An automated production device and preparation method for porous polymer microspheres

Abstract

The present invention discloses an automated production device and preparation method for porous polymer microspheres. This production device includes: a sample pre-processing module, including a continuous phase circulation reactor and a discrete phase liquid storage tank; a microsphere generation module, including an assembly substrate, a continuous phase delivery pipeline, a discrete phase delivery pipeline, a buffer sleeve, a coaxial inner pipeline, and a coaxial outer pipeline; the assembly substrate is provided with a first card slot and a second card slot; a microsphere post-processing module, including a double T-shaped material bottle, a light source, a microsphere receiving container, and a vacuum pump. The device of the present invention generates and transports microspheres in coaxial microchannels through the negative pressure drive of a vacuum pump, solidifies and collects the microspheres during the delivery process, and simultaneously completes the filtration and recycling of the continuous phase solution, thereby realizing automated batch production of polymer microspheres; the system has a simple and reliable structure, and the required accessories are low in cost, which is conducive to the commercial application of polymer microspheres.

Description

Technical Field

[0001] The present invention relates to the field of utilization of porous polymer microspheres, and in particular to an automated production device and a preparation method for porous polymer microspheres. Background Art

[0002] Porous polymer microspheres are a type of spherical microparticles formed by the aggregation of natural and / or synthetic polymers. They have a diameter of 100-300 microns and have a three-dimensional spatial structure inside, which can provide effective space for cell growth and proliferation.

[0003] Stem cell therapy currently holds great promise for disease treatment. Stem cell therapy products have been clinically used for a variety of conditions, including Alzheimer's disease, myocardial infarction, and osteoarthritis. However, large-scale production and acquisition of high-quality stem cells to meet clinical application needs remain a significant challenge for the stem cell industry.

[0004] Currently, the primary method for stem cell culture still relies on traditional two-dimensional culture techniques in culture flasks. This involves directly seeding stem cells into culture flasks or dishes for adherent culture to achieve cell expansion. However, these methods suffer from shortcomings such as a limited cell growth area, large footprint of the cell culture vessels, significant variability in cell quality between batches, high labor and time consumption, susceptibility to contamination during the culture process, and a lack of automated control.

[0005] Suspension culture technology based on porous polymer microspheres involves seeding stem cells onto microspheres, which are then suspended to culture the cell-laden microspheres. This technology combines the advantages of both adherent and suspension cultures, facilitating cell expansion and collection, and can address the current difficulties and pain points in large-scale stem cell culture. Compared with traditional two-dimensional culture processes, porous polymer microsphere culture offers advantages such as a large effective surface area for cell growth, higher cell yield per unit volume of culture fluid, easier observation of cell growth, good functional reproducibility between cell batches, a simple cell harvesting process, automated control of the entire process, and reduced labor costs and the risk of cell contamination. It is one of the most promising large-scale stem cell culture technologies.

[0006] At present, there are more than a dozen microsphere products on the international market, among which the commonly used commercial microsphere carriers are: Cytodex1, 2, 3 (dextran matrix), Cytopore (cellulose matrix) and Cytoline (polyethylene matrix). However, these commercial microcarriers have disadvantages such as non-degradability, solid sphere structure resulting in reduced surface area, porous microspheres requiring pore-forming agents to prepare, or the need to add protein coating to promote cell adhesion. Patent CN113651989A discloses a 3D gelatin microslide (3DTableTrix), which is a porous microcarrier whose core component is animal-derived protein and formaldehyde crosslinker solidified into porous microspheres with an appearance of 100-200μm. The patented technology provides a method for preparing microcarrier particles, including allowing the dispersed phase liquid to pass through a porous plate at low temperature to form liquid microspheres in the continuous phase, and allowing the artificial synthetic polymers and / or natural biomacromolecules in the liquid microspheres to undergo a solidification reaction at low temperature to form particles. However, when the dispersed phase liquid passing through the porous plate forms droplet microspheres in the continuous phase, it is affected by the pressure drop in the upstream and downstream, as well as the flow field interference between adjacent droplets, resulting in uneven droplet diameters (particle size 50-500μm, uniformity ≤100μm). Therefore, an additional filtration and screening process is required, resulting in a reduced effective utilization rate of materials and reagents. In addition, patent CN113651989A uses toxic cross-linking agents such as formaldehyde, which is not conducive to the non-toxic control of the production process and has potential toxic effects on cell culture in the later stage; and the conditions described in patent CN113651989A require production at a low temperature below -10°C, which greatly increases energy consumption and production costs. Patent CN113801367A discloses a method of dissolving a free radical cross-linkable water-soluble polymer, a sacrificial component polymer, and an initiator in water to prepare a precursor solution; the above-mentioned precursor solution is added to the oil phase, stirred to form oil-in-water droplets, and then subjected to a photocrosslinking reaction, and post-treated to obtain porous hydrogel microspheres. Patent CN113801367A uses photocrosslinking technology to partially solve the problem of using toxic crosslinkers in the microsphere production process. However, this patent uses traditional stirred droplet generation technology to produce oil-in-water droplets, which has disadvantages such as uneven diameter of the resulting microspheres, cumbersome procedures, and low efficiency. Summary of the Invention

[0007] In order to overcome the problems of uneven particle size and low efficiency of porous microspheres prepared in the prior art, one of the objects of the present invention is to provide an automated production device for porous polymer microspheres, a second object of the present invention is to provide a method for preparing porous polymer microspheres, and a third object of the present invention is to provide porous polymer microspheres.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is:

[0009] The first aspect of the present invention provides an automated production device for porous polymer microspheres, comprising:

[0010] A sample pre-processing module, comprising a continuous phase circulation reactor and a discrete phase liquid storage tank;

[0011] A microsphere generation module is provided inside the continuous phase circulation reactor, and includes an assembly base, a continuous phase delivery pipe, a discrete phase delivery pipe, a buffer sleeve, a coaxial inner pipe, and a coaxial outer pipe; the assembly base is provided with a first card slot and a second card slot;

[0012] The discrete phase delivery pipeline is connected to the discrete phase liquid storage tank; the output end of the continuous phase delivery pipeline is embedded in the input end of the buffer sleeve, and the input end of the coaxial outer pipeline is embedded in the output end of the buffer sleeve, and the first clamping groove fixes the buffer sleeve; the input end of the coaxial inner pipeline is embedded in the output end of the discrete phase delivery pipeline, and the output end of the coaxial inner pipeline is embedded in the input end of the coaxial outer pipeline; the second clamping groove fixes the coaxial outer pipeline and the coaxial inner pipeline;

[0013] A microsphere post-processing module includes a double T-shaped material bottle, a light source, a microsphere receiving container and a vacuum pump; the coaxial outer pipe is connected to the double T-shaped material bottle, and the vacuum pump is connected to the double T-shaped material bottle.

[0014] Preferably, in this automated production device for porous polymer microspheres, the discrete phase liquid storage tank is arranged inside the continuous phase circulation reactor; the discrete phase liquid storage tanks are all wrapped in the continuous phase solution. As long as the temperature of the continuous phase is controlled, the overall ambient temperature of the system can be kept consistent, which can improve the uniformity of the microsphere size.

[0015] Preferably, in this automated production device for porous polymer microspheres, the materials of the continuous phase conveying pipe, the discrete phase conveying pipe, the buffer sleeve, and the coaxial outer pipe are each individually selected from commercial elastic pipes; the material of the coaxial inner pipe is selected from commercial rigid pipes; further preferably, the materials of the continuous phase conveying pipe, the discrete phase conveying pipe, the buffer sleeve, and the coaxial outer pipe are each individually selected from one of polyimide, polyetheretherketone, silicone rubber, polypropylene, polyvinyl chloride, polycarbonate, polytetrafluoroethylene, fluorinated ethylene propylene copolymer, and polyurethane; the material of the coaxial inner pipe is selected from one of quartz glass, stainless steel, polyetheretherketone, polypropylene, and polycarbonate.

[0016] Preferably, in the automated production device for porous polymer microspheres, the inner diameter of the coaxial inner pipe is 0.05 mm to 1 mm; and the wall thickness of the coaxial inner pipe is 0.01 mm to 0.1 mm.

[0017] Preferably, in the automated production device for porous polymer microspheres, the inner diameter of the coaxial outer tube is 0.1 mm to 2 mm; and the wall thickness of the coaxial outer tube is 0.01 mm to 4 mm.

[0018] Preferably, in this automated production device for porous polymer microspheres, the spacing of the second slots is smaller than the outer diameter of the coaxial outer pipe. The device of the present invention controls the spacing of the second slots so that the spacing of the second slots is smaller than the outer diameter of the coaxial outer pipe, squeezes the coaxial outer pipe to produce deformation, and keeps the inner and outer pipes coaxial within the range of the second slots, which can effectively improve the stability of the droplet size, eliminates the need for additional filtering and screening processes, and further saves time and cost.

[0019] Preferably, in this automated production device for porous polymer microspheres, a potting compound is used to seal the pipeline after the pipeline is set up; further preferably, the potting compound includes one of organic silica gel, ethylene-vinyl acetate copolymer, polyacrylate, epoxy resin, and special glue for a specific material; the viscosity of the potting compound is adjusted to reduce its fluidity in the micro-pipeline, so that the pipeline connection port is sealed without blocking the internal flow channel.

[0020] A second aspect of the present invention provides a method for preparing porous polymer microspheres, which uses the above-mentioned automated production device for porous polymer microspheres to produce porous polymer microspheres, and specifically comprises the following steps:

[0021] 1) Turn on the vacuum pump, the discrete phase solution in the discrete phase storage tank passes through the discrete phase delivery pipe and then enters the coaxial inner pipe; the continuous phase solution in the continuous phase circulation reactor passes through the continuous phase delivery pipe and then enters the buffer sleeve and the coaxial outer pipe; microspheres are formed at the outlet of the coaxial inner pipe;

[0022] 2) turning on a light source for curing to obtain the porous polymer microspheres;

[0023] The discrete phase solution includes artificial synthetic polymers and / or natural biomacromolecules, and a curing agent;

[0024] The continuous phase solution includes an organic solvent and a non-ionic surfactant.

[0025] The method for preparing porous polymer microspheres of the present invention utilizes the principle of photocuring hydrogels and combines it with coaxial microchannel-type microfluidic droplet generation technology to prepare porous polymer microspheres. The present invention mainly uses natural and / or synthetic polymers modified with acryloyl or methacryloyl groups, mixed with a photoinitiator as the discrete phase, and an oily solvent containing an emulsifier as the continuous phase. Through negative or positive pressure driving, the discrete phase and the continuous phase are passed through the droplet generation microfluidic device designed by the present invention at a certain flow ratio to form droplets of uniform size and controllable size. Finally, the droplets are cross-linked under photocuring to spontaneously form porous polymer microspheres.

[0026] Preferably, in this preparation method, the synthetic polymer is selected from water-soluble polymers that can be cross-linked by free radicals, including acryloyl or methacryloyl modified products of polyglutamic acid, acryloyl or methacryloyl modified products of polylysine, acryloyl or methacryloyl modified products of hydroxymethyl cellulose, acryloyl or methacryloyl modified products of polyethylene glycol, acryloyl or methacryloyl modified products of polyethylene glycol derivatives, acryloyl or methacryloyl modified products of polyvinyl alcohol, acryloyl or methacryloyl modified products of polyethylene glycol diacrylate, acryloyl or methacryloyl modified products of polyacrylamide, or at least one of methacryloyl-modified products, acryloyl- or methacryloyl-modified products of polylactic acid, acryloyl- or methacryloyl-modified products of polyhydroxy acid, acryloyl- or methacryloyl-modified products of polylactic acid alkyd copolymer, acryloyl- or methacryloyl-modified products of polyanhydride, acryloyl- or methacryloyl-modified products of polyamide, acryloyl- or methacryloyl-modified products of polyamino acid, acryloyl- or methacryloyl-modified products of polyacetal, acryloyl- or methacryloyl-modified products of polyethylene, and acryloyl- or methacryloyl-modified products of polyethylene oxide.

[0027] Preferably, in this preparation method, the natural biomacromolecules include acryloyl or methacryloyl modified gelatin, acryloyl or methacryloyl modified collagen, acryloyl or methacryloyl modified dextran, acryloyl or methacryloyl modified silk fibroin, acryloyl or methacryloyl modified proteoglycan, acryloyl or methacryloyl modified glycoprotein, acryloyl or methacryloyl modified chitosan, acryloyl or methacryloyl modified alginic acid, at least one of an acryloyl- or methacryloyl-modified product of acryloyl- or methacryloyl-modified product of agar, an acryloyl- or methacryloyl-modified product of fibrinogen, an acryloyl- or methacryloyl-modified product of extracellular matrix, an acryloyl- or methacryloyl-modified product of hyaluronic acid, an acryloyl- or methacryloyl-modified product of chondroitin sulfate, an acryloyl- or methacryloyl-modified product of laminin, and an acryloyl- or methacryloyl-modified product of fibronectin.

[0028] Preferably, in this preparation method, the curing agent includes at least one of phenyl-2,4,6-trimethylbenzoyl lithium phosphite (LAP), 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone (Irgacure 2959) and a free radical photoinitiator, and the free radical photoinitiator can be at least one of a free radical type I photoinitiator and a free radical type II photoinitiator.

[0029] Preferably, in this preparation method, the curing agent is first added to a PBS solution to prepare a curing agent solution, and then the artificial synthetic polymer and / or natural biological macromolecule is added to the curing agent solution to prepare a discrete phase solution; further preferably, the curing agent concentration in the curing agent solution is 0.25%-4% (w / v), and the mass volume ratio of the artificial synthetic polymer and / or natural biological macromolecule to the curing agent solution is 1g:(8-12)mL.

[0030] Preferably, in this preparation method, the curing method is: light curing, and the wavelength of the light source of the light curing is 359-405nm.

[0031] Preferably, in this preparation method, the organic solvent includes at least one of paraffin oil, vegetable oil, mineral oil, fluorinated oil, silicone oil, olive oil, soybean oil, dichloromethane, petroleum ether, cyclohexane and tetrachloroethylene.

[0032] Preferably, in this preparation method, the nonionic surfactant includes at least one of ethylphenyl polyethylene glycol, sorbitan fatty acid ester, fatty acid glyceride, lauric acid ester, alkylphenol polyoxyethylene ether, high carbon fatty alcohol polyoxyethylene ether, Span, PO-500, monooleate and Teflon.

[0033] Preferably, in this preparation method, the volume ratio of the organic solvent to the non-ionic surfactant in the continuous phase solution is (5-20):1.

[0034] Preferably, in this preparation method, the temperature of the continuous phase solution is controlled at 35-55°C; further preferably, the temperature of the continuous phase solution is controlled at 40°C.

[0035] Preferably, in this preparation method, the temperature of the discrete phase solution is controlled at 35-55°C; further preferably, the temperature of the discrete phase solution is controlled at 40°C.

[0036] Preferably, in this preparation method, the organic solvent and the nonionic surfactant are mixed and stirred to obtain a continuous phase solution; further preferably, the stirring speed is 1600-2000 rpm, and the stirring time is 0.5-2h; even more preferably, the stirring speed is 1700-1900 rpm, and the stirring time is 2h.

[0037] Preferably, the preparation method further comprises the step of washing the porous polymer microspheres; further preferably, the cleaning agent used for washing comprises at least one of n-hexadecane, acetone, Tween, ethanol and deionized water.

[0038] The third aspect of the present invention provides a porous polymer microsphere prepared by the above-mentioned preparation method.

[0039] The beneficial effects of the present invention are:

[0040] The present invention uses the negative pressure drive of a vacuum pump to generate microspheres from a continuous phase solution and a discrete phase solution in a coaxial microchannel, and crosslinks and solidifies the microspheres during transportation, thereby realizing automated mass production of porous polymer microspheres. The system has a simple and reliable structure and requires low-cost accessories, which is conducive to the commercial application of polymer microspheres. The final size and yield of the polymer microspheres are collaboratively controlled by regulating multiple parameters such as driving pressure, channel structure (channel inner diameter, wall thickness), and external load (forced deformation applied to the external channel, including pressure and displacement).

[0041] For temperature-sensitive polymer materials, changes in viscosity can lead to larger size deviations in the generated microspheres when the ambient temperature fluctuates (for example, when the solution is transported over long distances and the ambient temperature varies between the fluid and air domains). The automated porous polymer microsphere production device of the present invention encloses both the discrete phase reservoir and the microsphere generation module in a continuous phase solution. By simply controlling the temperature of the continuous phase, the ambient temperature of the entire system can be maintained consistent, thereby improving the uniformity of microsphere size. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 This is a schematic diagram of the functional modules of the automated production device for porous polymer microspheres;

[0043] Figure 2 This is a schematic diagram of the structure of the microsphere generation module in the automated production device for porous polymer microspheres;

[0044] Figure 3 Provide a cross-sectional view of the pipelines of the microsphere generation module in the automated production device for porous polymer microspheres;

[0045] Figure 4 This is a schematic diagram of the coaxial pipeline cross section of the microsphere generation module in the automated production device for porous polymer microspheres;

[0046] Figure 5 A view of the porous polymer microspheres prepared in the embodiment in an aqueous solution;

[0047] Figure 6 This is a scanning electron micrograph of the freeze-dried porous polymer microspheres prepared in Example;

[0048] Attachment Figure 1-4 mark:

[0049] Sample pre-processing module 100, continuous phase circulation reactor 110, continuous phase solution 111, discrete phase liquid storage tank 120, discrete phase solution 121, microsphere generation module 200, assembly substrate 210, continuous phase delivery pipeline 220, discrete phase delivery pipeline 230, buffer sleeve 240, coaxial inner pipeline 250, coaxial outer pipeline 260, first card slot 211, second card slot 212, microsphere post-processing module 300, double T-shaped material bottle 310, filter 311, sealing plug 312, valve 313, light source 320, microsphere receiving container 330, vacuum pump 340, porous polymer microspheres 350. DETAILED DESCRIPTION

[0050] The embodiments of the present invention are described in detail below. The embodiments described with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and should not be understood as limiting the present invention.

[0051] The present invention is further described in detail below through specific examples.

[0052] In the description of the present invention, it should be noted that, unless otherwise expressly specified or limited, the terms "connected" and "connection" should be understood in a broad sense. For example, they can refer to fixed connection, detachable connection, or integral connection; mechanical connection, electrical connection; direct connection, indirect connection through an intermediate medium, or internal communication between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0053] Unless otherwise specified, the raw materials, reagents, or devices used in the examples can be obtained from conventional commercial sources or by existing methods. Unless otherwise specified, the experiments or testing methods are conventional methods in the art.

[0054] like Figure 1-4 As shown, the automated production device for porous polymer microspheres of the present invention includes a sample pre-processing module 100 , a microsphere generating module 200 , and a microsphere post-processing module 300 .

[0055] The sample pre-processing module 100 includes a continuous phase circulation reactor 110 and a discrete phase liquid storage tank 120;

[0056] The microsphere generation module 200 includes an assembly base 210, a continuous phase delivery pipe 220, a discrete phase delivery pipe 230, a buffer sleeve 240, a coaxial inner pipe 250, and a coaxial outer pipe 260; the assembly base 210 is provided with a first card slot 211 and a second card slot 212;

[0057] Among them, the assembly base 210 is used to set the continuous phase conveying pipe 220, the discrete phase conveying pipe 230, the buffer sleeve 240, the coaxial inner pipe 250, and the coaxial outer pipe 260; the first card groove 211 is used to fix the buffer sleeve 240; the output end of the continuous phase conveying pipe 220 is embedded in the input end of the buffer sleeve 240, and the input end of the coaxial outer pipe 260 is embedded in the output end of the buffer sleeve 240; the input end of the coaxial inner pipe 250 is embedded in the output end of the discrete phase conveying pipe 230, and the output end of the coaxial inner pipe 250 is embedded in the input end of the coaxial outer pipe 260; a second card groove 212 is provided on one side of the assembly base 210, and the spacing of the second card groove 212 is smaller than the outer diameter of the coaxial outer pipe 260, which is used to squeeze the coaxial outer pipe 260 to produce deformation, so that the inner and outer pipes remain coaxial within the range of the second card groove 212. After the above pipelines are set up, they are sealed with potting glue. The viscosity of the potting glue is adjusted to reduce its fluidity in the micro-pipeline, so that the pipeline connection port is sealed without blocking the internal flow channel.

[0058] The microsphere post-processing module 300 includes a double T-shaped material bottle 310 , a light source 320 , a microsphere receiving container 330 , and a vacuum pump 340 .

[0059] The double T-shaped material bottle 310 includes a filter 311 , a sealing plug 312 , and a valve 313 .

[0060] The microsphere post-processing module 300 further includes a microsphere delivery pipeline, the input end of which is connected to the output end of the coaxial outer pipe 260. The output end of the microsphere delivery pipeline passes through the sealing plug 312 of the double T-shaped material bottle 310, allowing the cured or to-be-cured polymer microspheres to freely settle to the bottom of the container in the oil phase. The microsphere post-processing module 300 also includes an exhaust pipeline, which also passes through the sealing plug 312 of the double T-shaped material bottle 310 and is connected to a vacuum pump 340. The light source 320 is a UV light source, located outside the double T-shaped material bottle 310, and is used to irradiate the generated microspheres, causing the polymer inside the droplets to undergo a cross-linking reaction and solidify into porous solid particles. The vacuum pump 340 provides a power source for the system, causing the atmospheric pressure of the external environment of the system to drive the oil phase and water phase in the sample pre-processing module 100 into the microsphere generation module 200 and generate microspheres at the intersection of the coaxial pipes. The generation rate and size of the microspheres are controlled by adjusting the negative pressure inside the double T-shaped material bottle 310.

[0061] Reference below Figure 1-4 The method for using the automated production device for porous polymer microspheres according to an embodiment of the present invention is described.

[0062] A continuous phase circulation reactor 110 is used to prepare a continuous phase solution 111, and a discrete phase liquid storage tank 120 is used to prepare a discrete phase solution 121. The vacuum pump 340 is turned on, and the continuous phase solution 111 and the discrete phase solution 121 are automatically driven by the pressure difference between the inside and outside of the system to flow through the microsphere generation module 200, and microspheres are generated in the coaxial pipe. The light source 320 is turned on, and the microspheres and the continuous phase flow from the outlet of the microsphere delivery pipeline connected to the coaxial outer pipe 260 into the first material bottle of the double T-shaped material bottle 310. The microspheres solidify and accumulate at the bottom of the bottle during sedimentation. Due to the large proportion of the continuous phase, when the liquid level exceeds the T-shaped connecting port, it will pass through the filter 311 and be stored in the material bottle on the other side. When the microsphere accumulation height reaches near the T-shaped connecting port, the batch production is terminated. The valve 313 at the bottom of the material bottle is opened, and the microspheres are collected in the microsphere receiving container 330. The filtered continuous phase solution flows into the reactor for recycling.

[0063] Application Examples

[0064] This embodiment provides a method for preparing porous polymer microspheres, comprising the following steps:

[0065] 1) Place the cleaned, dried, and sterilized continuous phase circulating reactor on a constant temperature stirring device. Add 9 mL of paraffin oil and 1 mL of surfactant to the continuous phase circulating reactor. Set the temperature to 40°C and the stirrer speed to 1800 rpm. Stir for 2 hours to prepare an oil phase solution.

[0066] 2) Weigh 1 g of methacrylated gelatin and place it into a cleaned, dried, and sterilized discrete phase reservoir. Add 10 mL of PBS solution containing 0.25% (w / v) LAP photoinitiator to the container and dissolve at 40°C for 30 minutes to prepare an aqueous phase solution.

[0067] 3) Fix the microsphere generation module below the liquid level inside the continuous phase circulation reactor. The coaxial inner pipe of the microsphere generation module has an inner diameter of 0.1mm and a wall thickness of 0.1mm. The coaxial outer pipe has an inner diameter of 0.3mm and a wall thickness of 0.3mm. The first slot spacing is 4mm, and the second slot spacing is 0.5mm. Immerse the input end of the continuous phase delivery pipe in the prepared continuous phase solution; connect the input end of the discrete phase delivery pipe to the discrete phase liquid storage tank, and immerse the bottom of the discrete phase liquid storage tank below the continuous phase liquid level;

[0068] 4) Place a double T-shaped material bottle connected by a filter in the middle into the continuous phase solution. Connect the two sealing plugs at the inlet of the double T-shaped material bottle to the microsphere delivery line and the exhaust line, respectively. Connect the exhaust line to a vacuum pump. Keep the two valves at the outlet of the double T-shaped material bottle closed, and place a 5 mL glass dish as a microsphere receiving container at the outlet of the double T-shaped material bottle;

[0069] 5) Turn on the vacuum pump and adjust the pressure regulating valve to stabilize the pressure at -10 kPa. Then, open the air inlet valve of the gas pipeline to allow the continuous phase and dispersed phase solutions to automatically flow through the microsphere generation module driven by the pressure difference between the inside and outside of the system, and generate microspheres in the coaxial pipeline.

[0070] 6) Turn on the ultraviolet light source of the light curing device and adjust the angle so that it irradiates the outlet of the microsphere delivery pipe in the material bottle, so that the microspheres are free to settle in the continuous phase, complete crosslinking, and solidify into porous polymer microspheres;

[0071] 7) After the system has been running for 24 hours, close the exhaust line valve and the vacuum pump, then open the two valves at the outlet of the double T-shaped material bottle to allow the solidified porous polymer microspheres to fall into the microsphere receiving container. At the same time, the filtered continuous phase solution enters the continuous phase circulation reactor for continued recycling;

[0072] 8) The collected microspheres were eluted with n-hexadecane, Tween and deionized water to obtain purified polymer microspheres. The diameter of the microspheres was measured by optical microscopy to be 266.52±2.08 μm, with a CV value of 0.78% and a range of 4.84 μm. Figure 5 As shown;

[0073] 9) Place the eluted porous polymer microspheres in a freeze dryer at -80°C for 72 hours;

[0074] 10) The freeze-dried porous microspheres were observed under a scanning electron microscope. It was found that the freeze-dried microspheres had a certain degree of shrinkage and deformation. The average particle size of the freeze-dried porous polymer microspheres was 150±5μm, and the average pore size was about 20-40μm. The electron microscope image of the freeze-dried porous microspheres is shown in Figure 1. Figure 6 shown.

[0075] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention are intended to be within the scope of protection of the present invention.

Claims

1. An automated production device for porous polymer microspheres, characterized in that: include: A sample pre-processing module (100), comprising a continuous phase circulation reactor (110) and a discrete phase liquid storage tank (120); A microsphere generation module (200) is provided inside the continuous phase circulation reactor (110), and comprises an assembly base (210), a continuous phase delivery pipe (220), a discrete phase delivery pipe (230), a buffer sleeve (240), a coaxial inner pipe (250), and a coaxial outer pipe (260); the assembly base (210) is provided with a first card slot (211) and a second card slot (212); The discrete phase delivery pipeline (230) is connected to the discrete phase liquid storage tank (120); the output end of the continuous phase delivery pipeline (220) is embedded in the input end of the buffer sleeve (240), the input end of the coaxial outer pipeline (260) is embedded in the output end of the buffer sleeve (240), and the first clamping groove (211) fixes the buffer sleeve (240); the input end of the coaxial inner pipeline (250) is embedded in the output end of the discrete phase delivery pipeline (230), and the output end of the coaxial inner pipeline (250) is embedded in the input end of the coaxial outer pipeline (260); the second clamping groove (212) fixes the coaxial outer pipeline (260) and the coaxial inner pipeline (250); A microsphere post-processing module (300), comprising a double T-shaped material bottle (310), a light source (320), a microsphere receiving container (330), a vacuum pump (340), and a microsphere transport pipeline and an exhaust pipeline; The double T-shaped material bottle (310) comprises a filter (311), a sealing plug (312), a valve (313), and a first material bottle and a material bottle on the other side located on both sides of the filter (311). The two material bottles are interconnected and have the same sealing plug (312) on the top. The valve (313) is provided below the first material bottle. The input end of the microsphere delivery pipeline is connected to the output end of the coaxial outer pipeline (260); the output end of the microsphere delivery pipeline passes through the sealing plug (312) of the double T-shaped material bottle (310) and is inserted into the first material bottle of the double T-shaped material bottle (310); one end of the exhaust pipeline is connected to the vacuum pump (340), and the other end passes through the sealing plug (312) of the double T-shaped material bottle (310) and is inserted into the other side material bottle of the double T-shaped material bottle (310).

2. The automated production device for porous polymer microspheres according to claim 1, characterized in that: The materials of the continuous phase conveying pipe (220), the discrete phase conveying pipe (230), the buffer sleeve (240), and the coaxial outer pipe (260) are each selected from commercial elastic pipes; the material of the coaxial inner pipe (250) is selected from commercial rigid pipes.

3. The automated production device for porous polymer microspheres according to claim 1, characterized in that: The inner diameter of the coaxial inner pipe (250) is 0.05 mm to 1 mm; the wall thickness of the coaxial inner pipe (250) is 0.01 mm to 0.1 mm.

4. The automated production device for porous polymer microspheres according to claim 1, characterized in that: The inner diameter of the coaxial outer pipe (260) is 0.1 mm to 2 mm; the wall thickness of the coaxial outer pipe (260) is 0.01 mm to 4 mm.

5. A method for preparing porous polymer microspheres, characterized in that: The porous polymer microspheres are produced using the automated production device for porous polymer microspheres according to any one of claims 1 to 4, specifically comprising the following steps: 1) Turn on the vacuum pump, the discrete phase solution in the discrete phase storage tank passes through the discrete phase delivery pipe and then enters the coaxial inner pipe; the continuous phase solution in the continuous phase circulation reactor passes through the continuous phase delivery pipe and then enters the buffer sleeve and the coaxial outer pipe; microspheres are formed at the outlet of the coaxial inner pipe; 2) turning on a light source for curing to obtain the porous polymer microspheres; The discrete phase solution includes artificial synthetic polymers and / or natural biomacromolecules, and a curing agent; The continuous phase solution includes an organic solvent and a non-ionic surfactant.

6. The preparation method according to claim 5, characterized in that The synthetic polymer is selected from water-soluble polymers that can be cross-linked by free radicals, including acryloyl or methacryloyl modified products of polyglutamic acid, acryloyl or methacryloyl modified products of polylysine, acryloyl or methacryloyl modified products of hydroxymethyl cellulose, acryloyl or methacryloyl modified products of polyethylene glycol, acryloyl or methacryloyl modified products of polyethylene glycol derivatives, acryloyl or methacryloyl modified products of polyvinyl alcohol, acryloyl or methacryloyl modified products of polyethylene glycol diacrylate, acryloyl or methacryloyl modified products of polyacrylamide, At least one of acyl-modified products, acryloyl- or methacryloyl-modified products of polylactic acid, acryloyl- or methacryloyl-modified products of polyhydroxy acid, acryloyl- or methacryloyl-modified products of polylactic acid alkyd copolymer, acryloyl- or methacryloyl-modified products of polyanhydride, acryloyl- or methacryloyl-modified products of polyamide, acryloyl- or methacryloyl-modified products of polyamino acid, acryloyl- or methacryloyl-modified products of polyacetal, acryloyl- or methacryloyl-modified products of polyethylene, and acryloyl- or methacryloyl-modified products of polyethylene oxide.

7. The preparation method according to claim 5, characterized in that The natural biomacromolecules include at least one of acryloyl or methacryloyl-modified gelatin, acryloyl or methacryloyl-modified collagen, acryloyl or methacryloyl-modified dextran, acryloyl or methacryloyl-modified silk fibroin, acryloyl or methacryloyl-modified proteoglycan, acryloyl or methacryloyl-modified glycoprotein, acryloyl or methacryloyl-modified chitosan, acryloyl or methacryloyl-modified alginate, acryloyl or methacryloyl-modified agar, acryloyl or methacryloyl-modified fibrinogen, acryloyl or methacryloyl-modified extracellular matrix, acryloyl or methacryloyl-modified hyaluronic acid, acryloyl or methacryloyl-modified chondroitin sulfate, acryloyl or methacryloyl-modified laminin, and acryloyl or methacryloyl-modified fibronectin.

8. The preparation method according to claim 5, characterized in that The organic solvent includes at least one of paraffin oil, vegetable oil, mineral oil, fluorinated oil, silicone oil, olive oil, soybean oil, dichloromethane, petroleum ether, cyclohexane and tetrachloroethylene.

9. The preparation method according to claim 5, characterized in that The volume ratio of the organic solvent to the non-ionic surfactant in the continuous phase solution is (5-20):1.

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