3D printing photocured gradient release drug hydrogel and preparation method and application thereof

Abstract

This invention provides a 3D-printable photocurable drug-releasing hydrogel, its preparation method, and its applications. A hydrogel system is prepared by forming a dynamic and covalent network, carrying a poorly water-soluble drug, and then photocuring. The hydrogel of this invention possesses excellent photocrosslinking ability, as well as outstanding printability and shape adaptability, supporting 3D printing and can be used to construct personalized drug-releasing scaffolds. Furthermore, as the hydrogel slowly degrades, the 3D scaffold collapses layer by layer, allowing for sustained, long-term, and gradual release of the drug at the lesion site, effectively inhibiting tumor growth. The hydrogel has a loose, porous network structure and excellent adhesion, enabling it to conform to postoperative bleeding sites, absorb wound exudate, and prevent postoperative massive hemorrhage. The hydrogel of this invention can serve as a novel drug delivery carrier for inhibiting postoperative recurrence in various cancers.

Description

Technical Field

[0001] This invention belongs to the field of polymer materials technology, specifically relating to the preparation of a 3D-printed photocrosslinked hydrogel for graded drug release and its application in inhibiting postoperative recurrence of cancer. Background Technology

[0002] Prostate cancer is one of the most common malignant tumors threatening men's health, with a persistently high incidence rate, seriously threatening patients' lives and health. Surgical treatment has always been the preferred treatment for early-stage prostate cancer patients; however, factors such as poor patient condition, incomplete surgical resection, and preoperative micrometastases significantly increase the risk of postoperative recurrence. Therefore, chemotherapy is usually administered postoperatively to eliminate residual tumor cells and reduce the risk of tumor recurrence and metastasis. In recent years, active ingredients from natural Chinese herbal medicines have received widespread attention due to their multi-target and low-toxicity anti-tumor properties. Curcumin and luteolin, as typical extracts of traditional Chinese medicine, can effectively inhibit the proliferation of prostate cancer cells and induce apoptosis. However, these two drugs have poor water solubility, rapid metabolism in vivo, and low bioavailability, severely limiting their clinical application. Therefore, developing a drug delivery system capable of carrying poorly water-soluble drugs and releasing them locally at the lesion site in a long-term, stepped manner has become a key challenge in inhibiting postoperative recurrence of prostate cancer.

[0003] Hydrogels, as functional materials with a three-dimensional network structure, have shown great potential in inhibiting postoperative tumor recurrence due to their excellent biocompatibility, tunable mechanical properties, and superior drug loading capacity, attracting the attention of numerous researchers. It is worth noting that...

[0004] Considering the side effects of systemic chemotherapy, a hydrogel capable of controlled, long-term drug release at the lesion site is urgently needed. Therefore, loading drugs onto a biodegradable hydrogel network can respond to the tumor microenvironment, enabling prolonged local drug release at the lesion site, enhancing therapeutic efficacy, reducing the side effects of systemic administration, and achieving long-term drug release. Summary of the Invention

[0005] The purpose of this invention is to provide a 3D-printed photocrosslinked hydrogel for the stepwise release of drugs and its application in inhibiting postoperative recurrence of prostate cancer.

[0006] Photocurable hydrogels can be rapidly cured by ultraviolet light, forming in situ to better adapt to irregular postoperative wounds, achieving close adhesion with tissue and enhancing therapeutic efficacy. Furthermore, photocurable hydrogels possess excellent printability and shape adaptability. By adding material layer by layer, a digital model is fabricated into a 3D solid scaffold, with poorly soluble drugs embedded between each layer. As the hydrogel degrades, the 3D scaffold collapses layer by layer, releasing the drug in stages, extending the drug release cycle and creating a personalized drug release system. Adding a dynamic network layer on top of the covalent network preserves the hydrogel's mechanical stability, preventing excessively rapid drug release, while also enhancing degradation controllability and preventing drug release difficulties. A mixture of monomers, the photoinitiator LAP, and poorly water-soluble chemotherapy drugs is injected into the lesion site after tumor resection, and then photocured layer by layer under ultraviolet light using 3D printing to form a double-network hydrogel structure. As the hydrogel slowly degrades in vivo, the drug is released layer by layer in stages, achieving long-term slow drug release and effectively inhibiting postoperative cancer recurrence. This invention not only provides a new solution for poorly water-soluble drug delivery systems, but also offers a new strategy for inhibiting postoperative recurrence of prostate cancer.

[0007] The three-dimensional porous structure of the hydrogel endows it with excellent hemostatic properties; the abundant functional groups on its surface endow it with excellent adhesion; the addition of natural polymers endows it with excellent biocompatibility and biodegradability; the photocrosslinking properties of the hydrogel make 3D printing possible, and the desired shape can be printed according to the local spatial structure of the lesion site where drug release is needed. The construction of the covalent network endows the hydrogel with good stability, which is conducive to long-term and sustained drug release, and the construction of the dynamic network endows the hydrogel with a controllable degradation rate, which is conducive to the layer-by-layer controlled release of drugs. This 3D printed photocurable hydrogel can effectively load poorly water-soluble drugs at high concentrations. As the hydrogel degrades, the 3D scaffold collapses layer by layer, and the drug is released locally at the lesion site in a long-term, step-by-step manner. Cell experiments and animal experiments further demonstrate that this 3D printed photocrosslinked hydrogel step-release system can be used to inhibit postoperative recurrence of prostate cancer. Combining the above characteristics, this novel multifunctional hydrogel with adhesion, hemostasis, biocompatibility, and 3D printability has great potential as a novel drug delivery material and has important application value in biomedical fields such as inhibiting tumor recurrence.

[0008] The preparation method of a 3D-printed photocurable graded-release chemotherapy drug hydrogel according to the present invention comprises the following specific steps:

[0009] (1) Preparation of oxidized konjac polysaccharide: 2.0 g of konjac polysaccharide was dissolved in 200 mL of deionized water, and 0.6 g of sodium periodate was added. The mixture was stirred in the dark for 12 h. After the reaction was completed, 4 mL of ethylene glycol was added to terminate the reaction, and stirring was continued for 2 h. The resulting reaction solution was then poured into a dialysis bag and dialyzed with deionized water for 3 days. The oxidized konjac polysaccharide was obtained by freeze-drying.

[0010] (2) Preparation of methacrylamide gelatin: 10.0 g gelatin was dissolved in 100 mL PBS, 1.0 mL methacrylic anhydride was added, and the mixture was stirred for 3 h. The reaction solution was diluted with PBS, then poured into a dialysis bag and dialyzed with deionized water for 7 days. The methacrylamide gelatin was obtained by freeze drying.

[0011] (3) Preparation of hydrogel: Prepare a 2% carboxymethyl chitosan aqueous solution, add 10-20 mg / mL of poorly water-soluble chemotherapeutic drug M, stir for 20 min and sonicate for 20 min until the chemotherapeutic drug is evenly dispersed in the solution and there are no obvious large particles precipitated. Then, add 2% methacrylamide gelatin and 0.5% photoinitiator LAP in sequence, and stir for 20 min to mix evenly. Subsequently, mix the above mixed solution with 2% oxidized konjac polysaccharide solution at a volume ratio of 1:1 and then inject it layer by layer into the lesion site after tumor resection using 3D printing. Cure and form a double cross-linked hydrogel by irradiation under a 405nm ultraviolet lamp for 0.5-3 minutes.

[0012] In embodiments of the present invention, the poorly water-soluble chemotherapeutic drug M loaded on the hydrogel can be one or a mixture of several of the following drugs: luteolin, TGF-β inhibitor (A-77-01), curcumin, and doxorubicin; thereby achieving localized, layer-by-layer, stepwise release at the tumor lesion site and effectively exerting the therapeutic effect of the drug.

[0013] For example, the photoinitiator is LAP (lithium phenyl-2,4,6-trimethylbenzoylphosphonite). Photocuring is performed under a 405 nm UV lamp.

[0014] An embodiment of the present invention also provides a 3D-printed photocurable step-release chemotherapy drug hydrogel prepared by the above method.

[0015] Embodiments of the present invention also provide a hydrogel with excellent photocrosslinking ability, as well as excellent printability and shape adaptability, supporting rapid 3D printing and can be used to construct personalized drug delivery scaffolds; by utilizing the hydrogel to load drugs and biodegrade, the 3D scaffold collapses layer by layer, and the drug is slowly released from the hydrogel scaffold, avoiding the drug burst effect.

[0016] Embodiments of the present invention also provide a drug delivery method using a 3D-printed photocurable stepwise release chemotherapy drug hydrogel. The 3D-printed photocurable stepwise release chemotherapy drug hydrogel is used as a drug delivery carrier to directly apply the drug to the tumor lesion site and release the drug locally at the lesion site. Furthermore, as the hydrogel slowly degrades, the drug is released layer by layer, thereby achieving long-term and sustained drug release.

[0017] Embodiments of the present invention also provide a hydrogel with a loose and porous structure and excellent adhesion, which can adhere to the postoperative bleeding site, absorb the exudate from the wound, and achieve the effects of adhesion and hemostasis.

[0018] Embodiments of the present invention also provide a 3D-printed photocurable stepwise release chemotherapy drug hydrogel, which directly injects the drug into a gel form and applies it to the lesion site, and its application in the preparation of drugs to inhibit the recurrence of tumors such as prostate cancer.

[0019] Compared with the prior art, the present invention has the following advantages and technical effects:

[0020] 1. The hydrogel constructed in this invention can load poorly water-soluble drugs and release them locally at the lesion site;

[0021] 2. Hydrogels have excellent photocrosslinking capabilities and can be rapidly cured by ultraviolet light, forming in situ, better adapting to irregular postoperative wounds, and achieving close adhesion with tissues;

[0022] 3. The hydrogel adopts a dual network structure of dynamic covalent bonds and covalent photocrosslinking, which has both good mechanical stability and controllable degradation, enabling long-term, stepwise, and slow release of drugs, avoiding burst release effects and prolonging the duration of drug action;

[0023] 4. The material has excellent printability and shape adaptability, supporting 3D printing and can be used to construct personalized drug delivery scaffolds. As the hydrogel degrades, the 3D scaffold collapses layer by layer, allowing for long-term, stepped release of drugs at the lesion site.

[0024] 5. The hydrogel has a porous internal structure, which has high liquid absorption and plays a role in exudate management and physical barrier, thus assisting in postoperative hemostasis;

[0025] 6. The raw materials are widely available and have good biocompatibility. The preparation process is mild, green and environmentally friendly, making it suitable for large-scale production and clinical translation.

[0026] 7. The novel 3D-printed, adhesive, and hemostatic multifunctional hydrogel of the present invention has great potential as a new drug delivery material. It is not only suitable for the prevention and control of postoperative recurrence of prostate cancer, but can also be extended to the treatment of other malignant tumors. It has broad application prospects in the fields of precision medicine and tissue engineering. Attached Figure Description

[0027] Figure 1 : Phase transition diagram of the 3D-printed photocurable step-release chemotherapy drug hydrogel prepared in Example 1. By comparing the changes in the substances in the tilted reagent bottle before and after, it can be seen that the hydrogel precursor solution is a homogeneous liquid, which becomes a gel solid after UV curing.

[0028] Figure 2 The image shows a scanning electron microscope (SEM) image of the 3D-printed photocurable, step-release chemotherapy drug hydrogel prepared in Example 1 after lyophilization. The image reveals a loose, porous structure within the hydrogel, which is beneficial for the loading and release of chemotherapy drugs and for absorbing postoperative wound exudate.

[0029] Figure 3 The image shows the 3D printing photocuring result of the hydrogel prepared in Example 1. As can be seen from the image, after the injection and photocuring processes, a hydrogel solid of the desired shape can be formed.

[0030] Figure 4 This image shows the in vitro drug release of the 3D-printed, photocurable, step-release chemotherapy drug hydrogel prepared in Example 1. The image shows that the chemotherapy drug loaded in the hydrogel is released slowly and continuously, with 70% of the drug being released over a period of more than 12 days.

[0031] Figure 5 This is a graph showing the in vitro release of the drug in its pure free form. The graph shows that the free drug is released relatively quickly, with 70% of the drug being released in just 3 days.

[0032] Figure 6 The image shows the effect of 3D-printed photocurable stepwise release chemotherapy drug hydrogel prepared in Example 1 on inhibiting postoperative recurrence of prostate cancer in mice. As can be seen from the image, after treatment, the residual tumor volume in the control group mice reached the maximum ethical limit for tumor recurrence; while the residual tumor in the mice in the photocurable stepwise release chemotherapy drug hydrogel group was significantly inhibited, indicating that the stepwise release of drugs by the drug-loaded hydrogel group can effectively inhibit postoperative recurrence of prostate cancer. Detailed Implementation

[0033] Example 1

[0034] A method for preparing a 3D-printed photopolymerized, step-release chemotherapy drug hydrogel:

[0035] (1) Preparation of oxidized konjac polysaccharide: 2.0 g of konjac polysaccharide was dissolved in 200 mL of deionized water, and 0.6 g of sodium periodate was added. The mixture was stirred in the dark for 12 h. After the reaction was completed, 4 mL of ethylene glycol was added to terminate the reaction, and stirring was continued for 2 h. The resulting reaction solution was then poured into a dialysis bag and dialyzed with deionized water for 3 days. The oxidized konjac polysaccharide was obtained by freeze-drying.

[0036] (2) Preparation of methacrylamide gelatin: 10.0 g gelatin was dissolved in 100 mL PBS, 1.0 mL methacrylic anhydride was added, and the mixture was stirred for 3 h. The reaction solution was diluted with PBS, then poured into a dialysis bag and dialyzed with deionized water for 7 days. The methacrylamide gelatin was obtained by freeze drying.

[0037] (3) Preparation of hydrogel: Prepare a 2% carboxymethyl chitosan aqueous solution, add 10 mg / mL luteolin, stir for 20 min, and sonicate for 20 min until the chemotherapeutic drug is evenly dispersed in the solution without obvious large particle precipitation. Then, add 2% methacrylamide gelatin and 0.5% photoinitiator LAP in sequence, and stir for 20 min to mix evenly. Subsequently, mix the above mixed solution with 2% oxidized konjac polysaccharide solution at a volume ratio of 1:1, and then inject it layer by layer into the lesion site after tumor resection using 3D printing. Cure the gel by irradiation under a 405nm ultraviolet lamp for 1.5 minutes to form a double crosslinked hydrogel.

[0038] Example 2

[0039] A method for preparing a 3D-printed photopolymerized, step-release chemotherapy drug hydrogel:

[0040] (1) Preparation of oxidized konjac polysaccharide: 2.0 g of konjac polysaccharide was dissolved in 200 mL of deionized water, and 0.6 g of sodium periodate was added. The mixture was stirred in the dark for 12 h. After the reaction was completed, 4 mL of ethylene glycol was added to terminate the reaction, and stirring was continued for 2 h. The resulting reaction solution was then poured into a dialysis bag and dialyzed with deionized water for 3 days. The oxidized konjac polysaccharide was obtained by freeze-drying.

[0041] (2) Preparation of methacrylamide gelatin: 10.0 g gelatin was dissolved in 100 mL PBS, 1.0 mL methacrylic anhydride was added, and the mixture was stirred for 3 h. The reaction solution was diluted with PBS, then poured into a dialysis bag and dialyzed with deionized water for 7 days. The methacrylamide gelatin was obtained by freeze drying.

[0042] (3) Preparation of hydrogel: Prepare a 2% carboxymethyl chitosan aqueous solution, add 10 mg / mL curcumin, stir for 20 min, and sonicate for 20 min until the chemotherapeutic drug is evenly dispersed in the solution and there are no obvious large particles precipitated. Then, add 2% methacrylamide gelatin and 0.5% photoinitiator LAP in sequence, and stir for 20 min to mix evenly. Subsequently, mix the above mixed solution with 2% oxidized konjac polysaccharide solution at a volume ratio of 1:1, and then inject it layer by layer into the lesion site after tumor resection using 3D printing. Cure the gel by irradiation under a 405nm ultraviolet lamp for 3.0 min to form a double crosslinked hydrogel.

[0043] Example 3

[0044] A method for preparing a 3D-printed photopolymerized, step-release chemotherapy drug hydrogel:

[0045] (1) Preparation of oxidized konjac polysaccharide: 2.0 g of konjac polysaccharide was dissolved in 200 mL of deionized water, and 0.6 g of sodium periodate was added. The mixture was stirred in the dark for 12 h. After the reaction was completed, 4 mL of ethylene glycol was added to terminate the reaction, and stirring was continued for 2 h. The resulting reaction solution was then poured into a dialysis bag and dialyzed with deionized water for 3 days. The oxidized konjac polysaccharide was obtained by freeze-drying.

[0046] (2) Preparation of methacrylamide gelatin: 10.0 g gelatin was dissolved in 100 mL PBS, 1.0 mL methacrylic anhydride was added, and the mixture was stirred for 3 h. The reaction solution was diluted with PBS, then poured into a dialysis bag and dialyzed with deionized water for 7 days. The methacrylamide gelatin was obtained by freeze drying.

[0047] (3) Preparation of hydrogel: Prepare a 2% carboxymethyl chitosan aqueous solution, add 20 mg / mL doxorubicin, stir for 20 min, and sonicate for 20 min until the chemotherapeutic drug is evenly dispersed in the solution and there are no obvious large particles precipitated. Then, add 2% methacrylamide gelatin and 0.5% photoinitiator LAP in sequence, and stir for 20 min to mix evenly. Subsequently, mix the above mixed solution with 2% oxidized konjac polysaccharide solution at a volume ratio of 1:1, and then inject it layer by layer into the lesion site after tumor resection using 3D printing. Cure the gel by irradiation under a 405nm ultraviolet lamp for 0.5 min to form a double crosslinked hydrogel.

[0048] Figure 4 and Figure 5 The comparison results show that the drug is released relatively quickly when it is free, with 70% of the drug released in just 3 days. In contrast, the 3D-printed photocurable cascade-release chemotherapy drug hydrogel prepared in Example 1 releases the chemotherapy drug slowly and continuously because the drug is supported by the hydrogel, with 70% of the drug released in more than 12 days.

[0049] The 3D-printed, photocurable, step-release chemotherapeutic drug hydrogel prepared in Example 1 was used to inhibit postoperative recurrence in mice with prostate cancer. The specific procedure was as follows: each mouse was subcutaneously injected with 1×10⁻⁶ treasury cells in its right chest. 6 Mouse prostate cancer cells were used to establish a prostate cancer xenograft model. When the average tumor volume reached 100 mm... 3After isoflurane inhalation anesthesia, 90% tumor resection was performed on all mice. For the remaining tumor sites, each group received corresponding treatment: the control group received 0.1 mL of PBS solution, the hydrogel group received 0.1 mL of hydrogel, the drug-only group received 0.1 mL of a drug-water mixture (containing 10 mg luteolin per mL of physiological saline), and the drug-loaded hydrogel group received 0.1 mL of a photocurable, stepwise release chemotherapy hydrogel containing luteolin. The skin was then closed using interrupted sutures. Postoperatively, each mouse received an intramuscular injection of 8 units of penicillin sodium (dissolved in 0.1 wt% physiological saline) to prevent infection. Mouse weight and tumor volume were monitored daily. When the tumor volume reached 2000 mm², the tumor was treated accordingly. 3 The mice were euthanized. By observing and comparing the changes in tumor volume and weight in each group, the effect of the hydrogel in inhibiting tumor recurrence could be determined. Figure 6 The image shows a 3D-printed, photocurable, step-release chemotherapeutic drug hydrogel prepared in Example 1 that inhibits postoperative recurrence of prostate cancer in mice. It can be seen that after 16 days of treatment, the residual tumor volume in the control group and the hydrogel group reached the maximum value for tumor ethics; the residual tumor volume in the drug-only group decreased to some extent, but was still relatively large; while the residual tumor volume in the drug-loaded hydrogel group was the smallest, and the tumor was significantly inhibited. This indicates that the 3D-printed, photocurable, step-release chemotherapeutic drug hydrogel of the present invention can effectively inhibit postoperative recurrence of prostate cancer.

Claims

1. A method for preparing a 3D-printed photocurable graded-release drug hydrogel, the specific steps of which are as follows: (1) Preparation of oxidized konjac polysaccharide: 2.0 g of konjac polysaccharide was dissolved in 200 mL of deionized water, and 0.6 g of sodium periodate was added. The mixture was stirred in the dark for 12 h. After the reaction was completed, 4 mL of ethylene glycol was added to terminate the reaction and the mixture was stirred for another 2 h. The resulting reaction solution was then poured into a dialysis bag and dialyzed with deionized water for 3 days. The oxidized konjac polysaccharide was obtained by freeze drying. (2) Preparation of methacrylamide gelatin: 10.0 g gelatin was dissolved in 100 mL PBS, 1.0 mL methacrylic anhydride was added, and the mixture was stirred for 3 h. The reaction solution was diluted with PBS, and then poured into a dialysis bag. The solution was dialyzed with deionized water for 7 days and then freeze-dried to obtain methacrylamide gelatin. (3) Preparation of hydrogel: Prepare a 2% carboxymethyl chitosan aqueous solution, add 10-20 mg / mL of poorly water-soluble chemotherapeutic drug M, stir for 20 min and sonicate for 20 min until the chemotherapeutic drug is evenly dispersed in the solution and there are no obvious large particles precipitated; then, add 2% methacrylamide gelatin and 0.5% photoinitiator LAP in sequence, stir for 20 min to mix evenly; then, mix the above mixed solution with 2% oxidized konjac polysaccharide solution at a volume ratio of 1:1 evenly, and then inject it layer by layer into the lesion site after tumor resection using 3D printing method, and cure it under 405nm ultraviolet light for 0.5-3 minutes to form a double crosslinked hydrogel.

2. A 3D-printed photocurable graded-release drug hydrogel, characterized in that: The hydrogel prepared according to the method described in claim 1 can load a poorly water-soluble chemotherapeutic drug M, which is one or a mixture of several of the following drugs: luteolin, TGF-β inhibitor (A-77-01), curcumin, and doxorubicin; thereby achieving local release at the lesion site and effectively exerting the therapeutic effect of the drug.

3. The hydrogel according to claim 2, characterized in that: Hydrogels possess excellent photocrosslinking capabilities, as well as outstanding printability and shape adaptability, supporting rapid 3D printing and enabling the construction of personalized drug delivery scaffolds. By utilizing hydrogels to load drugs and biodegrade, the 3D scaffold collapses layer by layer, allowing the drug to be slowly released from the hydrogel scaffold, avoiding the drug burst effect.

4. The hydrogel according to claim 2, characterized in that: Hydrogels have a loose and porous structure and excellent adhesion, which can fit the postoperative bleeding site, absorb the exudate from the wound, and achieve the effects of adhesion and hemostasis.

5. The application of the hydrogel according to any one of claims 2-4, characterized in that, Using the hydrogel as a drug delivery carrier, the drug is directly applied to the tumor lesion site and released locally at the lesion site; and as the hydrogel slowly degrades, the drug is released layer by layer, thereby achieving long-term sustained drug release.

6. The application of the hydrogel according to any one of claims 2-4, characterized in that, Used to prepare drugs that inhibit the recurrence of prostate cancer tumors.

Images