A cell-derived nanocarrier capable of enhancing the effect of photodynamic therapy and its application

Abstract

The present invention provides a cell-derived nanocarrier capable of enhancing the efficacy of photodynamic therapy and its application. The cell-derived nanocarrier is prepared by a method comprising the following steps: culturing and collecting cells, ultrasonically disrupting them, and homogenizing the particles using a liposome extruder. The cell-derived nanocarrier provided by the present invention is simple to obtain, has high yield, and is amenable to mass production; it is targeted to tumor sites, has good drug delivery, and does not involve the use of other exogenous synthetic compounds, exhibits low immunogenicity, and is highly safe.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine and relates to a cell-derived nanocarrier capable of enhancing the effect of photodynamic therapy and an application thereof. Background Art

[0002] Malignant tumors are still major diseases that seriously endanger human health and life safety. Cancer prevention and treatment are major health and scientific issues facing the world. Research on precise targeted treatment of tumors is particularly important and has always been an important research direction in the medical field at home and abroad.

[0003] Photodynamic therapy (PDT) uses light of a specific wavelength to irradiate the target area, which uses photosensitizers and oxygen molecules to generate reactive oxygen species (ROS), especially singlet oxygen ( 1 O2) induces cell apoptosis and necrosis to achieve the purpose of inhibiting tumor growth. As a new type of tumor treatment, PDT has attracted widespread attention due to its high selectivity, minimally invasive, non-invasive and repeatable treatment, and can be used to treat various types of solid tumors. Compared with surgery, chemotherapy and radiotherapy, the advantages of PDT are that it is minimally invasive, non-invasive, and can be completed locally with less damage to healthy tissue. In addition, PDT can improve the survival of patients, has lower treatment costs, usually does not require follow-up, and can be repeated in the case of recurrence, which has important practical application value. Therefore, the research on tumor photodynamic therapy has important theoretical and practical application value, and the research on targeted drugs, namely photosensitizers (photodynamic therapy drugs), is the key to affecting the prospects of photodynamic therapy. As the core substance of PDT, the important characteristic of photosensitizers is that they can be targeted and aggregated in tumor tissues and produce specific biological effects, while having little or no effect on surrounding normal tissues.

[0004] Currently, photodynamic therapy for cancer still has shortcomings, mainly in the following aspects: (1) Commonly used photosensitizers lack targeting to tumor tissues, making it difficult to enrich and reach effective concentrations, which affects the therapeutic effect; (2) Most photosensitizers are hydrophobic molecules that easily aggregate and are difficult to deliver to the tumor site in vivo. These shortcomings greatly limit the clinical application of photodynamic therapy.

[0005] In recent years, the rapid development of drug delivery systems has provided new ideas and methods for solving the above-mentioned problems. Exosomes (EXOs) are vesicles secreted by cells in the body, and their size (40-100 nm) is similar to that of nanomaterials. As a naturally derived drug carrier, EXOs have the advantages of low toxicity, non-immunogenicity, and good permeability, and therefore have great application prospects in the field of drug delivery. However, in actual applications, it has been found that the extraction process of EXOs is complex and the yield is low, which greatly limits the application of EXOs in the field of drug delivery.

[0006] In addition, some literature uses the cell membrane of tumor cells to deliver drugs. However, the nanoparticles obtained by directly wrapping drugs (photosensitizers) with cell membranes not only have poor stability, but also have relatively poor photodynamic therapy effects. Therefore, in order to improve the stability and delivery effect of nanoparticles, researchers generally use cell membranes + chemically synthesized nanoparticles (such as gold nanoparticles) as carriers to encapsulate drugs. However, on the one hand, this method is more complicated to synthesize and the cost is higher. More importantly, due to the use of chemically synthesized substances, the safety of the obtained delivery materials is difficult to guarantee and is prone to immunogenicity.

[0007] Therefore, how to provide a carrier to load photosensitizers to obtain a photodynamic therapy drug system that can simultaneously achieve the following effects: simple and easy to obtain, high yield, and easy to mass produce; have targeted effects on tumor sites; have good photodynamic efficacy; and, in addition to the photosensitizer, do not involve other exogenous synthetic compounds, have low immunogenicity, and have good safety, has become a problem that needs to be urgently solved by those skilled in the art. Summary of the Invention

[0008] In view of the shortcomings of the prior art, the purpose of the present invention is to provide a cell-derived nanocarrier and its application that can enhance the effect of photodynamic therapy.

[0009] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0010] In a first aspect, the present invention provides a cell-derived nanocarrier that can enhance the effect of photodynamic therapy. The cell-derived nanocarrier is prepared by a preparation method comprising the following steps: culturing and collecting cells, ultrasonically disrupting, and performing particle size homogenization treatment using a liposome extruder.

[0011] If the liposome extruder step is not performed, the average particle size of the cell-derived nanocarrier becomes larger and the particle size is uneven, resulting in poor stability. When used to encapsulate drugs, it is not conducive to the cellular uptake of the drugs, thereby affecting the photodynamic therapy effect.

[0012] Preferably, the cells comprise cancer cells.

[0013] Preferably, the ultrasonication is performed using an ultrasonic disruptor.

[0014] Preferably, the ultrasonic power is set to 10-20%, and the ultrasonic time is 30-200s.

[0015] The specific values ​​of the above 10-20% include 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, etc.

[0016] Specific values ​​in the above 30-200s include, for example, 30s, 40s, 50s, 60s, 70s, 80s, 90s, 100s, 120s, 140s, 150s, 160s, 180s, 200s, etc.

[0017] The power and time of ultrasound are conventional settings in the art and will not particularly affect the stability and drug loading effect of the cell-derived nanocarriers.

[0018] Preferably, after the particle size homogenization treatment, the cells are filtered using filter membranes with successively decreasing pore sizes.

[0019] Preferably, the pore sizes of the filter membranes are 0.6 μm, 0.4 μm, 0.2 μm and 0.1 μm, respectively;

[0020] Alternatively, the pore sizes of the filter membrane are 0.4 μm, 0.2 μm and 0.1 μm, respectively.

[0021] In a second aspect, the present invention provides use of the cell-derived nanocarrier capable of enhancing the photodynamic therapy effect as described in the first aspect in the preparation of a drug carrier, wherein the drug includes a photosensitizer.

[0022] In a third aspect, the present invention provides a cell-derived photosensitizer nano-delivery system, which comprises a photosensitizer and a cell-derived nano-carrier capable of enhancing the photodynamic therapy effect as described in the first aspect.

[0023] Preferably, the cell-derived photosensitizer nano-delivery system uses cell-derived nano-carriers that can enhance the effect of photodynamic therapy as photosensitizer carriers.

[0024] In a fourth aspect, the present invention provides a method for preparing the cell-derived photosensitizer nano-delivery system as described in the third aspect, the preparation method comprising: mixing a cell-derived nano-carrier capable of enhancing the photodynamic therapy effect, a photosensitizer and a solvent to obtain the system.

[0025] Preferably, the solvent comprises water and / or an organic solvent.

[0026] Preferably, the organic solvent includes any one of dimethyl sulfoxide, methanol or ethanol, or a combination of at least two thereof.

[0027] Preferably, the volume percentage of the organic solvent in the cell-derived photosensitizer nanodelivery system is 1%-15%, for example, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, etc.

[0028] Preferably, the mixing temperature is 15-40°C, for example, 15°C, 17°C, 20°C, 22°C, 25°C, 27°C, 30°C, 32°C, 35°C, 37°C, 40°C, etc.

[0029] Preferably, the mixing time is 1-3 h, for example, 1 h, 1.2 h, 1.5 h, 1.7 h, 2 h, 2.2 h, 2.5 h, 2.7 h, 3 h, etc.

[0030] The numerical range described in the present invention includes not only the point values ​​listed above, but also any point values ​​between the above numerical ranges that are not listed. Due to space limitations and for the sake of simplicity, the present invention no longer exhaustively lists the specific point values ​​included in the range.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] Currently, existing technologies generally use three strategies to deliver drugs: (1) using exosomes as carriers; (2) directly using cell membranes as carriers; and (3) using cell membranes + chemically synthesized nanoparticles (such as gold nanoparticles) as carriers.

[0033] The present invention has developed a cellular nano-delivery carrier (Cellular Vesicles, CVs) that can simulate exosomes (directly using cell fragments as carriers), and prepared a cellular nano-delivery system ZnPC-CVs (ZnPC is a photosensitizer and can be replaced at will). Compared with the three existing strategies, these have obvious advantages:

[0034] (1) CVs have similar particle size and composition to exosomes (EXOs). CVs can achieve the delivery effect of EXOs. The advantages of CVs are that CVs are produced by continuous extrusion of cells through microporous and nanoporous filters, while EXOs are secreted by cells. Therefore, CVs are significantly superior to exosomes in terms of yield and the difficulty of the extraction process.

[0035] (2) Compared with cell membrane vesicles (CMVs), the delivery system obtained by using CVs as a carrier to encapsulate drugs has better stability and is more easily taken up by cells. In particular, when the drug is a photosensitizer, compared with the free photosensitizer ZnPC and the nanodelivery system ZnPC-CMVs prepared using cell membrane nanocarriers (CMVs), the cell-derived nanodelivery system ZnPC-CV of the present invention has significantly better photodynamic effect. Therefore, the present invention is of great significance to the research on the targeted delivery of a new generation of photosensitizers and the improvement of the photodynamic therapy effect.

[0036] (3) Cell membrane + chemically synthesized nanoparticles (e.g., gold nanoparticles) as carriers to encapsulate drugs. On the one hand, this method is relatively complex to synthesize and has high costs. More importantly, due to the use of chemically synthesized substances, the safety of the resulting delivery material is difficult to guarantee. Therefore, compared with this strategy, the present invention not only has a good delivery effect, but also has significant advantages in terms of simplicity of preparation, cost, and safety.

[0037] In summary, the cell-derived nanocarriers provided by the present invention simultaneously achieve the advantages of simple preparation method, high yield, and easy mass production; they are targeted to the tumor site; have good drug loading effect (especially significantly enhanced photodynamic therapy effect when loaded with photosensitizers); do not involve other exogenous synthetic compounds, have low immunogenicity, and have good safety. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 It is the stability result diagram of CVs under different conditions.

[0039] Figure 2 This is a comparison of the particle sizes of ZnPC-CVs and ZnPC-CMVs.

[0040] Figure 3 This is a graph showing the cellular uptake results of free ZnPC, ZnPC-CVs, and ZnPC-CMVs.

[0041] Figure 4 This is a diagram showing the photodynamic effect of free ZnPC, ZnPC-CVs and ZnPC-CMVs.

[0042] Figure 5 This is the dark toxicity assessment result of ZnPC-CVs. DETAILED DESCRIPTION

[0043] The technical solution of the present invention is further described below by way of specific embodiments. It should be understood by those skilled in the art that the embodiments are merely to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0044] In the following examples, unless otherwise specified, all reagents and consumables were purchased from conventional reagent manufacturers in the field; unless otherwise specified, the experimental methods and technical means used were conventional methods and means in the field.

[0045] Example 1-Preparation of CVs

[0046] This example provides a method for preparing cell fragments as nano-delivery carriers (CVs), taking cholangiocarcinoma cell TFK-1 as an example, as follows:

[0047] Culture TFK-1 cholangiocarcinoma cells in RPMI medium supplemented with 10% fetal bovine serum at 37°C and 5% CO2, changing the medium every other day. When the cells in the T175 flask reach 90%-95% confluency, discard the cell culture medium, wash the cells twice with PBS, digest the cells with Tripsin (trypsin), and transfer the cells to a centrifuge tube. Centrifuge at 300 × g for 5 minutes and wash twice with HEPES buffer.

[0048] The cells were resuspended in 10 mL of HEPES buffer and disrupted by sonication using the following parameters: ultrasonic disruptor power was set to 10%, total sonication duration was set to 2 minutes, and each cycle was set to 10 seconds on and 10 seconds off. After sonication, the cell suspension was homogenized using a liposome extruder. The sonicated cell suspension was then filtered through membranes with pore sizes of 0.6 μm, 0.4 μm, 0.2 μm, and 0.1 μm, in that order. TFK-1 cell-derived CVs were obtained.

[0049] Example 2 - Characterization of CVs

[0050] (1) Particle size potential investigation

[0051] The experiment was repeated three times according to the method of Example 1 to prepare sample solutions CVs1, CVs2, and CVs3, respectively. One mL of the prepared CVs sample solution was diluted 10-fold with deionized water. The particle size and zeta potential were measured using a particle size potential meter. The results are shown in Table 1. The average particle size was 134.3 nm, the surface charge was negative, and the average potential was -16.1 mV.

[0052] Table 1

[0053] Sample name Average particle size (nm) Zeta potential (mV) CVs1 135.1 -16.2 CVs2 134.3 -17 CVs3 133.4 -15.2

[0054] (2) Determination of total protein concentration in CVs

[0055] The protein content of CVs was determined using a MicroBCA kit according to the kit instructions.

[0056] The average total protein concentration of the final CVs was 399.56 μg / mL.

[0057] (3) Determination of total phospholipid content in CVs

[0058] The total phospholipid content of CVs was determined using the classic Rouser Assay method, and the result was 498 μM.

[0059] Example 3-Study on the stability of CVs in different solvents

[0060] Since drug delivery using CVs often requires dissolution of the drug in an organic solvent, to assess the potential interference of different solvents on CV stability, this example evaluated the effects of varying concentrations (1%-30%) of DMSO (dimethyl sulfoxide), ethanol, and methanol on CV particle size stability. CV solutions were mixed with DMSO, anhydrous methanol, and anhydrous ethanol at 1%, 5%, 10%, 15%, 20%, and 30% by volume (the volume percentage of the organic solvent in the total solution), respectively. After shaking at 4°C for 18 hours, the particle size change was measured using a particle size potentiometer.

[0061] The results are as follows Figure 1 As shown in the figure, RT in the figure represents the test result after the CVs solution is not mixed with the organic solvent and shaken at room temperature (25°C) for 18 hours; 4°C represents the test result after the CVs solution is not mixed with the organic solvent and shaken at 4°C for 18 hours; the results show that room temperature or 4°C does not particularly affect the stability of CVs, and the effects of different concentrations of solvents on CVs have different results. In order to minimize the effect of organic solvents on the stability of CVs, the addition amount of each solvent is selected as much as possible: dimethyl sulfoxide (1%-15%), methanol (1%-10%) and ethanol (1%-10%).

[0062] Comparative Example 1-Preparation of CMVs

[0063] This comparative example provides a preparation method using cell membrane as a nano-delivery carrier (CMVs), taking bile duct cancer cell TFK-1 as an example, as follows:

[0064] Prepare CMVs extract: weigh 85.575 g sucrose, 0.37 g EDTA, and 4.766 g Hepes and dissolve them in 1 L deionized water. Adjust the pH to 7.4.

[0065] Culture TFK-1 cholangiocarcinoma cells in RPMI medium supplemented with 10% fetal bovine serum at 37°C and 5% CO2, changing the medium every other day. When the cells in the T175 flask reach 90%-95% confluency, discard the cell culture medium, wash the cells twice with PBS, digest the cells with Tripsin (trypsin), and transfer the cells to a centrifuge tube. Centrifuge at 300 × g for 5 minutes and wash twice with HEPES buffer.

[0066] After washing with HEPES buffer, add 5 mL of the prepared CMVs extract to the cells. Shake at 4°C for 30 minutes. Set the ultrasonic disruptor power to 10% and the total sonication time to 2 minutes, with each cycle consisting of 10 seconds on and 10 seconds off. After sonication, centrifuge at 4000 rpm for 10 minutes. Transfer the supernatant to a new centrifuge tube and centrifuge at 15000 rpm for 20 minutes. Wash twice with HEPES buffer. The cell membranes are precipitated at the bottom, representing the cell membranes. Finally, add 1 mL of HEPES buffer to the cell membranes, pipet and mix thoroughly. Filter the cell membrane suspension through filters with pore sizes of 0.6 μm, 0.4 μm, 0.2 μm, and 0.1 μm, in that order. This yields TFK-1 cell-derived CMVs.

[0067] Application Example 1

[0068] The CVs of Example 1 were used to load a photosensitizer (taking zinc phthalocyanine ZnPC as an example) to obtain a cell-derived photosensitizer nano-delivery system ZnPC-CVs.

[0069] The CMVs of Comparative Example 1 were loaded with photosensitizer to obtain ZnPC-CMVs for comparison.

[0070] (1) The preparation method is as follows:

[0071] 300 μg / mL CVs solution (or CMVs solution) was mixed with 109 μM ZnPC / DMSO solution at a volume ratio of 9:1, sonicated on ice for 3 min, and incubated with shaking at room temperature for 2 h to obtain ZnPC-CVs (or ZnPC-CMVs).

[0072] (2) Particle size characterization:

[0073] The particle size was measured using a particle size potential meter. Figure 2 shown.

[0074] The results showed that the particle size of ZnPC-CVs increased slightly after drug loading, but was still around 200 nm, which is still a relatively ideal particle size range. However, the particle size of ZnPC-CMVs increased significantly compared to before drug loading, approaching 300 nm, so it would be less conducive to cell uptake than ZnPC-CVs.

[0075] (3) Cellular uptake effect of ZnPC-CVs in TFK-1

[0076] The uptake of ZnPC in cells was observed using a laser confocal scanning microscope. 100 μL of TFK-1 cells were seeded in a 96-well plate (flat bottom transparent, black polystyrene) at a density of 1.5×10 4 / well. The cells were incubated overnight until 60-70% confluence. The cells were then incubated with free ZnPC, ZnPC-CVs and ZnPC-CMVs in RPMI for 15 minutes, 30 minutes and 60 minutes, respectively. The final ZnPC concentration in each group was 3 μM. After incubation, the cells were washed once with room temperature PBS at a volume of 100 μL / well, and then fixed with 2% paraformaldehyde in the dark for 15 minutes, washed once with PBS, and then 100 μL of diluted Hoechst33342 (a fluorescent dye, 10 μg / mL, dissolved in PBS) was added to each well and incubated at room temperature for 3 minutes. After aspirating the Hoechst33342, 100 μL of room temperature PBS was added to each well and photographed. The imaging parameters are as follows: Hoechst (λex = 405 nm; λem = 479 nm), ZnPC-CVs (λex = 660 nm; λem = 790 nm), ZnPC-CMVs (λex = 660 nm; λem = 790 nm). A 40× objective lens was used. The results are shown in Figure 2. Figure 3 As shown in the figure, ZnPC-CVs have better cellular uptake than free ZnPC at the 60-min time point, demonstrating the good delivery performance of CVs. However, ZnPC-CMVs still cannot be effectively taken up by cells at 60 minutes, indicating that the delivery effect of CMVs is poor.

[0077] (4) Photodynamic effect of ZnPC-CVs

[0078] TFK-1 cells 1.5×10 4 Cells were seeded into 96-well plates at 100 μL per well. After overnight plating, free ZnPC, ZnPC-CVs, and ZnPC-CMVs were added at final ZnPC concentrations of 0.25 μM, 0.5 μM, 1.0 μM, and 1.5 μM. Three replicate wells were set up for each group. After incubation for 6 hours, the cells were washed twice with PBS and 100 μL of RPMI was added to each well. 671 nm diode laser (CNI) was used for irradiation at 500 mW for 25 s per well, so that all cells received 15 J / cm 2 After irradiation, the cells were returned to the incubator and assayed for cell proliferation using the CellTiter 96 Aqueous One Solution (Promega) kit overnight.

[0079] The results are as follows Figure 4As shown in the figure, the IC50 values ​​of free ZnPC and ZnPC-CVs are 0.33μM and 0.24μM, respectively. It can be seen that the photodynamic effect of ZnPC-CVs is significantly better than that of free ZnPC. However, ZnPC-CMVs can only kill about 50% of cells even at the maximum concentration of 1.5μM, indicating a poor photodynamic effect. This is consistent with the result of poor intracellular uptake of ZnPC-CMVs confirmed in step (3).

[0080] (5) Safety of ZnPC-CVs (dark toxicity)

[0081] TFK-1 cells were cultured at 1.5×10 4 Cells were seeded in a 96-well plate at a volume of 100 μL per well. After overnight plating, ZnPC-CVs were added at final concentrations of 0.25 μM, 0.5 μM, 1.0 μM, and 1.5 μM, respectively. Three replicates were set up for each group. A blank control (cells not treated with other methods) and a positive control (1% Triton treatment) were also set up. After incubation for 24, 48, and 72 hours, cell proliferation was detected using the CellTiter 96Aqueous One Solution (Promega) kit. The results are shown in Figure 2. Figure 5 As shown in the results, ZnPC-CVs showed no cytotoxicity from 24 to 72 hours after administration, confirming that ZnPC-CVs are non-toxic and safe to cells in the unstimulated state.

[0082] In summary, the present invention has developed a cell-derived nanocarrier (CV) that overcomes the shortcomings of commonly used carriers (exosomes, cell membranes, and cell membranes combined with compound nanoparticles) in existing technologies. Using this carrier, a cell-derived nanocarrier (ZnPC-CV) was developed to enhance the photodynamic effect and effectively deliver the photosensitizer ZnPC. This delivery system has also been shown to exhibit excellent delivery efficacy, photodynamic effect, and safety.

[0083] The applicant states that while the present invention uses the aforementioned embodiments to illustrate a cell-derived nanocarrier capable of enhancing the efficacy of photodynamic therapy and its applications, the present invention is not limited to these embodiments, nor does it necessarily rely on these embodiments for implementation. Persons skilled in the art will appreciate that any improvements to the present invention, equivalent substitutions for various raw materials in the present invention's products, addition of auxiliary components, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present invention.

[0084] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0085] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A cell-derived nanocarrier capable of enhancing the effect of photodynamic therapy, characterized in that: The cell-derived nanocarrier is prepared by the following preparation method: culturing and collecting cells, ultrasonically disrupting, and performing particle size homogenization treatment using a liposome extruder to obtain the obtained nanocarrier; After the particle size homogenization treatment, the method further includes filtering the cells with filter membranes of decreasing pore sizes; The pore sizes of the filter membranes are 0.6 μm, 0.4 μm, 0.2 μm and 0.1 μm, respectively; Alternatively, the pore sizes of the filter membrane are 0.4 μm, 0.2 μm, and 0.1 μm, respectively; The cell-derived nanocarrier does not involve other exogenous synthetic compounds; The cells include cancer cells.

2. The cell-derived nanocarrier capable of enhancing the effect of photodynamic therapy according to claim 1, wherein: The ultrasonication is performed using an ultrasonic disruptor.

3. The cell-derived nanocarrier capable of enhancing the effect of photodynamic therapy according to claim 2, wherein: The ultrasonic power is set to 10-20%, and the ultrasonic time is 30-200 s.

4. Use of the cell-derived nanocarrier capable of enhancing the photodynamic therapy effect according to any one of claims 1 to 3 in the preparation of a drug carrier, characterized in that: The drug includes a photosensitizer.

5. A cell-derived photosensitizer nano-delivery system, characterized in that: The cell-derived photosensitizer nano-delivery system comprises a photosensitizer and a cell-derived nano-carrier capable of enhancing the photodynamic therapy effect according to any one of claims 1 to 3.

6. The method for preparing the cell-derived photosensitizer nano-delivery system according to claim 5, wherein: The preparation method comprises: mixing a cell-derived nanocarrier capable of enhancing the photodynamic therapy effect, a photosensitizer and a solvent to obtain the nanocarrier.

7. The method for preparing the cell-derived photosensitizer nano-delivery system according to claim 6, wherein: The solvent includes water and / or an organic solvent.

8. The method for preparing the cell-derived photosensitizer nano-delivery system according to claim 7, wherein: The organic solvent includes any one of dimethyl sulfoxide, methanol or ethanol, or a combination of at least two of them.

9. The method for preparing the cell-derived photosensitizer nano-delivery system according to claim 8, wherein: The volume percentage of the organic solvent in the cell-derived photosensitizer nano-delivery system is 1%-15%.

10. The method for preparing the cell-derived photosensitizer nano-delivery system according to claim 6, wherein: The mixing temperature is 15-40°C; The mixing time is 1-3 h.

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