Metal marker loaded with sensibilization and immune drug coating and preparation method of metal marker

Abstract

The invention relates to a metal marker loaded with a sensitizing and immune drug coating and a preparation method thereof, and the preparation method comprises the following steps: S1, preparing a micron-sized texture on the surface of the metal marker by using nanosecond laser to improve the surface specific surface area and surface energy of a matrix; s2, preparing an immunotherapy drug-loaded polymer coating on the micro-textured surface of the metal marker through an electrophoretic deposition method; s3, preparing a nanoscale texture on the polymer coating by using femtosecond laser to realize a super-hydrophilic surface; s4, loading a radiotherapy sensitizer through a liquid film layer-by-layer adsorption method, and completing preparation of the metal marker of the micro-nano textured drug-loaded coating with radiotherapy sensitization and immunotherapy performance. According to the invention, a metal marker for radiotherapy is used as a matrix, an electrophoretic deposition method and a liquid film layer-by-layer adsorption method are used for preparing the sensitization and immune drug-loaded coating, and the sensitization and immune drug-loaded coating has the functions of radiotherapy positioning, radiotherapy sensitization and immunotherapy, and can be used for radiation / immunotherapy guided by the metal marker.

Description

Technical Field

[0001] The invention relates to the technical field of tumor treatment, and in particular to a metal marker carrying a sensitization and immunity drug coating and a preparation method thereof. Background Art

[0002] Cancer is one of the major health threats facing mankind. Surgery, radiotherapy, chemotherapy and immunotherapy are currently common methods in clinical tumor treatment. Intensity modulated radiotherapy, as a modern digital precision radiotherapy technology, is one of the important ways to treat tumors. The use of metal marking to mark tumor lesions can guide the delineation of the target area before intensity modulated radiotherapy and accurately locate the radiotherapy target area, which is of great significance for tumor treatment. Among the metal markers commonly used in clinical practice, titanium, stainless steel, etc. have the characteristics of high hardness, corrosion resistance, and good biocompatibility, and play a pivotal role in the radiotherapy positioning of tumors. However, the metal markers currently used in radiotherapy are all bare metal markers, which can only play a role in guiding the delineation of the target area of ​​postoperative intensity modulated radiotherapy. Although radiotherapy technology has made significant progress, the law of radiation transfer in physics means that normal tissues will inevitably be irradiated to a certain extent. Malignant tumors will still encounter problems such as radiotherapy resistance and radiation damage to normal tissues around tumor lesions during radiotherapy. Therefore, it is necessary to increase the sensitivity of cancer cells to radiation through radiosensitizers to improve the effect of radiotherapy.

[0003] As a systemic treatment, immunotherapy cannot target the tumor site and has significant side effects on human tissues. Nanomaterials can be used to target cancer. Currently, there are many different nanomaterials combined with immunotherapy drugs. However, due to differences in the microenvironment within the tumor, nanomaterials often have limitations. Both radiosensitizers and immunotherapy drugs are currently delivered to the tumor site by intravenous injection, which is very complex and unknown.

[0004] By preparing a polymer coating on the surface of the implant as a carrier of radiosensitizers / immunotherapeutic drugs, the drug can be accurately delivered to the tumor location. At the same time, as a carrier of drugs, the coating can control the release behavior of the drugs carried on the surface of the implant at the tumor. At present, the most widely used drug-loaded coating is a degradable polymer drug-loaded coating, such as polylactic acid and its copolymers, and polylactic acid-glycolic acid is commonly used. Polylactic acid-glycolic acid has good biocompatibility and biodegradability. Polylactic acid-glycolic acid undergoes hydrolysis or enzymatic reaction in the body to generate mainly CO 2 and H 2O products are then excreted from the body. The drugs mixed in the polymer matrix can be gradually released as the polymer dissolves in the tissue. In addition, the coating and the implant matrix should have good bonding to avoid rupture and shedding, otherwise it will cause persistent inflammation. However, the huge difference in mechanical properties and physicochemical properties between polymers and metals leads to a very low membrane-matrix bonding strength, and the implant is prone to drug shedding before reaching the tumor lesion. Therefore, improving the membrane-matrix bonding strength of the drug-loaded coating is the key to achieving implant drug loading.

[0005] At present, in the art, the drug-loaded coating can be prepared on the surface of a bare metal substrate by dip coating, and the drug-loaded coating can be prepared on the surface of a bare metal substrate by electrophoretic deposition. However, these two methods still have the problem of insufficient membrane-substrate bonding strength, which ultimately leads to low drug loading, early burst release, and difficulty in sustained release. In addition, the prior art usually only includes a single coating for carrying radiotherapy sensitizers, chemotherapeutic drugs, or immunotherapy drugs, and the drug-loaded coating has a single function. Summary of the invention

[0006] The purpose of the present invention is to overcome the problems existing in the prior art and to provide a metal marker carrying a sensitizing and immune drug coating and a preparation method thereof. The immunotherapy drug coating is used as the first coating layer on the surface of the metal marker, and the radiotherapy sensitizer coating is used as the second coating layer. The metal marker can not only be used to mark the radiotherapy target area and accurately locate the radiotherapy target area, but also radiotherapy sensitization can be achieved and immunotherapy can be activated at the same time, which is of great significance for tumor treatment.

[0007] In order to achieve the above technical objectives and the above technical effects, the present invention is implemented through the following technical solutions: The present invention provides a method for preparing a metal marker carrying a sensitization and immune drug coating, the preparation method comprising the following steps: Step S1: using nanosecond laser to prepare micron-scale texture on the surface of the metal marker to increase the specific surface area and surface energy of the substrate surface; Step S2: preparing a polymer coating carrying an immunotherapeutic drug on the microtextured surface of the metal marker by electrophoretic deposition; Step S3: using a femtosecond laser to prepare a nanoscale texture on the polymer coating to achieve a super-hydrophilic surface; Step S4: Loading the radiosensitizer by liquid film layer-by-layer adsorption method to complete the preparation of the metal marker of the micro-nano textured drug-loaded coating with radiosensitization and immunotherapy properties.

[0008] Furthermore, in the step S1, before the nanosecond laser processing, the surface of the metal marker is first ground and polished, and then ultrasonically cleaned in ethanol to clean the surface.

[0009] Furthermore, in step S1, the nanosecond laser processing parameters are: laser pulse power is 1-10W, frequency is 100-500KHz, pulse width is 10-15ns, scanning speed is 50-200mm / s, and a micro-texture morphology of a groove structure is formed on the surface of the substrate, wherein the depth of the micro-groove is 10-50μm, the width is 50-150μm, and the spacing is 100-200μm.

[0010] Furthermore, in step S2, a colloidal liquid containing immunotherapeutic drug nanoparticles is first prepared by an emulsified solvent volatilization method, and the colloidal liquid is diluted by a certain proportion to obtain a suspension; then, an electrophoretic deposition platform is used to connect the micro-textured metal marker in step S1 to the positive electrode of a DC power supply, and a platinum electrode to the negative electrode of the DC power supply, and the obtained suspension is used as an electrolyte for electrophoretic deposition to obtain a metal marker carrying an immunotherapeutic drug coating.

[0011] Furthermore, in step S2, the preparation process of the colloidal liquid containing immunotherapy drug nanoparticles is as follows: 60 mg-75 mg of polylactic acid-glycolic acid particles and 60 mg-90 mg of PD-1 antibody or PD-L1 antibody are added to a volumetric flask containing 3 ml of dichloromethane solution and uniformly dispersed to obtain a uniform dispersion containing polylactic acid-glycolic acid, PD-1 antibody or PD-L1 antibody and dichloromethane; the obtained dispersion is added dropwise to the polyacrylic acid aqueous solution; at the same time, ultrasonic emulsification is performed using an ultrasonic cell disruptor; the emulsified solution is magnetically stirred, and the dichloromethane is evaporated to obtain a colloidal liquid containing immunotherapy drug nanoparticles.

[0012] Furthermore, the concentration of the suspension is 1 mg / ml-1.5 mg / ml, and the electrophoretic deposition is carried out at a voltage of 5-12 V for 15-40 min.

[0013] Furthermore, the volume ratio of the polyacrylic acid aqueous solution is 0.2%-0.5% polyacrylic acid solution and 99.5%-99.9% water, and the volume ratio of the polyacrylic acid aqueous solution to dichloromethane is 5:1-20:1.

[0014] Furthermore, in step S3, the femtosecond laser processing parameters are: using an objective lens with a numerical aperture of 0.8 to focus a linearly polarized femtosecond laser with a wavelength of 800nm ​​onto the surface of the polymer coating carrying immunotherapy drugs, the femtosecond pulse energy is 0.5-2μJ, the frequency is 500-1000Hz, the scanning number is 1-2 times, and the movement of the three-dimensional mobile platform is controlled by a computer to process a nanotexture morphology of a corrugated structure on the surface of the polymer coating, wherein the groove width is 20-50nm, the groove depth is 10-30nm, and the spacing is 20-50nm.

[0015] Furthermore, in step S4, the micro-nano textured coated metal marker obtained in step S3 is placed in a dichloromethane solution containing a sensitizing drug, and magnetic stirring is performed under a certain temperature environment. After being taken out, the dichloromethane is caused to volatilize to form a single liquid film, and the loading and drying processes are repeated until the sensitizing drug content in the micro-nano texture reaches the standard.

[0016] The present invention provides a metal marker loaded with a sensitization and immunity drug coating prepared according to the preparation method in the above scheme.

[0017] The beneficial effects of the present invention are: 1. The present invention uses metal markers for radiotherapy as a matrix, and utilizes electrophoretic deposition and liquid film layer-by-layer adsorption to prepare a radiotherapy sensitization / immunotherapy drug-loaded coating, which has radiotherapy positioning, radiotherapy sensitization and immunotherapy functions, and can be used for radiotherapy / immunotherapy guided by metal markers.

[0018] 2. Microscale textures and nanoscale textures are constructed on the surface of the metal marker substrate to enhance the bonding strength between the drug-loaded coating and the metal substrate.

[0019] 3. The “gaps” in the micro-nano composite texture units can be directly used for drug loading to increase the drug loading amount. This loading method is convenient and has a high loading rate.

[0020] 4. The drug-loaded micro-nano texture is encapsulated using an organic solution containing radiotherapy sensitizing drugs, so that a layer of organic film is formed on the surface of the micro-nano texture, which can play a role in controlling the release of drugs. The prepared metal marker has the functions of radiotherapy sensitization, radiotherapy positioning and immunotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 Schematic diagram of surface micro-texturing treatment of metal markers in the present invention; Figure 2 Schematic diagram of the preparation of the colloid liquid and suspension in the present invention; Figure 3 This is a schematic diagram of the preparation of an immunotherapy drug-carrying coating using an electrophoretic deposition platform according to the present invention; Figure 4 The surface morphology of the metal marker carrying the immunotherapy drug coating is taken under a super-depth microscope when the concentration of the suspension of the present invention is 1.25 mg / ml and the DC power supply voltage is 6 V, wherein the deposition time of a is 20 min, the deposition time of b is 30 min, and the deposition time of c is 40 min; Figure 5The surface morphology of the metal marker carrying the immunotherapy drug coating is taken under a super-depth microscope when the concentration of the suspension of the present invention is 1 mg / ml and the DC power supply voltage is 6 V, wherein the deposition time of a is 20 min, the deposition time of b is 30 min, and the deposition time of c is 40 min; Figure 6 Schematic diagram of the preparation of nano-scale texture on the surface of the polymer coating carrying immunotherapeutic drugs in the present invention; Figure 7 The invention relates to the preparation of the coating layer carrying the radiotherapy sensitizing drug; Figure 8 It is a schematic diagram of the overall preparation process of the drug-loaded coating in the present invention. DETAILED DESCRIPTION

[0022] The present invention will be described in detail below with reference to the accompanying drawings and in combination with embodiments.

[0023] A method for preparing a metal marker loaded with a sensitizing and immune drug coating, such as Figure 8 As shown, the preparation method comprises the following steps: Step S1: using nanosecond laser to prepare micron-scale texture on the surface of the metal marker to increase the specific surface area and surface energy of the substrate surface; Step S2: preparing a polymer coating carrying an immunotherapeutic drug on the microtextured surface of the metal marker by electrophoretic deposition; Step S3: using a femtosecond laser to prepare a nanoscale texture on the polymer coating to achieve a super-hydrophilic surface; Step S4: Loading the radiosensitizer by liquid film layer-by-layer adsorption method to complete the preparation of the metal marker of the micro-nano textured drug-loaded coating with radiosensitization and immunotherapy properties.

[0024] In the step S1, before the nanosecond laser processing, the surface of the metal marker is first ground and polished, and then ultrasonically cleaned in ethanol to clean the surface. The specific steps of the surface cleaning treatment are: first, the metal marker is placed on 80#, 200#, 500#, 800#, 1500#, and 2000# sandpaper for grinding until no obvious scratches are observed on the surface of the sample; then, a polishing cloth is fixed on the grinding and polishing machine, and a diamond polishing agent is sprayed, and the sample is ground and polished until the surface presents a mirror-like smooth effect, and surface oxides and pollutants are removed to obtain a substrate surface with a surface roughness of less than 10 nm; then, the ground and polished metal marker is placed in ethanol for cleaning for 15 minutes using ultrasonic cleaning.

[0025] In step S1, Figure 1As shown, the ultraviolet nanosecond laser is focused by an objective lens onto the surface of the metal marker after grinding, polishing and surface cleaning pretreatment, and the scanning process path and laser processing parameters of the laser processor are set. The nanosecond laser processing parameters are: laser pulse power of 1 to 10 W, frequency of 100 to 500 KHz, pulse width of 10 to 15 ns, scanning speed of 50 to 200 mm / s, and a micro-texture morphology of a groove structure is formed on the surface of the substrate, wherein the depth of the micro-groove is 10 to 50 μm, the width is 50 to 150 μm, and the spacing is 100 to 200 μm.

[0026] In step S2, a colloidal solution containing immunotherapy drug nanoparticles is first prepared by an emulsified solvent evaporation method, and the colloidal solution is diluted by a certain ratio to obtain a suspension; then, an electrophoretic deposition platform is used, such as Figure 3 As shown, the micro-textured metal marker in step S1 is connected to the positive electrode of a DC power supply, and the platinum electrode is connected to the negative electrode of the DC power supply. The obtained suspension is used as an electrolyte for electrophoretic deposition to obtain a metal marker carrying an immunotherapy drug coating.

[0027] In step S2, Figure 2 As shown, the loaded immunotherapeutic drug is PD-1 antibody or PD-L1 antibody, the carrier is polylactic acid-glycolic acid copolymer, and the preparation process of the colloidal liquid containing immunotherapeutic drug nanoparticles is as follows: first, the colloidal emulsion of PLGA nanoparticles loaded with immunotherapeutic drugs is prepared by the emulsification solvent volatilization method, 60mg-75mg of polylactic acid-glycolic acid particles and 60mg-90mg of PD-1 antibody or PD-L1 antibody are added to a volumetric flask containing 3ml of dichloromethane solution and evenly dispersed to obtain a uniform dispersion containing polylactic acid-glycolic acid, PD-1 antibody or PD-L1 antibody and dichloromethane; the obtained dispersion is added dropwise to the polyacrylic acid aqueous solution; at the same time, ultrasonic emulsification is performed using an ultrasonic cell disruptor, the power of ultrasonic emulsification is 200W, and the emulsification time is 5min; the emulsified solution is magnetically stirred, and dichloromethane is evaporated, the magnetic stirring speed is 200rpm, and the stirring time is 2h, to obtain a colloidal liquid containing immunotherapeutic drug nanoparticles.

[0028] The concentration of the suspension is 1 mg / ml-1.5 mg / ml, and the electrophoretic deposition is carried out under a voltage of 5-12 V for 15-40 min. The metal marker is taken out and placed in a fume hood to dry, thereby obtaining an immunotherapy drug-carrying coating. Figure 4 and Figure 5 Shown is the surface morphology of the metal marker coated with immunotherapy drugs, taken under an ultra-depth-of-field microscope.

[0029] The volume ratio of the polyacrylic acid aqueous solution components is 0.2%-0.5% polyacrylic acid solution and 99.5%-99.9% water, and the volume ratio of the polyacrylic acid aqueous solution to dichloromethane is 5:1-20:1.

[0030] In step S3, a single beam of femtosecond laser is used to irradiate the surface of the polymer coating carrying the immunotherapy drug, thereby inducing a periodic nanoscale texture on the surface, such as Figure 6 As shown in the figure, the femtosecond laser processing parameters are: using an objective lens with a numerical aperture of 0.8 (magnification of 80 times) to focus a linearly polarized femtosecond laser with a wavelength of 800nm ​​onto the surface of the polymer coating carrying immunotherapy drugs, the femtosecond pulse energy is 0.5-2μJ, the frequency is 500-1000Hz, the scanning number is 1-2 times, and the movement of the three-dimensional mobile platform is controlled by a corresponding computer to process a corrugated nanotexture morphology on the surface of the polymer coating, wherein the groove width is 20-50nm, the groove depth is 10-30nm, and the spacing is 20-50nm.

[0031] In step S4, Figure 7 As shown, the micro-nano textured coated metal marker obtained in step S3 is placed in a dichloromethane solution containing a sensitizing drug, such as sodium glycidyl bisazole (Simena) and polylactic acid-glycolic acid, wherein the concentration of the sensitizing drug is 1.5 mg / ml, the volume of dichloromethane is 10 ml, and magnetic stirring is performed at room temperature for 15-20 minutes. After being taken out, a hot dryer is used to cause the dichloromethane to volatilize to form a single liquid film, and the loading and drying process is repeated until the sensitizing drug content in the micro-nano texture reaches the standard, and the sensitizing drug content is 5-8 mg.

[0032] The present invention provides a metal marker loaded with sensitization and immune drug coatings prepared according to the preparation method in the above scheme, and the prepared metal marker has the functions of radiotherapy sensitization, radiotherapy positioning and immunotherapy.

[0033] Example 1 Step S1: firstly, pretreatment is performed, the surface of the metal marker titanium plate is ground and polished, and the metal marker is placed on 80#, 200#, 500#, 800#, 1500#, and 2000# sandpaper for grinding until no obvious scratches are observed on the surface of the sample; then, a polishing cloth is fixed on the grinding and polishing machine, and a diamond polishing agent is sprayed, and the sample is ground and polished until the surface presents a mirror-like smooth effect. After grinding and polishing, the metal marker is ultrasonically cleaned in ethanol for 15 minutes to perform surface cleaning; Then, nanosecond laser texturing was performed to prepare micron-scale texture on the surface of the metal marker titanium plate. The nanosecond laser processing parameters were: laser pulse power of 4W, frequency of 250KHz, pulse width of 13n, scanning speed of 20mm / s, and micron groove structure with a depth of 10μm, a width of 100μm, and a spacing of 125μm was induced on the substrate surface. Step S2: preparing a colloidal solution containing immunotherapeutic drug nanoparticles by an emulsified solvent evaporation method: adding 75 mg of polylactic acid-glycolic acid particles and 60 mg of PD-1 antibody to a volumetric flask containing 3 ml of dichloromethane solution and uniformly dispersing them, a uniform dispersion containing polylactic acid-glycolic acid, PD-1 and dichloromethane was obtained; the obtained dispersion was added dropwise to 30 ml of polyacrylic acid aqueous solution (the volume ratio of the polyacrylic acid aqueous solution was 0.3% polyacrylic acid solution and 99.7% water, respectively), and the volume ratio of polyacrylic acid aqueous solution to dichloromethane was 10:1; at the same time, ultrasonic emulsification was performed using an ultrasonic cell disruptor, the power of the ultrasonic emulsification was 200 W, and the emulsification time was 5 min; the emulsified solution was magnetically stirred, and dichloromethane was evaporated, and the magnetic stirring speed was 200 rpm, and the stirring time was 2 h to obtain a colloidal solution containing immunotherapeutic drug nanoparticles, and the colloidal solution and deionized water were mixed and diluted to obtain a suspension with a concentration of 1 mg / ml; Then, using an electrophoretic deposition platform, the micro-textured metal marker titanium plate in step S1 was connected to the positive electrode of a DC power supply, and the platinum electrode was connected to the negative electrode of the DC power supply. The obtained 1 mg / ml suspension was used as an electrolyte for electrophoretic deposition. The metal marker carrying the immunotherapy drug coating was deposited at a voltage of 6 V for 20 min. Step S3: irradiating the surface of the immunotherapy drug-loaded coating of step S2 with a single beam of femtosecond laser to induce a periodic nanoscale texture on the surface, and the processing parameters of the femtosecond laser are as follows: focusing a linearly polarized femtosecond laser with a wavelength of 800nm ​​onto the surface of the drug-loaded coating using an objective lens with a numerical aperture of 0.8, with a femtosecond pulse energy of 1μJ, a frequency of 500Hz, and a scanning number of 1 times; controlling the movement of the three-dimensional mobile platform by a computer to process a nanotexture morphology of a corrugated structure on the surface of the coating; wherein the groove width is 20nm, the groove depth is 10nm, and the spacing is 20nm; Step S4: Use the liquid film layer-by-layer adsorption method to load the radiotherapy sensitizer on the surface of the micro-nano textured coated metal marker in step S3, place the micro-nano textured coated metal marker in a dichloromethane solution containing sensitizing drugs sodium glycidyl bisazole and polylactic acid-hydroxyacetic acid, wherein the sensitizing drug concentration is 1.5 mg / ml, the dichloromethane volume is 10 ml, and magnetic stirring is performed at room temperature for 15 minutes. After taking out, use a hot dryer to cause the dichloromethane to evaporate to form a single liquid film, and repeat the loading and drying process until the sensitizing drug content in the micro-nano texture is 5 mg; the preparation of the micro-nano textured drug-loaded coated metal marker with radiotherapy sensitization and immunotherapy properties is completed.

[0034] Example 2 Step S1: firstly, pretreatment is performed, the surface of the metal marker titanium plate is ground and polished, and the metal marker is placed on 80#, 200#, 500#, 800#, 1500#, and 2000# sandpaper for grinding until no obvious scratches are observed on the surface of the sample; then, a polishing cloth is fixed on the grinding and polishing machine, and a diamond polishing agent is sprayed, and the sample is ground and polished until the surface presents a mirror-like smooth effect. After grinding and polishing, the metal marker is ultrasonically cleaned in ethanol for 15 minutes to perform surface cleaning; Then, nanosecond laser texturing was performed to prepare micron-scale texture on the surface of the metal marker titanium plate. The nanosecond laser processing parameters were: laser pulse power of 4W, frequency of 250KHz, pulse width of 13n, scanning speed of 20mm / s, and micron groove structure with a depth of 10μm, a width of 100μm, and a spacing of 125μm was induced on the substrate surface. Step S2: preparing a colloidal solution containing immunotherapeutic drug nanoparticles by an emulsified solvent evaporation method: adding 75 mg of polylactic acid-glycolic acid particles and 90 mg of PD-L1 to a volumetric flask containing 3 ml of dichloromethane solution and uniformly dispersing them, a uniform dispersion containing polylactic acid-glycolic acid, PD-L1 and dichloromethane was obtained; the obtained dispersion was added dropwise to 60 ml of polyacrylic acid aqueous solution (the volume ratio of the polyacrylic acid aqueous solution was 0.3% polyacrylic acid solution and 99.7% water, respectively), and the volume ratio of polyacrylic acid aqueous solution to dichloromethane was 20:1; at the same time, ultrasonic emulsification was performed using an ultrasonic cell disruptor, the power of ultrasonic emulsification was 200 W, and the emulsification time was 5 min; the emulsified solution was magnetically stirred, and dichloromethane was evaporated, and the magnetic stirring speed was 200 rpm, and the stirring time was 2 h to obtain a colloidal solution containing immunotherapeutic drug nanoparticles, and the colloidal solution and deionized water were mixed and diluted to obtain a suspension with a concentration of 1.25 mg / ml; Then, using an electrophoretic deposition platform, the micro-textured metal marker titanium plate in step S1 was connected to the positive electrode of a DC power supply, and the platinum electrode was connected to the negative electrode of the DC power supply. The obtained 1.25 mg / ml suspension was used as an electrolyte for electrophoretic deposition. The metal marker carrying the immunotherapy drug coating was deposited at a voltage of 6 V for 20 min. Step S3: irradiating the surface of the immunotherapy drug-loaded coating of step S2 with a single beam of femtosecond laser to induce a periodic nanoscale texture on the surface, and the processing parameters of the femtosecond laser are as follows: focusing a linearly polarized femtosecond laser with a wavelength of 800nm ​​onto the surface of the drug-loaded coating using an objective lens with a numerical aperture of 0.8, with a femtosecond pulse energy of 1μJ, a frequency of 500Hz, and a scanning number of 1 times; controlling the movement of the three-dimensional mobile platform by a computer to process a nanotexture morphology of a corrugated structure on the surface of the coating; wherein the groove width is 20nm, the groove depth is 10nm, and the spacing is 20nm; Step S4: Use the liquid film layer-by-layer adsorption method to load the radiotherapy sensitizer on the surface of the micro-nano textured coated metal marker in step S3, place the micro-nano textured coated metal marker in a dichloromethane solution containing sensitizing drugs sodium glycidyl bisazole and polylactic acid-hydroxyacetic acid, wherein the sensitizing drug concentration is 1.5 mg / ml, the dichloromethane volume is 10 ml, and magnetic stirring is performed at room temperature for 20 minutes. After taking out, a hot dryer is used to volatilize the dichloromethane to form a single liquid film, and the loading and drying process is repeated until the sensitizing drug content in the micro-nano texture is 8 mg; the preparation of the micro-nano textured drug-loaded coated metal marker with radiotherapy sensitization and immunotherapy properties is completed.

[0035] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing a metal marker with a sensitizing and immune drug coating, characterized in that: The preparation method comprises the following steps: Step S1: using nanosecond laser to prepare micron-scale texture on the surface of the metal marker to increase the specific surface area and surface energy of the substrate surface; Step S2: preparing a polymer coating carrying an immunotherapeutic drug on the microtextured surface of the metal marker by electrophoretic deposition; Step S3: using a femtosecond laser to prepare a nanoscale texture on the polymer coating to achieve a super-hydrophilic surface; Step S4: Loading the radiosensitizer by liquid film layer-by-layer adsorption method to complete the preparation of the metal marker of the micro-nano textured drug-loaded coating with radiosensitization and immunotherapy properties.

2. The method for preparing a metal marker carrying a sensitizing and immune drug coating according to claim 1, characterized in that: In the step S1, before the nanosecond laser processing, the surface of the metal marker is first ground and polished, and then ultrasonically cleaned in ethanol to clean the surface.

3. The method for preparing a metal marker carrying a sensitizing and immune drug coating according to claim 2, characterized in that: In the step S1, the nanosecond laser processing parameters are: laser pulse power of 1 to 10 W, frequency of 100 to 500 kHz, pulse width of 10 to 15 ns, scanning speed of 50 to 200 mm / s, and a micro-texture morphology of a groove structure is formed on the surface of the substrate, wherein the depth of the micro-groove is 10 to 50 μm, the width is 50 to 150 μm, and the spacing is 100 to 200 μm.

4. The method for preparing a metal marker carrying a sensitizing and immune drug coating according to claim 1, characterized in that: In the step S2, a colloidal liquid containing immunotherapeutic drug nanoparticles is first prepared by an emulsified solvent volatilization method, and the colloidal liquid is diluted by a certain proportion to obtain a suspension; then, an electrophoretic deposition platform is used to connect the micro-textured metal marker in step S1 to the positive electrode of a DC power supply, and a platinum electrode to the negative electrode of the DC power supply, and the obtained suspension is used as an electrolyte for electrophoretic deposition to obtain a metal marker carrying an immunotherapeutic drug coating.

5. The method for preparing a metal marker carrying a sensitizing and immune drug coating according to claim 4, characterized in that: In step S2, the preparation process of the colloidal liquid containing immunotherapy drug nanoparticles is as follows: 60 mg-75 mg of polylactic acid-glycolic acid particles and 60 mg-90 mg of PD-1 antibody or PD-L1 antibody are added to a volumetric flask containing 3 ml of dichloromethane solution and uniformly dispersed to obtain a uniform dispersion containing polylactic acid-glycolic acid, PD-1 antibody or PD-L1 antibody and dichloromethane; the obtained dispersion is added dropwise to a polyacrylic acid aqueous solution; ultrasonic emulsification is performed using an ultrasonic cell disruptor at the same time; the emulsified solution is magnetically stirred, and dichloromethane is evaporated to obtain a colloidal liquid containing immunotherapy drug nanoparticles.

6. The method for preparing a metal marker carrying a sensitizing and immune drug coating according to claim 4 or 5, characterized in that: The concentration of the suspension is 1 mg / ml-1.5 mg / ml, and the electrophoretic deposition is carried out at a voltage of 5-12 V for 15-40 min.

7. The method for preparing a metal marker carrying a sensitizing and immune drug coating according to claim 5, characterized in that: The volume ratio of the polyacrylic acid aqueous solution components is 0.2%-0.5% polyacrylic acid solution and 99.5%-99.9% water, and the volume ratio of the polyacrylic acid aqueous solution to dichloromethane is 5:1-20:

1.

8. The method for preparing a metal marker carrying a sensitizing and immune drug coating according to claim 1, characterized in that: In step S3, the femtosecond laser processing parameters are: using an objective lens with a numerical aperture of 0.8 to focus a linearly polarized femtosecond laser with a wavelength of 800nm ​​onto the surface of the polymer coating carrying the immunotherapy drug, the femtosecond pulse energy is 0.5-2μJ, the frequency is 500-1000Hz, the scanning number is 1-2 times, and the movement of the three-dimensional mobile platform is controlled by a computer to process a nanotexture morphology of a corrugated structure on the surface of the polymer coating, wherein the groove width is 20-50nm, the groove depth is 10-30nm, and the spacing is 20-50nm.

9. The method for preparing a metal marker carrying a sensitizing and immune drug coating according to claim 1, characterized in that: In step S4, the micro-nano textured coating metal marker obtained in step S3 is placed in a dichloromethane solution containing a sensitizing drug, and magnetic stirring is performed under a certain temperature environment. After being taken out, the dichloromethane is caused to volatilize to form a single liquid film, and the loading and drying processes are repeated until the sensitizing drug content in the micro-nano texture meets the standard.

10. A metal marker coated with sensitizing and immune drugs prepared according to the preparation method according to any one of claims 1 to 5.

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