CN110384654B - Preparation method of controlled-release hydrogel with photothermal treatment and wound repair functions - Google Patents
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Abstract
The invention relates to the field of medical macromolecules, and discloses a preparation method of controlled-release hydrogel with photothermal therapy and wound repair functions, wherein dopamine is modified on alginic acid, so that a molecular chain of the dopamine has an o-diphenol side group, and copper sulfide @ melanin-PEG @ dopamine nanoparticles with good photothermal property and radiotherapy effect are prepared; then adding the copper sulfide @ dopamine nano particles into a dopamine modified alginic acid solution, simultaneously adding astragaloside with the effect of promoting wound healing into the solution, and then forming gel through a catalytic crosslinking system of horseradish peroxidase and hydrogen peroxide. The hydrogel has good controlled release performance, and can play a role in reducing the administration dosage and relieving or avoiding toxic and side effects. The hydrogel prepared by the method has good biocompatibility, treats cancer cells by using the action of photothermal property and radiotherapy, and simultaneously combines the action of astragaloside to promote wound healing after operation.
Description
Technical Field
The invention relates to the field of medical high polymer materials, in particular to a preparation method of controlled-release hydrogel with photothermal treatment and wound repair functions.
Background
Cancer, a serious disease threatening human life and health, has long been low in cure rate and high in recurrence rate and death rate. At present, the tumor treatment method in China is mainly surgical resection, but most of the surgeries can not completely remove the tumor, so that the tumor is relapsed. After surgical removal of the tumor, a gentle treatment with a small local effect is needed to remove the residual tumor tissue. Photothermal therapy, which can be targeted for treatment in a specific area and has mild conditions, is undoubtedly a relatively suitable choice, and slow healing of postoperative wounds is one of the problems to be solved urgently in the field.
The hydrogel is a tissue engineering material with good prospect, is a polymer material which is formed by crosslinking linear polymer chains to form a three-dimensional network space structure, has excellent water absorption capacity and is insoluble in water. It has the features of environment sensitivity, excellent biocompatibility, biodegradability, etc. In addition, the hydrogel has little surface protein and cell adhesion, contains a large amount of water very similar to body tissues filled with a large amount of aqueous liquid, and has a soft and moist surface and affinity with the tissues, so that the irritation to the human body is greatly reduced. These properties make it possible to load a series of tumor therapeutic agents, thus obtaining tumor therapeutic and repair materials with good biocompatibility.
Disclosure of Invention
In order to solve the technical problems, the invention provides a preparation method of controlled-release hydrogel with photothermal treatment and wound repair functionsThe preparation method is that firstly dopamine is modified on alginic acid to make the molecular chain have an o-diphenol side group, and CuCl2 & 2H is used2O and Na2S·9H2The copper sulfide @ melanin nano particles with good photo-thermal performance are prepared by taking O and melanin as raw materials, and meanwhile, the particles also have good radiotherapy effect. Then using PEG-NH2Modifying the copper sulfide @ melanin nano particles, and then coating the particles with polydopamine to obtain the copper sulfide @ melanin-PEG @ dopamine nano particles, wherein the surface of the copper sulfide @ melanin-PEG @ dopamine nano particles is provided with an o-diphenol group while the photo-thermal performance is enhanced; then adding the copper sulfide @ dopamine nano particles into a dopamine modified alginic acid solution, simultaneously adding astragaloside with the effect of promoting wound healing into the solution, and then forming gel through a catalytic crosslinking system of horseradish peroxidase and hydrogen peroxide. Meanwhile, the hydrogel has good controlled release performance, and can play a role in reducing the dosage and relieving or avoiding toxic and side effects. The crosslinking degree of the hydrogel can be changed by changing the dosage of the peroxidase and the hydrogen peroxide so as to change the controlled release capacity of the hydrogel; by this method, new routes of administration can be established, and new routes of administration can be established. The hydrogel prepared by the method has good biocompatibility, treats cancer cells by utilizing the photothermal property and the radiotherapy effect of polydopamine and copper sulfide particles, and is combined with the effect of promoting postoperative wound healing of astragaloside.
The specific technical scheme of the invention is as follows: a preparation method of controlled release hydrogel with photothermal therapy and wound repair functions comprises the following steps of measuring mu g, mg, g, mu L and mL:
1) adding 1-1.5g alginic acid into 47-48ml MES buffer solution, and stirring for 1.5-2.5 h.
Alginic acid is a biodegradable natural polymer, which solves the problem of environmental pollution. The good biocompatibility of the material ensures that the material can be taken out again when being used as a tissue engineering material, thereby reducing the pain of patients.
2) 85-90mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 51-55mg of N-hydroxysuccinimide were dissolved in 1-1.5mL of MES buffer, respectively.
3) Putting the solution obtained in the step 1) into a water bath, heating to 65-65 ℃, adding the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride solution and the N-hydroxysuccinimide solution obtained in the step 2) under the protection of nitrogen, and stirring for 1-1.5 h.
In step 3), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide are used to activate the carboxyl groups in alginic acid, while in MES buffer to prevent the pH of the solution from changing too much during the reaction.
4) Then 550-600mg of dopamine is added into the solution obtained in the step 3) and reacts for 12-16h under stirring.
In step 4), the activated carboxyl in alginic acid and the amino group on dopamine are subjected to an amide reaction under acidic conditions, so that the alginic acid molecular chain has an o-diphenol side group, and the acidic conditions are also used for preventing self-polymerization of dopamine.
5) Dialyzing the solution obtained in the step 4) for 2-3d, and drying by using a freeze-drying method to obtain alginic acid-dopamine powder.
6) Dissolving melanin 0.4-0.6g in deionized water 8-12mL to obtain melanin water solution, and collecting CuCl 1-1.5g2·2H2Dissolving 350mg of polyvinylpyrrolidone O and 300-150 mL of deionized water, then combining the two solutions, stirring for 2-3h, and adding 200mg of Na2S·9H2O, then placing the mixture in a water bath, heating the mixture to 85-90 ℃, and stirring the mixture for 15-20min to obtain dark green copper sulfide nano suspension; and dialyzing in deionized water for 3-4 days, centrifuging to obtain precipitate, and vacuum drying at 55-65 ℃ overnight to obtain the copper sulfide @ melanin nanoparticles.
In the step 6), the melanin is used as a template, and the copper sulfide @ melanin nanoparticles are synthesized in an aqueous solution, so that a layer of melanin is coated outside the copper sulfide, and the nanoparticles can be well dispersed in water. Meanwhile, the surface of the nanoparticle contains dihydroxyindole to prepare for the subsequent modification of PEG.
7) 0.5-0.8g of the copper sulfide @ melanin particles are dispersed in 0.5-0.8mL of water solution, uniformly dispersed by ultrasonic treatment for 25-35min, and then dropwise added to18-22mg/mL PEG5000-NH2Vigorously stirring in water solution for 10-14h, washing with deionized water for several times, and washing to remove unreacted PEG5000-NH2(ii) a And removing water by a freeze-drying method to obtain the copper sulfide @ melanin-PEG nano particles.
In step 7), by PEG5000-NH2The amino group in the PEG modified copper sulfide @ melanin nanoparticle is covalently combined with dihydroxyindole in melanin on the surface of the copper sulfide @ melanin nanoparticle, so that the PEG modified copper sulfide @ melanin nanoparticle is obtained, and meanwhile, the PEG modified copper sulfide @ melanin nanoparticle has good biocompatibility and dispersibility.
8) And (2) dispersing 0.4-0.6g of the copper sulfide @ melanin-PEG nano particles into 120mL of 100-mL buffer solution, adding 0.4-0.6g of dopamine, stirring at room temperature for 12-14h, washing with deionized water for multiple times, and removing water by a freeze-drying method to obtain the copper sulfide @ melanin-PEG @ polydopamine nano particles.
In the step 8), a layer of polydopamine is coated on the surface of the copper sulfide @ melanin-PEG nanoparticle by self-polymerization of the dopamine on the surface of the copper sulfide @ melanin-PEG nanoparticle, and the polydopamine also has good photo-thermal properties, so that the photo-thermal properties of the polydopamine are enhanced, and meanwhile, the surface of the polydopamine is provided with abundant o-diphenol side groups.
9) Dissolving 0.8-1.2mg of astragaloside in 0.8-1.2mL of PBS buffer solution.
10) Dissolving 25-35mg of alginic acid-dopamine powder obtained in the step 5) in 0.8-1.2mL of deionized water, adding 100-200 μ g of copper sulfide @ melanin-PEG @ polydopamine nanoparticles obtained in the step 8) and 10-20 μ L of the astragaloside IV solution obtained in the step 9), stirring for 2-3h at 35-39 ℃, adding 10-20 μ L of 1000U/mL horseradish peroxidase aqueous solution, mixing uniformly, adding 10-20 μ L of 0.4-0.6% hydrogen peroxide solution, mixing uniformly, transferring the mixture to a PTFE (polytetrafluoroethylene) mold by using a liquid transfer gun, and standing for 3-4h at 35-39 ℃ for molding.
In the step 10), copper sulfide @ melanin-PEG @ polydopamine nanoparticles and a matrix material alginic acid-dopamine are mutually crosslinked through the catalytic crosslinking effect of horseradish peroxidase and hydrogen peroxide on o-diphenol, so that the nano-particles have good mechanical properties, and meanwhile, astragaloside is embedded in hydrogel.
11) Demolding the alginic acid hydrogel loaded with astragaloside IV and copper sulfide @ melanin-PEG @ polydopamine formed in the step 10) to obtain the controlled-release hydrogel with photothermal therapy and wound repair functions.
Preferably, in step 3) and step 4), the solution pH is maintained between 5 and 6 during stirring.
Preferably, in step 5), the dialysis process is: the solution was dialyzed in 8000Da dialysis bag in deionized water in acidic environment for 2-3 d.
Preferably, in step 6), the polyvinylpyrrolidone has a molecular weight of 24000.
Preferably, in step 6), the aqueous melanin solution is slowly added dropwise to the CuCl over 30min under the action of ultrasound2·2H2O and polyvinylpyrrolidone in water solution.
Preferably, in step 6), the dialysis process is: the solution was dialyzed in deionized water for 3-4d in a 8000Da dialysis bag.
Preferably, in the step 6), the centrifugal rotating speed is 12000-16000r/m, and the separation time is 10-15 mm.
Preferably, in step 7), 25 to 30 wt% NH is added during stirring4Adjusting the pH value of the OH solution to 8.8-9.2.
Preferably, in step 7) and step 8), the washing process is as follows: centrifuging, redispersing and centrifuging the solution after reaction, and repeating the centrifuging for three times; the rotation speed of each centrifugation is 12000-16000r/m, and the time is 10-15 min.
Preferably, in the steps 5), 6), 7), 8) and 9), the obtained product is sealed and placed in an environment with the temperature of 1-5 ℃ for refrigeration for standby.
Compared with the prior art, the invention has the beneficial effects that:
the invention utilizes alginic acid-dopamine with good biocompatibility to load copper sulfide @ melanin-PEG @ dopamine nano particles with good photothermal property and radiotherapy function and astragaloside with the effect of promoting wound healing, and the obtained hydrogel can be used for photothermal treatment and radiotherapy of postoperative tumor parts and can also play a role in accelerating healing and repairing of postoperative wound surfaces. Meanwhile, the hydrogel has good controlled release performance, and can play a role in reducing the dosage and relieving or avoiding toxic and side effects. The stability of the medicine is improved, and the medicine is convenient to store; several new routes of administration can be established. Meanwhile, the hydrogel is a catalytic crosslinking system utilizing horseradish peroxidase and hydrogen peroxide, so that the cells are not obviously influenced in the later experimental process, the scientificity of experimental results is not influenced, and the experimental operation process is simple, non-toxic, harmless, green and environment-friendly.
Detailed Description
The present invention will be further described with reference to the following examples.
Example 1:
1) adding 1g of alginic acid into 47mL of MES buffer solution, and stirring for 2 h;
2) respectively dissolving 85mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 51 mg of N-hydroxysuccinimide in 1mL of MES buffer solution;
3) putting the solution obtained in the step 1) into a water bath, heating to 60 ℃, and adding the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride solution and the N-hydroxysuccinimide solution obtained in the step 2) under the protection of nitrogen. Stirring for 1h, and maintaining the pH of the solution at 5 by using hydrochloric acid or sodium hydroxide during stirring;
4) adding 550mg of dopamine into the solution obtained in the step 3), reacting for 12 hours under the stirring action, and maintaining the pH of the solution to be 5 by using hydrochloric acid or sodium hydroxide during stirring;
5) putting the solution obtained in the step 4) into a 8000Da dialysis bag, dialyzing in deionized water in an acid environment for 2d, freezing in an ultra-low temperature refrigerator overnight, and drying in a freeze dryer for 3 d. Drying to obtain alginic acid-dopamine powder, sealing, and refrigerating at 1 deg.C;
6) 0.5g of melanin is dissolved in 10mL of deionized water, and 1g of CuCl is taken2·2H2Dissolving O and 300mg polyvinylpyrrolidone with molecular weight of 24000 in 100mL deionized water, slowly adding melanin solution dropwise into the solution within 30min under the action of ultrasound, and stirring for 2 h. After stirring, 150mg of Na was added2S·9H2And O. Then theAnd (3) placing the solution in a water bath kettle, heating to 85 ℃, and stirring for 15min to obtain the dark green copper sulfide nano suspension. Then putting the solution into a 8000Da dialysis bag, dialyzing the solution in deionized water for 3d, centrifuging the solution for 10min at a centrifugal rotating speed of 12000r/m, removing supernatant, taking precipitate, drying the precipitate in a vacuum drying oven at 60 ℃ overnight to obtain copper sulfide @ melanin nanoparticles, sealing the copper sulfide @ melanin nanoparticles and refrigerating the copper sulfide @ melanin nanoparticles in an environment at 1 ℃ for later use;
7) and (3) dispersing 0.5g of the copper sulfide @ melanin particles obtained in the step 6) in 0.5mL of aqueous solution, and performing ultrasonic treatment for 30min to uniformly disperse the copper sulfide @ melanin particles. The mixed solution was added dropwise to 20mg/mL PEG5000-NH2In aqueous solution. Stirring vigorously for 12h, during which NH is added4OH solution (28 wt%) adjusted the pH of the solution to 9. Then the solution after reaction is centrifuged/redispersed/centrifuged, and the process is repeated for three times; the rotation speed of each centrifugation is 12000r/m, and the centrifugation treatment time is 10 min. Unreacted PEG5000-NH2The deionized water was washed several times. Freezing in an ultra-low temperature refrigerator overnight, drying in a freeze dryer for 3d, removing the hydrosolvent to obtain copper sulfide @ melanin-PEG nanoparticles, sealing, and refrigerating at 1 deg.C;
8) taking 0.5g of the copper sulfide @ melanin-PEG nanoparticles obtained in the step 7), dispersing in 100mL of Tirs buffer solution (ph is 8.5), adding 0.5g of dopamine into the solution, stirring at room temperature for 12h, centrifuging/redispersing/centrifuging the reacted solution, and repeating for three times; the rotation speed of each centrifugation is 12000r/m, and the centrifugation treatment time is 10 min. Freezing in an ultra-low temperature refrigerator overnight, drying in a freeze dryer for 3d, removing the hydrosolvent to obtain copper sulfide @ melanin-PEG @ polydopamine nanoparticles, sealing, and refrigerating at 1 deg.C;
9) dissolving 1mg of astragaloside IV in 1mL of PBS buffer solution, sealing, and refrigerating at 1 deg.C;
10) dissolving 30mg of alginic acid-dopamine powder obtained in the step 5) in 1mL of deionized water, adding 100 μ g of copper sulfide @ melanin-PEG @ polydopamine nanoparticles obtained in the step 8) and 10 μ L of astragaloside IV solution obtained in the step 9) into the solution, stirring at 37 ℃ for 2 hours, adding 10 μ L of 1000U/mL horseradish peroxidase aqueous solution, mixing uniformly, and adding 10 μ L of 0.5% hydrogen peroxide. Then, uniformly mixing the materials by using a liquid transfer gun, adding the mixture into a PTFE (polytetrafluoroethylene) mold, and standing the mixture for 3 hours at the temperature of 37 ℃ for molding;
11) and (3) taking the alginate hydrogel loaded with astragaloside IV and copper sulfide @ melanin-PEG @ polydopamine formed in the step 10) out of the mold, thus obtaining the hydrogel with the controlled release function for radiotherapy combined with photothermal therapy and wound repair after tumor operation.
In the embodiment, the copper sulfide @ melanin-PEG @ polydopamine nanoparticles are used in a small amount, the photo-thermal effect of the prepared hydrogel can reach about 55 ℃ in 5min, the dosages of horseradish peroxidase and hydrogen peroxide are also slightly low, the drug release is fast due to the low crosslinking degree, the release rate reaches 63% in 12h, and the release rate reaches 85% in 10 d.
Example 2:
1) adding 1.25g of alginic acid into 47.5mL of MES buffer solution, and stirring for 2 h;
2) respectively dissolving 87.5mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 53mg of N-hydroxysuccinimide in 1.25mL of MES buffer solution;
3) putting the solution obtained in the step 1) into a water bath, heating to 60 ℃, and adding the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride solution and the N-hydroxysuccinimide solution obtained in the step 2) under the protection of nitrogen. Stirring for 1.25h, and maintaining the pH of the solution at 5 with hydrochloric acid or sodium hydroxide during stirring;
4) adding 570mg of dopamine into the solution obtained in the step 3), reacting for 14 hours under the stirring action, and maintaining the pH of the solution to be 5 by using hydrochloric acid or sodium hydroxide during stirring;
5) putting the solution obtained in the step 4) into a 8000Da dialysis bag, dialyzing in deionized water in an acid environment for 2d, freezing in an ultra-low temperature refrigerator overnight, and drying in a freeze dryer for 3 d. Drying to obtain alginic acid-dopamine powder, sealing, and refrigerating at 4 deg.C;
6) 0.5g of melanin is dissolved in 10mL of deionized water, and 1.25g of CuCl is taken2·2H2Dissolving O and 325mg polyvinylpyrrolidone with molecular weight of 24000 in 125mL deionized water, and treating with ultrasound at 30%Slowly dripping melanin solution into the solution within min, and stirring for 2-3 h. After stirring, 175mg of Na was added2S·9H2And O. Then the solution is placed in a water bath to be heated to 87 ℃, and stirred for 17min, so as to obtain dark green copper sulfide nano suspension. Then putting the solution into a 8000Da dialysis bag, dialyzing the solution in deionized water for 4 days, centrifuging the solution for 13min at the centrifugal rotating speed of 14000r/m, removing supernatant, taking precipitate, drying the precipitate in a vacuum drying oven at 60 ℃ overnight to obtain copper sulfide @ melanin nanoparticles, sealing the copper sulfide @ melanin nanoparticles and refrigerating the nanoparticles in an environment at 4 ℃ for later use;
7) and (3) dispersing 0.7g of the copper sulfide @ melanin particles obtained in the step 6) in 0.7mL of aqueous solution, and performing ultrasonic treatment for 30min to uniformly disperse the copper sulfide @ melanin particles. The mixed solution was added dropwise to 20mg/mL PEG5000-NH2In aqueous solution. Stirring vigorously for 12h, during which NH is added4OH solution (28 wt%) adjusted the pH of the solution to 9. Then the solution after reaction is centrifuged/redispersed/centrifuged, and the process is repeated for three times; the rotation speed of each centrifugation is 14000r/m, and the centrifugation treatment time is 13 min. Unreacted PEG5000-NH2The deionized water was washed several times. Freezing in an ultra-low temperature refrigerator overnight, drying in a freeze dryer for 3d, removing the hydrosolvent to obtain copper sulfide @ melanin-PEG nanoparticles, sealing, and refrigerating at 4 deg.C;
8) taking 0.5g of the copper sulfide @ melanin-PEG nanoparticles obtained in the step 7), dispersing in 110mL of Tirs buffer solution (ph is 8.5), adding 0.5g of dopamine into the solution, stirring at room temperature for 13h, centrifuging/redispersing/centrifuging the reacted solution, and repeating for three times; the rotation speed of each centrifugation is 14000r/m, and the centrifugation treatment time is 13 min. Freezing in an ultra-low temperature refrigerator overnight, drying in a freeze dryer for 3d, removing the hydrosolvent to obtain copper sulfide @ melanin-PEG @ polydopamine nanoparticles, sealing, and refrigerating at 4 deg.C;
9) dissolving 1mg of astragaloside IV in 1mL of PBS buffer solution, sealing, and refrigerating at 1-5 deg.C;
10) dissolving 30mg of alginic acid-dopamine powder obtained in the step 5) in 1mL of deionized water, adding 150 μ g of copper sulfide @ melanin-PEG @ polydopamine nanoparticles obtained in the step 8) and 15 μ L of astragaloside IV solution obtained in the step 9) into the solution, stirring at 37 ℃ for 2 hours, adding 15 μ L of 1000U/mL horseradish peroxidase aqueous solution, mixing uniformly, and adding 15 μ L of 0.5% hydrogen peroxide. Then, uniformly mixing the materials by using a liquid transfer gun, adding the mixture into a PTFE (polytetrafluoroethylene) mold, and standing the mixture for 3.5 hours at the temperature of 37 ℃ for molding;
11) and (3) taking the alginate hydrogel loaded with astragaloside IV and copper sulfide @ melanin-PEG @ polydopamine formed in the step 10) out of the mold, thus obtaining the hydrogel with the controlled release function for radiotherapy combined with photothermal therapy and wound repair after tumor operation.
In the embodiment, the copper sulfide @ melanin-PEG @ polydopamine nanoparticles are moderate in dosage, the prepared hydrogel is relatively good in photo-thermal effect, the temperature can reach about 64 ℃ in 5min, the dosage of horseradish peroxidase and hydrogen peroxide is moderate, the crosslinking degree is low, the drug release is smooth, the release rate reaches 56% in 12h, and the release rate reaches 78% in 10 d.
Example 3:
1) adding 1.5g of alginic acid into 48mL of MES buffer solution, and stirring for 2 h;
2) respectively dissolving 90mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 55mg of N-hydroxysuccinimide in 1.5ml of a local buffer solution;
3) putting the solution obtained in the step 1) into a water bath, heating to 60 ℃, and adding the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride solution and the N-hydroxysuccinimide solution obtained in the step 2) under the protection of nitrogen. Stirring for 1.5h, and maintaining the pH of the solution at 6 with hydrochloric acid or sodium hydroxide during stirring;
4) then 600mg of dopamine is added into the solution obtained in the step 3), and then the solution reacts for 16 hours under the stirring action, and hydrochloric acid or sodium hydroxide is used for maintaining the pH value of the solution to be 6 during the stirring;
5) putting the solution obtained in the step 4) into a 8000Da dialysis bag, dialyzing in deionized water in an acid environment for 3d, freezing in an ultra-low temperature refrigerator overnight, and drying in a freeze dryer for 3 d. Drying to obtain alginic acid-dopamine powder, sealing, and refrigerating at 5 deg.C;
6) dissolving 0.5g melanin in 10mL deionized water, and collecting1.5g CuCl2·2H2O and 300-350mg polyvinylpyrrolidone with molecular weight of 24000 are dissolved in 150mL deionized water, and then under the action of ultrasound, the melanin solution is slowly dripped into the solution within 30min and stirred for 3 h. After stirring, 200mg of Na was added2S·9H2And O. Then the solution is placed in a water bath to be heated to 90 ℃, and stirred for 20min, so as to obtain the dark green copper sulfide nano suspension. Then putting the solution into a 8000Da dialysis bag, dialyzing the solution in deionized water for 4d, centrifuging the solution for 15min at a centrifugal rotating speed of 16000r/m, removing supernatant, taking precipitate, drying the precipitate in a vacuum drying oven at 60 ℃ overnight to obtain copper sulfide @ melanin nanoparticles, and hermetically storing the nanoparticles in an environment at 1-5 ℃ for refrigeration for later use;
7) and (3) dispersing 0.5-0.8g of the copper sulfide @ melanin particles obtained in the step 6) in 0.8mL of aqueous solution, and performing ultrasonic treatment for 30min to uniformly disperse the copper sulfide @ melanin particles. The mixed solution was added dropwise to 20mg/mL PEG5000-NH2In aqueous solution. Stirring vigorously for 12h, during which NH is added4OH solution (28 wt%) adjusted the pH of the solution to 9. Then the solution after reaction is centrifuged/redispersed/centrifuged, and the process is repeated for three times; the rotation speed of each centrifugation is 16000r/m, and the centrifugation treatment time is 15 min. Unreacted PEG5000-NH2The deionized water was washed several times. Freezing in an ultra-low temperature refrigerator overnight, drying in a freeze dryer for 3d, removing the hydrosolvent to obtain copper sulfide @ melanin-PEG nanoparticles, sealing, and refrigerating at 5 deg.C;
8) taking 0.5g of the copper sulfide @ melanin-PEG nanoparticles obtained in the step 7), dispersing in 120mL of Tirs buffer solution (ph is 8.5), adding 0.5g of dopamine into the solution, stirring at room temperature for 14h, centrifuging/redispersing/centrifuging the reacted solution, and repeating for three times; the rotation speed of each centrifugation is 16000r/m, and the centrifugation treatment time is 15 min. Freezing in an ultra-low temperature refrigerator overnight, drying in a freeze dryer for 3d, removing the hydrosolvent to obtain copper sulfide @ melanin-PEG @ polydopamine nanoparticles, sealing, and refrigerating at 5 deg.C;
9) dissolving 1mg of astragaloside IV in 1mL of PBS buffer solution, sealing, and refrigerating at 5 deg.C;
10) dissolving 30mg of alginic acid-dopamine powder obtained in the step 5) in 1mL of deionized water, adding 200 μ g of copper sulfide @ melanin-PEG @ polydopamine nanoparticles obtained in the step 8) and 20 μ L of astragaloside IV solution obtained in the step 9) into the solution, stirring the solution at 37 ℃ for 3 hours, adding 20 μ L of 1000U/mL horseradish peroxidase aqueous solution, mixing the solution uniformly, and adding 20 μ L of 0.5% hydrogen peroxide. Then, uniformly mixing the materials by using a liquid transfer gun, adding the mixture into a PTFE (polytetrafluoroethylene) mold, and standing the mixture for 4 hours at the temperature of 37 ℃ for molding;
11) and (3) taking the alginate hydrogel loaded with astragaloside IV and copper sulfide @ melanin-PEG @ polydopamine formed in the step 10) out of the mold, thus obtaining the hydrogel with the controlled release function for radiotherapy combined with photothermal therapy and wound repair after tumor operation.
In the embodiment, the copper sulfide @ melanin-PEG @ polydopamine nanoparticles are used in a large amount, the prepared hydrogel is good in photo-thermal effect, can reach 72 ℃ in 5min, simultaneously is large in the amount of horseradish peroxidase and hydrogen peroxide, and due to the fact that the crosslinking degree of the hydrogel is large, the drug release is slow, the release rate reaches 52% in 12h, and the release rate reaches 73% in 10 d.
The raw materials and equipment used in the invention are common raw materials and equipment in the field if not specified; the methods used in the present invention are conventional in the art unless otherwise specified.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, alterations and equivalents of the above embodiments according to the technical spirit of the present invention are still within the protection scope of the technical solution of the present invention.
Claims (10)
1. A preparation method of controlled release hydrogel with photothermal therapy and wound repair functions is characterized in that: in μ g, mg, g, μ L and mL, comprising the steps of:
1) adding 1-1.5g of alginic acid into 47-48mL of MES buffer solution, and stirring for 1.5-2.5 h;
2) respectively dissolving 85-90mg of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride and 51-55mg of N-hydroxysuccinimide in 1-1.5mL of MES buffer solution;
3) putting the solution obtained in the step 1) in a water bath, heating to 60 ℃, adding the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride solution and the N-hydroxysuccinimide solution obtained in the step 2) under the protection of nitrogen, and stirring for 1-1.5 h;
4) adding 550-600mg of dopamine into the solution obtained in the step 3), and reacting for 12-16h under stirring;
5) dialyzing the solution obtained in the step 4) for 2-3d, and drying by using a freeze-drying method to obtain alginic acid-dopamine powder;
6) dissolving melanin 0.4-0.6g in deionized water 8-12mL to obtain melanin water solution, and collecting CuCl 1-1.5g2·2H2Dissolving 350mg of polyvinylpyrrolidone O and 300-150 mL of deionized water, then combining the two solutions, stirring for 2-3h, and adding 200mg of Na2S·9H2O, then placing the mixture in a water bath, heating the mixture to 85-90 ℃, and stirring the mixture for 15-20min to obtain dark green copper sulfide nano suspension; then dialyzing in deionized water for 3-4d, centrifuging to obtain precipitate, and vacuum drying at 55-65 ℃ overnight to obtain copper sulfide @ melanin nanoparticles;
7) 0.5-0.8g of the copper sulfide @ melanin particles are dispersed in 0.5-0.8mL of water solution, uniformly dispersed by ultrasonic treatment for 25-35min, and then dropwise added to 18-22mg/mL of PEG5000-NH2Vigorously stirring in water solution for 10-14h, washing with deionized water for several times, and washing to remove unreacted PEG5000-NH2(ii) a Removing water by a freeze-drying method to obtain copper sulfide @ melanin-PEG nano particles;
8) dispersing 0.4-0.6g of the copper sulfide @ melanin-PEG nanoparticles into 120mL of 100-mL buffer solution, adding 0.4-0.6g of dopamine, stirring at room temperature for 12-14h, washing with deionized water for multiple times, and removing water by a freeze-drying method to obtain copper sulfide @ melanin-PEG @ polydopamine nanoparticles;
9) dissolving 0.8-1.2mg of astragaloside in 0.8-1.2mL of PBS buffer solution;
10) dissolving 25-35mg of alginic acid-dopamine powder obtained in the step 5) in 0.8-1.2mL of deionized water, adding 100-200 μ g of copper sulfide @ melanin-PEG @ polydopamine nanoparticles obtained in the step 8) and 10-20 μ L of the astragaloside IV solution obtained in the step 9), stirring for 2-3h at 35-39 ℃, adding 10-20 μ L of 1000U/mL horseradish peroxidase aqueous solution, mixing uniformly, adding 10-20 μ L of 0.4-0.6% hydrogen peroxide solution, mixing uniformly, transferring the mixture to a PTFE (polytetrafluoroethylene) mold by using a liquid transfer gun, and standing for 3-4h at 35-39 ℃ for molding;
11) demolding the alginic acid hydrogel loaded with astragaloside IV and copper sulfide @ melanin-PEG @ polydopamine formed in the step 10) to obtain the controlled-release hydrogel with photothermal therapy and wound repair functions.
2. The method for preparing controlled-release hydrogel with photothermal therapy and wound healing functions as claimed in claim 1, wherein in step 3) and step 4), the solution pH is maintained at 5-6 during stirring.
3. The method for preparing controlled-release hydrogel with photothermal therapy and wound healing functions as claimed in claim 1, wherein in step 5), the dialysis process is: the solution was dialyzed in 8000Da dialysis bag in deionized water in acidic environment for 2-3 d.
4. The method for preparing controlled-release hydrogel for photothermal therapy and wound healing according to claim 1, wherein in step 6), the molecular weight of polyvinylpyrrolidone is 24000.
5. The method for preparing controlled-release hydrogel with photothermal therapy and wound healing effects as claimed in claim 1, wherein in step 6), aqueous solution of melanin is slowly added dropwise to CuCl within 30min under the action of ultrasound2·2H2O and polyvinylpyrrolidone in water solution.
6. The method for preparing controlled-release hydrogel with photothermal therapy and wound healing functions as claimed in claim 1, wherein in step 6), the dialysis process is: the solution was dialyzed in deionized water for 3-4d in a 8000Da dialysis bag.
7. The method as claimed in claim 1, wherein in step 6), the centrifugation speed is 12000-16000r/m, and the separation time is 10-15 min.
8. The method for preparing controlled-release hydrogel with photothermal therapy and wound healing functions as claimed in claim 1, wherein in step 7), 25-30 wt% of NH is added during stirring4Adjusting the pH value of the OH solution to 8.8-9.2.
9. The method for preparing controlled-release hydrogel with photothermal therapy and wound healing functions as claimed in claim 1, wherein in step 7) and step 8), the washing process is as follows: centrifuging, redispersing and centrifuging the solution after reaction, and repeating the centrifuging for three times; the rotation speed of each centrifugation is 12000-16000r/m, and the time is 10-15 min.
10. The method for preparing controlled-release hydrogel with photothermal therapy and wound healing functions as claimed in claim 1, wherein in step 5), step 6), step 7), step 8) and step 9), the obtained product is hermetically stored in an environment of 1-5 ℃ for later use.
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| CN114632057A (en) * | 2022-03-09 | 2022-06-17 | 福建卫生职业技术学院 | Astragaloside IV diatomic ethosome gel transdermal drug delivery preparation and preparation method thereof |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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Non-Patent Citations (4)
| Title |
|---|
| Dopamine-Melanin Colloidal Nanospheres： An Efficient Near-Infrared Photothermal Therapeutic Agent for In Vivo Cancer;Yanlan Liu等;《Advanced Materials》;20121220;第25卷(第9期);第1353-1359页 * |
| Polydopamine-mediated bio-inspired synthesis of copper sulfide nanoparticles for T1-weighted magnetic resonance imaging guided photothermal cancer therapy;Yuxuan Xiong等;《Colloids and Surfaces B: Biointerfaces》;20181010;第19卷;第607-615页 * |
| Topical Astragaloside IV-Releasing Hydrogel Improves Healing of Skin Wounds in Vivo;Li-Hua Peng等;《Biol. Pharm. Bull.》;20120630;第35卷(第6期);第881-888页 * |
| 有机光热转换材料及其在光热疗法中的应用;陈瑞等;《化学进展》;20170324;第29卷;第329-336页 * |
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