Aerogel vascular tumor model and application thereof

Abstract

The invention discloses an aerogel vascular tumor model and application thereof. According to the method, a tubular aerogel stent attached with cells is utilized and integrated into a perfusion culture device, and through a highly porous structure of the aerogel stent and nutrient substance exchange in the perfusion process, a three-dimensional model of large-size (millimeter-level) tumor infringement great vessels is constructed in vitro for the first time; the technical difficulty that the model cannot be constructed in the prior art is overcome.

Description

Technical Field

[0001] The invention relates to the technical field of medical simulation models, and in particular to an aerogel vascular tumor model and application thereof. Background Art

[0002] Liver cancer is one of the most common malignant tumors worldwide. Studies have shown that by 2025, the number of new cases of liver cancer worldwide will exceed 1 million per year. Among the many types of liver cancer, hepatocellular carcinoma (HCC) is the most common type of liver cancer, which mainly occurs in patients with chronic liver disease and cirrhosis. Due to the high invasiveness and early metastasis ability of HCC, its treatment still faces huge challenges. Vascular invasion is one of the most important factors for the poor prognosis of HCC. Since HCC can cause cancer cells to invade blood vessels, especially the portal vein and hepatic vein, it generally leads to intrahepatic spread and distant metastasis. Therefore, vascular invasion caused by HCC often leads to postoperative recurrence and a significant shortening of patient survival.

[0003] In the prior art, although the mouse model can show the vascular invasion characteristics of tumor cells, due to the small size of mouse tumors and blood vessels, as well as ethical restrictions, it has many deficiencies in completely replicating human diseases. Although the traditional tumor organ chip model can simulate the tumor environment supported by microvessels, it is still insufficient in reproducing the invasion of tumors on large blood vessels. Moreover, for the general method of constructing microvascular networks in vitro, due to the difficulty in excreting metabolic products inside the tissue and the lack of sufficient nutrient supply, necrosis can easily occur inside the tissue, thereby greatly reducing the success rate of construction. It can be seen that there are still huge challenges in constructing large blood vessel channels in vitro and maintaining their structure and function.

[0004] Therefore, developing a 3D HCC model with vascular invasion characteristics that allows researchers to better observe the interactions between human hepatocytes and endothelial cells and the physiological response to drugs will be of great significance for researchers' understanding of HCC treatment. Summary of the invention

[0005] The present invention aims to solve at least one of the above-mentioned technical problems existing in the prior art. To this end, the purpose of the present invention is to provide an aerogel vascular tumor model and its application. The present invention utilizes a tubular aerogel scaffold to which cells are attached and integrates it into a perfusion culture device. Through the highly porous structure of the aerogel scaffold and the exchange of nutrients during the perfusion process, a three-dimensional model of large-scale (millimeter-level) tumors invading large blood vessels can be constructed in vitro.

[0006] A first aspect of the present invention provides an aerogel scaffold, wherein raw materials for preparing the aerogel scaffold include lactide-caprolactone copolymer and 1,4-dioxane, wherein the mass volume ratio of the lactide-caprolactone copolymer to 1,4-dioxane is 1-3:100.

[0007] In some embodiments of the present invention, the mass volume ratio of the lactide-caprolactone copolymer to 1,4-dioxane is 1.5-3:100.

[0008] In some embodiments of the present invention, the mass volume ratio of the lactide-caprolactone copolymer to 1,4-dioxane is 2:100.

[0009] In some embodiments of the present invention, the raw materials for preparing the aerogel scaffold are only lactide-caprolactone copolymer and 1,4-dioxane.

[0010] In some embodiments of the present invention, the aerogel scaffold is hydrophilically modified.

[0011] In some embodiments of the present invention, the hydrophilic modification is achieved by plasma treatment. Of course, those skilled in the art can select other hydrophilic modification methods to treat the aerogel scaffold according to actual use requirements, including but not limited to plasma treatment, hydrophilic layer (coupling agent, surfactant or hydrophilic monomer or polymer) coating, ultraviolet irradiation, high energy radiation and ozone method, etc.

[0012] In some embodiments of the present invention, the aerogel support has no specific limitation on shape and can be designed into different tubular structures such as round tubes, square tubes, rectangular tubes, elliptical tubes, hexagonal tubes, etc. as needed.

[0013] In some embodiments of the present invention, the aerogel scaffold has a porous structure with a pore size of 30-80 μm, a Young's modulus of 0.5-2 kPa, and a tensile modulus of 115-260%.

[0014] The second aspect of the present invention provides a method for preparing the aerogel scaffold described in the above aspect, comprising the following steps:

[0015] A mixed solution of lactide-caprolactone copolymer and 1,4-dioxane is used to cast a mold, and the mold is obtained after freeze-drying.

[0016] In some embodiments of the present invention, the mold can be prepared by any method in the art, including but not limited to 3D printing.

[0017] In some embodiments of the present invention, the mold is obtained by 3D printing, and the 3D printing material is 8200pro resin material. Of course, those skilled in the art can also choose other materials that are conducive to demolding for printing according to actual needs.

[0018] In some embodiments of the present invention, the lactide-caprolactone copolymer and 1,4-dioxane are fully mixed and completely dissolved.

[0019] In some embodiments of the present invention, the freeze drying is performed using a freeze dryer under vacuum conditions.

[0020] In some embodiments of the present invention, the lactide-caprolactone copolymer and 1,4-dioxane are mixed and dissolved by stirring at a rate of 1000 rpm for more than 2 hours, and then the mixed solution is introduced into a mold and frozen at -20°C / -80°C for more than 1 hour, and then the frozen mold is placed in a freeze dryer for freeze drying overnight.

[0021] In some embodiments of the present invention, the aerogel scaffold is subjected to a hydrophilic treatment after vacuum freeze drying.

[0022] In some embodiments of the present invention, the hydrophilic modification is achieved by plasma treatment. Of course, those skilled in the art can select other hydrophilic modification methods to treat the aerogel scaffold according to actual use requirements, including but not limited to plasma treatment, hydrophilic layer (coupling agent, surfactant or hydrophilic monomer or polymer) coating, ultraviolet irradiation, high energy radiation and ozone method, etc.

[0023] In some embodiments of the present invention, the hydrophilic modification is carried out by plasma treatment, the plasma irradiation power is 120 W, and the irradiation time is 1 min.

[0024] The third aspect of the present invention provides the use of the aerogel scaffold described in the above aspects in preparing a medical simulation model.

[0025] In some embodiments of the present invention, the medical simulation model is a blood vessel model.

[0026] In some embodiments of the present invention, the medical simulation model is a tumor invasion blood vessel model.

[0027] In some embodiments of the present invention, the medical simulation model is a HCC invasion of blood vessels model.

[0028] A fourth aspect of the present invention provides a vascular tumor simulation model, which includes the aerogel scaffold of the above aspect, endothelial cells attached to the inner surface of the aerogel scaffold, and tumor cells attached to the outer surface of the aerogel scaffold.

[0029] In some embodiments of the invention, the endothelial cells and tumor cells are not cross-attached at the same site.

[0030] A fifth aspect of the present invention provides a method for constructing a vascular tumor simulation model, comprising the following steps:

[0031] Endothelial cells and tumor cells are respectively planted on the inner surface and outer surface of the aerogel scaffold described in the above aspect. After the cells are fixed on the aerogel scaffold, perfusion culture is performed. After 1-6 days of culture, a vascular tumor simulation model is obtained.

[0032] In some embodiments of the present invention, the aerogel scaffold is continuously rotated during implantation.

[0033] In some embodiments of the present invention, the cell concentration of endothelial cells and tumor cells used for implantation is 3-5×10 7 / mL.

[0034] Of course, those skilled in the art can reasonably adjust the cell concentrations of endothelial cells and tumor cells used for implantation based on actual conditions.

[0035] In some embodiments of the present invention, the endothelial cells may include HUVECs, and the tumor cells may include liver cancer cells, such as Hep3B cells. Of course, it should be understood by those skilled in the art that the tumor invasion of large blood vessels described in the present invention is not limited to liver cancer, but other types of tumors may also have such a phenomenon, such as pancreatic cancer, renal cell carcinoma and lung cancer. Therefore, the situation of tumor invasion of large blood vessels caused by other tumors can also be simulated using the simulation model of the present invention.

[0036] A sixth aspect of the present invention provides the use of the vascular tumor simulation model described in the above aspect or the vascular tumor simulation model obtained by the construction method described in the above aspect in an in vitro disease simulation model.

[0037] In some embodiments of the invention, the disease comprises a tumor.

[0038] In some embodiments of the invention, the disease is angioinvasive tumor.

[0039] In some embodiments of the invention, the disease is HCC.

[0040] The seventh aspect of the present invention provides the use of the vascular tumor simulation model described in the above aspect or the vascular tumor simulation model obtained by the construction method described in the above aspect in constructing an in vitro drug screening platform.

[0041] The beneficial effects of the present invention are:

[0042] 1. This invention proposes for the first time a simulation model of tumor invasion of large blood vessels constructed by using a tubular aerogel scaffold with cell attachment, which solves the gap of no in vitro model in the prior art. At the same time, compared with conventional mouse tumor models, this model can realize the construction of large blood vessels and large-sized tumors. On the one hand, the tumors and blood vessels of mice cannot grow to this size level, and on the other hand, it is difficult to effectively simulate this process due to ethical restrictions.

[0043] 2. The model construction method of the present invention is simple, fast and easy to implement. Moreover, by combining the tubular aerogel scaffold with the perfusion culture device, it meets multiple requirements such as highly porous structure and nutrient exchange, thereby overcoming the technical defects that cannot be constructed in the prior art and achieving the technical effect of constructing a large-scale (millimeter level) three-dimensional model in vitro. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 A complete flowchart for preparing aerogel vascular tumor models.

[0045] Figure 2 This is a physical picture of the 3D printed bracket mold.

[0046] Figure 3 The hydrophilicity and hydrophobicity changes of the scaffold surface before and after plasma treatment.

[0047] Figure 4 This is a physical picture of the tubular aerogel scaffold.

[0048] Figure 5 The scanning electron microscope images of Examples 1-4 are shown.

[0049] Figure 6 Schematic diagram of the stretching of tubular aerogel scaffold.

[0050] Figure 7 Schematic diagram of the perfusion device.

[0051] Figure 8 The infiltration of cells on the scaffold after one day of perfusion (DAPI was used for cell nucleus staining, and Cy5 was used for scaffold staining) (scale bar, 200 μm).

[0052] Fig. 9Images of the cell distribution of HUVEC cells and Hep3B cells at the front, middle and back ends of the scaffold (scale bar: 500 μm).

[0053] Fig.10 The immunostaining images of VE-Cadherin and CD31 of the scaffolds are shown (scale bar: 200 μm).

[0054] Fig.11 The image shows the cell distribution when there are only Hep3B cells on the scaffold (scale bar is 500 μm).

[0055] Fig.12 Ki67 staining images of cells on scaffolds under three culture conditions (scale bar is 200 μm).

[0056] Fig.13 Live / dead staining images of the scaffolds after co-culture with three drugs (sorafenib, lenvatinib, and LNP-PLK1) for 48 h (scale bar, 200 μm).

[0057] Fig.14 Ki67 immunofluorescence staining images after the scaffolds were co-cultured with three drugs (sorafenib, lenvatinib and LNP-PLK1) for 48 hours (scale bar is 200 μm). DETAILED DESCRIPTION

[0058] The present invention is further described in detail below by specific examples. Unless otherwise specified, the raw materials, reagents or devices used in the examples and comparative examples can be obtained from conventional commercial sources or can be obtained by prior art methods. Unless otherwise specified, the experiments or test methods are conventional methods in the art.

[0059] Example 1

[0060] This embodiment provides a method for constructing an aerogel vascular tumor model, the process of which is as follows: Figure 1 As shown, the specific steps include:

[0061] (1) Construction of aerogel scaffold:

[0062] According to the required size, a mold for preparing tubular stents is designed using a 3D printing method. Then, the lactide-caprolactone copolymer (PLCL) powder is fully added to the 1,4-dioxane solution, and stirred at room temperature at 1000 rpm for more than 2 hours using a magnetic stirrer to fully dissolve it. After forming a mixed solution, the mixed solution is de-bubbled and poured into the prepared tubular stent mold. The mold is placed at -20℃ / -80℃, frozen for more than 1 hour, completely frozen, and then transferred to a freeze dryer for overnight freeze drying to obtain a tubular aerogel stent with a porous structure.

[0063] Among them, in 3D printing, SolidWorks can be used to realize the mold design for the tubular bracket. The 3D printing material selected is 8200pro resin material.

[0064] The mass volume ratio of PLCL powder to 1,4-dioxane solution is 1.5:100.

[0065] The mold of the obtained tubular support is as follows Figure 2 shown.

[0066] The prepared tubular aerogel scaffold with a porous structure is a super-hydrophobic material. In order to facilitate cell adhesion and growth, the prepared tubular aerogel scaffold is placed in a vacuum plasma pump and treated with a power of 120W for 1 minute to complete the surface modification of the tubular aerogel scaffold. After the modification, the tubular aerogel scaffold is transformed into a super-hydrophilic material, which is beneficial to cell adhesion and culture medium penetration (such as Figure 3 shown).

[0067] (2) Construction of liver cancer model:

[0068] Take 50 μL of 5×10 7 / mL of human hepatoma cell line Hep3B and 30μL of 5×10 7 / mL of human umbilical vein endothelial cells (HUVEC) were planted on the outer wall surface and inner wall surface of the tubular aerogel scaffold obtained in step (1), first placed in a cell culture incubator at 37°C for overnight culture, and then connected to a peristaltic pump for perfusion culture. The culture method adopted was perfusion culture. Under the conditions of a 37°C, 5% CO2 cell culture incubator, DMEM culture medium: ECM culture medium = 1:1 (volume ratio) was cultured for 5 days to form dense tumor tissue. Among them, based on the differences in the planted cells, a layer of endothelial cell barrier will be formed on the inner wall of the tubular aerogel scaffold, while the outer wall will be tightly surrounded by liver cancer cells, thereby forming a structure for vascular invasion and obtaining an aerogel vascular tumor model.

[0069] The tubular aerogel scaffold with a porous structure in step (1) is as follows Figure 4 shown.

[0070] Example 2

[0071] This embodiment provides a method for constructing an aerogel vascular tumor model, and the process is the same as that of Example 1, except that the mass volume ratio of PLCL powder to 1,4-dioxane solution is 2:1.

[0072] Example 3

[0073] This embodiment provides a method for constructing an aerogel vascular tumor model, and the process is the same as that of Example 1, except that the mass volume ratio of PLCL powder to 1,4-dioxane solution is 2.5:100.

[0074] Example 4

[0075] This embodiment provides a method for constructing an aerogel vascular tumor model, and the process is the same as that of Example 1, except that the mass volume ratio of PLCL powder to 1,4-dioxane solution is 3:100.

[0076] Example 5

[0077] At an accelerating voltage of 10 kV, a scanning electron microscope (SU8220, Hitachi, Japan) was used to characterize the pore sizes of the surface and cross section of the aerogel vascular tumor model prepared in Examples 1-4.

[0078] The results are as follows Figure 5 shown.

[0079] It can be found that the aerogel vascular tumor models prepared in Examples 1-4 all have a porous structure of uniform size, which is conducive to the connection and communication between cells. In addition, as the PLCL content increases, the pore size of the tubular aerogel scaffold with a porous structure gradually decreases.

[0080] Example 6

[0081] A tubular stent was prepared using a mold having a length of 25 mm, a width of 10 mm, and a thickness of 3 mm according to the method of Examples 1-4, and then a tensile test (such as Figure 6 The tensile test was performed using a universal testing system (Instron 68TM-5, USA) with a travel speed of 0.5 mm min -1 .

[0082] The Young's modulus and tensile modulus of the tubular scaffold are shown in Table 1.

[0083] Table 1 Young's modulus and tensile modulus of tubular scaffolds

[0084] project Example 1 Example 2 Example 3 Example 4 Pore ​​size (μm) 77.75 68.20 45.31 35.83 Young's modulus(kPa) 0.548 0.889 1.564 1.880 Tensile modulus (%) 118.98 250.82 189.44 183.78

[0085] It can be found that as the PLCL content increases, the Young's modulus of the material also increases, and the aerogel material of Example 2 has the best ductility.

[0086] Example 7

[0087] In this example, when the preparation is carried out according to the method in Example 1-4, in step (1), after obtaining a mixed solution of PLCL and 1,4-dioxane, Cy5 with a final concentration of 10 μg / mL is added, and after completing the subsequent steps, a tubular aerogel scaffold with Cy5 fluorescence can be obtained. 7 HUVEC cell suspension and 5×10 7 After mixing the Hep3B cell suspension of 100 μL / mL in a ratio of 1:1, 50 μL of the mixed cell suspension was applied to the outer surface of the tubular aerogel scaffold, and 30 μL of the mixed cell suspension was applied to the inner surface of the tubular aerogel scaffold, and then placed in a cell culture incubator for 40 minutes. During this period, the culture medium (DMEM culture medium: ECM culture medium = 1:1) was continuously removed and added to maintain cell viability. Finally, a sufficient amount of culture medium was added to culture it overnight. Among them, the culture was perfusion culture, and the perfusion culture device used was as follows Figure 7 shown.

[0088] After the model was constructed, the model was fixed with 4% paraformaldehyde for 2-24 hours, then placed in OCT compound (Sakura Finetek Japan Co., Ltd, Tokyo, Japan), and frozen to obtain 100 μm sections. Cell infiltration was observed under a confocal microscope (Nikon, A1R, Japan).

[0089] The results are as follows Figure 8 shown.

[0090] It can be found that after one day of perfusion, the models in Example 1 and Example 2 achieved complete cell infiltration, while in Example 3 and Example 4, only part of the cells were infiltrated due to the pore size, and the infiltration was not complete. In addition, combined with the Young's modulus and ductility, among the four embodiments, the scaffold produced in Example 2 is most advantageous for the subsequent construction of the liver cancer model. However, for other embodiments, based on the differences in the cells used and the length of the perfusion time, it can be used for the planting of other cells and model construction.

[0091] Example 8

[0092] The method in the above embodiment is used to treat the stent in Example 2, wherein human liver cancer cell line Hep3B and human umbilical vein endothelial cells HUVEC are respectively planted on the outer surface and inner surface of the tubular stent, and then perfusion culture is performed for five days to form dense tumor tissue, while a layer of endothelial barrier can be formed inside the stent, and its outside is tightly surrounded by liver cancer cells, forming a structure for vascular invasion.

[0093] The specific steps are as follows:

[0094] (1) In order to observe the location and distribution of HUVEC and Hep3B cells, membrane dyes DiO and DiD were diluted at a ratio of 1:1000 and added to the corresponding cell culture medium 2 h before digestion of the two cells to mark the two cells. Then, HUVEC and Hep3B cells were digested with 0.25% trypsin, and the cells were collected in 1.5 mL centrifuge tubes and placed on ice. The control cell density was 5×10 7 / Tube.

[0095] (2) When the scaffold is being seeded with cells, the scaffold is rotated continuously. The specific operation is as follows: While the scaffold is being rotated, 50 μL of Hep3B cell suspension is first added to the outside of the scaffold, and then 30 μL of HUVEC cell suspension is added to the inside. The scaffold that has been seeded with cells is placed in a cell culture incubator at 37°C for 40 minutes, during which the culture medium is continuously removed and replenished.

[0096] (3) After culturing overnight in a cell culture incubator, the scaffolds were perfused using a peristaltic pump, wherein the culture medium was a mixed culture medium of DMEM and ECM (V:V = 1:1).

[0097] (4) After five days of perfusion culture, the scaffolds were removed, and the anterior, middle, and posterior segments were frozen and observed using a confocal microscope.

[0098] The results are as follows Fig. 9 shown.

[0099] It can be found that the three slice images of the front, middle and back sections show the uniformity of cell distribution inside the scaffold. HUVEC cells form an endothelial-like structure inside the scaffold, while Hep3B cells tightly surround the outside of the endothelial layer, together forming a structure of vascular invasion.

[0100] VE-Cadherin and CD31 are markers for angiogenesis and tight junctions of endothelial cells. By further immunostaining the sections with VE-Cadherin and CD31, it can be found that endothelial cells can form tight cell junctions on the inner wall of the stent, forming a vascular barrier and promoting the formation of microvessels ( Fig.10 ).

[0101] In order to further verify the effectiveness of the scaffold in modeling, the inventors also tested the situation when only Hep3B cells were used for planting according to the above method. It was found that when only Hep3B cells were planted on the scaffold and perfused, cells could only be found on the outside of the scaffold, while no cells were detected inside the scaffold ( Fig.11 ). This indicates that when Hep3B cells exist alone, the cells can only grow outside the scaffold but cannot penetrate into the scaffold.

[0102] Ki67 is a positive marker for cell proliferation. The inventors designed three different culture conditions (HUVEC cells and Hep3B cells coexist on the scaffolds in perfusion culture; only Hep3B cells coexist on the scaffolds in perfusion culture; and HUVEC cells and Hep3B cells coexist on the scaffolds in static culture), and then stained the scaffolds.

[0103] The results are as follows Fig.12 shown.

[0104] Under perfusion culture conditions, when Hep3B and HUVEC were cultured on the scaffold at the same time, the cells at the edge and inside of the scaffold showed strong cell proliferation activity. When only Hep3B cells were present, the cells at the edge of the scaffold also continued to proliferate. However, when Hep3B and HUVEC were statically co-cultured, the cells could not enter the scaffold and the cell proliferation activity was low. This shows that perfusion culture can continuously supply nutrients to the cells on the scaffold and remove metabolic waste to promote cell proliferation, while static culture limits the metabolic activity and material exchange between cells.

[0105] Example 9

[0106] Sorafenib and lenvatinib are currently the most effective monotherapy for patients with advanced hepatocellular carcinoma, and siRNA-based lipid nanoparticles (LNPs) are emerging drugs that show broad application prospects in the field of cancer treatment. Therefore, by comparing the effects of sorafenib, lenvatinib and siRNA-encapsulated LNPs on the model, it can be used to verify the model's ability to predict the effectiveness of drug treatment.

[0107] Polo-like kinase 1 (PLK1) protein is highly expressed in a variety of cancers and plays a key role in cell cycle control and DNA damage response. It is one of the molecules necessary for the rapid proliferation of tumor cells. In addition, PLK1 is also significantly overexpressed in patients with hepatocellular carcinoma with a poor prognosis. Therefore, LNP-encapsulated siRNA-PLK1 was compared with sorafenib and lenvatinib to verify the ability of this model in drug screening.

[0108] The sense strand of siRNA-PLK1 is 5'-UGAAGAAGAUCACCCUCCUUAdTdT-3' (SEQ ID NO: 1), and the antisense strand is 5'-UAAGGAGGGUGAUCUUCUUCAdTdT-3' (SEQ ID NO: 2), wherein d represents a deoxyribonucleotide.

[0109] According to the operation of the above embodiment, the model after five days of perfusion culture was co-cultured with 2.5 μg / mL sorafenib, 2.5 μg / mL lenvatinib and 150 nM LNP-PLK1 for 48 hours, and the cells in the model were subjected to live / dead staining analysis.

[0110] The results are as follows Fig.13 shown.

[0111] It can be found that cells in the three experimental groups all underwent significant cell death (red fluorescence).

[0112] Since these drugs mainly inhibit disease progression by inhibiting cell proliferation, the inventors further conducted comparative tests on the changes in cell proliferation after drug treatment, wherein Ki67 immunofluorescence staining in the above examples was used to characterize the changes in cell proliferation.

[0113] The results are as follows Fig.14 shown.

[0114] The results showed that all three drugs had a certain inhibitory effect, and sorafenib and lenvatinib had a more significant inhibitory effect. The inventors believe that this difference may be due to a variety of factors, such as sorafenib and lenvatinib as multi-target kinase inhibitors, which can directly block multiple cancer-related signaling pathways, and lipid nanoparticles have low stability.

[0115] The prior art (El Dika, I, et al. "An Open-Label, Multicenter, Phase I, DoseEscalation Study with Phase II Expansion Cohort to Determine the Safety, Pharmacokinetics, and Preliminary Antitumor Activity of Intravenous TKM-080301 in Subjects with Advanced Hepatocellular Carcinoma." The Oncologist 24.6 (2019): theoncologist. 2018-0838-.) confirmed that si-PLK1 has a significant inhibitory effect on cell proliferation in two-dimensional cultured cells and in mouse subcutaneous tumor models, but has limited anti-tumor effects in clinical human trials and has no significant tumor inhibitory effect, thereby confirming the accuracy of the above conclusions. It can be shown that compared with the traditional mouse model, the model in the present invention can be used as a more effective, more accurate, and more convenient drug evaluation platform that is closer to clinical results, for screening and evaluation of different types of drugs, and has a high application potential.

[0116] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. An aerogel scaffold, characterized in that: The raw materials for preparing the aerogel scaffold include lactide-caprolactone copolymer and 1,4-dioxane, wherein the mass volume ratio of the lactide-caprolactone copolymer to 1,4-dioxane is 1-3:100, preferably 1.5-3:

100.

2. The aerogel scaffold according to claim 1, characterized in that: The aerogel scaffold is hydrophilically modified.

3. The method for preparing the aerogel scaffold according to claim 1 or 2, comprising the following steps: A mixed solution of lactide-caprolactone copolymer and 1,4-dioxane is used to cast a mold, and the mold is obtained after freeze-drying.

4. The preparation method according to claim 3, characterized in that: The freezing temperature is between -20 and -80°C, and the freezing time is at least 1 hour.

5. Use of the aerogel scaffold according to claim 1 or 2 in preparing a medical simulation model; preferably, the medical simulation model is a blood vessel model.

6. A vascular tumor simulation model, characterized in that: The vascular tumor simulation model comprises the aerogel scaffold according to claim 1 or 2, endothelial cells attached to the inner surface of the aerogel scaffold, and tumor cells attached to the outer surface of the aerogel scaffold.

7. A method for constructing a vascular tumor simulation model, comprising the following steps: The endothelial cells and the tumor cells are respectively planted on the inner surface and the outer surface of the aerogel scaffold according to claim 1 or 2, After the cells are fixed on the aerogel scaffold, perfusion culture is performed, and a vascular tumor simulation model is obtained after 1-6 days of culture; Preferably, the aerogel scaffold is continuously rotated during planting.

8. The construction method according to claim 7, characterized in that: The cell concentration of endothelial cells and tumor cells used for seeding was 3-5×10 7 / mL.

9. Use of the vascular tumor simulation model described in claim 6 or the vascular tumor simulation model obtained by the construction method of any one of claims 7 to 8 in an in vitro disease simulation model, preferably, the disease includes a tumor.

10. Use of the vascular tumor simulation model described in claim 6 or the vascular tumor simulation model obtained by the construction method according to any one of claims 7 to 8 in constructing an in vitro drug screening platform.

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