A polymer microsphere integrated preparation device and method based on microfluidic technology

Abstract

The application belongs to the technical field of microsphere preparation, and specifically discloses a polymer microsphere integrated preparation device and method based on microfluidic technology. The preparation device comprises a microfluid driving device, a phase separation device and a collection device. The microfluid driving device comprises a dispersed phase fluid propeller, a continuous phase fluid propeller and a collection phase fluid propeller. The phase separation device integrates a phase change channel, a continuous phase channel, a collection phase channel and a flow rate controller. The continuous phase channel is connected with the flow rate controller. The collection device comprises a continuous phase collector and a microsphere collection device. The microsphere collection device comprises a microsphere collector, a phase filtration membrane and a collection phase collector. The dispersed phase fluid propeller, the continuous phase fluid propeller and the collection phase fluid propeller are connected with the inlet of the phase separation device. One end of the flow rate controller is connected with the outlet of the continuous phase channel. The other end of the flow rate controller is connected with the continuous phase collector. The outlet of the collection phase channel is connected with the microsphere collector.

Description

Technical Field

[0001] This invention belongs to the field of microsphere preparation technology, specifically relating to an integrated polymer microsphere preparation device and preparation method based on microfluidic technology. Background Technology

[0002] Due to their tiny size and unique properties, microspheres are widely used in the biomedical field for drug delivery systems, diagnostic reagents, tissue engineering, and gene therapy. As a core component of drug carriers, they are precisely embedded or firmly adsorbed into polymer matrices to construct microparticle systems, demonstrating outstanding application potential. In the field of dye adsorption, some novel microsphere materials, such as hydrophilic-hydrophobic heterospheres, have been developed and successfully applied to the treatment of dye wastewater. Integrating the preparation process and improving production efficiency can further enhance their influence in the field.

[0003] Currently, there are numerous technical pathways for preparing these high-performance polymer microspheres, encompassing diverse methods such as emulsion polymerization, dispersion polymerization, seed polymerization, suspension polymerization, micro-suspension polymerization, spray drying, and template methods. However, traditional preparation equipment is often limited by challenges such as high reagent consumption, complex and lengthy process flows, difficulty in perfectly controlling microsphere sphericity, and uneven particle size distribution. These challenges significantly restrict further improvements in production efficiency and product quality. Therefore, developing an innovative integrated polymer microsphere preparation device has become a critical task urgently needing to be addressed by the industry. This device aims to significantly improve preparation efficiency through technological innovation, while substantially reducing reagent consumption and optimizing the entire preparation process to ensure that microsphere products meet higher quality standards and consistency requirements.

[0004] Microfluidic technology, with its unique advantages, has gradually become a significant force driving the upgrading of polymer microsphere preparation technology. Its core advantage lies in the ingenious application of microchannel design, enabling efficient and precise microsphere preparation through precise control of fluid behavior at the microscale. However, traditional microfluidic devices are often limited by the design of a single preparation channel, resulting in cumbersome oil-water phase separation processes and low microsphere collection efficiency, thus restricting their potential for large-scale production and industrial applications. Therefore, it is essential to develop a new microsphere preparation device to address the problems of existing oil-water phase separation, integrated preparation, and low production efficiency. Summary of the Invention

[0005] The purpose of this invention is to overcome the defects of the existing technology and provide an integrated polymer microsphere preparation device and method based on microfluidic technology. The device has the advantages of integration and miniaturization. By designing three channels, it achieves controllable separation of oil and water phases based on the phase density itself. Under the condition of ensuring the sphericity and size uniformity of microspheres, it greatly improves the microsphere preparation efficiency and realizes highly controllable large-scale and continuous production.

[0006] To achieve the above objectives, one of the technical solutions of the present invention is: an integrated polymer microsphere preparation device based on microfluidic technology, comprising a microfluidic driving device, a phase separation device, and a collection device. The microfluidic driving device includes a dispersed phase fluid propeller, a continuous phase fluid propeller, and a collecting phase fluid propeller. The phase separation device integrates a phase change channel, a continuous phase channel, a collecting phase channel, and a flow rate controller. The continuous phase channel is connected to the flow rate controller. The collection device includes a continuous phase collector and a microsphere collecting device. The microsphere collecting device includes a microsphere collector, a phase filter membrane, and a collecting phase collector. The inlet end of the phase separation device is connected to the dispersed phase fluid propeller, the continuous phase fluid propeller, and the collecting phase fluid propeller. The outlet end of the continuous phase channel is connected to one end of the flow rate controller, the other end of the flow rate controller is connected to the continuous phase collector, and the outlet end of the collecting phase channel is connected to the microsphere collector.

[0007] The aforementioned collected phase and continuous phase determine their channels based on their own density. By adjusting the flow rate controller to adapt to the liquid flow rate of the three-phase fluid propulsion device, the preparation process becomes controllable, thereby realizing the integrated preparation of microfluidic microspheres.

[0008] In a preferred embodiment of the present invention, the phase change channel, continuous phase channel, and collection phase channel in the phase separation device are arranged asymmetrically. The three channels are arranged in a single-channel structure and are not on the same horizontal plane. The phase change channel is not fixed, but is determined based on the density of the phase itself. When the density of the continuous phase fluid is greater than that of the collection phase fluid, the collection phase channel is closed, and the phase change channel becomes the collection phase channel. When the density of the collection phase is greater than that of the continuous phase, the continuous phase channel is closed, and the phase change channel becomes the continuous phase channel. The flow rate controller is connected to the continuous phase channel and has a multi-level flow rate adjustment function. By finely adjusting the flow rate of the continuous phase fluid, the phase separation process can be ensured to operate stably in a fully controllable automated mode.

[0009] In a preferred embodiment of the present invention, the microsphere collection device further includes a phase filter membrane and a microsphere collector together to achieve efficient and precise collection and purification of the target microspheres.

[0010] In use, the integrated microsphere preparation device involves placing the prepared dispersed phase fluid and continuous phase fluid into the dispersed phase fluid propeller and continuous phase fluid propeller, respectively, and using the propellers to drive the liquid flow. By adjusting and proportioning the flow rate of the propellers, the dispersed phase fluid is sheared into monodisperse microspheres by the continuous phase fluid at the outlet of the connecting channel. These microspheres then flow out of the continuous phase channel with the continuous phase fluid into the phase separation device. The channel is determined based on the density of the phase itself, and the flow rate controller is adjusted to adapt to the three-phase fluid flow rate, allowing the microspheres to flow into the microsphere collector. Finally, qualified microspheres are obtained through freeze drying.

[0011] In a preferred embodiment of the present invention, the microspheres are polymer porous microspheres with loading function, and the target loading material is loaded in the pore structure of the polymer porous microspheres. The target loading material includes, but is not limited to, organic small molecules and inorganic nanoparticles, and the particle size of the microspheres is 300-500 μm.

[0012] More preferably, the polymer porous microspheres are made of one or more of the following: hyaluronic acid and its derivatives, chondroitin sulfate and its derivatives, alginate and its derivatives, chitosan and its derivatives, and polylactic acid and its derivatives.

[0013] More preferably, the polymer porous microspheres are made of one or more of hyaluronic acid, chondroitin sulfate, alginate, chitosan, and polylactic acid.

[0014] In a further preferred embodiment, the polymer porous microspheres are made of one or more of hyaluronic acid and chondroitin sulfate.

[0015] To achieve the above objectives, the second technical solution of the present invention is: a method for preparing polymer microspheres based on an integrated polymer microsphere preparation device, comprising the following steps:

[0016] S1: The continuous phase fluid and the dispersed phase fluid are injected into the microfluidic device and mixed. Under the surface tension and shearing action of the continuous phase fluid, polymer microspheres are obtained. The polymer microspheres are freeze-dried to obtain the polymer porous microspheres.

[0017] S2: The obtained microspheres, continuous phase fluid, and collecting phase fluid are injected together into the separation phase device for mixing. The polymer microspheres are separated from the continuous phase fluid and enter the collecting phase fluid. When the density of the continuous phase fluid is greater than that of the collecting phase fluid, the collecting phase channel is closed, and the polymer microspheres enter the microsphere collector from the phase change channel (collecting phase channel) along with the collecting phase fluid, while the continuous phase fluid is discharged from the continuous phase channel. When the density of the collecting phase fluid is greater than that of the continuous phase fluid, the continuous phase channel is closed, and the polymer microspheres enter the microsphere collector from the collecting phase channel along with the collecting phase fluid, while the continuous phase fluid is discharged from the phase change channel (continuous phase channel).

[0018] In a preferred embodiment of the present invention, the dispersed phase fluid includes a modified polymer, a water-soluble salt, a photocuring agent, and an inorganic solvent, wherein the inorganic solvent includes, but is not limited to, water and phosphate-buffered saline (PBS); the continuous phase fluid includes, but is not limited to, organic solvents such as ethyl acetate and dichloromethane (DCM); and the collected phase fluid includes, but is not limited to, inorganic solvents such as water and PBS.

[0019] More preferably, the photocuring agent is lithium phenyl (2,4,6-trimethylbenzoyl) phosphate, and the water-soluble salt preferably includes one of sodium selenite, sodium permanganate, and potassium permanganate, more preferably sodium selenite and sodium permanganate; the modified polymer preferably includes one or more of hyaluronic acid and its derivatives, chondroitin sulfate and its derivatives, alginate and its derivatives, chitosan and its derivatives, and polylactic acid and its derivatives; more preferably, it includes one or more of hyaluronic acid, chondroitin sulfate, alginate, chitosan, and polylactic acid; and even more preferably, it includes one or more of hyaluronic acid and chondroitin sulfate.

[0020] More preferably, the mass ratio of the modified polymer, water-soluble salt, photocuring agent and inorganic solvent is 0.8-1.8:0.15-0.3:0.4-0.6:160-200.

[0021] More preferably, the method for preparing the dispersed phase includes the following steps: dissolving the modified polymer and the water-soluble salt in water respectively and then mixing them, and then adding a photocuring agent to obtain a dispersed phase solution.

[0022] In a preferred embodiment of the present invention, the injection rate of the dispersed phase fluid is 0.02-0.05 mL / min; the injection rate of the continuous phase fluid is 1.50-2.50 mL / min; and the injection rate of the collecting phase fluid is 1.50-2.50 mL / min.

[0023] In a preferred embodiment of the present invention, the mixing temperature of the preparation steps S1 and S2 is 20-30°C.

[0024] More preferably, the mixing temperature of the preparation steps S1 and S2 is 25-30℃.

[0025] To achieve the above objectives, the third technical solution of the present invention is: a method for preparing polymer microspheres to obtain polymer microspheres.

[0026] To achieve the above objectives, the fourth technical solution of the present invention is: the application of the above-mentioned polymer microspheres in biomedicine and dye adsorption.

[0027] Compared with the prior art, the present invention has the following beneficial effects:

[0028] The microfluidic integrated microsphere preparation device designed in this invention can be used to prepare polymer porous microspheres with loading functions. By adjusting the flow rate controller to adapt to the liquid flow rate of the three-phase fluid propulsion device, the preparation process is made controllable, thereby realizing the integrated preparation of microfluidic microspheres. Compared with traditional microfluidic microsphere preparation devices, it simplifies the device structure, improves the flexibility and adaptability of the preparation process, ensures the high efficiency and precision of microsphere preparation, and ensures the uniformity and stability of microsphere products. Attached Figure Description

[0029] Figure 1 This is a flowchart of the integrated fabrication process of polymer microspheres based on microfluidics.

[0030] Figure 2 This is a schematic diagram of a phase separation device;

[0031] Figure 3 This is a schematic diagram of the cross-section of the flow rate controller;

[0032] Figure 4 This is a schematic diagram of a microsphere collection device;

[0033] Figure 5 A schematic diagram of a usable molded microfluidic chip;

[0034] Figure 6 Optical and SEM images of the GLP-MnO2@ChSMA microspheres prepared in Example 2;

[0035] Figure 7 Optical and SEM images of the HAMA / CS-Se@PDA microspheres prepared in Example 3;

[0036] Figure 8 Optical images of the HAMA / PEP@MnO2 microspheres prepared in Example 4, and SEM images of the HAMA / PEP@MnO2 microspheres and the HAMA / PEP@MnO2 / BC composite film.

[0037] Figure 9 This is a diagram showing the results of co-culturing GLP-MnO2@ChSMA microspheres prepared in Example 2 with chondrocytes.

[0038] Figure 10 This is a graph showing the biocompatibility analysis results of the HAMA / ChS-Se@PDA microspheres prepared in Example 3;

[0039] Figure 11 To analyze the adsorption principle and mechanism obtained by fitting the kinetic model and the isothermal model;

[0040] In the diagram: 1. Microfluidic drive device, 1-1. Dispersed phase fluid thruster, 1-2. Continuous phase fluid thruster, 1-3. Collecting phase fluid thruster, 2. Phase separation device, 2-1. Phase change channel, 2-2. Continuous phase channel, 2-3. Collecting phase channel, 2-4. Flow rate controller, 2-4-1. Main fluid channel, 2-5. Pressure balance port, 2-6. Collection device, 3. Continuous phase collector, 3-1. Microsphere collection device, 3-2. Microsphere collector, 3-2-1. Phase filter membrane, 3-2-2. Collecting phase collector, 3-2-3. Fluid communication pipe, 4. Microfluidic chip, 5. Detailed Implementation

[0041] To make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be described in more detail below with reference to the accompanying drawings and specific embodiments, but the scope of protection of the present invention is not limited to these embodiments.

[0042] Example 1

[0043] An integrated fabrication device for polymer microspheres based on microfluidics, such as Figure 1 As shown, it includes a microfluidic driving device 1, a phase separation device 2, a collection device 3, and a fluid communication pipe 4. The microfluidic driving device 1 includes a dispersed phase fluid thruster 1-1, a continuous phase fluid thruster 1-2, and a collection phase fluid thruster 1-3. The collection device 3 includes a continuous phase collector 3-1 and a microsphere collection device 3-2.

[0044] like Figure 2 As shown, the phase separation device 2 integrates a phase change channel 2-1, a continuous phase channel 2-2, a collection phase channel 2-3, and a flow rate controller 2-4. The continuous phase channel 2-2 is connected to the flow rate controller 2-4. It also includes a main fluid channel 2-5 and a pressure balance port 2-6. The three channels, phase change channel 2-1, continuous phase channel 2-2, and collection phase channel 2-3, are arranged in a single-channel structure and are not on the same horizontal plane. The phase change channel 2-1 is not fixed, but its phase is determined based on its own density. When the density of the continuous phase fluid is greater than that of the collection phase fluid, the collection phase channel 2-3 is closed, and the phase change channel 2-1 becomes the collection phase channel 2-3. When the density of the collection phase is greater than that of the continuous phase, the continuous phase channel 2-2 is closed, and the phase change channel 2-1 becomes the continuous phase channel 2-2.

[0045] Figure 3 The diagram shows a cross-sectional view of the flow rate controller 2-4. The four flow rate controller switches 2-4-1 can meet the different requirements of the liquid flow rate in the propeller during different microsphere preparation processes, ensuring that the liquid flow rate in the three-phase fluid propeller can be accurately matched and meet the specific requirements of each microsphere preparation process.

[0046] Figure 4The diagram shows the microsphere collection device 3-2, which includes a microsphere collector 3-2-1, a phase filter membrane 3-2-2, and a phase collector 3-2-3.

[0047] The inlet end of the phase separation device 2 is connected to the dispersed phase fluid thruster 1-1, the continuous phase fluid thruster 1-2 and the collecting phase fluid thruster 1-3. The outlet end of the continuous phase channel 2-2 is connected to one end of the flow rate controller 2-4. The other end of the flow rate controller 2-4 is connected to the continuous phase collector 2-1. The outlet end of the collecting phase channel 2-3 is connected to the microsphere collector 3-2-1.

[0048] Figure 5 The diagram shows possible fabricated microfluidic chips, including (a) a glass chip with integrated microchannels, (b) a membrane emulsion chip, (c) a simple conduit chip, and (d) a cross-type fabrication chip.

[0049] Example 2

[0050] A polymer microsphere fabrication device based on microfluidics, comprising the following steps:

[0051] (1) At room temperature, weigh a certain amount of Ganoderma lucidum spore powder after cell wall disruption, add ethanol at a ratio of 1:30 (g / mL), stir evenly for 30 min, let stand, filter, and continue to add solvent at a ratio of 1:30 (g / mL) for 3-4 times. Then place in an oven and dry at 60℃ for later use. Weigh 10g of defatted Ganoderma lucidum spore powder, mix thoroughly with distilled water at a ratio of 1:30 (g / mL), then treat with ultrasound for 30 min, keep at 60℃ in a water bath for 2 h, and finally collect the filtrate using a centrifuge (8000r / min, 15 min), and concentrate under reduced pressure to one-third of the original volume. Add 10 times the volume of anhydrous ethanol to the concentrated polysaccharide solution for alcohol precipitation, and finally obtain the precipitate using a G4 sintered glass funnel. Dialyze it for 1-2 days, and then freeze-dry under vacuum for 48 h to obtain Ganoderma lucidum polysaccharide (GLP) for later use.

[0052] (2) At room temperature, 2 g of chondroitin sulfate (ChS) was dissolved in 50 mL of phosphate-buffered saline (PBS; pH = 7.4). Then, 16.9 mL of methacrylic anhydride (MA) was added to the above solution. The pH was adjusted to about 8.0 with 5 M NaOH solution. After the reaction, the solution was magnetically stirred at 4 °C for 24 hours. The solution was then precipitated with ethanol. After centrifugation (10000 rpm, 10 min), the precipitate was obtained. The precipitate was washed with ultrapure water and centrifuged again. The precipitate was dissolved in ultrapure water and dialyzed for three days. Finally, it was lyophilized to obtain methacryloyl chondroitin sulfate (ChSMA).

[0053] (3) At room temperature, prepare 0.01M NaMnO4·H2O solution and 20mg / mL GLP solution, and prepare 5mL solution by mixing them in a 1:1 ratio. Stir with magnetic force to make it react evenly. After 24h, the reaction is complete and GLP-MnO2 is obtained. Then add 60mg ChSMA and 10mg phenyl-2,4,6-trimethylbenzoyl phosphate lithium salt (LAP), stir evenly and keep away from light for later use.

[0054] (4) Using the mixed solution as the dispersed phase, draw 5 mL into a syringe, connect the microfluidic syringe, and set the flow rate to 0.02 mL / min; using a mixture of 85 wt% oil and 15 wt% Span 80 as the continuous phase, draw 50 mL into a syringe, connect the microfluidic syringe, and set the flow rate to 1.50 mL / min; using PBS as the collection phase, draw 50 mL into a syringe, connect the microfluidic syringe, and set the flow rate to 1.50 mL / min. Turn on the microfluidic syringe and use the molded microfluidic chip ( Figure 5 (a) Preparation of photocured GLP-MnO2@ChSMA microspheres. Since the density of the continuous phase mixed oil is less than that of the collected phase water, the continuous phase channel is closed within the phase separation device. The GLP-MnO2@ChSMA microspheres enter the microsphere collection device along with the collected phase channel, while the continuous phase mixed oil is discharged from the phase change channel (i.e., the continuous phase channel) and enters the continuous phase collector. Subsequently, the microspheres are washed with isopropanol to remove the oil and then rinsed five times with PBS to obtain the prepared microspheres.

[0055] The GLP-MnO2@ChSMA microspheres prepared in this embodiment were characterized by optical and SEM analysis, and the results are as follows: Figure 6 As shown. Figure 6 (a) and (b) show that the microspheres appear as uniform spheres under a light microscope, with a size of approximately 150-200 μm; Figure 6 (b) and (c) show that the pores of the microspheres are similar in size and uniformly distributed. By magnifying and observing the structure inside the pores, it can be seen that the elliptical GLP-MnO2 NPs are uniformly distributed on the surface of the microspheres.

[0056] Example 3

[0057] A polymer microsphere fabrication device based on microfluidics, comprising the following steps:

[0058] (1) 100 mL of sodium selenite (Na2SeO3, 10 mM) solution was mixed evenly with 100 mL of ascorbic acid (VC, 20 mM) solution in an ice bath environment. After stirring and reacting in the dark for 12 h, the solution was centrifuged (12000 rpm, 10 min) to concentrate it. Finally, the reaction product selenium nanoparticles (Se NPs) were obtained by freeze drying and stored in a drying oven.

[0059] (2) At room temperature, 250 mg ChS was added to 100 mL of Na2SeO3 (10 mM) solution. After it was uniformly dissolved, the solution was transferred to an ice bath environment, and then 100 mL of VC (20 mM) solution was added. The reaction was stirred in the dark for 12 h, and then the solution was centrifuged (12000 rpm, 10 min) to concentrate. Finally, the reaction product chondroitin sulfate grafted selenium nanoparticles (ChS-Se NPs) were obtained by freeze drying and stored in a drying oven.

[0060] (3) At room temperature, 80 mg of the ChS-Se NPs prepared by the above reaction were mixed evenly with 50 mL of Tris-HCl (20 mM, pH = 8.8) solution, and then 50 mL of dopamine hydrochloride solution (1.6 mg / mL) was added dropwise. The mixture was stirred vigorously for 12 h with the container open, and then the solution was centrifuged (12000 rpm, 10 min) to concentrate. Finally, the product was freeze-dried to obtain polydopamine-coated chondroitin sulfate-grafted selenium nanoparticles (ChS-Se@PDANPs), which were stored in a drying oven.

[0061] (4) Prepare 100 mL of ultrapure water and N,N-dimethylformamide in a 2:1 ratio, then add hyaluronic acid (HA) to prepare a 1 wt% HA solution. Stir at 37°C until completely dissolved. Add 4 mL of MA to the HA solution and prepare 3M NaOH to maintain the pH at 8-9. Stir continuously at 4°C for 24 h to obtain methacryloyl hyaluronic acid (HAMA). Dialyze with ultrapure water for three days, then freeze-dry to obtain HAMA, and store in a sealed, dry container.

[0062] (5) Dissolve 80 mg ChS-Se@PDANPs in 10 mL of deionized water, then add 125 mg HAMA and stir until completely dissolved. After the HAMA is completely dissolved, add 25 mg LAP and stir until dissolved evenly at room temperature in the dark to obtain a microfluidic dispersion solution. Aspirate a 5 mL syringe, connect the microfluidic syringe, and set the flow rate to 0.04 mL / min. Using CH2Cl2 as the continuous phase, aspirate a 50 mL syringe, connect the microfluidic syringe, and set the flow rate to 2.50 mL / min. Using H2O as the collecting phase, aspirate a 50 mL syringe, connect the microfluidic syringe, and set the flow rate to 2.50 mL / min. Turn on the microfluidic syringe and use the molded microfluidic chip (…). Figure 5(b) Preparation: Photocured HAMA / ChS-Se@PDA microspheres were obtained. Since the density of the continuous phase CH2Cl2 is greater than that of the collected phase H2O, the collected phase channel is closed in the phase separation device. The HAMA / ChS-Se@PDA microspheres enter the microsphere collection device from the phase change channel (collected phase channel) along with the collected phase, while the continuous phase CH2Cl2 is discharged from the continuous phase channel and enters the continuous phase collector.

[0063] The HAMA / ChS-Se@PDA microspheres prepared in this embodiment were characterized and analyzed by optical and SEM methods, and the results are as follows: Figure 7 As shown. Figure 7 (a) The microspheres appear as uniform spheres under a light microscope, with a size of approximately 300 μm; Figure 7 (b) It can be seen that the prepared HAMA / ChS-Se@PDA microspheres are approximately spherical with many void structures on them, which serve as drug loading sites.

[0064] Example 4

[0065] A polymer microsphere fabrication device based on microfluidics, comprising the following steps:

[0066] (1) Prepare HAMA using the same method as described above, and store it in a sealed, dry place.

[0067] (2) Dissolve 24 mg of purified *Ulva prolifera* polysaccharide (PEP) in 4.7 mL of ultrapure water. After dissolution, add 300 μL of 0.1 M NaMnO4 solution until the reaction is complete to obtain a PEP@MnO2 solution. Add 62.5 mg of HAMA and 13 mg of LAP to the solution, stir well, and set aside.

[0068] (3) Using the mixed solution as the dispersed phase, draw 5 mL into a syringe, connect it to a microfluidic injector, and set the flow rate to 0.05 mL / min; using CH2Cl2 as the continuous phase, draw 50 mL into a syringe, connect it to a microfluidic injector, and set the flow rate to 2.00 mL / min; using H2O as the collecting phase, draw 50 mL into a syringe, connect it to a microfluidic injector, and set the flow rate to 2.00 mL / min. Turn on the microfluidic injector and use a simple microfluidic chip ( Figure 5 (c) Preparation: Photocured HAMA / PEP@MnO2 microspheres were obtained at the collection port. Since the density of the continuous phase CH2Cl2 is greater than that of the collected phase water, the collection phase channel is closed in the phase separation device. The HAMA / PEP@MnO2 microspheres enter the microsphere collection device from the phase change channel (i.e., the collection phase channel) along with the collected phase, while the continuous phase CH2Cl2 is discharged from the continuous phase channel and enters the continuous phase collector.

[0069] (4) The prepared HAMA / PEP@MnO2 microspheres and BC were mixed evenly at a ratio of 1:10. After filtration, a uniformly mixed HAMA / PEP@MnO2 / BC membrane (PMM) was obtained to verify the adsorption performance of the prepared HAMA / PEP@MnO2 microspheres.

[0070] The HAMA / PEP@MnO2 microspheres and PMM prepared in this embodiment were characterized by optical and SEM analysis, and the results are as follows: Figure 8 As shown. Figure 8 (a) The microspheres appear as uniform spheres under a light microscope, with a size of approximately 450 μm; Figure 8 (b) It can be seen that the prepared PEP@MnO2 microspheres are approximately spherical with many porous structures, which can increase the adsorption sites for methylene blue; through Figure 8 (c) It can be observed that microspheres are embedded on both the surface and inside of the PMM, and the microspheres are distributed evenly in all parts of the membrane.

[0071] Example 5

[0072] Application of GLP-MnO2@ChSMA microspheres prepared in Example 2 in biomedicine:

[0073] like Figure 9 As shown in (a), after co-culturing freeze-dried GLP-MnO2@ChSMA microspheres with cells, the cells grew uniformly on the surface of the microspheres and showed good survival rate, compared to Figure 9 As shown in (b), more cells grew on the freeze-dried microspheres, possibly due to the porous structure of the microspheres after freeze-drying, which provided more sites for cell growth. Simultaneously, the recruited cells were reconstructed in three dimensions. The results showed that cells were able to attach and spread on the GLP-MnO2@ChSMA microspheres, indicating that the microspheres can provide cells with a three-dimensional network structure similar to cartilage ECM. Figure 9 (c)).

[0074] Application of HAMA / ChS-Se@PDA microspheres prepared in Example 3 in biomedicine:

[0075] The biocompatibility of HAMA / ChS-Se@PDA microspheres was assessed using the CCK8 assay and cell viability / death staining. Figure 10 As shown, after Calcein / PI staining, some dead cells were present in the component with the maximum concentration of ChS-Se@PDA at 20 μg / mL, while no obvious cell-killing effect was observed in the components with concentrations of 0-10 μg / mL. This indicates that HAMA / ChS-Se@PDA microspheres do not have obvious cytotoxicity and have good biocompatibility.

[0076] Application of HAMA / PEP@MnO2 microspheres prepared in Example 4 in dye adsorption:

[0077] The adsorption processes of bacterial cellulose membrane (BCM) and HAMA / PEP@MnO2 / BC membrane (PMM) in MB solution were fitted using pseudo-first-order and pseudo-second-order kinetic models to explore the adsorption mechanism of MB dye in the two membranes.

[0078] ln(Q.-Qt)-lnQe-k(1)

[0079]

[0080] Q e (mg / g) and Q t (mg / g) represents the adsorption capacity at equilibrium and at time t (h), respectively. k1 and k2 represent the pseudo-first-order kinetic rate constants (h). -1 ) and pseudo-second-order kinetic rate constant (g / (mg·h)).

[0081] The experimental data were fitted with pseudo-first-order and pseudo-second-order dynamic models to obtain... Figure 11 (a), (b) and Table 1. It can be found that the correlation coefficient of pseudo-first-order kinetics is higher than that of pseudo-second-order kinetics for both BCM and PMM, indicating that the pseudo-first-order kinetic equation is closer to the experimental results and can better represent the adsorption process of MB by BCM and PMM.

[0082] from Figure 11 As can be seen in (a), the slope of the BCM fitting curve is less than that of PMM, indicating that the adsorption rate constant k1 of BCM is less than that of PMM, which shows that the addition of microspheres improves the adsorption rate of the membrane.

[0083] In addition, the thermodynamics of the membrane was studied using two adsorption isotherm models, Langmuir and Freundlich, to explore its surface properties and adsorption mechanism.

[0084]

[0085]

[0086] Q e (mg / g) represents the amount of MB adsorbed by the sample at equilibrium. Q m (mg / g) indicates the maximum adsorption capacity of the sample. K L and K F These are the Langmuir and Freundlich adsorption constants, respectively. C e (mg / L) represents the mass concentration of the equilibrium solution. n is the dimensionless exponent of the Freundlich equation.

[0087] By fitting the Langmuir and Freundlich isotherm models, we obtain Figure 11 (c), (d) and Table 1. It can be observed that the correlation coefficient of the Freundlich isotherm model is higher than that of the Langmuir isotherm model for both BCM and PMM, indicating that the Freundlich isotherm model is closer to the experimental results and better represents the adsorption process of MB by BCM and PMM. The K0 of BCM... F Both n and n are less than the value of PMM, indicating that the adsorption performance of PMM is better than that of BCM, which further proves that the addition of microspheres is beneficial to the adsorption of MB solution by the membrane.

[0088] Table 1 shows the kinetics and isothermal adsorption modeling parameters of the PMM prepared in Example 4. From the results in Table 1, it can be seen that:

[0089] For both BCM and PMM, the correlation coefficients of pseudo-first-order kinetics are higher than those of pseudo-second-order kinetics, indicating that the pseudo-first-order kinetic equations are closer to the experimental results. The correlation coefficients of the Freundlich isotherm model are higher than those of the Langmuir isotherm model, indicating that the Freundlich isotherm model is closer to the experimental results. Both results demonstrate that the addition of microspheres is beneficial to the adsorption of MB solution by the membrane.

[0090] Table 1. Parameter results related to the kinetics and isothermal adsorption modeling of PMM prepared in Example 2.

[0091]

[0092] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features therein. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A device for integrated preparation of polymer microspheres based on microfluidic technology, comprising a microfluidic driving device, a phase separation device and a collection device, wherein the microfluidic driving device comprises a dispersed phase fluid propeller, a continuous phase fluid propeller and a collection phase fluid propeller; the phase separation device integrates a phase change channel, a continuous phase channel, a collection phase channel and a flow rate controller, and the continuous phase channel is connected to the flow rate controller; the collection device comprises a continuous phase collector and a microsphere collection device, and the microsphere collection device comprises a microsphere collector, a phase filter membrane and a collection phase collector; the dispersed phase fluid propeller, the continuous phase fluid propeller and the collection phase fluid propeller are connected to the inlet of the phase separation device; one end of the continuous phase channel is connected to one end of the flow rate controller, and the other end of the flow rate controller is connected to the continuous phase collector; and the outlet of the collection phase channel is connected to the microsphere collector; the phase change channel, the continuous phase channel and the collection phase channel in the phase separation device adopt an asymmetric layout, and the three channels are arranged in a single-channel structure and are not in the same horizontal plane, wherein the phase change channel is not fixed, but is determined based on the phase itself; when the density of the continuous phase fluid is greater than that of the collection phase fluid, the collection phase channel is closed, and the phase change channel is the collection phase channel; when the density of the collection phase fluid is greater than that of the continuous phase fluid, the continuous phase channel is closed, and the phase change channel is the continuous phase channel; and the flow rate controller is connected to the continuous phase channel.

2. The integrated polymer microsphere production device of claim 1, wherein The microspheres are polymer porous microspheres with a loading function, and the target load is loaded in the pore structure of the polymer porous microspheres; the particle size of the microspheres is 300-500 μm.

3. The integrated polymer microsphere production device of claim 2, wherein the polymerization reactor is a microfluidic device. The material of the polymer porous microspheres comprises one or more of hyaluronic acid and its derivatives, chondroitin sulfate and its derivatives, alginic acid and its derivatives, chitosan and its derivatives, and polylactic acid and its derivatives.

4. A method for preparing polymer microspheres based on the integrated polymer microsphere preparation device according to any one of claims 1 to 3, characterized in that The device comprises the following steps: S1: the continuous phase fluid and the dispersed phase fluid are separately injected into the microfluidic device for mixing, and polymer microspheres are obtained under the action of surface tension and shearing of the continuous phase fluid; the polymer microspheres are freeze-dried to obtain polymer porous microspheres; S2: the microspheres, the continuous phase fluid and the collection phase fluid are jointly injected into the phase separation device for mixing, and the polymer microspheres are separated from the continuous phase fluid and enter the collection phase fluid; when the density of the continuous phase fluid is greater than that of the collection phase fluid, the collection phase channel is closed, and the polymer microspheres enter the microsphere collector with the collection phase fluid from the phase change channel, i.e. the collection phase channel; and the continuous phase fluid is discharged from the continuous phase channel; when the density of the collection phase fluid is greater than that of the continuous phase fluid, the continuous phase channel is closed, and the polymer microspheres enter the microsphere collector with the collection phase fluid from the collection phase channel; and the continuous phase fluid is discharged from the phase change channel, i.e. the continuous phase channel.

5. The method of claim 4, wherein the polymer microspheres are prepared by a process comprising: The dispersed phase fluid comprises modified polymer, water-soluble salt, photocuring agent and inorganic solvent in a mass ratio of 0.8-1.8:0.15-0.3:0.4-0.6:160-200, wherein the inorganic solvent comprises water and phosphate buffered saline; the continuous phase fluid comprises ethyl acetate and dichloromethane; and the collection phase fluid comprises water and PBS.

6. The method of claim 5, wherein the polymer microspheres are prepared by a process comprising: The photocuring agent is lithium phenylphosphate, the water-soluble salt comprises one of ammonium selenite, sodium permanganate and potassium permanganate, and the modified polymer comprises one or more of hyaluronic acid and its derivatives, chondroitin sulfate and its derivatives, alginic acid and its derivatives, chitosan and its derivatives, and polylactic acid and its derivatives.

7. The method of claim 4, wherein the polymer microspheres are prepared by the process of claim 1 or 2. The injection speed of the dispersed phase fluid is 0.02-0.05 mL / min, the injection speed of the continuous phase fluid is 1.50-2.50 mL / min, and the injection speed of the collection phase fluid is 1.50-2.50 mL / min. 8.A polymer microsphere prepared by the method of any one of claims 4-7. 9.Use of the polymer microsphere of claim 8 in biomedicine and dye adsorption.

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