Microneedle patch for treating malignant melanoma and preparation method thereof

Abstract

The application discloses a microneedle patch for treating malignant melanoma and a preparation method thereof, and relates to the biomedical technical field.The preparation method comprises the following steps: preparing nitroprusside nanoparticles by using nitroprusside solid particles;compounding 0.01-0.15 g / ml of the nitroprusside nanoparticles with equimolar ferrous lactate nanoparticles, dissolving 0.08 g / ml of sodium carboxymethyl cellulose solution in deionized water, and uniformly mixing to form a gel-like mixture; adding the gel-like mixture into a mold, manufacturing a microneedle patch by using a centrifugal method; and performing centrifugation on the microneedle patch under vacuum conditions and drying the microneedle patch.The preparation method process for treating malignant melanoma provided by the embodiment of the application is clear, the dual effects of gas treatment and photothermal treatment can remove residual tumors after operation and repair defects after operation.Meanwhile, the microneedle patch prepared by the method has a reasonable structure, is convenient for self-administration, can cover and protect the skin, and avoids secondary trauma when the dressing is replaced.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biomedicine, and in particular to a microneedle patch for treating malignant melanoma and a preparation method thereof. BACKGROUND

[0002] Malignant melanoma has strong invasion ability, easy metastasis and recurrence, and occurs in the maxillofacial region, which can cause serious structural damage, facial deformity and dysfunction. Malignant melanoma is a common disease, and the main cause is not clear, which may be related to genetics, ultraviolet rays, color nevus and immune abnormalities. The main clinical symptoms of malignant melanoma are skin black spots, nodules, ulceration and pain, which can cause infection and metastasis complications.

[0003] The current treatment plan for malignant melanoma is mainly surgical resection, supplemented by radiotherapy or chemotherapy. Malignant melanoma is treated by surgical resection of superficial tumors and surrounding skin tissue, but the remaining asymptomatic but infiltrative tumor tissue cannot be completely removed, even if the range of the surgical area is expanded, it is still easy to relapse and accompanied by large area skin defects, difficult to repair and other deficiencies. Chemotherapy is a common postoperative adjuvant therapy, but it is limited by the easy tolerance of chemotherapy drugs and obvious systemic side effects, and its efficacy is easy to reach a bottleneck period. As another adjuvant therapy, radiotherapy can reduce the recurrence rate to a certain extent after surgery, but it has a large side effect, which can seriously damage the patient's immune system. SUMMARY

[0004] The microneedle patch for treating malignant melanoma provided by the embodiments of the present application solves the technical problems of incomplete removal and high recurrence rate of malignant melanoma in the prior art.

[0005] In a first aspect, the embodiments of the present application provide a preparation method of a microneedle patch for treating malignant melanoma, comprising: preparing nitroprusside nanoparticles by using nitroprusside solid particles; dissolving 0.01-0.15 g / ml of the nitroprusside nanoparticles, an equimolar ratio of ferrous lactate nanoparticles, 0.08 g / ml of sodium carboxymethyl cellulose solution in deionized water, and uniformly mixing into a gel-like mixture; adding the gel-like mixture into a mold, and manufacturing a microneedle patch by a centrifugal method; and centrifuging the microneedle patch under vacuum conditions and drying.

[0006] In combination with the first aspect, in a possible implementation manner, the nitroprusside nanoparticles are 0.03 g / ml.

[0007] In combination with the first aspect, in a possible implementation manner, the centrifugal method is a negative pressure centrifugal casting method.

[0008] In combination with the first aspect, in a possible implementation manner, the centrifuging of the microneedle patch comprises:

[0009] The microneedle patch is centrifuged at -0.08 MPa using a suction pump.

[0010] With reference to the first aspect, in a possible implementation manner, the drying temperature is 20-25 DEG C.

[0011] With reference to the first aspect, in a possible implementation manner, the nitroprusside nanoparticles are prepared from the nitroprusside solid particles, including: completely dissolving the nitroprusside solid particles in deionized water and ethanol to form a mixed solution; dispersing the mixed solution using ultrasonic vibration; keeping at -120 DEG C to -70 DEG C for more than 24 hours; and obtaining the nitroprusside nanoparticles through freeze-drying.

[0012] With reference to the first aspect, in a possible implementation manner, the material of the mold is polydimethylsiloxane.

[0013] The second aspect, the embodiment of the present application provides a microneedle patch prepared by using the method of the first aspect or any possible implementation manner of the first aspect, the microneedle patch includes a substrate, a microneedle arranged on the substrate, and a release layer loaded on the microneedle and the substrate;Wherein, the release layer is formed by the gel-like mixture in the mold.

[0014] With reference to the second aspect, in a possible implementation manner, the substrate is circular, and the microneedles are distributed radially along the substrate.

[0015] With reference to the second aspect, in a possible implementation manner, the diameter of the substrate is 17.5 mm, and the number of microneedles is 385.

[0016] The one or more technical solutions provided in the embodiment of the present application have at least the following technical effects:

[0017] The preparation method of the microneedle patch for treating malignant melanoma provided in the embodiment of the application, the SNP-Fe@MNs (sodium nitroprusside and Fe ions loaded microneedles) patch prepared by the method is activated under ultraviolet light or natural light, changes from dark brown to Prussian blue, releases a large amount of NO, is used for gas treatment, and can promote neovascularization; further reacts with ferrous ions to generate in-situ photothermal products, generates a photothermal effect under near-infrared light irradiation, and has high photothermal conversion efficiency. Then the SNP-Fe@MNs patch is attached to the postoperative resection site, can release NO at a low concentration for a long time, and thus can slow-release repair the wound. The heat sensitivity of tumor cells is higher than that of normal body cells, and the intratumoral delivery of the near-infrared laser responsive biomaterials realizes photothermal treatment. The application is directed to the characteristics of strong invasion and metastasis of malignant melanoma cells, and the gas treatment is used in cooperation to inhibit tumor invasion activity, and further ensures that the tumor is completely removed. The application is directed to the skin defect repair problem of the patient after surgical resection, and then the early healing of the wound can be realized by the low-concentration NO gas molecules. Therefore, the preparation method process for treating malignant melanoma provided in the embodiment of the application is clear, has low requirements on instruments, low cost, and is easy to realize, and through the dual effects of gas treatment and photothermal treatment, the residual tumor after operation and the defect after operation can be removed. At the same time, the SNP-Fe@MNs patch prepared by the method has a reasonable structure, is convenient for self-administration, can cover and protect the skin, and avoids secondary trauma when the dressing is changed. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the application, the drawings needed to be used in the description of the embodiments of the application will be briefly introduced. Obviously, the drawings in the following description are some embodiments of the application, and other drawings can be obtained by those skilled in the art without creative effort on the basis of these drawings.

[0019] Figure 1 The flowchart of the preparation method of the microneedle patch for treating malignant melanoma provided in the embodiment of the application is shown in the figure.

[0020] Figure 2 The structure schematic diagram of the microneedle patch provided in the embodiment of the application is shown in the figure.

[0021] Figure 3 The photothermal and NO release performance diagram of the microneedle patch with different concentrations provided in the embodiment of the application is shown in the figure.

[0022] Figure 4 The SEM images of the microneedle patch provided in the embodiment of the application before and after pressing on the mouse skin for 1 min, 5 min, 15 min and 30 min are shown in the figures.

[0023] Figure 5 Fluorescence top view and cross-sectional view images of mouse skin treated with rhodamine B-loaded microneedle patches for 1 minute according to an embodiment of the present application;

[0024] Figure 6 Biocompatibility of microneedle patches according to an embodiment of the present application;

[0025] Figure 7 In vivo tissue regeneration performance of microneedle patches according to an embodiment of the present application;

[0026] Figure 8 Flowchart of a preparation method for preparing nitroprusside nanoparticles using nitroprusside solid particles according to an embodiment of the present application.

[0027] Reference numerals: 1 - substrate; 2 - microneedle. DETAILED DESCRIPTION

[0028] The technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are part of, but not all of the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of the present application.

[0029] As shown in Figure 1 The present application provides a microneedle patch preparation method for treating malignant melanoma, which comprises S101 to S104:

[0030] S101: preparing nitroprusside nanoparticles using nitroprusside solid particles;

[0031] S102: dissolving 0.01-0.15 g / ml nitroprusside nanoparticles, an equimolar ratio of ferrous lactate nanoparticles, and 0.08 g / ml sodium carboxymethyl cellulose solution in deionized water, and mixing to form a gel-like mixture;

[0032] S103: adding the gel-like mixture to a mold to manufacture microneedle patches by centrifugation;

[0033] S104: centrifuging and drying the microneedle patches under vacuum conditions.

[0034] It should be noted that all the above manufacturing and storage processes are carried out in the dark or low light and dry conditions to avoid the degradation of the microneedle patch. The SNP-Fe@MNs (sodium nitroprusside and Fe ions loaded microneedles) patch prepared by the above method is activated under ultraviolet light or natural light irradiation, changes from dark brown to Prussian blue, releases a large amount of NO, which is used for gas therapy and can promote neovascularization; further reacts with ferrous ions to generate in-situ photothermal products, which produce photothermal effect under near-infrared light irradiation, and the photothermal conversion efficiency is high. Then the SNP-Fe@MNs patch is attached to the postoperative resection site, which can release NO at a low concentration for a long time, so as to release and repair the wound. The heat sensitivity of tumor cells is higher than that of normal body cells, and the intratumoral delivery of near-infrared laser responsive biomaterials can achieve photothermal therapy. In view of the strong invasion and metastasis ability of malignant melanoma cells, the gas therapy is used in cooperation to inhibit the tumor invasion activity, and further ensure the complete removal of the tumor. In view of the skin defect repair problem of the patient after surgical resection, the early healing of the wound can be achieved by low-concentration NO gas molecules. Therefore, the preparation method for treating malignant melanoma provided in the embodiments of the present application has clear process flow, low requirement for instruments, low cost, and is easy to realize. Through the dual action of gas therapy and photothermal therapy, the residual tumor after operation and the defect after operation can be removed. At the same time, the SNP-Fe@MNs prepared by this method has a reasonable structure, is convenient for self-administration, can cover and protect the skin, and avoids secondary trauma when changing the dressing.

[0035] In the embodiments of the present application, the sodium nitroprusside nanoparticles are 0.03 g / ml.

[0036] It should be noted that, since sodium nitroprusside has a potential antihypertensive effect and a cumulative hazard of nitrite, the drug dose of each SNP-Fe@MNs package is mainly based on the median lethal dose (rhodamine b50) of intravenous injection 11.2 mg / kg and subcutaneous injection 9.3-13.9 mg / kg, and the gradient concentration of loaded sodium nitroprusside. Therefore, in order to balance between treatment effect and drug safety, we set the concentration of SNP-Fe@MNs to be 0.01, 0.03, 0.05, 0.10 and 0.15 g / ml of sodium nitroprusside and equimolar ratio of ferrous lactate. Figure 3 Figure a of the present application shows the heating curve of SNP-Fe@MNs loaded with 0.01, 0.03, 0.05, 0.10 and 0.15 g / ml of sodium nitroprusside under 808 nm near-infrared irradiation at 1 W / cm2. As can be seen from the figure, the highest temperature of the 0.15 g / ml group is about 42.4℃, and the highest temperature of the 0.01 g / ml group is about 31.6℃. Figure 3Fig. 1c is a heating curve of SNP-Fe@MNs loaded with 0.03 g / ml SNP under 808 nm near-infrared irradiation at 1.5 W / cm2. 2 irradiation, the heating curves of SNP-Fe@MNs loaded with 0.01, 0.03, 0.05, 0.10 and 0.15 g / ml SNP, the highest temperature of the 0.15 g / ml group was about 86.6℃, and the highest temperature of the 0.01 g / ml group was about 51.8℃. The temperature of the 0.03 g / ml group reached about 54.2℃ within 10 min. Although a large number of reports have shown that high temperature above 43℃ can induce tumor cell apoptosis, in vivo experiments have shown that 52℃ is a lower threshold. Implantable antitumor biomaterials may face problems such as skin barrier, location and volume of tumor, depth and size of implant, which may affect heat distribution, resulting in insufficient heating area. However, excessive in situ thermal ablation may cause irreversible heat, leading to extensive overheating of the epidermis, dermis and subcutaneous tissue. Among them, SNP-Fe@MNs loaded with 0.03 g / ml have controllable near-infrared absorbance and heat production capacity, which can reach the threshold temperature of the lowest concentration. Therefore, the subsequent experiment selected SNP-Fe@MNs loaded with 0.03 g / ml SNP, and recorded the temperature curve under different power densities. Figure 3 Fig. 1c is a heating curve of SNP-Fe@MNs loaded with 0.03 g / ml SNP under 808 nm near-infrared irradiation at different power densities. In addition, Figure 3 Fig. 1d is a repeated thermal stability of SNP-Fe@MNs loaded with 0.03 g / ml SNP by 5 cycles of heating-cooling irradiation. Therefore, SNP-Fe@MNs loaded with 0.03 g / ml SNP has the most stable photothermal cycle, and it can reach a photothermal temperature of 54.2℃, which can not only ablate and kill tumors, but also not cause burns to the surrounding normal tissues.

[0037] In addition, the NO gas release capacity in the synergistic therapy was also tested in the embodiments of the present application. Since the reaction is initiated by ultraviolet light, SNP-Fe@MNs loaded with 0.01, 0.03, 0.05, 0.10 and 0.15 g / ml SNP were respectively soaked in 1 ml polybutylene succinate, and then irradiated with and without ultraviolet light for 1 min, and then the solution was extracted into the well plate for comparison. Figure 3 Fig. 1e is a NO release curve of SNP-Fe@MNs loaded with 0.01, 0.03, 0.05, 0.10 and 0.15 g / ml SNP under ultraviolet light activation, Figure 3Figure f shows the NO release curves of SNP-Fe@MNs loaded with 0.01, 0.03, 0.05, 0.10, and 0.15 g / ml sodium nitroprusside without UV activation. As can be seen from the figure, in the UV-irradiated group, there is an inflection point at 60 min, after which the curve begins to decline or the rate of increase tends to flatten. After the removal of UV irradiation, the curve appears to flatten, but a small amount of NO is still released because it cannot completely avoid the light exposure of the microneedles in the environment.

[0038] Figure 3 The g-figure used a fluorescent NO probe to assess the actual NO uptake by cells in B16 (a highly metastatic melanoma cell line) and HUVECs (human umbilical vein endothelial cells). It was confirmed that high doses of NO can induce tumor cell apoptosis and inhibit NO-mediated angiogenesis, and vice versa. Therefore, HUVECs' SNP-Fe@MNs, after 60 minutes of UV irradiation, avoided the gas explosion of B16 cells, based on the previous NO release curve. Figure 3 The h-th image shows the intracellular fluorescent probe intensities of B16 and HUVECs. Analysis of the intracellular fluorescent probe intensities revealed that intracellular NO gradually accumulated from 15 to 60 minutes, exhibiting a dose-time proportional relationship. The intensity difference between HUVEC-MNS-N0(+) and HUVEC-control (control group) was smaller than that in the B16 group, consistent with a normal ablation-repair process.

[0039] like Figure 6 As shown, the biocompatibility of SNP-Fe@MNs was investigated through in vitro cell experiments. Under light-free conditions, the biocompatibility of SCMC@MNs (carboxymethylcellulose sodium-loaded microneedles) and SNP-Fe@MNs loaded with 0.01, 0.03, 0.05, 0.10, and 0.15 g / ml sodium nitroprusside was evaluated using the CCK-8 assay. The control group consisted of SNP-Fe@MNs loaded with sodium carboxymethyl cellulose microneedles, the 0.01 group consisted of SNP-Fe@MNs loaded with 0.01 g / ml sodium nitroprusside, the 0.03 group consisted of SNP-Fe@MNs loaded with 0.03 g / ml sodium nitroprusside, the 0.05 group consisted of SNP-Fe@MNs loaded with 0.05 g / ml sodium nitroprusside, the 0.10 group consisted of SNP-Fe@MNs loaded with 0.10 g / ml sodium nitroprusside, and the 0.15 group consisted of SNP-Fe@MNs loaded with 0.15 g / ml sodium nitroprusside. Figure 6Figure 1a shows that the relative cell viability of L929 mouse fibroblasts after co-culturing with the microneedle patches for 24 hours and 48 hours had no statistical difference between the control group and the 0.03 group, and the activity decreased from 0.05 g / ml to 0.15 g / ml as the concentration gradually increased. The above results show that the increase in concentration indicates the increase in biological toxicity from the 0.05 g / ml group. Therefore, the critical 0.03 g / ml group is selected for subsequent experiments, which is consistent with the results of the above photothermal test screening.

[0040] Figure 6 Figure 1b is a graph showing the biological compatibility of the control group (control group), SCMC@MNs group, SNP-Fe@MNs-NO(-) group (without NO release) and SNP-Fe@MNs-NO(+) group (with NO release) using Calcein AM / PI kit to stain HUVECs, and the results are arranged in Figure 6 Figure 1c. There is no statistical difference between the experimental group and the control group whether NO exists in the 0.03 g / ml group or not, which shows that the biological compatibility of SNP-Fe@MNs meets the basic requirements for further research.

[0041] Figure 7 Figure 2a is a conceptual design of the postoperative full-thickness defect model; Figure 7 Figure 2b is a photograph of the control group, SCMC@MNs group, SNP-Fe@MNs-NO(-) group (without NO release) and SNP-Fe@MNs-NO(+) group (with NO release); Figure 7 Figure 2c is a simulated change image of the wound shape and size at the relevant five time points; Figure 7 Figure 2d is a statistical curve of the wound closure rate of the control (control group), SCMC@MNs group, SNP-Fe@MNs-NO(-) group (without NO release) and SNP-Fe@MNs-NO(+) group (with NO release) (n = 5); Figure 7 Figure 2e is a H&E (hematoxylin-eosin staining) immunohistochemical staining image of the wound anatomical tissue of the control group, SCMC@MNs group, SNP-Fe@MNs-NO(-) group (without NO release) and SNP-Fe@MNs-NO(+) group (with NO release) after 16 days; Figure 7 Figure 2f is a Masson (trichrome staining) immunohistochemical staining image of the wound anatomical tissue of the control group, SCMC@MNs group, SNP-Fe@MNs-NO(-) group (without NO release) and SNP-Fe@MNs-NO(+) group (with NO release) after 16 days; Figure 7Image g shows the CD31 (platelet-endothelial cell adhesion molecule) immunohistochemical staining images of wound anatomical tissues from the control group, SCMC@MNs group, SNP-Fe@MNs-NO(-) group (no NO release), and SNP-Fe@MNs-NO(+) group (NO release) 16 days later. NO, as an endothelial cell survival factor, can inhibit apoptosis and promote endothelial cell proliferation.

[0042] Early healing is beneficial for reducing infection and restoring function. Therefore, promoting skin tissue regeneration in wound patches is crucial for early healing and avoiding prolonged disease progression in postoperative wound patients. A full-thickness skin defect model was used on the left side of SD rats to simulate a large open wound after conventional prolonged surgery to remove melanoma. Microneedle patches with a diameter of 17.5 mm were fabricated, and 17 mm diameter live circular dermatomes were extracted using a skin drill. Custom-designed silicone rings were then sutured around the wound to prevent skin stretching or contraction. After wound establishment, microneedle patches were applied to the wounds of the corresponding groups: SCMC@MNs, SNP-Fe@MNs-NO(-), and SNP-Fe@MNs-NO(+). Figure 7 As shown in Figure a, from day 0 to day 16, considering the long-term metabolism and degradation of the biomaterials, the wound dressing was changed every 4 days to ensure overall therapeutic efficacy, and this was synchronized with the frequency of photography during the healing process. Figure 7 As shown in image b, this is to track changes in wound size. In addition to visually appealing photographs, such as... Figure 7 As shown in Figure c, simulated wound size and morphology images at five time points demonstrate the effect of SNP-Fe@MNs-NO(+) in promoting wound closure and healing. Furthermore, by calculating the wound healing rate, the NO-containing microneedle patch exhibited a unique ability to promote early wound healing, particularly in the early stages, where the wound healing rate was significantly higher than in other groups. This helped reduce the occurrence of adverse reactions. At day 16, as shown... Figure 7 As shown in Figure d, the wound closure rate was almost 100%. At the end of day 16, the fully healed wound was dissected and analyzed using H&E staining, Masson staining, and CD31 immunohistochemical staining. H&E staining results showed that the control group, SCMC@MNs group, and SNP-Fe@MNs-NO(-) group had significant scarring under immature granulation tissue of lengths of 5683 μm, 5287 μm, and 4434 μm, respectively. Conversely, during the 16-day treatment period, the SNP-Fe@MNs-NO(+) group exhibited minimal immature granulation tissue or complete recovery, with new functional epidermis and dermis tissue, well-defined boundaries, and skin appendages such as hair follicles, glands, and capillaries. Figure 7 As shown in the e-th image, instead of scar tissue healing. Figure 8As shown in FIG. 1f, consistent with H&E staining, a dense layer of activated collagen fibers can be seen in the SNP-Fe@MNs-NO(+) group by Masson staining. It is known that revascularization of the wound bed is essential for skin regeneration, so, for example, Figure 2 As shown in FIG. 1g, by analyzing CD31 by immunohistochemical staining, it can be seen that the SNP-Fe@MNs-NO(+) group has the most CD31 positive blood vessels, confirming that NO plays an important role in the formation of new blood vessels in vivo.

[0043] In the embodiments of the present application, the centrifugal method is a negative pressure centrifugal casting method. The centrifugal casting method is to fill and solidify the gel-like mixture under the action of centrifugal force. The feeding effect of the gel-like mixture is very good, and the outer layer of the tissue is dense, without molding and core making, thereby saving related materials and equipment investment. Of course, the present application is not limited to the negative pressure centrifugal casting method, and other centrifugal methods such as high-speed centrifugation can also be used.

[0044] In the embodiments of the present application, S104 centrifuges the microneedle patch, including: using a suction pump to centrifuge the microneedle patch at-0.08 MPa.

[0045] In the embodiments of the present application, the drying temperature is 20-25℃, i.e. room temperature.

[0046] In the embodiments of the present application, as shown in FIG. 1a, S101 specifically includes S801-S804. The preparation of nitroprusside nanoparticles using nitroprusside solid particles includes: Figure 4

[0047] S801: completely dissolving the nitroprusside solid particles in deionized water and ethanol to form a mixed solution.

[0048] S802: dispersing the mixed solution using ultrasonic vibration.

[0049] S803: keeping at-120℃ to-70℃ for more than 24 hours.

[0050] S804: obtaining nitroprusside nanoparticles by freeze-drying.

[0051] In the embodiments of the present application, the material of the mold is polydimethylsiloxane, and the polydimethylsiloxane mold prepared microneedle type edge is smooth, and the adhesion between the microneedle film is small, and easy to demold. Of course, the present application is not limited to the polydimethylsiloxane mold, and the material of the mold can also be stainless steel, silicone, silicon wafer, etc.

[0052] As shown in FIG. 1a, S101 specifically includes S801-S804. The preparation of nitroprusside nanoparticles using nitroprusside solid particles includes: Figure 5 ​As shown, the microneedle patch prepared using the above method in this application embodiment includes a substrate 1, microneedles 2 disposed on the substrate 1, and a release layer loaded on the microneedles 2 and the substrate 1. The release layer is a gel-like mixture formed in a mold. Microneedles (MNs) patches are an effective minimally invasive drug delivery platform that can penetrate the stratum corneum with minimal loss to reach the dermis, delivering drugs to the target site of skin malignancies. Furthermore, compared to other drug delivery routes, microneedle patches have significant advantages, including painlessness, easily permissible drug delivery, simplicity, good reproducibility, ideal tissue integration, biocompatibility, and mature clinical applications, and have been integrated into various cancer treatment strategies. Therefore, microneedle patches have been prepared as a novel alternative treatment strategy for head and neck malignant melanoma to prevent tumor recurrence and metastasis and promote early wound healing, contributing to the restoration of potential facial deformities.

[0053] In this embodiment of the application, the substrate 1 is circular, and the microneedles 2 are distributed radially along the substrate 1.

[0054] In this embodiment, the diameter of the base 1 is 17.5 mm, and the number of microneedles 2 is 385. Specifically, each microneedle 2 has a height of approximately 500 μm to ensure penetration of the dermis. To evaluate the puncture capability and tissue interaction performance of the microneedles 2, the changes in the microneedles after compressing the skin at different times were observed using scanning electron microscopy.

[0055] like ​ As shown, the microneedle patch was completely absorbed within approximately 30 minutes.

[0056] like ​ As shown, both the top vertical view and cross-section of the mouse skin indicate that the microneedle 2 effectively penetrated the skin early within 1 minute, facilitating self-administration. Simultaneously, after pressing the microneedle 2 onto the mouse skin for 1 minute, the effective puncture sites and micropores gradually closed within 20 minutes, demonstrating good integration with the surrounding tissue.

[0057] Therefore, the SNP-Fe@MNs patch prepared by the above method serves as a controllable photothermal synergy and sustained NO release platform for the ablation and tissue regeneration of malignant skin tumors. As a transdermal drug delivery system, the SNP-Fe@MNs patch can penetrate the dermis through relevant signaling pathways, exhibiting significant photothermal capabilities to achieve a burst release of NO and promote tumor cell apoptosis.

[0058] The various embodiments in this specification are described in a progressive manner. For the same or similar parts between the various embodiments, please refer to each other. Each embodiment focuses on describing the differences from other embodiments.

[0059] The above examples are only used to illustrate the technical solutions of the present application, and are not intended to limit the present application; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that the technical solutions recorded in the foregoing examples can still be modified, or some or all of the technical features thereof can be replaced by equivalents; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the present application.

Claims

1. A method for preparing a microneedle patch for treating malignant melanoma, characterized in that, include: Nitroprusside nanoparticles were prepared using solid nitroprusside particles. The preparation of sodium nitroprusside nanoparticles using sodium nitroprusside solid particles includes: The sodium nitroprusside solid particles were completely dissolved in deionized water and ethanol to form a mixed solution; The mixed solution was dispersed using ultrasonic vibration; Keep at -120℃ to -70℃ for more than 24 hours; The sodium nitroprusside nanoparticles were obtained by freeze-drying. The 0.01-0.15 g / ml nitroprusside nanoparticles were combined with ferrous lactate nanoparticles in an equimolar ratio, and dissolved in deionized water with a 0.08 g / ml sodium carboxymethyl cellulose solution, and mixed to form a gel-like mixture. The gel-like mixture is added to a mold, and microneedle patches are manufactured by centrifugation. The centrifugal method is a negative pressure centrifugal casting method; The microneedle patch was centrifuged and dried under vacuum conditions.

2. The method for preparing a microneedle patch for treating malignant melanoma according to claim 1, characterized in that, The sodium nitroprusside nanoparticles have a concentration of 0.03 g / ml.

3. The method for preparing a microneedle patch for treating malignant melanoma according to claim 1, characterized in that, The centrifugation of the microneedle patch includes: The microneedle patch was centrifuged using a suction pump at -0.08 MPa.

4. The method for preparing a microneedle patch for treating malignant melanoma according to claim 1, characterized in that, The drying temperature is 20-25℃.

5. The method for preparing a microneedle patch for treating malignant melanoma according to claim 1, characterized in that, The mold is made of polydimethylsiloxane.

6. A microneedle patch prepared using the method described in any one of claims 1-5, characterized in that, It includes a substrate, microneedles disposed on the substrate, and a release layer loaded on the microneedles and the substrate; The release layer is formed in the mold by the gel-like mixture.

7. The microneedle patch according to claim 6, characterized in that, The substrate is circular, and the microneedles are distributed radially along the substrate.

8. The microneedle patch according to claim 7, characterized in that, The substrate has a diameter of 17.5 mm and the number of microneedles is 385.

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