Moisturizing hydrogel living body material and preparation method thereof
By introducing 2-methacryloyloxyethylphosphate choline or its derivatives into the hydrogel living material, the problem of hydrogel dehydration is solved by using solvent replacement method, achieving the effect of maintaining moisturization and functionality for a long time, and is suitable for the creation of microbial growth environment.
Patent Information
- Application Number
- CN202510289013.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-12
- Publication Date
- 2025-06-20
AI Technical Summary
Hydrogel living materials are prone to dehydration, shrinkage and deformity in open systems, and lose their functionality, and traditional solvents have compatibility and biocompatibility problems in biological applications.
2-methacryloyloxyethylphosphate choline or its derivatives are used as the moisturizing functional material, and are introduced into the hydrogel network through solvent replacement method to form a moisturizing hydrogel living material.
It realizes the long-term moistness of the internal environment of the hydrogel, provides a suitable microbial growth environment, and maintains the functionality of the hydrogel and has good biocompatibility.
Smart Images
Figure CN120173931A_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the field of biomaterials, and particularly to a moisturizing hydrogel living material and a preparation method thereof. Background Art
[0002] A hydrogel living material is a novel dynamic composite system formed by encapsulating living microbial cells (such as bacteria, fungi) in a programmable hydrogel matrix. It endows the material with functions such as sensing and chemical production through cells, and at the same time precisely regulates cell behavior by using the chemical gradient, mechanical restriction and 3D spatial structure of the hydrogel, breaking through the limitations of the disordered structure of the traditional liquid culture system and the static function of ordinary hydrogels. The hydrogel living material realizes the two-way interaction between cells and materials (such as the dynamic regulation of material properties by metabolites), shows the unique advantages of "living materials" in the fields of biomanufacturing, environmental remediation and intelligent medicine, and has become a frontier direction of the cross-integration of synthetic biology and materials science.
[0003] Although hydrogel living materials have great potential in biomedicine and wearable flexible devices, when the hydrogel is applied in an open system, a key problem commonly existing in the industry is that the moisture inside the hydrogel will inevitably evaporate, causing it to dehydrate, shrink and deform, and even lose its functionality. If traditional solvents such as lithium chloride solution, glycerol, dimethyl sulfoxide, etc. are introduced into the hydrogel network, there are certain limitations in the biological application process. These solvents will reduce the compatibility with the hydrogel or deteriorate the biocompatibility, making it difficult to create a microenvironment suitable for the growth and reproduction of microorganisms. Summary of the Invention
[0004] Aiming at the deficiencies of the prior art, the invention object of the present application is to provide a moisturizing hydrogel living material and a preparation method thereof. The moisturizing hydrogel living material has good biocompatibility, and at the same time can maintain the internal environment of the hydrogel moist for a long time, providing a suitable environment for the growth of microorganisms.
[0005] To achieve the above invention object, in one aspect of the present application, there is provided a moisturizing hydrogel living material, including: a matrix material with the hydrogel living material as the first component and a moisturizing functional material with 2-methacryloyloxyethyl phosphorylcholine or its derivative as the second component.
[0006] Among them, 2-methacryloyloxyethyl phosphorylcholine is an amphoteric ionic compound with excellent water solubility. The phosphorylcholine group in its molecular structure is similar to the phospholipids of biological cell membranes, capable of constructing a biomimetic membrane structure and effectively preventing water loss. In addition, it has good biocompatibility and its chemical properties are stable, not affected by environmental factors. Due to its amphiphilic structure, 2-methacryloyloxyethyl phosphorylcholine has good solubility in water and some organic solvents, which enables it to easily blend into various formulations and fully exert its moisturizing effect. Therefore, 2-methacryloyloxyethyl phosphorylcholine or its derivatives can be used as a moisturizing functional component and introduced into the hydrogel network through a solvent replacement method to achieve the moisturizing function.
[0007] Optionally, in combination with any of the above aspects, in another implementation manner of this aspect, the second component is dispersed in the network skeleton of the first component.
[0008] Optionally, in combination with any of the above aspects, in another implementation manner of this aspect, it further includes microorganisms dispersed in the matrix material.
[0009] Optionally, in combination with any of the above aspects, in another implementation manner of this aspect, the microorganisms include natural microorganisms and / or engineered microorganisms. The natural microorganisms include one or more combinations of bacteria, fungi, and algae, and the engineered microorganisms include one or more combinations of engineered fungi and engineered bacteria.
[0010] Optionally, in combination with any of the above aspects, in another implementation manner of this aspect, the matrix material is selected from one or more combinations of methacrylated gelatin, methacrylated sodium alginate, methacrylated hyaluronic acid, and methacrylated glycidyl esterified polyvinyl alcohol.
[0011] Another aspect of the present application provides a method for preparing a moisturizing hydrogel living material, including the following steps:
[0012] (1) Synthesize a hydrogel living material containing microorganisms;
[0013] (2) Immerse the hydrogel living material in a moisturizing functional material for solvent replacement to obtain a moisturizing hydrogel living material; wherein, the moisturizing functional material includes 2-methacryloyloxyethyl phosphorylcholine or its derivatives.
[0014] Optionally, in combination with any of the above aspects, in another implementation manner of this aspect, the step (1) includes:
[0015] Synthesize the hydrogel components used for the living material;
[0016] Dissolve the hydrogel component and photoinitiator in water, add microorganisms and mix evenly to obtain a hydrogel precursor solution containing microorganisms;
[0017] Carry out photocuring on the hydrogel precursor solution containing microorganisms by one or a combination of spin coating method, mold forming method or 3D printing method to obtain the hydrogel living material.
[0018] Optionally, in combination with any of the above aspects, in another implementation manner of this aspect, in step (2), the derivative is a different functionalized product obtained by chemically modifying or synthesizing 2-methacryloyloxyethyl phosphorylcholine.
[0019] Optionally, in combination with any of the above aspects, in another implementation manner of this aspect, the derivative uses 2-methacryloyloxyethyl phosphorylcholine as a monomer and is prepared by atom transfer radical polymerization.
[0020] Optionally, in combination with any of the above aspects, in another implementation manner of this aspect, the atom transfer radical polymerization uses 2,2′-bipyridine as a ligand, methyl 2-bromopropionate as a chain initiator, and cuprous bromide as a catalyst.
[0021] Optionally, in combination with any of the above aspects, in another implementation manner of this aspect, the moisturizing hydrogel living material has the following functions:
[0022] (1) Endow the hydrogel living material with a variety of unique functions that simulate the life system, including biosensing, biomanufacturing, and bioremediation;
[0023] (2) Endow the hydrogel living material with hydrophobic hydration function, keep the internal environment of the hydrogel living material moist, ensure a suitable growth environment for microorganisms while maintaining the functionality of the hydrogel.
[0024] As described above, the present application has the following beneficial effects: Through the hydrophobic hydration function of the moisturizing functional unit 2-methacryloyloxyethyl phosphorylcholine or its derivative, the present application promotes water molecules to form a cage-like water structure, and uses the solvent replacement method to replace the solvent of the moisturizing functional unit with that of the hydrogel living material to form a moisturizing hydrogel living material. It not only has good biocompatibility, but also can create a long-term moisturizing growth microenvironment for microorganisms, ensure the stability of the humidity inside the wet gel, effectively resist the risk of dehydration, and provide a new idea for the preparation of hydrogel living materials.
[0025] The above invention content is provided to introduce some concepts in a simplified form, which will be further described in detail in the following specific implementation manners. The above invention content is neither intended to identify the key features or essential features of the claimed subject matter, nor is it intended to assist in determining the scope of the claimed subject matter. The claimed subject matter of the present invention is not limited to embodiments that solve any or all of the disadvantages pointed out in the background art. BRIEF DESCRIPTION OF THE DRAWINGS
[0026] The drawings herein are incorporated into and constitute a part of this specification, showing embodiments consistent with the present application, and are used together with the specification to explain the principles of the present application. To more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, for those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings. These drawings and textual descriptions are not intended to limit the scope of the concept of the present application in any way, but to illustrate the concept of the present application to those skilled in the art by referring to specific embodiments.
[0027] Figure 1 Schematic diagram for preparing a hydrogel living material with moisture retention in the present application;
[0028] Figure 2 1H NMR spectrum of poly(2-methacryloyloxyethyl phosphorylcholine) in Example 1 of the present application;
[0029] Figure 3 Moisture retention performance diagram of the hydrogel in Example 2 of the present application;
[0030] Figure 4 Moisture retention performance diagrams of 4 different hydrogels with 40% PMPC introduced respectively in Examples 2, 3, 4, and 5 of the present application;
[0031] Figure 5 Diagram of a hydrogel living material with moisture retention containing Escherichia coli in Example 6 of the present application;
[0032] Figure 6 Sensing diagram of different hydrogel living materials with moisture retention containing Escherichia coli in Example 7 of the present application;
[0033] Figure 7 Growth diagram of a hydrogel living material with moisture retention containing Chlorella prepared by 3D printing in Example 8 of the present application;
[0034] Figure 8 Growth diagram of different hydrogel living materials with moisture retention containing Chlorella prepared by the template method in Example 9 of the present application. DETAILED DESCRIPTION OF THE EMBODIMENTS
[0035] The following further describes the present application in detail with specific embodiments.
[0036] 2-Methacryloyloxyethyl phosphorylcholine is a methacrylate monomer containing a phosphorylcholine group and is the monomer most suitable for mimicking the polar groups of cell membrane phospholipids. It has special biological inertness, can construct molecular structures with adjustable properties, and has characteristics such as anti-cell adhesion, anticoagulation, and anti-nonspecific protein adsorption. In addition, it also has good biocompatibility and chemical stability, is not affected by environmental factors, and has good solubility in water and some organic solvents, which enables it to easily integrate into various formulations and fully exert its moisturizing effect.
[0037] The present application innovatively utilizes the moisturizing ability and biocompatibility of this substance to improve the moisturizing ability of hydrogel living materials. Its moisturizing mechanism is mainly based on its hydrophobic hydration function. Through the hydrophobic hydration function of the three methyl groups in the trimethylammonium group, it promotes water molecules to form an ice-like cage water structure, effectively locking in water molecules. At the same time, the inner salt structure formed by the cation and anion in the phosphorylcholine group can maintain the stability of the surrounding water molecules. Since this substance has weak interaction with other molecules, in a biological environment, it will not destroy the water structure around itself due to interaction with other molecules, thus continuously promoting water molecules to form a cage water structure and achieving efficient moisturization.
[0038] In terms of the synthesis process, the solvent replacement method is used to replace the solvents of the moisturizing functional material and the hydrogel living material to form a moisturizing hydrogel living material, thereby achieving the purpose of anti-dehydration. This method is simpler and more efficient, effectively overcoming the compatibility and biocompatibility problems caused by traditional moisturizing solvents, and providing an effective way to solve the anti-dehydration and biocompatibility problems of living materials in related fields.
[0039] When using the moisturizing hydrogel living material in biosensing, such as Figure 1 shown, N-(3-oxododecanoyl)-L-homoserine lactone (3O-C 12 -HSL), which belongs to the N-acyl homoserine lactone (AHL) family, is a class of special small molecule water-soluble compounds. The LasR protein is the sensing factor of AHL, belongs to the LuxR family of transcriptional regulators, and is also a DNA-binding transcriptional activation element. When the concentration of 3O-C 12 -HSL accumulates to a certain threshold, it binds to the LasR protein. The LasR-3O-C 12 -HSL complex is a complex of a protein and a signaling molecule used for bacterial communication and regulation of gene expression. The bound complex can activate the transcriptional expression of the luciferase gene and secrete GFP fluorescent protein, thereby making it emit light.
[0040] Based on the above characteristics, the moisturizing hydrogel living material provided by the embodiments of the present application has the following functions:
[0041] (1) Endow the hydrogel living material with a variety of unique functions that simulate the life system, including biosensing, biomanufacturing, and bioremediation;
[0042] (2) Endow the hydrogel living material with hydrophobic hydration function, keep the internal environment of the hydrogel living material moist, ensure a suitable growth environment for microorganisms, and maintain the functionality of the hydrogel at the same time.
[0043] Next, the technical solutions of the present invention will be described clearly and completely in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.
[0044] Example 1
[0045] Preparation of poly(2-methacryloyloxyethyl phosphorylcholine) by atom transfer radical polymerization
[0046] Please refer to Figure 1 , which shows the reaction formula for preparing poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) by ATRP polymerization of 2-methacryloyloxyethyl phosphorylcholine (MPC) as a monomer, 2,2′-bipyridine (BPY) as a ligand, methyl 2-bromopropionate (MBP) as a chain initiator, and copper(I) bromide (CuBr) as a catalyst in anhydrous methanol at 30 °C. Among them, the molar ratio of [MPC]:[MBP] is 5:1, and the molar ratio of [CuBr] / [MBP] / [BPY] is 1:1:2. During purification, the obtained reactant is passed through a silica column with anhydrous ethanol as the eluent, and then precipitated with an excess of tetrahydrofuran, and dried after recrystallization three times to obtain poly(2-methacryloyloxyethyl phosphorylcholine). Figure 2 shows the 1H NMR spectra of 2-methacryloyloxyethyl phosphorylcholine and poly(2-methacryloyloxyethyl phosphorylcholine) in Example 1. The complete consumption of the vinyl group in the MPC chain participating in the reaction is indicated by the chemical shift of 5.70 - 6.30 ppm, and the grafted methyl 2-bromopropionate block appears at the chemical shift of 0.80 - 1.20 ppm, indicating the successful synthesis of PMPC.
[0047] Example 2
[0048] Preparation of GelMA hydrogel containing poly(2-methacryloyloxyethyl phosphorylcholine) by solvent replacement method
[0049] Weigh 45 mg of double-bonded gelatin (GelMA) and 1.5 mg of photoinitiator lithium phenyl(2,4,6-trimethylbenzoyl)phosphate and dissolve them in 300 μL of deionized water to prepare a 15% (w / v) GelMA solution (the content of the photoinitiator is 0.5% (w / v)). Transfer it to a cylindrical mold with a diameter of 11 mm and a height of 5 mm, and irradiate it with 405 nm blue light for 5 min to carry out free radical copolymerization reaction. After demolding, a cylindrical GelMA hydrogel block with a diameter of 11 mm and a height of 5 mm is obtained.
[0050] Weigh 400 mg of PMPC and 800 mg of PMPC respectively and dissolve them in 3 mL of deionized water to prepare 20% (w / v) PMPC solution and 40% (w / v) PMPC solution. Immerse two GelMA hydrogel blocks with the same weight into 20% (w / v) PMPC and 40% (w / v) PMPC solutions respectively, and carry out solvent replacement at room temperature for 24 h to replace PMPC into the hydrogel network.
[0051] After the solvent replacement is completed, dry the water on the surface of the GelMA hydrogel block, and add another GelMA hydrogel block that has not undergone solvent replacement as a comparison. Place them in an environment of 25 °C and R.H = 54% to dry, and weigh every 2 h to obtain the moisture retention rate. The moisture retention rate of the hydrogel at different times is calculated by the following formula: Moisture retention rate = m t / m0 * 100%, where m0 is the initial weight of the hydrogel, and m t is the mass of the hydrogel at t hours. Figure 3 The moisture retention performance graph of the hydrogel as a comparison is shown. The moisture retention performance of the GelMA hydrogel block after PMPC solvent replacement starts to stand out after 4 hours, and the GelMA hydrogel block with a higher PMPC solution concentration obviously has higher moisture retention performance.
[0052] Example 3
[0053] Preparation of SA-MA hydrogel containing poly(2-methacryloyloxyethyl phosphorylcholine) by solvent replacement method
[0054] Weigh 6 mg of methacrylated sodium alginate (MA) and 1.5 mg of photoinitiator lithium phenyl(2,4,6-trimethylbenzoyl)phosphate and dissolve them in 300 μL of deionized water to prepare a 2% (w / v) SA-MA solution (the content of the photoinitiator is 0.5% (w / v)). Transfer it to a cylindrical mold with a diameter of 11 mm and a height of 5 mm, and irradiate it with 405 nm blue light for 5 min to carry out free radical polymerization. After demolding, a cylindrical SA-MA hydrogel block with a diameter of 11 mm and a height of 5 mm is obtained.
[0055] Weigh 800 mg of PMPC and dissolve it in 3 mL of deionized water to prepare a 40% (w / v) PMPC solution. Immerse the SA-MA hydrogel block in the 40% (w / v) PMPC solution and perform solvent replacement at room temperature for 24 h to replace PMPC into the hydrogel network.
[0056] After the solvent replacement is completed, dry the surface moisture of the SA-MA hydrogel block and place it in an environment of 25 °C and R.H = 54% for drying. Weigh it every 2 h to obtain the moisture retention rate. The moisture retention rate of the hydrogel at different times is calculated by the following formula: Moisture retention rate = m t / m0 * 100%, where m0 is the initial weight of the hydrogel and m t is the mass of the hydrogel at t hours.
[0057] Example 4
[0058] Preparation of SH-MA hydrogel containing poly(2-methacryloyloxyethyl phosphorylcholine) by solvent replacement method
[0059] Weigh 6 mg of methacrylated hyaluronic acid and 1.5 mg of photoinitiator lithium phenyl(2,4,6-trimethylbenzoyl)phosphate and dissolve them in 300 μL of deionized water to prepare a 2% (w / v) SH-MA solution (the content of the photoinitiator is 0.5% (w / v)). Transfer it to a cylindrical mold with a diameter of 11 mm and a height of 5 mm, and irradiate it with 405 nm blue light for 5 min for free radical polymerization. After demolding, a cylindrical SH-MA hydrogel block with a diameter of 11 mm and a height of 5 mm is obtained.
[0060] Weigh 800 mg of PMPC and dissolve it in 3 mL of deionized water to prepare a 40% (w / v) PMPC solution. Immerse the SH-MA hydrogel block in the 40% (w / v) PMPC solution and perform solvent replacement at room temperature for 24 h to replace PMPC into the hydrogel network.
[0061] After the solvent replacement is completed, dry the surface moisture of the SH-MA hydrogel block and place it in an environment of 25 °C and R.H = 54% for drying. Weigh it every 2 h to obtain the moisture retention rate. The moisture retention rate of the hydrogel at different times is calculated by the following formula: Moisture retention rate = m t / m0 * 100%, where m0 is the initial weight of the hydrogel and m t is the mass of the hydrogel at t hours.
[0062] Example 5
[0063] Preparation of PVA-GMA hydrogel containing poly(2-methacryloyloxyethyl phosphorylcholine) by solvent replacement method
[0064] Weigh 6 mg of methacrylic acid glycidyl esterified polyvinyl alcohol and 1.5 mg of photoinitiator lithium phenyl(2,4,6-trimethylbenzoyl)phosphate and dissolve them in 300 μL of deionized water to prepare a 2% (w / v) PVA-GMA solution (the content of the photoinitiator is 0.5% (w / v)). Transfer it to a cylindrical mold with a diameter of 11 mm and a height of 5 mm, and irradiate it with 405 nm blue light for 5 min for free radical polymerization. After demolding, a cylindrical PVA-GMA hydrogel block with a diameter of 11 mm and a height of 5 mm is obtained.
[0065] Weigh 800 mg of PMPC and dissolve it in 3 mL of deionized water to prepare a 40% (w / v) PMPC solution. Immerse the PVA-GMA hydrogel block in the 40% (w / v) PMPC solution and perform solvent replacement at room temperature for 24 h to replace PMPC into the hydrogel network.
[0066] After the solvent replacement is completed, dry the surface moisture of the PVA-GMA hydrogel block and place it in an environment of 25 °C and R.H = 54% for drying. Weigh it every 2 h to obtain the moisture retention rate. The moisture retention rate of the hydrogel at different times is calculated by the following formula: Moisture retention rate = m t / m0 * 100%, where m0 is the initial weight of the hydrogel and m t is the mass of the hydrogel at t hours.
[0067] Figure 4 The comparison chart of the moisture retention performance of 4 different hydrogels respectively introducing 40% PMPC in Examples 2-5 is shown. It can be seen that although the same concentration of PMPC moisture retention unit components are used for solvent replacement, choosing different hydrogel matrix materials will also lead to different moisture retention performance differences. However, all hydrogel materials can maintain a moisture retention rate of more than 30% after 24 h after being replaced by the moisture retention material. Among them, the GelMA hydrogel material has the most excellent moisture retention performance.
[0068] Example 6
[0069] Preparation of hydrogel living materials loaded with Escherichia coli by 3D printing
[0070] Place Escherichia coli (E. coli pLuxR-GFP) in LB medium and activate it for 12 h at 37 °C. Dilute the bacterial liquid concentration to OD 600The value is between 0.6 and 0.8; aspirate 250 μL of Escherichia coli solution in the logarithmic growth phase and centrifuge it at 3000 rpm for 2 min. After centrifugation, remove the supernatant and leave the precipitate for later use; aspirate 5 mL of 15% (w / v) GelMA solution (the photoinitiator content is 0.5% (w / v)) and mix it evenly with the Escherichia coli precipitate. Load it into a 3D printing syringe, fix it on the nozzle, turn on the computer, set the parameters, the syringe heating temperature is 20 °C, the platform temperature is 4 °C, adjust the distance between the nozzle and the platform to 0.5 mm, the printing speed is 800 mm / min, select the pre-set grid pattern, the specification is set to 4×4 mm, the length and width are set to 16 mm, and the number of layers is set to 10 layers. After setting, first manually adjust the air pressure to extrude evenly and stably. After the extrusion is stable, turn off the air pressure and click print. The machine will automatically print until it is completed. The printed GelMA hydrogel living material loaded with Escherichia coli is as Figure 5 shown.
[0071] Example 7
[0072] Preparation of Different Moisture-Retaining Hydrogel Living Materials Loaded with Escherichia coli by 3D Printing, Cultivation and Biosensing
[0073] Place Escherichia coli (E.coli pLuxR-GFP) in LB medium and activate it at 37 °C for 12 h. Dilute the bacterial solution concentration to OD 600The value is between 0.6 and 0.8; 250 μL of Escherichia coli solution in the logarithmic growth phase was centrifuged at 3000 rpm for 2 min. After centrifugation, the supernatant was removed and the precipitate was reserved for use. 5 mL of 15% (w / v) GelMA solution, 2% (w / v) SA-MA solution, 2% (w / v) SH-MA, and 2% (w / v) PVA-GMA solution were respectively pipetted. The photoinitiator content of the above hydrogel solution was 0.5% (w / v). It was mixed evenly with the Escherichia coli precipitate, loaded into a 3D printing syringe, fixed on the nozzle, the computer was turned on, the parameters were set, the syringe heating temperature was 20 °C, the platform temperature was 4 °C, the distance between the nozzle and the platform was adjusted to 0.5 mm, the printing speed was 800 mm / min, a pre-set grid pattern was selected, the specification was set to 4×4 mm, the length and width were set to 16 mm. For the convenience of observing green fluorescence, the number of layers was set to 2 layers. After setting, first manually adjust the air pressure to extrude evenly and stably. After the extrusion was stable, the air pressure was turned off, and the print button was clicked. The machine would automatically print until completion. The printed different hydrogel living materials loaded with Escherichia coli were respectively soaked in poly(2-methacryloyloxyethyl phosphorylcholine) for 24 h for solvent replacement, so that PMPC was introduced into the hydrogel network to obtain a moisturizing hydrogel living material. After the replacement, N-acyl homoserine lactone (AHL) was dropped onto the moisturizing hydrogel living material containing Escherichia coli and cultured for 4 h or more under dark conditions at room temperature. The Escherichia coli in the moisturizing hydrogel living material responded to AHL and expressed green fluorescent protein. Green fluorescence could be clearly observed at an excitation wavelength of 488 nm. In the initial stage of culture, the green fluorescence in the moisturizing hydrogel living material was not obvious; but when cultured for 4 h, the green fluorescence in the moisturizing hydrogel living material could be clearly observed. As Figure 6 shown, it shows the fluorescence images of different moisturizing hydrogel living materials at different culture durations. After culturing for 8 h, the fluorescence tended to be stable.
[0074] Example 8
[0075] Preparation and culture of hydrogel living materials loaded with Chlorella by 3D printing
[0076] Chlorella vulgaris (GY-D27 Chlorella vulgaris) was placed in BG11 medium and enriched for 5 days at 25 °C. The algal solution concentration was diluted to OD 680The value is between 0.8 and 1.0; 250 μL of the Chlorella solution in the logarithmic growth phase was taken and centrifuged at 3000 rpm for 2 min. After centrifugation, the supernatant was removed and the precipitate was reserved for use; 15% (w / v) GelMA solution (photoinitiator content 0.5% (w / v)) was taken and mixed evenly with the Chlorella precipitate, filled into a 3D printing syringe, fixed on the nozzle, the computer was turned on, the parameters were set, the syringe heating temperature was 20 °C, the platform temperature was 4 °C, the distance between the needle and the platform was adjusted to 0.5 mm, the printing speed was 800 mm / min, a pre-set equilateral triangle pattern was selected, the base was set to 18 mm, and the number of layers was set to 5 layers. After setting, first manually adjust the air pressure to extrude evenly and stably. After the extrusion was stable, the air pressure was turned off, and the print button was clicked, and the machine would automatically print until completion.
[0077] The hydrogel living material loaded with Chlorella was soaked in poly(2-methacryloyloxyethyl phosphorylcholine) for 24 h for solvent replacement, so that PMPC was introduced into the hydrogel network to obtain a moisturizing hydrogel living material. After the replacement, the moisturizing hydrogel living material encapsulating Chlorella was placed in an incubator. The culture conditions were: temperature 25 °C, light intensity 8100 Lux, light-dark time 12 h:12 h, rotation speed 100 rpm / min. As time went by, the growth of Chlorella could be clearly seen, as Figure 7 shown.
[0078] Example 9
[0079] Preparation and culture of a moisturizing hydrogel living material loaded with Chlorella by the template method
[0080] Chlorella (GY-D27 Chlorella vulgaris) was placed in BG11 medium and enriched for 5 days at 25 °C, and the algal solution concentration was diluted to OD 680The value is between 0.8 and 1.0; 250 μL of the Chlorella solution in the logarithmic growth phase was taken and centrifuged at 3000 rpm for 2 min. After centrifugation, the supernatant was removed and the precipitate was reserved for use; A certain amount of 2% (w / v) SA-MA solution, 2% (w / v) SH-MA and 2% (w / v) PVA-GMA solution were respectively aspirated with a 1 mL syringe into a template with a fixed shape and cured under blue light at 405 nm to obtain different hydrogel living materials encapsulating Chlorella. The different hydrogel living materials encapsulating Chlorella were respectively soaked in poly(2-methacryloyloxyethyl phosphorylcholine) for solvent replacement for 24 h to introduce PMPC into the hydrogel network, and a moisturizing hydrogel living material was obtained. After the replacement, the moisturizing hydrogel living material loaded with Chlorella was placed in an incubator, and the culture conditions were: temperature 25 °C, light intensity 8100 Lux, light-dark time 12 h:12 h, and rotation speed 100 rpm / min. The growth of Chlorella could be clearly seen over time. As Figure 8 shown, it presents the diagrams of hydrogel living materials at different culture durations.
[0081] The serial numbers of the embodiments of the present application above are only for description and do not represent the superiority or inferiority of the embodiments.
[0082] In the present application, for the description of the same or similar term concepts, technical solutions and / or application scenarios, generally only the first occurrence is described in detail. When it appears repeatedly later, for the sake of brevity, it is generally not described again. When understanding the technical solutions and other contents of the present application, for the same or similar term concepts, technical solutions and / or application scenarios that are not described in detail later, reference can be made to their previous relevant detailed descriptions.
[0083] In the present application, the descriptions of each embodiment have their own emphases. For the parts not detailed or recorded in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.
[0084] The technical features of the technical solutions of the present application can be combined arbitrarily. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as the combinations of these technical features do not conflict, they should be considered to be within the scope recorded in the present application.
[0085] The above are only the preferred embodiments of the present application, and do not limit the patent scope of the present application accordingly. Any equivalent structural or equivalent process transformation made by using the content of the specification and drawings of the present application, or directly or indirectly applied in other related technical fields, shall be similarly included in the patent protection scope of the present application.
Claims
1. A moisturizing hydrogel living material, characterized in that: include: The invention discloses a base material with a hydrogel living material as the first component and a moisturizing functional material with 2-methacryloyloxyethyl phosphorylcholine or its derivative as the second component.
2. A moisturizing hydrogel living material according to claim 1, characterized in that: The second component is dispersed in the network framework of the first component.
3. The moisturizing hydrogel living material according to claim 1, characterized in that: Also included are microorganisms dispersed within the matrix material.
4. The moisturizing hydrogel living material according to claim 3, characterized in that: The microorganisms include natural microorganisms and / or engineered microorganisms. The natural microorganisms include a combination of one or more of bacteria, fungi and algae, and the engineered microorganisms include a combination of one or more of engineered fungi and engineered bacteria.
5. The moisturizing hydrogel living material according to claim 1, characterized in that: The matrix material is selected from one or more combinations of methacrylated gelatin, methacrylated sodium alginate, methacrylated hyaluronic acid, and methacrylic glycidyl esterified polyvinyl alcohol.
6. A method for preparing a moisturizing hydrogel living material, characterized in that: The following steps are involved: (1) Synthesizing hydrogel living materials containing microorganisms; (2) Immersing the hydrogel living material in a moisturizing functional material to perform solvent replacement to obtain a moisturizing hydrogel living material; wherein the moisturizing functional material comprises 2-methacryloyloxyethyl phosphorylcholine or a derivative thereof.
7. The preparation method according to claim 6, characterized in that: The step (1) comprises: Hydrogel components used in synthetic living materials; Dissolving the hydrogel component and the photoinitiator in water, adding microorganisms and mixing well to obtain a hydrogel precursor solution containing microorganisms; The hydrogel precursor solution containing the microorganisms is photocured by one or a combination of spin coating, mold forming or 3D printing to obtain the hydrogel living material.
8. The preparation method according to claim 6, characterized in that: In the step (2), the derivatives are different functionalized products obtained by polymerizing 2-methacryloyloxyethyl phosphorylcholine through chemical modification or synthesis methods.
9. The preparation method according to claim 8, characterized in that: The derivative is prepared by using 2-methacryloyloxyethyl phosphorylcholine as a monomer through an atom transfer free radical polymerization method.
10. The preparation method according to claim 9, characterized in that: The atom transfer free radical polymerization method uses 2,2'-bipyridine as a ligand, methyl 2-bromopropionate as a chain initiator, and cuprous bromide as a catalyst.