CN115252538A - Preparation method of photo-thermal controlled drug release poly-dopamine iron drug-loaded nanoparticle hydrogel - Google Patents

Preparation method of photo-thermal controlled drug release poly-dopamine iron drug-loaded nanoparticle hydrogel Download PDF

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CN115252538A
CN115252538A CN202210860912.0A CN202210860912A CN115252538A CN 115252538 A CN115252538 A CN 115252538A CN 202210860912 A CN202210860912 A CN 202210860912A CN 115252538 A CN115252538 A CN 115252538A
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dopamine
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朱晨辉
范代娣
李娉
任震
张梓萱
唐浩东
朱安妮
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Northwest University
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Abstract

The invention discloses a preparation method of a photo-thermal controlled release polydopamine iron drug-loaded nanoparticle hydrogel, which comprises the following raw materials of dopamine, ferric chloride hexahydrate, resveratrol ethanol solution, sodium alginate and 1-ethyl- (3-dimethyl ether)Aminopropyl) carbodiimide, N-hydroxysuccinimide, dopamine hydrochloride, hydrogen peroxide and horseradish peroxidase solution; the method comprises the following steps: providing polydopamine iron nanoparticles loaded with resveratrol; providing sodium alginate grafted with dopamine; and adding hydrogen peroxide and a horseradish peroxidase solution, standing and crosslinking to obtain the photo-thermal controlled drug release poly-dopamine-iron drug-loaded nanoparticle hydrogel. The hydrogel can be directly injected into tumor to catalyze H in tumor2O2Production of O2The hypoxia inside the tumor is relieved, and the hyperpyrexia is generated under the near-infrared laser auxiliary irradiation condition, so that the drug is released, and melanoma cells are killed.

Description

Preparation method of photo-thermal controlled drug release poly-dopamine iron drug-loaded nanoparticle hydrogel
Technical Field
The invention belongs to the technical field of hydrogel, and particularly relates to a preparation method of photo-thermal controlled drug release poly-dopamine iron drug-loaded nanoparticle hydrogel.
Background
Melanoma is a highly invasive skin cancer originating from melanocytes, and brings huge economic and psychological burdens to people, and the current clinical treatment of melanoma mainly has the following problems: (1) drug resistance, melanoma has significant resistance to chemotherapy/radiotherapy; (2) hypoxia, the therapeutic effect of hypoxia-limited oxygen intervention, is also an important factor in melanoma recurrence and metastasis; (3) inflammation, an important factor in the Tumor Microenvironment (TME) that promotes tumor development.
To overcome these obstacles, alternative therapies such as photodynamic therapy (PDT), photothermal therapy (PTT), gene therapy, immunotherapy, etc. have been developed for the treatment of melanoma, but the problems of hypoxia, metastasis and recurrence during the treatment of melanoma are not yet solved. Therefore, it is highly desirable to design an injectable hydrogel that can controllably release drugs, provide oxygen, and prevent melanoma metastasis and recurrence.
Disclosure of Invention
The technical problem to be solved by the invention is to solve the defects of the prior artThe preparation method of the photo-thermal controlled release polydopamine iron drug-loaded nanoparticle hydrogel is provided. The photo-thermal controlled-release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel obtained by the preparation method can be directly injected into a tumor to catalyze H in the tumor2O2Production of O2The hypoxia inside the tumor is relieved, and the hyperpyrexia generated by PDA-Fe @ Res is beneficial to the release of the medicine and kills melanoma cells under the condition of 808nm near-infrared laser auxiliary irradiation.
In order to solve the technical problems, the invention adopts the technical scheme that: a preparation method of a photo-thermal controlled release polydopamine iron drug-loaded nanoparticle hydrogel is characterized in that raw materials comprise dopamine, ferric chloride hexahydrate, resveratrol ethanol solution, sodium alginate, 1-ethyl- (3-dimethylaminopropyl) carbodiimide, N-hydroxysuccinimide, dopamine hydrochloride, hydrogen peroxide and horseradish peroxidase solution;
the method comprises the following steps:
providing polydopamine iron nanoparticles loaded with resveratrol;
providing sodium alginate grafted with dopamine;
adding hydrogen peroxide and a horseradish peroxidase solution into a system containing the dopamine-grafted sodium alginate and resveratrol-loaded poly-dopamine iron nanoparticle, and standing for crosslinking to obtain a photo-thermal controlled drug-release poly-dopamine iron drug-loaded nanoparticle hydrogel; the mass of the dopamine-grafted sodium alginate is 8-50 times of that of polydopamine iron nanoparticles loaded with resveratrol; the volume of the hydrogen peroxide is 53-133 times of the mass of the polydopamine iron nanoparticle loaded with the resveratrol, the unit of the volume of the hydrogen peroxide is mu L, the unit of the mass of the polydopamine iron nanoparticle loaded with the resveratrol is mg, and the concentration of the hydrogen peroxide is 0.1-0.5 wt%; the volume of the horseradish peroxidase solution is 44-133 times of the mass of the polydopamine iron nanoparticle loaded with the resveratrol, the unit of the volume of the horseradish peroxidase solution is mu L, the unit of the mass of the polydopamine iron nanoparticle loaded with the resveratrol is mg, and the concentration of the horseradish peroxidase solution is 0.5-4 mg/mL.
The preparation method of the photo-thermal controlled release poly-dopamine iron drug-loaded nanoparticle hydrogel is characterized by comprising the following steps:
adding ferric chloride hexahydrate into a dopamine aqueous solution, adjusting the pH value to 8.5, stirring for reaction for 0.5-1.5 h, and centrifugally drying to obtain poly-dopamine-iron nanoparticles;
dispersing the polydopamine iron nanoparticles obtained in the step one in water to obtain a system A, adding a resveratrol ethanol solution into the system A, stirring and reacting for 12-48 hours, and centrifugally drying to obtain polydopamine iron nanoparticles loaded with resveratrol;
placing sodium alginate in water to obtain a sodium alginate aqueous solution, adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide into the sodium alginate aqueous solution, activating for 20-45 min, adding dopamine hydrochloride powder, reacting for 12-18 h in a dark place to obtain a system B, placing the system B in deionized water, dialyzing for 48-96 h, and freeze-drying to obtain dopamine-grafted sodium alginate;
dissolving the dopamine-grafted sodium alginate in the step three in a solvent to obtain a system C, ultrasonically dispersing the resveratrol-loaded polydopamine iron nanoparticles in the step two in the system C to obtain a system D, adding hydrogen peroxide and a horseradish peroxidase solution into the system D, uniformly stirring at 25 ℃, standing for 200-430 s for crosslinking, and obtaining the photo-thermal controlled drug-release polydopamine iron drug-loaded nanoparticle hydrogel.
The preparation method of the photothermal controllable drug release poly-dopamine-iron drug-loaded nanoparticle hydrogel is characterized in that in the first step, the mass of ferric chloride hexahydrate is 0.04-0.1 times of the volume of a dopamine aqueous solution, the unit of the mass of ferric chloride hexahydrate is mg, the unit of the volume of the dopamine aqueous solution is mL, and the mass percentage of dopamine in the dopamine aqueous solution is 35-70%.
The preparation method of the photothermal controllable drug release polydopamine iron drug-loaded nanoparticle hydrogel is characterized in that in the second step, the volume of water is 0.5-1 times of the mass of the polydopamine iron nanoparticle, the unit of the volume of water is mL, and the unit of the mass of the polydopamine iron nanoparticle is mg.
The preparation method of the photo-thermal controlled release polydopamine iron drug-loaded nanoparticle hydrogel is characterized in that in the second step, the volume of the resveratrol ethanol solution is 0.3-1.2 times of the mass of the polydopamine iron nanoparticle, the unit of the volume of the resveratrol ethanol solution is mL, the unit of the mass of the polydopamine iron nanoparticle is mg, the resveratrol ethanol solution is a resveratrol ethanol solution obtained by dissolving resveratrol in absolute ethanol, and the concentration of resveratrol in the resveratrol ethanol solution is 1-3 mg/mL.
The preparation method of the photothermal controllable drug release poly-dopamine iron drug-loaded nanoparticle hydrogel is characterized in that in the third step, the cut-off molecular weight of the dialysis bag for dialysis is 8000-14000.
The preparation method of the photothermal controllable drug release poly-dopamine iron drug-loaded nanoparticle hydrogel is characterized in that in the third step, the volume of water is 0.1-0.2 times of the mass of sodium alginate, the volume unit of water is mL, and the mass unit of sodium alginate is mg.
The preparation method of the photo-thermal controlled release polydopamine iron drug-loaded nanoparticle hydrogel is characterized in that in the third step, the mass of the 1-ethyl- (3-dimethylaminopropyl) carbodiimide is 0.9-1 time of that of sodium alginate, the mass of the N-hydroxysuccinimide is 0.5-1.6 times of that of sodium alginate, and the mass of the dopamine hydrochloride is 1-2 times of that of sodium alginate.
The preparation method of the photo-thermal controlled release polydopamine iron drug-loaded nanoparticle hydrogel is characterized in that in the fourth step, the solvent is PBS buffer solution or deionized water.
The preparation method of the photo-thermal controlled release poly-dopamine-iron drug-loaded nanoparticle hydrogel is characterized in that in the fourth step, the horseradish peroxidase solution is obtained by dissolving horseradish peroxidase in water.
Compared with the prior art, the invention has the following advantages:
1. the preparation method of the photo-thermal controlled drug release polydopamine iron drug-loaded nanoparticle hydrogel comprises the step of crosslinking polydopamine iron nanoparticle (PDA-Fe @ Res Nps) loaded with resveratrol and sodium alginate grafted with dopamine in the presence of hydrogen peroxide and horseradish peroxidase to obtain the photo-thermal controlled drug release polydopamine iron drug-loaded nanoparticle (PDA-Fe @ Res) hydrogel. The photo-thermal controlled-release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel obtained by the preparation method can be directly injected into a tumor to catalyze H in the tumor2O2Production of O2The hypoxia inside the tumor is relieved, and the hyperpyrexia generated by PDA-Fe @ Res is favorable for releasing the medicine and killing melanoma cells under the near-infrared laser auxiliary irradiation condition of 808 nm.
2. The preparation method of the invention comprises the steps of taking horseradish peroxidase as a catalyst, promoting the formation of hydrogel, and catalyzing hydrogen peroxide to generate toxic hydroxyl free radicals (. OH) in tumors as peroxidase so as to assist in killing melanoma.
3. The preparation method comprises the step of preparing polydopamine iron nanoparticles loaded with the resveratrol by taking dopamine, ferric chloride and resveratrol ethanol solution as raw materials, wherein the polydopamine iron nanoparticles loaded with the resveratrol have the property similar to catalase and can catalyze H in tumors2O2Generating oxygen (O)2) And can relieve internal hypoxia of tumor.
4. Preferably, the preparation method of the invention comprises the step of obtaining the dopamine-grafted sodium alginate by taking sodium alginate, 1-ethyl- (3-dimethylaminopropyl) carbodiimide, N-hydroxysuccinimide and dopamine hydrochloride as raw materials to react in a dark place, wherein the dopamine-grafted sodium alginate is used as a hydrogel framework and has excellent adhesiveness, self-healing property, injectability and anti-inflammation property.
The technical solution of the present invention is further described in detail with reference to the accompanying drawings and embodiments.
Drawings
Fig. 1a is a schematic diagram of a preparation process of polydopamine iron nanoparticles (PDA-fe @ res Nps) loaded with resveratrol in step two of example 1.
FIG. 1b is the SEM image of polydopamine iron nanoparticle (PDA-Fe @ Res Nps) loaded with resveratrol in step two of example 1.
FIG. 1c is a transmission electron microscope image of polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol in step two of example 1.
FIG. 1d is the HAADF-STEM element distribution of polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol in step two of example 1.
FIG. 1e is a diagram of the distribution of the C element in the region shown in FIG. 1 d.
FIG. 1f is a graph showing the distribution of O elements in the region shown in FIG. 1 d.
FIG. 1g is a distribution diagram of N elements in the region shown in FIG. 1 d.
FIG. 1h is a distribution diagram of Fe element in the region shown in FIG. 1 d.
FIG. 1i is an XPS total spectrum of polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol in step two of example 1.
FIG. 1j is a C1s spectrum of polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol in step two of example 1.
FIG. 1k is the N1s spectrum of polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol in step two of example 1.
FIG. 2a is a schematic view of a process of obtaining photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel by crosslinking in example 1.
FIG. 2b is a schematic diagram of injection performance of the photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel.
FIG. 2c is a schematic diagram of rheological behavior of photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel obtained by crosslinking in example 1.
FIG. 2d is a graph showing the stability of the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel obtained by crosslinking in example 1.
FIG. 2e is a schematic diagram of a test of self-healing performance of a photo-thermal controlled drug release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel.
FIG. 2f is a schematic view showing the state of the photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel adhered to the skin.
FIG. 2g is a schematic view showing the state that the photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel of the present invention is adhered to the surface of different substrates.
FIG. 2h is an SEM picture of a dopamine grafted sodium alginate (SA-DA-EN) hydrogel of comparative example 1 and an SEM picture of a photothermal controlled release polydopamine iron drug loaded (PDA-Fe @ Res) nanoparticle hydrogel of example 1.
FIG. 2i shows the swelling ratio of the photo-thermal controlled release polydopamine iron drug loaded (PDA-Fe @ Res) nanoparticle hydrogel of example 1 and the sodium alginate (SA-DA-EN) hydrogel grafted with dopamine of comparative example 1.
FIG. 3a is a graph showing the peroxidase activity test results of dopamine-grafted sodium alginate (SA-DA) of example 1.
FIG. 3b is a graph showing peroxidase activity of the dopamine grafted sodium alginate (SA-DA-EN) hydrogel of comparative example 1 as a function of SA-DA concentration.
FIG. 3c shows peroxidase activity as H for the dopamine grafted sodium alginate (SA-DA-EN) hydrogel of comparative example 12O2Schematic diagram of concentration change.
FIG. 3d is a schematic diagram of the oxygen release capacity of the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel of example 1.
FIG. 3e is a graph showing the photo-thermal performance test results of the photo-thermal controlled release poly dopamine iron drug loaded (PDA-Fe @ Res) nanoparticle hydrogel of example 1 and the sodium alginate grafted with dopamine (SA-DA-EN) hydrogel of comparative example 1.
FIG. 3f is a schematic diagram showing a photo-thermal performance test result of the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel under near-infrared irradiation of different powers.
FIG. 3g is a schematic diagram of the photo-thermal stability test result of the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel of example 1.
FIG. 3h is a schematic diagram illustrating the single-cycle photothermal stability test result of the photothermal controllable drug release polydopamine iron drug loading (PDA-Fe @ Res) nanoparticle hydrogel of example 1.
FIG. 3i is a schematic diagram showing the negative natural logarithm relationship between the temperature change and the cooling time in the cooling stage of the single-cycle photothermal stability test.
FIG. 3j is a schematic view of the drug release performance of the photothermal controlled drug release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel in example 1.
FIG. 3k is a schematic view of the drug release performance of the photothermal controllable drug release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel of example 1 under the condition of 808nm near-infrared light irradiation.
FIG. 3m is a schematic diagram of the result of the blood compatibility test of the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel of example 1.
Detailed Description
Example 1
The embodiment provides a preparation method of a photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel, which comprises the following steps:
step one, 6.2mg ferric chloride hexahydrate (FeCl) was added to 130ml dopamine aqueous solution3·6H2O), adjusting the pH value to 8.5, stirring for reaction for 1h, and centrifugally drying to obtain poly-dopamine-iron (PDA-Fe) nanoparticles; the mass percentage of the dopamine in the dopamine aqueous solution is 35%, and the pH is adjusted by using Tris with the mass percentage of 2.25%; the temperature of the stirring reaction is 25 ℃; the drying temperature is 37 ℃;
step two, dispersing 10mg of the polydopamine iron (PDA-Fe) nano particles obtained in the step one into 10ml of water to obtain a system A, adding 9ml of resveratrol (Res) ethanol solution with the concentration of 1mg/ml into the system A, stirring for reaction for 24 hours, and centrifugally drying to obtain 13.9mg of polydopamine iron nano particles (PDA-Fe @ Res Nps) loaded with resveratrol; the temperature of the stirring reaction is 25 ℃, and the drying temperature is 37 ℃;
step three, placing 500mg of sodium alginate in 100ml of water to obtain a Sodium Alginate (SA) aqueous solution, adding 0.48g of 1-ethyl- (3-dimethylaminopropyl) carbodiimide (EDC) and 0.80g of N-hydroxysuccinimide (NHS) into the Sodium Alginate (SA) aqueous solution, activating for 30min, adding 500mg of dopamine hydrochloride (DA & HCl) powder, reacting for 12h in a dark place to obtain a system B, placing the system B in deionized water for dialysis for 72h, and freeze-drying to obtain dopamine-grafted sodium alginate (SA-DA); the activation is stirring activation at 25 ℃, and the cut-off molecular weight of the dialysis bag for dialysis is 8000-14000; the temperature of freeze-drying is-55 ℃;
step four, dissolving the dopamine-grafted sodium alginate (SA-DA) in the step three in a PBS buffer solution to obtain a system C with the dopamine-grafted sodium alginate mass percentage content of 4.0wt%, ultrasonically dispersing 1mg of the resveratrol-loaded polydopamine iron nanoparticles (PDA-Fe @ Res Nps) in the step two in 1mL of the system C to obtain a system D with the resveratrol-loaded polydopamine iron nanoparticle concentration of 1mg/mL, and adding 20 mu L of hydrogen peroxide (H) with the concentration of 0.5wt% into 150 mu L of the system D2O2) And 20 mu L of horseradish peroxidase (HRP) solution with the concentration of 3mg/ml, stirring uniformly at 25 ℃, standing for 240s for crosslinking to obtain the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel; the horseradish peroxidase (HRP) solution is obtained by dissolving horseradish peroxidase in water.
Comparative example 1
The present comparative example provides a method of preparing a dopamine grafted sodium alginate (SA-DA-EN) hydrogel, comprising the steps of:
step one, 500mg of sodium alginate is placed in 100ml of water to obtain a Sodium Alginate (SA) aqueous solution, 0.48g of 1-ethyl- (3-dimethylaminopropyl) carbodiimide (EDC) and 0.80g of N-hydroxysuccinimide (NHS) are added into the Sodium Alginate (SA) aqueous solution, 500mg of dopamine hydrochloride (DA & HCl) powder is added after activation is carried out for 30min, a dark reaction is carried out for 12h to obtain a system B, the system B is placed in deionized water for dialysis for 72h, and freeze drying is carried out to obtain the dopamine-grafted sodium alginate (SA-DA); the activation is stirring activation at 25 ℃, and the cut-off molecular weight of the dialysis bag for dialysis is 8000-14000; the temperature of the freeze-drying is-55 ℃;
step two, dissolving the dopamine-grafted sodium alginate (SA-DA) in the step one in a PBS buffer solution to obtain a system C with the dopamine-grafted sodium alginate mass percentage content of 4.0wt%, and adding 20 mu L of hydrogen peroxide (H) with the concentration of 0.5wt% into 150 mu L of the system C2O2) And 20 mu L of horseradish peroxidase (HRP) solution with the concentration of 3mg/ml, stirring uniformly at 25 ℃, standing for 200s for crosslinking to obtain dopamine-grafted sodium alginate (SA-DA-EN) hydrogel; the horseradish peroxidase (HRP) solution is obtained by dissolving horseradish peroxidase in water.
Example 2
The embodiment provides a preparation method of a photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel, which comprises the following steps:
step one, adding 12.4mg ferric chloride hexahydrate (FeCl) to 130ml dopamine aqueous solution3·6H2O), adjusting the pH value to 8.5, stirring for reacting for 1.5h, and centrifugally drying to obtain poly-dopamine-iron (PDA-Fe) nanoparticles; the mass percentage of the dopamine in the dopamine aqueous solution is 35%, and the pH is adjusted by using Tris with the mass percentage of 2.25%; the temperature of the stirring reaction is 25 ℃; the drying temperature is 37 ℃;
step two, dispersing 10mg of the polydopamine iron (PDA-Fe) nanoparticles obtained in the step one in 10ml of water to obtain a system A, adding 3ml of resveratrol (Res) ethanol solution with the concentration of 2mg/ml into the system A, stirring for reaction for 48 hours, and centrifugally drying to obtain 12.8mg of polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol; the temperature of the stirring reaction is 25 ℃, and the drying temperature is 37 ℃;
step three, placing 1g of sodium alginate in 100ml of water to obtain a Sodium Alginate (SA) aqueous solution, adding 0.96g of 1-ethyl- (3-dimethylaminopropyl) carbodiimide (EDC) and 1.60g of N-hydroxysuccinimide (NHS) into the Sodium Alginate (SA) aqueous solution, activating for 25min, adding 1g of dopamine hydrochloride (DA & HCl) powder, reacting for 15h in a dark place to obtain a system B, placing the system B in deionized water for dialysis for 96h, and freeze-drying to obtain dopamine-grafted sodium alginate (SA-DA); the activation is stirring activation at 25 ℃, and the cut-off molecular weight of the dialysis bag for dialysis is 8000-14000; the temperature of freeze-drying is-55 ℃;
step four, dissolving the dopamine-grafted sodium alginate (SA-DA) in the step three in a PBS buffer solution to obtain a system C with the dopamine-grafted sodium alginate mass percentage content of 4.0wt%, ultrasonically dispersing 2mg of the resveratrol-loaded polydopamine iron nanoparticles (PDA-Fe @ Res Nps) in the step two in 1mL of the system C to obtain a system D with the resveratrol-loaded polydopamine iron nanoparticle concentration of 2mg/mL, and adding 30 mu L of hydrogen peroxide (H) with the concentration of 0.5wt% into 150 mu L of the system D2O2) And 20 mu L of horseradish peroxidase (HRP) solution with the concentration of 3mg/ml, stirring uniformly at 25 ℃, standing for 200s for crosslinking to obtain the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel; the horseradish peroxidase (HRP) solution is obtained by dissolving horseradish peroxidase in water.
The photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel performance of the embodiment is basically consistent with that of the embodiment 1.
Example 3
The embodiment provides a preparation method of a photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel, which comprises the following steps:
step one, 6.2mg ferric chloride hexahydrate (FeCl) was added to 130ml dopamine aqueous solution3·6H2O), adjusting the pH value to 8.5, stirring for reaction for 0.5h, and centrifugally drying to obtain poly-dopamine-Fe (PDA-Fe) nanoparticles; the mass percentage of the dopamine in the dopamine aqueous solution is 70%, and the pH is adjusted by using Tris with the mass percentage of 2.25%; the temperature of the stirring reaction is 25 ℃; the drying temperature is 37 ℃;
step two, dispersing 10mg of the polydopamine iron (PDA-Fe) nanoparticles obtained in the step one in 10ml of water to obtain a system A, adding 3mg of resveratrol (Res) ethanol solution with the concentration of 1mg/ml into the system A, stirring for reaction for 24 hours, and centrifugally drying to obtain 11.2mg of polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol; the temperature of the stirring reaction is 25 ℃, and the drying temperature is 37 ℃;
step three, placing 500mg of sodium alginate in 100ml of water to obtain a Sodium Alginate (SA) aqueous solution, adding 0.48g of 1-ethyl- (3-dimethylaminopropyl) carbodiimide (EDC) and 0.58g of N-hydroxysuccinimide (NHS) into the Sodium Alginate (SA) aqueous solution, activating for 20min, adding 1g of dopamine hydrochloride (DA & HCl) powder, reacting for 18h in a dark place to obtain a system B, placing the system B in deionized water for dialysis for 72h, and freeze-drying to obtain dopamine-grafted sodium alginate (SA-DA); the activation is stirring activation at 25 ℃, and the cut-off molecular weight of the dialysis bag for dialysis is 8000-14000; the temperature of the freeze-drying is-55 ℃;
step four, dissolving the dopamine-grafted sodium alginate (SA-DA) in the step three in a PBS buffer solution to obtain a system C with the dopamine-grafted sodium alginate mass percentage content of 5.0wt%, ultrasonically dispersing 1mg of the resveratrol-loaded polydopamine iron nanoparticles (PDA-Fe @ Res Nps) in the step two in 1mL of the system C to obtain a system D with the resveratrol-loaded polydopamine iron nanoparticle concentration of 1mg/mL, and adding 20 mu L of hydrogen peroxide (H) with the concentration of 0.1 wt% into 150 mu L of the system D2O2) And 20 mul of Horse Radish Peroxidase (HRP) solution with the concentration of 0.5mg/ml, stirring uniformly at 25 ℃, standing for 340s for crosslinking to obtain photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel; the horseradish peroxidase (HRP) solution is obtained by dissolving horseradish peroxidase in water.
The photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel performance of the embodiment is basically consistent with that of the embodiment 1.
Example 4
The embodiment provides a preparation method of a photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel, which comprises the following steps:
step one, adding 12.4mg ferric chloride hexahydrate (FeCl) to 130ml dopamine aqueous solution3·6H2O), adjusting the pH value to 8.5, stirring for reaction for 1h, and centrifugally drying to obtain poly-dopamine-iron (PDA-Fe) nanoparticles; the mass percentage of the dopamine in the dopamine aqueous solution is 70%, and the pH value is adjusted by using Tris with the mass percentage of 22.5%; the temperature of the stirring reaction is 25 ℃; the drying temperature is 37 ℃;
step two, dispersing 10mg of the polydopamine iron (PDA-Fe) nano particles obtained in the step one in 5ml of water to obtain a system A, adding 12mg of resveratrol (Res) ethanol solution with the concentration of 1mg/ml into the system A, stirring for reaction for 12 hours, and centrifugally drying to obtain 12.4mg of polydopamine iron nano particles (PDA-Fe @ Res Nps) loaded with resveratrol; the temperature of the stirring reaction is 25 ℃, and the drying temperature is 37 ℃;
step three, placing 1g of sodium alginate in 100ml of water to obtain a Sodium Alginate (SA) aqueous solution, adding 968mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide (EDC) and 582mg of N-hydroxysuccinimide (NHS) into the Sodium Alginate (SA) aqueous solution, activating for 45min, adding 1.92g of dopamine hydrochloride (DA & HCl) powder, reacting for 15h in a dark place to obtain a system B, placing the system B in deionized water for dialysis for 96h, and freeze-drying to obtain dopamine-grafted sodium alginate (SA-DA); the activation is stirring activation at 25 ℃, and the cut-off molecular weight of the dialysis bag for dialysis is 8000-14000; the temperature of the freeze-drying is-55 ℃;
step four, dissolving the dopamine-grafted sodium alginate (SA-DA) in the step three in deionized water to obtain a system C with the dopamine-grafted sodium alginate mass percentage content of 2.5wt%, ultrasonically dispersing 6mg of the polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with the resveratrol in the step two in 2mL of the system C to obtain a system D with the polydopamine iron nanoparticle concentration of 3mg/mL loaded with the resveratrol, and adding 25 mu L of hydrogen peroxide (H) with the concentration of 0.5wt% into 150 mu L of the system D2O2) And 20 μ L of horseradish peroxidase (HRP) solution with concentration of 3mg/ml, stirring at 25 deg.C, standing for 430s to allowCrosslinking to obtain photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nano-particle hydrogel; the horseradish peroxidase (HRP) solution is obtained by dissolving horseradish peroxidase in water.
The property of the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel is basically consistent with that of the example 1.
Example 5
The embodiment provides a preparation method of photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel, which comprises the following steps:
step one, 6.2mg ferric chloride hexahydrate (FeCl) was added to 130ml dopamine aqueous solution3·6H2O), adjusting the pH value to 8.5, stirring for reacting for 1.5h, and centrifugally drying to obtain poly-dopamine-iron (PDA-Fe) nanoparticles; the mass percentage of the dopamine in the dopamine aqueous solution is 35%, and the pH is adjusted by using Tris with the mass percentage of 2.25%; the temperature of the stirring reaction is 25 ℃; the drying temperature is 37 ℃;
step two, dispersing 10mg of the polydopamine iron (PDA-Fe) nanoparticles obtained in the step one in 5ml of water to obtain a system A, adding 3ml of resveratrol (Res) ethanol solution with the concentration of 3mg/ml into the system A, stirring for reaction for 48 hours, and centrifugally drying to obtain 11.2mg of polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol; the temperature of the stirring reaction is 25 ℃, and the drying temperature is 37 ℃;
step three, placing 500mg of sodium alginate in 100ml of water to obtain a Sodium Alginate (SA) aqueous solution, adding 484mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide (EDC) and 582mg of N-hydroxysuccinimide (NHS) into the Sodium Alginate (SA) aqueous solution, activating for 40min, adding 500mg of dopamine hydrochloride (DA & HCl) powder, reacting for 18h in a dark place to obtain a system B, placing the system B in deionized water for dialysis for 48h, and freeze-drying to obtain the dopamine-grafted sodium alginate (SA-DA); the activation is stirring activation at 25 ℃, and the cut-off molecular weight of the dialysis bag for dialysis is 8000-14000; the temperature of the freeze-drying is-55 ℃;
step four, dissolving the dopamine-grafted sodium alginate (SA-DA) in the step three in deionized water to obtain a system C with the dopamine-grafted sodium alginate mass percentage content of 4.0wt%, ultrasonically dispersing 2.5mg of the resveratrol-loaded polydopamine iron nanoparticles (PDA-Fe @ Res Nps) in the step two in 1mL of the system C to obtain a system D with the resveratrol-loaded polydopamine iron nanoparticles concentration of 2.5mg/mL, and adding 20 mu L of hydrogen peroxide (H) with the concentration of 0.3wt% into 150 mu L of the system D2O2) And 20 mu L of horseradish peroxidase (HRP) solution with the concentration of 4mg/ml, stirring uniformly at 25 ℃, standing for 300s for crosslinking to obtain photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel; the horseradish peroxidase (HRP) solution is obtained by dissolving horseradish peroxidase in water.
The photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel performance of the embodiment is basically consistent with that of the embodiment 1.
Performance evaluation:
FIG. 1a is a schematic diagram of a preparation process of polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol in step two of example 1, and it can be known from FIG. 1a that Fe is in an alkaline condition3+Performing coordination polymerization with dopamine molecules to obtain polydopamine iron (PDA-Fe) nanoparticles, and then performing pi-pi accumulation with resveratrol to obtain polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol.
FIG. 1b is a scanning electron microscope image of polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol in step two of example 1, and it can be seen from FIG. 1b that the particles are spherical, the particle size is uniformly distributed around 95nm, and the structure is uniform.
Fig. 1c is a transmission electron microscope image of polydopamine iron nanoparticles (PDA-fe @ res Nps) loading resveratrol in step two of example 1, and fig. 1c further proves that the particles are in a uniform spherical shape.
Fig. 1d shows the HAADF-STEM element distribution of polydopamine iron nanoparticles (PDA-Fe @ res Nps) loaded with resveratrol in step two of example 1, fig. 1e shows the C element distribution map in the region shown in fig. 1d, fig. 1f shows the O element distribution map in the region shown in fig. 1d, fig. 1g shows the N element distribution map in the region shown in fig. 1d, fig. 1h shows the Fe element distribution map in the region shown in fig. 1d, and in combination with fig. 1d to 1h, it can be seen that C, O, N, and Fe elements are uniformly distributed in the particles.
Fig. 1i is an XPS total spectrum of the polydopamine iron nanoparticle loaded with resveratrol (PDA-fe @ res Nps) in step two of example 1, fig. 1j is a C1s spectrum of the polydopamine iron nanoparticle loaded with resveratrol (PDA-fe @ res Nps) in step two of example 1, and fig. 1k is an N1s spectrum of the polydopamine iron nanoparticle loaded with resveratrol (PDA-fe @ res Nps) in step two of example 1. According to fig. 1i, elements of C, O, N and Fe are present in polydopamine iron nanoparticles (PDA-Fe @ res Nps) loaded with resveratrol. FIG. 1j is a C1s spectrum with the main band centered at 284.6eV coinciding with C-C, the other two bands at 286.4eV and 288.9eV coinciding with C-O and C = O, respectively, FIG. 1k is a N1s spectrum with the two peaks at 399.7eV and 401.7eV corresponding to-NH in dopamine2And = N-H. As can be seen from the combination of FIGS. 1a to 1k, polydopamine iron nanoparticles (PDA-Fe @ Res Nps) loaded with resveratrol are successfully prepared.
FIG. 2a is a schematic state diagram of photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel obtained by crosslinking in example 1. As can be seen from fig. 2a, the nanoparticles are uniformly dispersed in the hydrogel.
FIG. 2b shows that the photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel is loaded into an injector and can smoothly flow out, which indicates that the photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel has injectability.
FIG. 2c is a schematic diagram of rheological behavior of photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel obtained by crosslinking in example 1. As can be seen, the storage modulus (G ') is greater than the loss modulus (G'), the hydrogel is in the solid state, and when the loss modulus (G ') is greater than the storage modulus (G') after reaching the stress critical point (500%), the hydrogel is in the liquid state, indicating that the hydrogel of the present invention has mechanical fluidity. Hydrogel rheological behavior was tested using a rheometer in a strain sweep mode.
FIG. 2d shows that the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel obtained by crosslinking in example 1 is subjected to stability test, according to the test in FIG. 2d, the hydrogel is destroyed under 500% of strain, and when the applied strain is recovered to 1%, the hydrogel is rapidly recovered to be close to the initial gel state, which indicates that the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel has self-healing property of rapid repair after fracture. The hydrogel stability test is a strain amplitude sweep pattern test.
FIG. 2e is a schematic diagram of a test on self-healing performance of a photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel. The hydrogel is adhered to the skin, the cut sections are contacted after cutting, the sections can be healed, and the hydrogel adhered to the skin after the finger is bent does not crack.
FIG. 2f is a schematic view of a state that the photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel is adhered to the skin, and FIG. 2g is a schematic view of a state that the photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel is adhered to the surfaces of different substrates, which shows that the photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel can be adhered to different substrates and has excellent tissue adhesion.
FIG. 2h is an SEM picture of a dopamine-grafted sodium alginate (SA-DA-EN) hydrogel in comparative example 1 and an SEM picture of a photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel in example 1, and as can be seen from the SEM pictures, the aperture of the dopamine-grafted sodium alginate (SA-DA-EN) hydrogel in comparative example 1 is 143 μ M, and the aperture of the photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel is 228 μ M, which indicates that the aperture of the hydrogel is increased.
FIG. 2i shows the swelling ratios of the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel in example 1 and the dopamine-grafted sodium alginate (SA-DA-EN) hydrogel in comparative example 1, and it can be seen from the graphs that the swelling ratio of the hydrogel disclosed by the invention is about 939%, which is significantly higher than that of the dopamine-grafted sodium alginate (SA-DA-EN) hydrogel, and the method disclosed by the invention can effectively improve the swelling ratio of the hydrogel, which indicates that the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel disclosed by the invention has higher nanoparticle release performance.
The photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nano-particle hydrogel is expected to be directly injected into the tumor in clinical application.
FIG. 3a is a graph showing the results of peroxidase activity test of dopamine-grafted sodium alginate (SA-DA) of example 1, the peroxidase activity test using 3,3', 5' -Tetramethyldiphenylamine (TMB) as a probe, and determining the peroxidase activity according to whether the probe is oxidized by OH to form blue oxTMB, the test method comprising: 0.01g of the lyophilized hydrogel, 100. Mu.M hydrogen peroxide, and a TMB solution (24 mg/ml) dissolved in dimethyl sulfoxide were added to an acetate buffer (pH = 4.0), and the mixture was maintained at room temperature for 5 minutes, and a full-scan spectrum of the solution was measured using a UV spectrophotometer, and the result is shown in FIG. 3 a. As can be seen from FIG. 3a, only H is contained2O2No blue reaction with TMB, sodium alginate (SA-DA) containing only grafted dopamine and no blue reaction with TMB, indicating no OH is generated, and H is contained2O2And the dopamine-grafted sodium alginate (SA-DA) in the system, TMB has blue reaction, which shows that OH is generated and TMB is oxidized, and shows that the dopamine-grafted sodium alginate (SA-DA) can catalyze H2O2OH was produced, indicating that the hydrogel of the invention can kill melanoma cells.
FIG. 3b is a graph showing peroxidase activity of the dopamine grafted sodium alginate (SA-DA-EN) hydrogel of comparative example 1 as a function of SA-DA concentration. As can be seen from FIG. 3b, the amount of OH, which is a hydroxyl radical, increases with the increase of the concentration of SA-DA, indicating that increasing the SA-DA content within a certain range can increase the peroxidase activity.
FIG. 3c shows peroxidase activity as a function of H for the dopamine-grafted sodium alginate (SA-DA-EN) hydrogel of comparative example 12O2Schematic diagram of concentration change. From FIG. 3c, it can be seen that following H2O2The increase in the concentration of the water-soluble polymer,the amount of OH, which is a hydroxyl radical, increases, indicating that H increases within a certain range2O2The content can improve the peroxidase activity.
FIG. 3d is a schematic diagram of the oxygen release capacity of the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel of example 1, and the test method includes: placing 1ml of the hydrogel in different pH buffers (pH =5.0, pH =6.4, pH = 7.4) containing hydrogen peroxide (100 μ M), and optionally performing 808nm near-infrared laser irradiation (1.0W-cm)-2)The content of oxygen generated was measured by a JPBJ-608 portable dissolved oxygen analyzer, and the value was recorded every 1min, and the result is shown in FIG. 3 d. As can be seen from the figure, the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel has certain catalase-like activity under acidic and neutral conditions, compared with pH =7.4, the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel catalase activity under acidic conditions is inhibited, and the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel has obviously increased catalase-like activity under 808nm laser irradiation, which indicates that laser irradiation can realize reactivation of the hydrogel and promote oxygen generation.
FIG. 3e is a graph showing the photo-thermal performance test results of the photo-thermal controlled release poly dopamine iron drug loaded (PDA-Fe @ Res) nanoparticle hydrogel of example 1 and the sodium alginate grafted with dopamine (SA-DA-EN) hydrogel of comparative example 1, the test method comprising: the hydrogel was placed in a glass vial containing 1mL of water at 808nm (1W/cm)2And 10 min), testing the temperature change of the photo-thermal controlled drug release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nano particle hydrogel with different qualities by using a thermodetector. According to the figure 3e, the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel under infrared irradiation can convert light energy into heat, and the temperature amplification is obvious along with the increase of the hydrogel mass, so that the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel has excellent photo-thermal performance.
FIG. 3f is a schematic diagram of a photo-thermal performance test result of the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel under the irradiation of near-infrared light with different powers. The test method is the same as that of fig. 3 e. As can be seen from fig. 3f, the temperature increase increases with increasing near infrared power. As can be seen from the combination of fig. 3e and fig. 3f, the photothermal performance of the photothermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel is more significantly influenced by the illumination power.
Fig. 3g is a schematic diagram showing a photo-thermal stability test result of the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-fe @ res) nanoparticle hydrogel of example 1, and the test method includes performing five on/off periodic irradiation of near infrared light of 808 nm. According to the figure 3g, in the whole period process, the hydrogel can be rapidly heated up to 47.7 ℃ when near-infrared illumination is carried out, and can be gradually cooled to the initial temperature when the near-infrared illumination is shut down, so that the photo-thermal controlled drug release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel has stable photo-thermal conversion performance.
FIG. 3h is a diagram showing the single-cycle photothermal stability test result of the photothermal controllable drug release polydopamine iron drug loading (PDA-Fe @ Res) nanoparticle hydrogel of example 1, under the condition of using 808nm near-infrared laser (1.0W cm)-210 min) cooling was performed with irradiation/off for one cycle. FIG. 3i is a schematic diagram showing the negative natural logarithm relationship between the temperature change and the cooling time in the cooling stage of the single-cycle photo-thermal stability test. According to the graphs of 3h and 3i, the photo-thermal controlled release poly-dopamine iron drug-loading (PDA-Fe @ Res) nano particle hydrogel can reach 47.7 ℃ within 10min, the tumor ablation requirement of the photo-thermal effect is met, and the photo-thermal conversion efficiency is as high as 42.7%.
Fig. 3j is a schematic view of drug release performance of the photothermal controlled drug release polydopamine iron drug-loaded (PDA-fe @ res) nanoparticle hydrogel of example 1, and the test method includes: the freeze-dried hydrogel was immersed in centrifuge tubes containing PBS phosphate buffer solutions with different pH values (5.0, 6.2, and 7.4), then placed in an incubator at 37 ℃ for a preset incubation time, 1mL of the leach solution was collected and an equal amount of fresh buffer solution (1 mL) was added, then absorbance at 306nm was detected using an ultraviolet-visible spectrophotometer, and the amount of drug released was calculated in conjunction with a resveratrol standard curve, with the results shown in fig. 3 j. As can be seen from FIG. 3j, the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel has a high drug release amount at pH 5.0 and pH 6.2, which is probably because the hydrogel can be induced to swell under an acidic pH condition, so that the drug outflow is accelerated.
FIG. 3k is a schematic view of drug release performance of the photo-thermal controlled release polydopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel in example 1 under 808nm near-infrared irradiation. According to the graph 3k, the hydrogel drug release capability is obviously improved under the near-infrared illumination condition, and the near-infrared triggered off-on characteristic is shown, so that the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel has the photo-thermal controlled release performance, which is probably attributed to the characteristic that pi-pi accumulation or hydrogen bonds are influenced by temperature.
FIG. 3m is a schematic diagram of the result of the blood compatibility test of the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nanoparticle hydrogel of example 1. In which the haemocompatibility was assessed with erythrocytes using pure water as a positive control. The test method comprises the following steps: different hydrogel samples were added to tubes containing PBS (0.8 mL), 0.2mL of fresh diluted blood was added to each tube, respectively, the mixture was incubated in a water bath environment at 37 ℃ for 60 minutes, followed by centrifugation at 3000rpm for 5 minutes, and absorbance at 540nm was measured using an ultraviolet spectrophotometer with deionized water and PBS as positive and negative controls, wherein the hemolysis rate = [ (ODh-ODn)/(ODp-ODn) ] × 100%. Wherein ODh, ODn and ODp represent absorbance values of the sample, PBS and DI water, respectively, and the results are shown in FIG. 3 m. The result shows that the hemolysis rate of the experimental group is in a safe range, namely less than 5%, and the photo-thermal controlled release poly-dopamine iron drug-loaded (PDA-Fe @ Res) nano-particle hydrogel has good in-vivo biosafety.
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, changes and equivalent structural changes made to the above embodiment according to the technical spirit of the present invention still fall within the protection scope of the technical solution of the present invention.
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, changes and equivalent structural changes made to the above embodiment according to the technical spirit of the present invention still fall within the protection scope of the technical solution of the present invention.

Claims (10)

1. A preparation method of a photo-thermal controlled release polydopamine iron drug-loaded nanoparticle hydrogel is characterized in that raw materials comprise dopamine, ferric chloride hexahydrate, resveratrol ethanol solution, sodium alginate, 1-ethyl- (3-dimethylaminopropyl) carbodiimide, N-hydroxysuccinimide, dopamine hydrochloride, hydrogen peroxide and horseradish peroxidase solution;
the method comprises the following steps:
providing polydopamine iron nanoparticles loaded with resveratrol;
providing sodium alginate grafted with dopamine;
adding hydrogen peroxide and a horseradish peroxidase solution into a system containing the dopamine-grafted sodium alginate and resveratrol-loaded polydopamine iron nanoparticle, and standing for crosslinking to obtain a photothermal controllable drug-release polydopamine iron drug-loaded nanoparticle hydrogel; the mass of the dopamine-grafted sodium alginate is 8-50 times of that of the polydopamine iron nanoparticle loaded with resveratrol; the volume of the hydrogen peroxide is 53-133 times of the mass of the polydopamine iron nanoparticle loaded with the resveratrol, the unit of the volume of the hydrogen peroxide is mu L, the unit of the mass of the polydopamine iron nanoparticle loaded with the resveratrol is mg, and the concentration of the hydrogen peroxide is 0.1-0.5 wt%; the volume of the horseradish peroxidase solution is 44-133 times of the mass of the polydopamine iron nanoparticle loaded with the resveratrol, the unit of the volume of the horseradish peroxidase solution is mu L, the unit of the mass of the polydopamine iron nanoparticle loaded with the resveratrol is mg, and the concentration of the horseradish peroxidase solution is 0.5-4 mg/mL.
2. The preparation method of the photothermal controlled release polydopamine iron drug-loaded nanoparticle hydrogel according to claim 1, which is characterized by comprising the following steps:
step one, adding ferric chloride hexahydrate into a dopamine aqueous solution, adjusting the pH value to 8.5, stirring for reaction for 0.5-1.5 h, and centrifugally drying to obtain poly-dopamine-iron nanoparticles;
dispersing the polydopamine iron nanoparticles obtained in the step one in water to obtain a system A, adding a resveratrol ethanol solution into the system A, stirring and reacting for 12-48 h, and centrifugally drying to obtain polydopamine iron nanoparticles loaded with resveratrol;
placing sodium alginate in water to obtain a sodium alginate aqueous solution, adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide into the sodium alginate aqueous solution, activating for 20-45 min, adding dopamine hydrochloride powder, reacting for 12-18 h in a dark place to obtain a system B, placing the system B in deionized water, dialyzing for 48-96 h, and freeze-drying to obtain the dopamine-grafted sodium alginate;
dissolving the dopamine-grafted sodium alginate in the step three in a solvent to obtain a system C, ultrasonically dispersing the resveratrol-loaded polydopamine iron nanoparticles in the step two in the system C to obtain a system D, adding hydrogen peroxide and a horseradish peroxidase solution into the system D, uniformly stirring at 25 ℃, standing for 200-430 s for crosslinking, and obtaining the photo-thermal controlled drug-release polydopamine iron drug-loaded nanoparticle hydrogel.
3. The preparation method of the photothermal controlled release poly-dopamine iron drug-loaded nanoparticle hydrogel according to claim 2, wherein in step one, the mass of ferric chloride hexahydrate is 0.04-0.1 times of the volume of the dopamine aqueous solution, the unit of the mass of ferric chloride hexahydrate is mg, the unit of the volume of the dopamine aqueous solution is mL, and the mass percentage of dopamine in the dopamine aqueous solution is 35% -70%.
4. The preparation method of the photothermal controlled release polydopamine iron drug-loaded nanoparticle hydrogel according to claim 2, wherein in the second step, the volume of the water is 0.5-1 times the mass of the polydopamine iron nanoparticle, the unit of the volume of the water is mL, and the unit of the mass of the polydopamine iron nanoparticle is mg.
5. The preparation method of the photo-thermal controlled release polydopamine iron drug-loaded nanoparticle hydrogel according to claim 2, wherein in the second step, the volume of the resveratrol ethanol solution is 0.3-1.2 times of the mass of the polydopamine iron nanoparticles, the unit of the volume of the resveratrol ethanol solution is mL, the unit of the mass of the polydopamine iron nanoparticles is mg, the resveratrol ethanol solution is a resveratrol ethanol solution obtained by dissolving resveratrol in absolute ethanol, and the concentration of resveratrol in the resveratrol ethanol solution is 1-3 mg/mL.
6. The preparation method of the photothermal controllable drug release poly-dopamine iron drug-loaded nanoparticle hydrogel according to claim 2, characterized in that in the third step, the cut-off molecular weight of the dialysis bag for dialysis is 8000-14000.
7. The preparation method of the photothermal controllable drug release poly-dopamine iron drug-loaded nanoparticle hydrogel according to claim 2, characterized in that in the third step, the volume of the water is 0.1-0.2 times of the mass of sodium alginate, the volume unit of the water is mL, and the unit of the mass of sodium alginate is mg.
8. The preparation method of the photothermal controlled release poly-dopamine iron drug-loaded nanoparticle hydrogel according to claim 2, characterized in that in step three, the mass of the 1-ethyl- (3-dimethylaminopropyl) carbodiimide is 0.9-1 time of the mass of sodium alginate, the mass of the N-hydroxysuccinimide is 0.5-1.6 times of the mass of sodium alginate, and the mass of the dopamine hydrochloride is 1-2 times of the mass of sodium alginate.
9. The preparation method of the photothermal controlled release poly-dopamine iron drug-loaded nanoparticle hydrogel according to claim 2, wherein in the fourth step, the solvent is PBS buffer solution or deionized water.
10. The preparation method of the photothermal controllable drug release poly-dopamine-iron-drug-loaded nanoparticle hydrogel according to claim 2, characterized in that in the fourth step, the horseradish peroxidase solution is obtained by dissolving horseradish peroxidase in water.
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CN117100716A (en) * 2023-08-29 2023-11-24 广东龄值生物科技有限公司 Nano microsphere containing iNKT cells and preparation method thereof

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117100716A (en) * 2023-08-29 2023-11-24 广东龄值生物科技有限公司 Nano microsphere containing iNKT cells and preparation method thereof
CN117100716B (en) * 2023-08-29 2024-03-12 广东龄值生物科技有限公司 Nano microsphere containing iNKT cells and preparation method thereof

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