Biological fluorescent intelligent film as well as preparation method and application thereof

Abstract

The invention provides a biological fluorescent intelligent film as well as a preparation method and application thereof. The preparation method comprises the following steps: (1) adding an isocyanate monomer and an amino compound into a first solvent, and stirring for reaction; (2) adding a biological fluorescent agent and a hydroxyl compound into the reaction system, and stirring for reaction to obtain a polyurea reactant; wherein the biological fluorescent agent is obtained through addition polymerization reaction by taking an isocyanate monomer and an amino compound as reaction raw materials; and (3) taking the polyurea reactant as a printing line, and carrying out printing film formation in a 4D printing mode to obtain the biological fluorescent intelligent film. According to the preparation method, the biological fluorescent agent is self-made, and the biological fluorescent agent and the hydroxyl compound are introduced into the polyurea material for polymerization reaction, so that the biological fluorescent polyurea intelligent membrane has excellent biological fluorescent performance, and the biological fluorescent intelligent membrane has no toxic or side effect on cells, has good biocompatibility, and can be used for preparing the biological fluorescent polyurea intelligent membrane. Meanwhile, the shape memory performance is good, and intelligent regulation and control can be achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of functional materials, and in particular to a bioluminescent intelligent film and a preparation method and application thereof. Background Art

[0002] With the development of science and technology, the demand for materials with special functions is increasing. In biomedical research, materials that can effectively label and detect biological molecules and cells are needed; in the field of environmental science, there is also an urgent need for sensor materials that can detect pollutants in real time and sensitively.

[0003] Traditional detection methods and materials often have some limitations, such as poor labeling specificity, low detection sensitivity, insufficient sensor stability, etc. At the same time, the development of smart materials provides new ideas for solving these problems. Polyurea materials have attracted attention due to their good mechanical properties and chemical stability. However, the existing polyurea materials have poor biocompatibility, and their comprehensive performance in bioluminescence and smart response needs to be improved.

[0004] Based on this, there is an urgent need to provide a bioluminescent smart membrane and a preparation method and application thereof. Summary of the invention

[0005] The embodiment of the present invention provides a bioluminescent smart membrane and a preparation method and application thereof, which can solve the problems of poor biocompatibility of traditional polyurea materials and poor fluorescence stability during the application process.

[0006] In a first aspect, the present invention provides a method for preparing a bioluminescent smart film, the preparation method comprising the following steps:

[0007] (1) adding an isocyanate monomer and an amino compound into a first solvent and stirring for reaction;

[0008] (2) adding a bioluminescent agent and a hydroxyl compound to a reaction system, stirring and reacting to obtain a polyurea reactant; wherein the bioluminescent agent is obtained by an addition polymerization reaction using an isocyanate monomer and an amino compound as reaction raw materials;

[0009] (3) Using the polyurea reactant as a printing solution to form a film using a 4D printing method, thereby obtaining the bioluminescent smart film.

[0010] Preferably, in step (2), the bioluminescent agent is prepared by the following method: adding the isocyanate monomer and the amino compound into a second solvent for mixed reaction to obtain the bioluminescent agent; wherein the second solvent is an acetone aqueous solution or a phosphate buffered saline solution.

[0011] Preferably, the content of the isocyanate monomer is 5-20 mmol, the content of the amino compound is 5-20 mmol, and the content of the second solvent is 85-95%.

[0012] Preferably, the reaction temperature is 28-32°C and the reaction time is 3-5h.

[0013] Preferably, the isocyanate monomer is toluene diisocyanate or isophorone diisocyanate; the amino compound is polyetheramine D-400, polyetheramine D-2000, diethyltoluenediamine or tetramethylethylenediamine.

[0014] Preferably, in step (1), the molar ratio of the isocyanate monomer to the amino compound is 10:(3-7).

[0015] More preferably, the first solvent is N,N'-dimethylformamide, N,N'-dimethylacetamide or dichloromethane, and the content of the first solvent is 10-30 mL.

[0016] Preferably, in step (2), the hydroxy compound is polycaprolactone diol-2000 or polycaprolactone diol-4000, and the molar ratio of the total amount of the hydroxy compound and the amino compound to the isocyanate monomer is 1:1.

[0017] Preferably, in the reaction system, the content of the bioluminescent agent is 20-50 mg / mL.

[0018] Preferably, in step (1), the reaction temperature is 20-30°C and the reaction time is 3-5h.

[0019] Preferably, in step (2), the reaction temperature is 50-60°C and the reaction time is 20-27h.

[0020] Preferably, in step (3), the printing temperature is 80-120°C.

[0021] In a second aspect, the present invention further provides a bioluminescent smart membrane, which is prepared by using any preparation method described in the first aspect above.

[0022] In a third aspect, the present invention further provides an application of the bioluminescent smart film described in the second aspect in biomedical materials.

[0023] Compared with the prior art, the present invention has at least the following beneficial effects:

[0024] In the present invention, a small molecular weight bioluminescent agent is first prepared using an isocyanate monomer and an amino compound as reaction raw materials, and then the bioluminescent agent and a hydroxyl compound are introduced into a polymerization reaction during the synthesis of a polyurea material to form a polyurea reactant, and finally the polyurea reactant is printed into a film using 4D printing to obtain a bioluminescent smart film; in the present invention, a small molecular weight bioluminescent agent is prepared by self-preparing the bioluminescent agent and the bioluminescent agent and a hydroxyl compound are introduced into a polyurea material for polymerization reaction, so that the bioluminescent polyurea smart film of the present invention has excellent bioluminescent properties, can efficiently emit fluorescence when excited by light of a specific wavelength, and the bioluminescent smart film of the present invention has no toxic side effects on cells, has good biocompatibility, and has good shape memory properties, and can achieve intelligent regulation. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0026] Figure 1 This is a luminescence image of a bioluminescent agent particle provided by an embodiment of the present invention;

[0027] Figure 2 is a particle size distribution diagram of the bioluminescent agent particles provided in Example 1 of the present invention;

[0028] Figure 3 This is the ultraviolet absorption spectrum of the bioluminescent agent particles provided in Example 1 of the present invention. DETAILED DESCRIPTION

[0029] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0030] The embodiment of the present invention provides a method for preparing a bioluminescent smart film, the method comprising the following steps:

[0031] (1) adding an isocyanate monomer and an amino compound into a first solvent and stirring for reaction;

[0032] (2) adding a bioluminescent agent and a hydroxyl compound to a reaction system, stirring and reacting to obtain a polyurea reactant; wherein the bioluminescent agent is obtained by an addition polymerization reaction using an isocyanate monomer and an amino compound as reaction raw materials;

[0033] (3) Using the polyurea reactant as a printing line to form a film using a 4D printing method, thereby obtaining the bioluminescent smart film.

[0034] In the embodiment of the present invention, firstly, an isocyanate monomer and an amino compound are used as reaction raw materials to prepare a bioluminescent agent with a smaller particle size, and then the bioluminescent agent and a hydroxyl compound are introduced into a polymerization reaction during the synthesis of a polyurea material to form a polyurea reactant, and finally the polyurea reactant is printed into a film using 4D printing to obtain a bioluminescent smart film; in the present invention, a bioluminescent agent with a small molecular weight is prepared by itself, and the bioluminescent agent and the hydroxyl compound are introduced into a polyurea material for polymerization reaction, so that the bioluminescent polyurea smart film of the present invention has excellent bioluminescent properties, and can efficiently emit fluorescence when excited by light of a specific wavelength, and the bioluminescent smart film of the present invention has no toxic side effects on cells, has good biocompatibility, and has good shape memory properties, and can achieve intelligent regulation.

[0035] According to some preferred embodiments, in step (2), the bioluminescent agent is prepared by the following method: adding the isocyanate monomer and the amino compound into a second solvent for mixed reaction to obtain the bioluminescent agent; wherein the second solvent is an acetone aqueous solution or a phosphate buffered saline solution.

[0036] In the embodiment of the present invention, an addition polymerization reaction is carried out in a second solvent system using isocyanate and an amino compound as raw materials to form a bioluminescent agent with a high fluorescence quantum yield and excellent fluorescence performance. The particle size of the bioluminescent agent particles is small (100-300nm), so that the particle size of the prepared solution is smaller, the bioluminescent agent can be evenly dispersed in the bioluminescent smart film, and the bioluminescent agent is very sensitive to changes in the polarity of the environment, and can show different fluorescence intensities and wavelengths in different environments, and no quenching occurs during the application process.

[0037] According to some preferred embodiments, the content of the isocyanate monomer is 5-20 mmol (for example, it can be 5 mmol, 8 mmol, 10 mmol, 12 mmol, 15 mmol, 18 mmol or 20 mmol), the content of the amino compound is 5-20 mmol (for example, it can be 5 mmol, 8 mmol, 10 mmol, 12 mmol, 15 mmol, 18 mmol or 20 mmol), and the content of the second solvent is 85-95% (for example, it can be 85%, 88%, 90%, 92% or 95%).

[0038] According to some preferred embodiments, the reaction temperature is 28-32°C (for example, 28°C, 29°C, 30°C, 31°C or 32°C), and the reaction time is 3-5h (for example, 3h, 4h or 5h).

[0039] In the embodiment of the present invention, during the reaction process, the type, content and reaction conditions of the isocyanate monomer, the amino compound and the second solvent are reasonably controlled, so as to prepare a bioluminescent agent with good solubility, good biocompatibility and excellent bioluminescent performance, and thus to prepare a bioluminescent smart film with stable fluorescence performance. The molar ratio of the isocyanate monomer to the amino compound is preferably 1:1. For example, if the content of the isocyanate monomer is too low, some amino groups may fail to react fully, forming incomplete intermediates or by-products, which will eventually have an adverse effect on the purity and fluorescence performance of the bioluminescent agent; and if the content of the isocyanate monomer is too high, more cross-linked structures will be formed, thereby affecting the molecular weight distribution and solubility of the bioluminescent agent.

[0040] It should be noted that, in the embodiment of the present invention, the second solvent is preferably an acetone aqueous solution, wherein the mass ratio of acetone to water is 3:7.

[0041] According to some preferred embodiments, the isocyanate monomer is toluene diisocyanate or isophorone diisocyanate; the amino compound is polyetheramine D-400, polyetheramine D-2000, diethyltoluenediamine or tetramethylethylenediamine.

[0042] In the embodiment of the present invention, by using the above-mentioned types of isocyanate monomers and amino compounds, toluene diisocyanate and isophorone diisocyanate can form a stable ring structure with diethyltoluenediamine or tetramethylethylenediamine, which not only improves the stability of the molecule but also helps to ensure the better fluorescence performance of the bioluminescent agent.

[0043] At the same time, in an embodiment of the present invention, the polyurea material is prepared with the above-mentioned isocyanate monomers and amino compounds, and the synthetic raw materials of the bioluminescent agent are preferably the same as the raw materials for synthesizing the polyurea matrix material, so that the bioluminescent agent can be fully dissolved and evenly dispersed in the polyurea material, which is beneficial to ensure the stable and uniform fluorescence performance of the bio-intelligent membrane; further, the above-mentioned isocyanate monomers have high reactivity and can quickly undergo addition reactions with amino compounds to form stable polyurea segments. At the same time, compared with other types of isocyanates, toluene diisocyanate and isophorone diisocyanate have lower toxicity under appropriate reaction conditions, and by selecting the above-mentioned amino compounds to react with them, the distribution and metabolic behavior of the polymer reactants in the organism can be optimized, so that the bio-intelligent membrane has excellent biocompatibility.

[0044] According to some preferred embodiments, in step (1), the molar ratio of the isocyanate monomer to the amino compound is 10:(3-7) (for example, it can be 10:3, 10:4, 10:5, 10:6 or 10:7); the first solvent is N,N'-dimethylformamide, N,N'-dimethylacetamide or dichloromethane, and the content of the first solvent is 10-30 mL (for example, it can be 10 mL, 20 mL or 30 mL).

[0045] In the embodiment of the present invention, in the process of synthesizing the polyurea reactant using isocyanate monomer and amino compound as raw materials in the above-mentioned first solvent, by reasonably controlling the ratio of isocyanate monomer and amino compound, it is beneficial to prepare a polyurea reactant with suitable crosslinking degree and high purity. If the content of isocyanate monomer is too high or too low, unreacted reactants will exist in the reaction system, thereby affecting the molecular weight of the polyurea reactant.

[0046] According to some preferred embodiments, in step (2), the hydroxy compound is polycaprolactone diol-2000 or polycaprolactone diol-4000; and the molar ratio of the total amount of the hydroxy compound and the amino compound to the isocyanate monomer is 1:1.

[0047] In the embodiment of the present invention, during the reaction of isocyanate monomer and amino compound, the isocyanate group compound containing benzene ring structure can be used as the hard segment of the polymer, and the amino compound can be used as the soft segment of the polymer. By further introducing a certain amount of hydroxyl compound, the hydroxyl compound can be used as a chain extender to control the ratio of hard and soft segments during the reaction, thereby regulating the thermodynamic properties and toughness of the polyurea reactant, so that the polyurea reactant has excellent shape memory properties.

[0048] According to some preferred embodiments, in the reaction system, the content of the bioluminescent agent is 20-50 mg / mL (for example, 20 mg / mL, 30 mg / mL, 40 mg / mL or 50 mg / mL).

[0049] In the embodiment of the present invention, during the synthesis of the polyurea reactant, the bioluminescent agent prepared as above is introduced, and the content of the bioluminescent agent is reasonably controlled, so that the bioluminescent agent can interact with the polyurea molecular chain through chemical bonding of the isocyanate component and the amino compound component, so that the bioluminescent agent can be uniformly dispersed in the polyurea reactant matrix without affecting the curing performance and shape memory performance of the polyurea reactant, so that the bioluminescent smart film has excellent bioluminescent performance; for example, in the reaction system of polyurea synthesis, if the content of the bioluminescent agent is too low, it is not conducive to significantly enhancing the fluorescence performance of the bio-intelligent film, and if the content of the bioluminescent agent is too high, not only will the printable polyurea reactant material be reduced, but also the subsequent curing effect will be affected.

[0050] According to some preferred embodiments, in step (1), the reaction temperature is 20-30°C (for example, 20°C, 22°C, 25°C, 28°C or 30°C), and the reaction time is 3-5h (for example, 3h, 4h or 5h); in step (2), the reaction temperature is 50-60°C (for example, 50°C, 52°C, 55°C, 58°C or 60°C), and the reaction time is 20-27h (for example, 20h, 22h, 24h or 27h).

[0051] In the embodiment of the present invention, during the synthesis of the polyurea reactant, it is first reacted at 20-30°C for 3-5h, and then the temperature of the reaction system is increased to 50-60°C, and then the bioluminescent agent and the hydroxyl compound are added in sequence and stirred to obtain a polyurea reactant having fluorescent properties and shape memory properties (glass transition temperature is 40-45°C).

[0052] According to some preferred embodiments, in step (3), the printing temperature is 80-120°C (for example, it can be 80°C, 90°C, 100°C, 110°C or 120°C).

[0053] In an embodiment of the present invention, the polyurea reactant obtained by the reaction is used as a printing solution, and a film with certain gaps is designed and printed on a suitable substrate (such as a glass sheet, a plastic sheet or a metal sheet, etc., selected according to the application scenario) using a 4D printing method. During the printing process, the printing temperature is controlled to be 80-120°C and the bottom temperature of the printing is controlled to be 55°C, thereby forming a bioluminescent smart membrane with a certain porosity. In this way, the bio-intelligent membrane can be infiltrated and exchanged substances during the application process, allowing cells to exchange necessary substances with the external environment, such as the entry and exit of oxygen, carbon dioxide, nutrients and metabolic waste.

[0054] It should be noted that in the embodiment of the present invention, after printing is completed, the printed bio-intelligent membrane needs to be dried, cured and other processing steps. Specifically, the drying temperature can be 60-80°C, the time is more than 48 hours, and the curing temperature can be 55-65°C, the time is 40 hours.

[0055] The embodiment of the present invention further provides a bioluminescent smart film, which is prepared by any of the preparation methods described above.

[0056] The bioluminescent smart film in the embodiment of the present invention has excellent fluorescence performance and biocompatibility, and has shape memory performance and thermal degradation performance, and can realize intelligent control of the bioluminescent film by regulating temperature.

[0057] The embodiment of the present invention also provides the application of the above-mentioned bioluminescent smart film in biomedical materials.

[0058] The bioluminescent smart film in the embodiment of the present invention can efficiently emit fluorescence (such as Figure 1 The fluorescence emission wavelength range is wide and can be adjusted according to the selected fluorescent substance, from ultraviolet light to visible light. It has high fluorescence intensity and good light stability, and can maintain the stability of the fluorescence signal for a long time, which is conducive to long-term detection and observation. The above-mentioned bioluminescent smart membrane can be used to mark and observe specific structures or molecules in living cells.

[0059] In order to more clearly illustrate the technical solution and advantages of the present invention, a preparation method and application of a bioluminescent smart film are described in detail below through several embodiments.

[0060] Embodiment 1:

[0061] Preparation of bioluminescent agent: 5 mmol of isocyanate monomer (toluene diisocyanate) and 5 mmol of amino compound (diethyltoluenediamine) were added into 20 mL of the first solvent (acetone and deionized water in a mass ratio of 3:7) and reacted at 30° C. for 5 h, and the bioluminescent agent was obtained after centrifugation;

[0062] (1) 10 mmol of isocyanate monomer (toluene diisocyanate) and 5 mmol of amino compound (polyetheramine D-400) were added to 30 mL of the first solvent (N,N'-dimethylformamide) and reacted at 30°C for 5 h;

[0063] (2) then adding 30 mg / mL of the bioluminescent agent prepared above and 5 mmol of the hydroxy compound (polycaprolactone diol-2000) to the reaction system of step (1) in sequence, stirring and mixing, and raising the temperature of the reaction system to 60° C., reacting for 25 hours to obtain a polyurea reactant;

[0064] (3) Using polyurea reactants as printing materials, a film was printed on a glass sheet using a 4D printing method. During the printing process, the printing temperature was controlled at 180°C to obtain a bioluminescent smart film.

[0065] Embodiment 2:

[0066] Preparation of bioluminescent agent: adding 20 mmol of isocyanate monomer (isophorone diisocyanate) and 20 mmol of amino compound (tetramethylethylenediamine) into 80 mL of the first solvent (acetone and deionized water in a mass ratio of 3:7) at 30° C. for reaction for 5 h, and obtaining the bioluminescent agent after centrifugation;

[0067] (1) 10 mmol of isocyanate monomer (isophorone diisocyanate) and 3 mmol of amino compound (polyetheramine D-2000) were added to 30 mL of the first solvent (N,N'-dimethylformamide) and reacted at 30°C for 4 h;

[0068] (2) then adding 50 mg / mL of the bioluminescent agent prepared above and 7 mmol of the hydroxy compound (polycaprolactone diol-4000) to the reaction system of step (1) in sequence, stirring and mixing, and raising the temperature of the reaction system to 55° C., reacting for 26 hours to obtain a polyurea reactant;

[0069] (3) Using the polyurea reactant as the printing line, a film was printed on a glass sheet using a 4D printing method. During the printing process, the printing temperature was controlled at 180°C to obtain a bioluminescent smart film.

[0070] Embodiment 3:

[0071] Preparation of bioluminescent agent: 15 mmol of isocyanate monomer (toluene diisocyanate) and 15 mmol of amino compound (diethyltoluenediamine) were added to 60 mL of the first solvent (acetone and deionized water in a mass ratio of 3:7) at 30° C. for reaction for 5 h, and the bioluminescent agent was obtained after centrifugation;

[0072] (1) adding 10 mmol of isocyanate monomer (toluene diisocyanate) and 7 mmol of amino compound (tetramethylethylenediamine) into 30 mL of the first solvent (dichloromethane) and reacting at 25° C. for 5 h;

[0073] (2) then adding 20 mg / mL of the bioluminescent agent prepared above and 3 mmol of the hydroxy compound (polycaprolactone diol-2000) to the reaction system of step (1) in sequence, stirring and mixing, and raising the temperature of the reaction system to 60° C., reacting for 25 hours to obtain a polyurea reactant;

[0074] (3) Using the polyurea reactant as the printing line, a film was printed on a glass sheet using a 4D printing method. During the printing process, the printing temperature was controlled at 180°C to obtain a bioluminescent smart film.

[0075] Embodiment 4:

[0076] Example 4 is substantially the same as Example 1, except that in step (1), the content of the isocyanate monomer is 15 mmol.

[0077] Embodiment 5:

[0078] Example 5 is basically the same as Example 1, except that in step (2), the content of the hydroxy compound is 15 mmol.

[0079] Embodiment 6:

[0080] Example 6 is basically the same as Example 1, except that in step (2), the hydroxy compound is replaced by the same content of polyetheramine-400.

[0081] Embodiment 7:

[0082] Example 7 is basically the same as Example 1, except that in step (1), the isocyanate monomer (isophorone diisocyanate) is replaced by the same amount of hexamethylene diisocyanate trimer, and the amino compound (diethyltoluenediamine) is replaced by the same amount of tris(2-aminoethyl)amine.

[0083] Embodiment 8:

[0084] Example 8 is substantially the same as Example 1, except that when preparing the bioluminescent agent, the content of the isocyanate monomer is 10 mmol.

[0085] Embodiment 9:

[0086] Example 9 is basically the same as Example 1, except that in step (2), the content of the bioluminescent agent is 60 mg / mL.

[0087] Comparative Example 1:

[0088] Comparative Example 1 is substantially the same as Example 1, except that in step (2), the bioluminescent agent is replaced with the same content of rhodamine B.

[0089] Comparative Example 2:

[0090] Comparative Example 2 is substantially the same as Example 1, except that in step (2), no hydroxy compound is added.

[0091] The bioluminescent smart membranes prepared in Examples 1 to 9 and Comparative Examples 1 to 2 (hereinafter referred to as samples) were subjected to performance tests. The test results are as follows: Figure 1 And as shown in Table 1.

[0092] Fluorescence test: The fluorescence properties of the sample are detected by ultraviolet light irradiation; Biocompatibility test: Cells are cultured on the surface of each sample and the growth of cells is observed after 7 days of cell culture to evaluate the biocompatibility of the sample.

[0093] Table 1

[0094]

[0095] From Table 1 and Figures 1 to 3 It can be seen that compared with the comparative example, the particle size of the bioluminescent agent in the embodiment of the present invention is smaller (particle size 100-300nm) and has excellent fluorescence performance, so that the particle size of the prepared solution is smaller, and the bioluminescent agent can be evenly dispersed in the bioluminescent smart film, so that the bioluminescent smart film prepared in the embodiment of the present invention has excellent fluorescence performance, and the bioluminescent smart film has good biocompatibility and shape memory performance, and is easy to print; in Example 4 and Example 5, if the content of the isocyanate monomer is too high or too low, the reactant will remain after the reaction and printing cannot be performed; in Example 6 If the hydroxyl compound adopts the material in the prior art, the glass transition temperature of the biological membrane will increase, and it will be difficult to have shape memory in the body temperature range. In Example 7, if the isocyanate monomer and the amino compound adopt the compounds commonly used in the prior art, not only will the material be unable to be printed, but it will also be difficult to prepare a fluorescent smart membrane without biological toxicity. In Example 8, if the amount of the bioluminescent agent added is too much, it will not only be unfavorable to enhance the fluorescence performance, but will inhibit cell growth. Furthermore, in Comparative Example 1, if the bioluminescent agent in the prior art is used, not only will the bio-intelligent membrane be unable to emit light stably, but it will also have toxic side effects on cells.

[0096] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing a bioluminescent smart film, characterized in that: The preparation method comprises the following steps: (1) adding an isocyanate monomer and an amino compound into a first solvent and stirring for reaction; (2) adding a bioluminescent agent and a hydroxyl compound to a reaction system, stirring the reaction to obtain a polyurea reactant; wherein the bioluminescent agent is obtained by an addition polymerization reaction using an isocyanate monomer and an amino compound as reaction raw materials; (3) Using the polyurea reactant as a printing solution to form a film using a 4D printing method, thereby obtaining the bioluminescent smart film.

2. The preparation method according to claim 1, characterized in that: In step (2), the bioluminescent agent is prepared by the following method: adding the isocyanate monomer and the amino compound into a second solvent for mixed reaction to obtain the bioluminescent agent; wherein the second solvent is an acetone aqueous solution or a phosphate buffered saline solution; Preferably, the content of the isocyanate monomer is 5-20 mmol, the content of the amino compound is 5-20 mmol, and the content of the second solvent is 85-95 wt %.

3. The preparation method according to claim 2, characterized in that: The reaction temperature is 28-32°C and the reaction time is 3-5h.

4. The preparation method according to claim 1 or 2, characterized in that: The isocyanate monomer is toluene diisocyanate or isophorone diisocyanate; the amino compound is polyetheramine D-400, polyetheramine D-2000, diethyltoluenediamine or tetramethylethylenediamine.

5. The preparation method according to claim 1, characterized in that: In step (1), the molar ratio of the isocyanate monomer to the amino compound is 10:(3-7); and / or The first solvent is N,N'-dimethylformamide, N,N'-dimethylacetamide or dichloromethane, and the content of the first solvent is 10-30 mL.

6. The preparation method according to claim 1, characterized in that: In step (2), the hydroxy compound is polycaprolactone diol-2000 or polycaprolactone diol-4000, and the molar ratio of the total amount of the hydroxy compound and the amino compound to the isocyanate monomer is 1:1; and / or In the reaction system, the content of the bioluminescent agent is 20-50 mg / mL.

7. The preparation method according to claim 1, characterized in that: In step (1), the reaction temperature is 20-30°C and the reaction time is 3-5h; and / or In step (2), the reaction temperature is 50-60°C and the reaction time is 20-27h.

8. The preparation method according to claim 1, characterized in that: In step (3), the printing temperature is 80-120°C.

9. A bioluminescent smart film, characterized in that: The preparation method is described in any one of claims 1 to 8.

10. Use of the bioluminescent smart film according to claim 9 in biomedical materials.

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