft group in size and arrangement; the electrogenesis enzyme cascade conductive microneedle patch is proved to have good effect on repairing the sciatic nerve. Animal experiments show that the self-generating enzyme-linked conductive microneedle patch provided by the invention has a good repairing effect on injury of peripheral nerves such as sciatic nerve and the like, and has an obvious curative effect.
Finally, it should be noted that: although the present description is described in terms of embodiments, not every embodiment includes only a single embodiment, and such description is for clarity only, and those skilled in the art should be able to integrate the description as a whole, and the embodiments can be appropriately combined to form other embodiments as will be understood by those skilled in the art.
Claims (8)
1. The self-electrogenesis enzyme cascade microneedle patch is characterized by consisting of a microneedle array consisting of a conductive substrate without a target drug and at least one needle body containing the target drug, wherein the substrate is prepared from polydopamine-modified polypyrrole, hyaluronic acid and polyvinyl alcohol, and the needle body is prepared from polydopamine-modified polypyrrole, hyaluronic acid and polyvinyl alcohol, glucose oxidase and horseradish peroxidase/catalase.
2. The self-electrogenic enzyme-linked microneedle patch according to claim 1, wherein glucose oxidase, horseradish peroxidase/catalase are immobilized on a metal-organic framework material ZIF-8.
3. The self-electrogenic enzyme-linked microneedle patch according to claim 2, wherein the substrate comprises dopamine-modified polypyrrole, hyaluronic acid and polyvinyl alcohol in a mass ratio of 0.01 to 1:1 to 100: 0.5-50, wherein in the needle body, the mass ratio of polypyrrole modified by dopamine, hyaluronic acid, polyvinyl alcohol, glucose oxidase, horseradish peroxidase/catalase and ZIF-8 is 0.01-1: 1 to 100:0.5 to 50: 0.005-0.5: 0.005-0.5.
4. The method for producing a self-electrogenic enzyme-linked microneedle patch according to any one of claims 1 to 3, comprising the steps of:
(1) Under the conditions of water bath and magnetic stirring, proportionally adding polydopamine-modified polypyrrole, hyaluronic acid and polyvinyl alcohol into deionized water to obtain microneedle patch substrate gel;
(2) Mixing the microneedle patch substrate gel prepared in the step (1) with glucose oxidase to obtain glucose oxidase needle body gel; mixing with horse radish peroxidase/catalase to obtain horse radish peroxidase/catalase needle gel;
(3) Absorbing 5-20ul of glucose oxidase needle gel prepared in the step (2) by using a pipette, dripping the glucose oxidase needle gel on a half pinhole of a PDMS micro needle template, and putting the PDMS micro needle template into a vacuum drying oven for vacuum negative pressure injection molding under the pressure of-1 to-0.8 MPa: repeatedly vacuumizing for 2-3 times, each time for 3-5 min, and naturally drying at room temperature to obtain a microneedle template carrying glucose oxidase;
(4) Sucking 5-20ul of the prepared horseradish peroxidase/catalase needle body gel in the step (2) by using a pipette, dripping the horseradish peroxidase/catalase needle body gel on the other half of the pinholes of the microneedle template in the step (3), and putting the microneedle template into a vacuum drying oven for vacuum negative pressure injection molding at-1 to-0.8 MPa: repeatedly vacuumizing for 2-3 times, each time for 3-5 min, and naturally drying at room temperature to obtain a glucose oxidase-horse radish peroxidase/catalase-loaded conductive microneedle patch needle body;
(5) Placing the substrate gel prepared in the step (1) on the microneedle body template obtained in the step (4), placing the microneedle body template in a vacuum drying box again, setting the vacuum to be-1 to-0.8 MPa, performing negative pressure injection molding and vacuumizing for 10-30 min, and naturally drying at room temperature to obtain the self-electrogenesis enzyme cascade microneedle patch needle body;
(6) And (3) paving the conductive hydrogel prepared in the step (1) on the microneedle template of which the needle body is filled with the conductive polymer matrix gel in the step (5), and naturally drying to obtain the self-electrogenesis enzyme-linked microneedle patch.
5. The method of claim 4, wherein the temperature of the water bath in step (1) is 0 to 90 ℃.
6. Use of the self-generating enzyme-linked microneedle patch as claimed in any one of claims 1 to 3, for preparing a medicament for treating a local disease.
7. Use of the self-generating enzyme-linked microneedle patch as claimed in any one of claims 1 to 3 for the preparation of a medicament for the treatment of skin injury.
8. Use of the self-generating enzyme-linked microneedle patch as claimed in any one of claims 1 to 3 for the preparation of a medicament for the treatment of nerve injury.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018202922A1 (en) * | 2017-05-05 | 2018-11-08 | University Of Ulster | A minimally invasive diagnostic device |
| US20190201675A1 (en) * | 2015-08-17 | 2019-07-04 | National Technology & Engineering Solutions Of Sandia, Llc | Electromolded microneedles and fabrication methods thereof |
| CN110123340A (en) * | 2019-04-12 | 2019-08-16 | 南通纺织丝绸产业技术研究院 | Fibroin albumen microneedle electrodes and its preparation and application |
| CN111343970A (en) * | 2017-09-13 | 2020-06-26 | 北卡罗莱纳州立大学 | Microneedle patch for locally inducing browning of adipose tissue and treating obesity |
| CN111479581A (en) * | 2017-10-10 | 2020-07-31 | 北卡罗莱纳州立大学 | For H2O2Core-shell microneedle patch for insulin delivery triggered by PH cascade |
| CN111643447A (en) * | 2020-06-10 | 2020-09-11 | 赵超超 | Drug-loaded microneedle, drug-loaded microneedle patch, electrically controlled drug release microneedle system and preparation method of drug-loaded microneedle |
| CN114795113A (en) * | 2022-03-24 | 2022-07-29 | 中山大学 | Microneedle electrode array sensor and preparation method and application thereof |
| CN114848577A (en) * | 2022-04-15 | 2022-08-05 | 兰州大学 | Double-layer conductive microneedle patch for treating subcutaneous tumors and preparation method and application thereof |
-
2022
- 2022-08-30 CN CN202211057304.2A patent/CN115337255A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190201675A1 (en) * | 2015-08-17 | 2019-07-04 | National Technology & Engineering Solutions Of Sandia, Llc | Electromolded microneedles and fabrication methods thereof |
| WO2018202922A1 (en) * | 2017-05-05 | 2018-11-08 | University Of Ulster | A minimally invasive diagnostic device |
| CN111343970A (en) * | 2017-09-13 | 2020-06-26 | 北卡罗莱纳州立大学 | Microneedle patch for locally inducing browning of adipose tissue and treating obesity |
| CN111479581A (en) * | 2017-10-10 | 2020-07-31 | 北卡罗莱纳州立大学 | For H2O2Core-shell microneedle patch for insulin delivery triggered by PH cascade |
| US20210186865A1 (en) * | 2017-10-10 | 2021-06-24 | North Carolina State University | Core-shell microneedle patch for h2o2 and ph cascade-triggered insulin delivery |
| CN110123340A (en) * | 2019-04-12 | 2019-08-16 | 南通纺织丝绸产业技术研究院 | Fibroin albumen microneedle electrodes and its preparation and application |
| CN111643447A (en) * | 2020-06-10 | 2020-09-11 | 赵超超 | Drug-loaded microneedle, drug-loaded microneedle patch, electrically controlled drug release microneedle system and preparation method of drug-loaded microneedle |
| CN114795113A (en) * | 2022-03-24 | 2022-07-29 | 中山大学 | Microneedle electrode array sensor and preparation method and application thereof |
| CN114848577A (en) * | 2022-04-15 | 2022-08-05 | 兰州大学 | Double-layer conductive microneedle patch for treating subcutaneous tumors and preparation method and application thereof |
Non-Patent Citations (4)
| Title |
|---|
| XIANGLI ZHANG等: "Implantable Nerve Conduit Made of a Self-Powered Microneedle Patch for Sciatic Nerve Repair", ADV. HEALTHCARE MATER., vol. 12, no. 31, 2 August 2023 (2023-08-02), pages 1 - 10 * |
| XIANGLI ZHANG等: "Self-powered enzyme-linked microneedle patch for scar-prevention healing of diabetic wounds", SCIENCE ADVANCES, vol. 9, no. 2, 14 July 2023 (2023-07-14), pages 1 - 13 * |
| 张向丽: "产电酶联微针贴片用于皮肤及神经缺损修复研究", 中国优秀硕士学位论文全文数据库 基础科学辑, no. 02, 15 February 2024 (2024-02-15), pages 006 - 577 * |
| 王婷婷等: "负载纳米银/石墨烯复合物的海藻酸钠水凝胶薄膜的制备及应用", 复合材料学报, vol. 38, no. 9, 30 September 2021 (2021-09-30), pages 3008 - 3017 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116059156A (en) * | 2023-04-06 | 2023-05-05 | 四川大学 | Double-layer network hydrogel microneedle and preparation method and application thereof |
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