CN116535728B - Porous polymer microsphere and preparation method thereof - Google Patents
Porous polymer microsphere and preparation method thereof Download PDFInfo
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- CN116535728B CN116535728B CN202310516758.XA CN202310516758A CN116535728B CN 116535728 B CN116535728 B CN 116535728B CN 202310516758 A CN202310516758 A CN 202310516758A CN 116535728 B CN116535728 B CN 116535728B
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- microspheres
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- 239000004005 microsphere Substances 0.000 title claims abstract description 137
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- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims abstract description 38
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- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 description 2
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Classifications
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
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- C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
- C08J9/26—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof by elimination of a solid phase from a macromolecular composition or article, e.g. leaching out
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- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
- A61K9/1605—Excipients; Inactive ingredients
- A61K9/1629—Organic macromolecular compounds
- A61K9/1641—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poloxamers
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- A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
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- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
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- C08J2405/04—Alginic acid; Derivatives thereof
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2429/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an alcohol, ether, aldehydo, ketonic, acetal, or ketal radical; Hydrolysed polymers of esters of unsaturated alcohols with saturated carboxylic acids; Derivatives of such polymer
- C08J2429/02—Homopolymers or copolymers of unsaturated alcohols
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- C—CHEMISTRY; METALLURGY
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- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2487/00—Characterised by the use of unspecified macromolecular compounds, obtained otherwise than by polymerisation reactions only involving unsaturated carbon-to-carbon bonds
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- Chemical & Material Sciences (AREA)
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- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
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Abstract
The invention provides a porous polymer microsphere and a preparation method thereof, and relates to the technical field of polymer materials. The microsphere is prepared by taking the micro-fluidic chip with the same-direction structure as a main micro-fluidic device, the size of the microsphere can be unified by the method, the size of the microsphere can be controlled by adjusting the flow rate ratio (emulsion/continuous phase), the applicability is stronger, and the microsphere preparation effect is better. The microsphere prepared by the method takes polyethylene glycol as a framework, hyaluronic acid as a monomer, forms a crosslinking substance together, takes emulsion formed by the macromolecular substance as an oil phase, takes sodium alginate solution as an internal water phase, and takes polyvinyl alcohol solution as a continuous phase to form the porous polymer microsphere with higher biocompatibility and cell compatibility in a matching way, so that the porous polymer microsphere can be used in the fields of plastic and cosmetic filling implants, drug carriers, cell culture scaffolds, tissue engineering, regenerative medicine, organoids and the like, and has higher use value.
Description
Technical Field
The invention relates to the technical field of high polymer materials, in particular to a porous high polymer microsphere and a preparation method thereof.
Background
The porous material is a material with a large number of gaps in the structure, and has the characteristics of low density, light weight, large specific surface area and the like compared with the common dense material. And can be classified into three types according to pore size: micropores with the pore diameter smaller than 2nm, mesopores with the pore diameter between 2nm and 50nm, and macropores with the pore diameter larger than 50 nm. Wherein, the micropore and mesoporous materials have high specific surface area and narrow aperture, and are widely applied in the fields of catalysis, adsorption separation, energy storage and the like.
Based on this, the porous microsphere is used in the biomedical field because it has biodegradability, biocompatibility, and cell compatibility, and is harmless to the human body.
Disclosure of Invention
The invention aims to provide a preparation method of porous polymer microspheres, which takes a microfluidic chip with a homodromous structure as a core to prepare microspheres with uniform particle size and smaller particle size, and can achieve the effect of controlling the size of the microspheres by adjusting the flow rate ratio of emulsion/continuous phase to phase, so that the preparation method has high use value.
Another object of the present invention is to provide a porous polymeric microsphere which has a good compatibility with cells and does not affect the growth metabolism of the cells.
The invention solves the technical problems by adopting the following technical scheme.
On one hand, the invention provides a preparation method of porous polymer microspheres, which mainly comprises the following steps:
mixing hyaluronic acid and salts thereof with polyethylene glycol, and heating in water bath to obtain oil phase; preparing sodium alginate solution by using water as a solvent to prepare an inner water phase; mixing the oil phase with the inner water phase to obtain emulsion; preparing a polyvinyl alcohol solution by taking water as a solvent to prepare a continuous phase;
pumping the emulsion and the continuous phase into a microfluidic device respectively, assembling a microfluidic chip with a homodromous structure in the microfluidic device to prepare microspheres, and then placing the microspheres in alkali liquor for soaking to prepare the porous microspheres.
In another aspect, the present invention provides a porous polymeric microsphere prepared by the above method.
The porous polymer microsphere and the preparation method thereof have at least the following beneficial effects:
on the one hand, in the application, the porous polymer microsphere is prepared by taking the microfluidic chip with the homodromous structure as a main microfluidic device, so that the sizes of the microspheres are unified, the sizes of the microspheres can be controlled by adjusting the flow rate ratio (emulsion/continuous phase), the applicability is higher, and meanwhile, the microsphere preparation effect is better.
On the other hand, the prepared porous polymer microsphere takes polyethylene glycol as a grafting framework, hyaluronic acid and salts thereof as grafting monomers, macromolecular substances are formed together, emulsion formed by the macromolecular substances is taken as an oil phase, sodium alginate solution is taken as an internal water phase, polyvinyl alcohol solution is taken as a continuous phase, and the porous polymer microsphere with higher biocompatibility and cell compatibility is formed by matching, and has no toxic effect on cell activity, so that the porous polymer microsphere can be used in the fields of plastic and cosmetic filling implants, drug carriers, cell culture scaffolds, tissue engineering, regenerative medicine, organoids and the like, and has higher use value.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a microfluidic chip used in an embodiment of the present invention;
FIG. 2 is a graph showing the percentage of microspheres of different particle sizes in example 1;
FIG. 3 is a graph showing the percentage of microspheres of different particle sizes in example 2
FIG. 4 is a graph showing the percentage of microspheres of different particle sizes in example 3
FIG. 5 is a graph showing the percentage of microspheres of different sizes in example 4
FIG. 6 is a graph showing the percentage of microspheres of different sizes in example 5
FIG. 7 is a graph showing the growth of cells in effect example 3;
FIG. 8 is a graph showing the comparison of the cell proliferation rate in effect example 3.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
It should be noted that, in the case of no conflict, the embodiments and features in the embodiments may be combined with each other. The present invention will be described in detail with reference to specific examples.
In the application, the preparation process of the porous polymer microsphere mainly adopts the following raw materials: polyvinyl alcohol solution, hyaluronic acid, sodium alginate solution, polyethylene glycol and acetic acid solution. The specific effects of the raw materials are as follows:
polyvinyl alcohol: polyvinyl alcohol is a high molecular organic matter with higher safety, is nontoxic to human body, has no side effect, and has good biocompatibility. Polyvinyl alcohol is used as one of the raw materials for preparing the porous polymer microsphere, and is mainly because the polyvinyl alcohol is hardly dissolved in an organic solvent, is not compatible with polyethylene glycol and is compatible with water, so that the polyvinyl alcohol is selected as a continuous phase to prepare the microsphere. Specifically, the continuous phase has better water solubility, and interfacial tension and shearing force are generated when the continuous phase is contacted with the oil phase substance in the application, so that emulsion drops with uniform size are formed, and the microspheres with uniform size are prepared, so that the expected effect is achieved.
It should be noted that in the present application, the continuous phase is polyvinyl alcohol, and the collecting phase is polyvinyl alcohol, that is, when forming emulsion droplets with uniform size, the emulsion droplets flow into the collecting phase along with the liquid, and precipitate to form microspheres, so as to achieve the desired effect.
Hyaluronic acid and salts thereof: hyaluronic acid, also known as hyaluronic acid, is a disaccharide unit glycosaminoglycan composed of D-glucuronic acid and N-acetylglucosamine. Because it belongs to high-grade polysaccharide, it has unique molecular structure, so that it has good characteristics in body, such as lubricating joint, regulating vascular wall permeability, regulating protein and promoting wound healing, etc.. On the other hand, hyaluronic acid stabilizes intercellular fiber and membrane protein structures, has good viscoelasticity, lubricity, biocompatibility, and can promote drug absorption, and thus, is used in surgery, drug carriers, and cosmetics.
The hyaluronate has the characteristic of easy dissolution in water, and when the hyaluronate is used as a raw material, the hyaluronate can be used as a solvent to participate in the reaction, so that the preparation effect is good.
However, hyaluronic acid has a short shelf life and is easily degraded because it has poor stability and strong sensitivity to strong acids, strong bases, etc.
Therefore, in the application, the hyaluronic acid is taken as a grafting monomer, so that the performance of the hyaluronic acid is improved, and the microsphere has better acid and alkali resistance and better biocompatibility and cell compatibility.
Sodium alginate: the sodium alginate is a polymer formed by connecting beta-D-mannuronic acid and alpha-L-Gu Luotang aldehyde acid, is a natural polysaccharide, and has stability, solubility, viscosity and safety.
In this application, regard as one of the raw materials with sodium alginate, form the emulsion droplet together with polyethylene glycol, hyaluronic acid to when separating out and forming the microballon, the accessible alkali lye dissolves the mode, dissolves out the sodium alginate and gets rid of, thereby make the microballon inside have the aperture, and the structure of microballon is 3D network structure and distributes, and then prepares a porous polymer microballon, and has better biocompatibility, cell compatibility and stronger security performance.
Polyethylene glycol: polyethylene glycol is used as a nonionic water-soluble polymer, and under the initiation condition, water can be used as a solvent to rapidly carry out a crosslinking reaction with hyaluronic acid and salts thereof to form a crosslinking substance. The cross-linked material formed by the method has good moisture retention and lubricity and has good inhibition effect on bacteria and the like.
When polyethylene glycol is used as a grafting framework and hyaluronic acid is used as a grafting monomer, an initiator can be added, so that the activation degree of carboxylic acid functional groups on the hyaluronic acid and salts thereof is better, the bonding effect between the functional groups is stronger, and further, the stability of the microsphere is better.
In this application, ammonium persulfate or potassium persulfate may be selected as the initiator, and after the oil phase is prepared, the initiator may be removed by removing the liquid, avoiding introduction of the initiator into the microspheres.
In conclusion, the porous polymer microsphere uses polyethylene glycol as a grafting framework, uses hyaluronic acid as a grafting monomer to jointly form a macromolecular substance, uses emulsion formed by the macromolecular substance as an oil phase, uses sodium alginate solution as an internal water phase, uses polyvinyl alcohol solution as a continuous phase, and cooperates with the sodium alginate solution to form the porous polymer microsphere with higher biocompatibility and cell compatibility, so that the expected effect is achieved.
In the invention, the concentration of the polyvinyl alcohol solution is 0.5-3 w/v%, and the concentration of the sodium alginate solution is 6-9 w/v%.
When the concentration of the polyvinyl alcohol solution is low, the polyvinyl alcohol solution cannot immediately generate strong interfacial tension and shear stress when being contacted with oil phase, and the formation of emulsion drops is not favored. On the contrary, when the concentration of the polyvinyl alcohol is higher, the viscosity of the polyvinyl alcohol correspondingly increases, so that when the polyvinyl alcohol flows as a continuous phase, the inner pipe wall of the injection pipe has larger influence on the polyvinyl alcohol, and turbulent flow movement is in a main flowing state, and at the moment, emulsion drops with different sizes are easily formed when the polyvinyl alcohol is mixed with oil phase, so that the uniformity of the microspheres is influenced, and the expected effect cannot be achieved.
In the invention, the concentration of the sodium alginate solution is 6-9 w/v%. Under the condition, the sodium alginate can form a colloid suspension with uniform texture with water, and the colloid suspension is uniformly distributed in a solvent. When the concentration of the sodium alginate is large, the sodium alginate cannot be inlaid and wrapped in the graft copolymerization product when being mixed with the graft copolymerization product, but is easy to wrap the graft copolymerization product in, so that the yield of the microsphere is low. And when the concentration of the sodium alginate is low, the micropore and mesopore quantity in the microsphere is small, so that the expected effect can not be achieved.
The mass ratio of the hyaluronic acid and the salt to the polyethylene glycol is (1.5-2.3): 2.
In the invention, an emulsifier can be added to enhance the emulsifying effect of the dispersed phase (i.e., oil phase). The emulsifier is added, so that the surface tension of two phases can be effectively reduced, the dispersion of the graft copolymer in the dispersion phase is more uniform, and meanwhile, the size of liquid drops formed by taking the graft copolymer as a core is smaller, so that the size of the microsphere is smaller, and the effect is better.
In the invention, the ratio of the emulsifier to the oil phase is (0.05-0.08): 1g/L, thereby achieving the emulsifying effect.
In the present invention, the emulsifier may be Span 80 or Tween 80. The Span 80 is totally called sorbitan monooleate, belongs to a lipophilic emulsifier, has better emulsification and dispersion characteristics, has the advantages of no peculiar smell, easy volatilization, no irritation and the like, and has higher use value. Tween 80 is fully called sorbitan monooleate, belongs to a lipophilic and hydrophilic nonionic surfactant, and has good emulsifying, stabilizing and dispersing effects.
The invention also provides a preparation method of the porous polymer microsphere, which mainly comprises the following steps:
mixing hyaluronic acid and salts thereof with polyethylene glycol, and heating in water bath to obtain oil phase; preparing sodium alginate solution by using water as a solvent to prepare an inner water phase; mixing the oil phase with the inner water phase to obtain emulsion; preparing a polyvinyl alcohol solution by taking water as a solvent to prepare a continuous phase;
pumping the emulsion and the continuous phase into a microfluidic device respectively, assembling a microfluidic chip with a homodromous structure in the microfluidic device to prepare microspheres, and then placing the microspheres in alkali liquor for soaking to prepare the porous microspheres.
Specifically, aqueous solutions of hyaluronic acid and salts thereof are gradually added to an aqueous polyethylene glycol solution and stirred during the mixing process at a speed of 50rpm to 70rpm for 24 hours to 48 hours. Stirring can lead the dispersing effect of hyaluronic acid and polyethylene glycol to be better, thereby leading the effect of crosslinking reaction to be better and the quality of crosslinking to be more uniform.
When the stirring speed is high, the interaction force between molecules is easily broken by the external force, thereby suppressing the formation of the copolymer. Specifically, when the carboxylate ions of hyaluronic acid and salts thereof and the hydroxyl groups of polyethylene glycol act on electron transfer, the position of the molecular action point is easily changed rapidly by external force of stirring, so that the reaction of the two ions is blocked, and the formation of the copolymer is further affected. When the stirring speed is low, the dispersion degree of the hyaluronic acid is poor, and the quality of the copolymer is easy to be poor.
In the application, the hyaluronic acid and the salt thereof and the polyethylene glycol carry out crosslinking reaction under the condition of water bath heating, and the temperature of the water bath heating is 50-60 ℃, so that the crosslinking reaction stably occurs, the process is controllable, and the preparation effect is better.
After the oil phase is prepared, the prepared internal aqueous phase can be mixed with the oil phase to prepare emulsion. When the inner water phase is prepared, the solvent can be heated to 60-70 ℃ to improve the solubility of sodium alginate, so that the sodium alginate can be dissolved in the solvent to form a homogeneous solution, namely the inner water phase. The internal aqueous phase is then mixed with the oil phase, at which point the internal aqueous phase is able to fill the network of the copolymer and form stable emulsion droplets.
It should be noted that after the emulsion is prepared, the emulsion may be subjected to ultrasonic treatment, so that the particle size of the droplets in the emulsion is smaller and the dispersion effect is better.
And the frequency of the ultrasonic wave is 50-55 kHZ, and the time of the ultrasonic wave is 1-1.5 h. Under the condition, the size of the liquid drops is smaller, the emulsifying effect is better, and meanwhile, the emulsion texture is more uniform. When the ultrasonic frequency is high, the energy provided by the ultrasonic is high, the structure of the liquid drops is easy to damage, and the yield is affected.
After the emulsion and continuous phase are prepared, the microsphere is prepared by adopting a microfluidic technology. The method comprises the following steps:
and selecting a needle head and a PVC pipe with specific specifications to construct a channel device, namely an emulsion channel and a continuous phase channel. And one end of the channel device, which is far away from the emulsion and the continuous phase, is assembled and connected with a graduated injector, so that the emulsion flows in an emulsion pipeline, the continuous phase flows in a continuous phase pipeline and is converged at a microfluidic chip to prepare emulsion drops, the emulsion drops continuously flow into a collecting phase along with the continuous phase, and microspheres are formed after precipitation.
In the invention, the structure of the microfluidic chip is a homodromous structure and is manufactured by adopting a 3D printing mode. The microfluidic chip is a PDMS microfluidic chip, has strong biological inertia and low cost, and is shown in fig. 1, wherein the fig. 1 shows the microfluidic chip with the same-direction structure, the middle part is an emulsion channel (the part pointed by the single-diagonal shear head), and the peripheral part is a continuous phase channel (the part pointed by the double-diagonal shear head).
In the invention, the flow rate of the emulsion in the emulsion channel is 0.03mL/min-0.08mL/min, and the flow rate of the continuous phase in the continuous phase channel is 1.5mL/min-3mL/min. In PDMS microfluidic chip with homodromous structure, the emulsion is intersected with continuous phase, and the droplets with consistent size are formed in the continuous phase, so as to achieve the expected preparation effect.
In the invention, two microfluidic injection pump devices are started to respectively control an emulsion pipeline and a continuous phase pipeline.
After passing through the microfluidic device, droplets are formed and precipitate out in the collection phase to form microspheres. At this time, after collecting the microspheres, washing with water, draining, and soaking in alkali solution to obtain porous microspheres.
In the invention, the pH value of the alkali liquor is 8.0-8.5, the sodium alginate colloid filled in the microspheres is dissolved by the alkali liquor, and the porous microspheres are prepared by washing for multiple times after the dissolution liquor is removed.
It should be noted that after dissolution, the washing should be stopped after a plurality of times until the washing liquid is neutral, specifically, the washing can be stopped when the pH value of the washing liquid is between 6.0 and 7.0, and the usable microspheres are prepared.
In the invention, the alkali liquor can be sodium hydroxide solution or sodium bicarbonate solution.
In the invention, the porous microspheres can be subjected to freeze drying after being washed with water to prepare the finished product, so that the porous structure of the finished product is firmer and firmer, and the preservation and transportation are facilitated.
It should be noted that in the present invention, the freeze-drying temperature is-25℃to-20℃and the drying time is 24 hours to 48 hours.
The features and capabilities of the present invention are described in further detail below in connection with the examples.
Example 1
The aim of the embodiment is to provide a porous polymer microsphere, which comprises the following raw materials:
polyvinyl alcohol solution, hyaluronic acid, sodium alginate solution and polyethylene glycol; wherein the concentration of the polyvinyl alcohol is 1w/v% when the polyvinyl alcohol is used as a continuous phase, and the concentration of the polyvinyl alcohol is 2.5w/v% when the polyvinyl alcohol is used as a collecting phase; the concentration of the sodium alginate solution is 8w/v%, and the mass ratio of hyaluronic acid to polyethylene glycol is 1.9:2.
The preparation method of the porous polymer microsphere comprises the following steps:
mixing hyaluronic acid and polyethylene glycol (stirring speed is 80rpm, stirring time is 36 h), heating in water bath at 55deg.C, and reacting to obtain oil phase; preparing sodium alginate solution by using water as a solvent to prepare an inner water phase; mixing the oil phase with the inner water phase to obtain emulsion; preparing a polyvinyl alcohol solution by taking water as a solvent to prepare a continuous phase;
pumping the emulsion and the continuous phase into a microfluidic device (two microfluidic injection pump devices are started), wherein the flow rate of the emulsion is 0.05mL/min, the flow rate of the continuous phase is 2.0mL/min, microspheres are prepared, and the microspheres are soaked in alkali liquor (alkali liquor is 0.01mmoL/L sodium hydroxide, and the pH value of the alkali liquor is 8.0), so that the porous microspheres are prepared. And then the porous microspheres are washed by ultrapure water until the pH value of the washing liquid is 6.0-6.5.
Example 2
The aim of the embodiment is to provide a porous polymer microsphere, which comprises the following raw materials:
polyvinyl alcohol solution, hyaluronic acid, sodium alginate solution, polyethylene glycol and emulsifying agent; wherein the concentration of the polyvinyl alcohol is 1w/v% when the polyvinyl alcohol is used as a continuous phase, and the concentration of the polyvinyl alcohol is 2.5w/v% when the polyvinyl alcohol is used as a collecting phase; the concentration of the sodium alginate solution is 8w/v%, and the mass ratio of hyaluronic acid to polyethylene glycol is 1.9:2; the emulsifier was Span 80 and the ratio of emulsifier to oil phase was 0.07:1g/L.
The preparation method of the porous polymer microsphere comprises the following steps:
mixing hyaluronic acid and polyethylene glycol (stirring speed is 80rpm, stirring time is 36 h), heating in water bath at 55deg.C, and reacting to obtain oil phase; preparing sodium alginate solution by using water as a solvent to prepare an inner water phase; mixing the oil phase with the emulsifier, and then mixing with the internal water phase to prepare emulsion; preparing a polyvinyl alcohol solution by taking water as a solvent to prepare a continuous phase;
pumping the emulsion and the continuous phase into a microfluidic device (two microfluidic injection pump devices are started), wherein the flow rate of the emulsion is 0.05mL/min, the flow rate of the continuous phase is 2.0mL/min, microspheres are prepared, and the microspheres are soaked in alkali liquor (alkali liquor is 0.01mmoL/L sodium hydroxide, and the pH value of the alkali liquor is 8.0), so that the porous microspheres are prepared. And then the porous microspheres are washed by ultrapure water until the pH value of the washing liquid is 6.0-6.5.
Example 3
The aim of the embodiment is to provide a porous polymer microsphere, which comprises the following raw materials:
polyvinyl alcohol solution, sodium hyaluronate, sodium alginate solution, polyethylene glycol and emulsifying agent; wherein the concentration of the polyvinyl alcohol is 1w/v% when the polyvinyl alcohol is used as a continuous phase, and the concentration of the polyvinyl alcohol is 2.5w/v% when the polyvinyl alcohol is used as a collecting phase; the concentration of the sodium alginate solution is 8w/v%, and the mass ratio of hyaluronic acid to polyethylene glycol is 1.9:2; the emulsifier was Span 80 and the ratio of emulsifier to oil phase was 0.07:1g/L.
The preparation method of the porous polymer microsphere comprises the following steps:
mixing hyaluronic acid and polyethylene glycol (stirring speed is 80rpm, stirring time is 36 h), heating in water bath at 55deg.C, and reacting to obtain oil phase; preparing sodium alginate solution by using water as a solvent to prepare an inner water phase; mixing the oil phase with an emulsifier, mixing with an internal water phase to prepare emulsion, and carrying out ultrasonic treatment on the emulsion for 2 hours under the condition of 50 kHZ; preparing a polyvinyl alcohol solution by taking water as a solvent to prepare a continuous phase;
pumping the emulsion and the continuous phase into a microfluidic device (two microfluidic injection pump devices are started), wherein the flow rate of the emulsion is 0.05mL/min, the flow rate of the continuous phase is 2.0mL/min, microspheres are prepared, and the microspheres are soaked in alkali liquor (alkali liquor is 0.01mmoL/L sodium hydroxide, and the pH value of the alkali liquor is 8.0), so that the porous microspheres are prepared. And then the porous microspheres are washed by ultrapure water until the pH value of the washing liquid is 6.0-6.5.
Example 4
The aim of the embodiment is to provide a porous polymer microsphere, which comprises the following raw materials:
polyvinyl alcohol solution, sodium hyaluronate, sodium alginate solution, polyethylene glycol and emulsifying agent; wherein the concentration of the polyvinyl alcohol is 0.5w/v% when the polyvinyl alcohol is used as a continuous phase, and the concentration of the polyvinyl alcohol is 3w/v% when the polyvinyl alcohol is used as a collecting phase; the concentration of the sodium alginate solution is 6w/v%, and the mass ratio of hyaluronic acid to polyethylene glycol is 1.5:2; the emulsifier was Span 80 and the ratio of emulsifier to oil phase was 0.05:1g/L.
The preparation method of the porous polymer microsphere comprises the following steps:
mixing hyaluronic acid and polyethylene glycol (stirring speed is 70rpm, stirring time is 48 h), heating in water bath at 50deg.C, and reacting to obtain oil phase; preparing sodium alginate solution by using water as a solvent to prepare an inner water phase; mixing the oil phase with an emulsifier, mixing with an internal water phase to prepare emulsion, and carrying out ultrasonic treatment on the emulsion for 1h under the condition of 55 kHZ; preparing a polyvinyl alcohol solution by taking water as a solvent to prepare a continuous phase;
pumping the emulsion and the continuous phase into a microfluidic device (two microfluidic injection pump devices are started), wherein the flow rate of the emulsion is 0.03mL/min, the flow rate of the continuous phase is 1.5mL/min, microspheres are prepared, and the microspheres are soaked in alkali liquor (alkali liquor is 0.01mmoL/L sodium hydroxide, and the pH value of the alkali liquor is 8.3), so that the porous microspheres are prepared. And then the porous microspheres are washed by ultrapure water until the pH value of the washing liquid is 6.0-6.5. After washing with water, the microspheres were placed in a freeze dryer for freeze drying under the following conditions: the temperature is minus 25 ℃ and the time is 24 hours.
Example 5
The aim of the embodiment is to provide a porous polymer microsphere, which comprises the following raw materials:
polyvinyl alcohol solution, sodium hyaluronate, sodium alginate solution, polyethylene glycol and emulsifying agent; wherein the concentration of the polyvinyl alcohol is 1.5w/v% when the polyvinyl alcohol is used as a continuous phase, and the concentration of the polyvinyl alcohol is 3w/v% when the polyvinyl alcohol is used as a collecting phase; the concentration of the sodium alginate solution is 9w/v%, and the mass ratio of hyaluronic acid to polyethylene glycol is 2.3:2; the emulsifier is Tween 80, and the ratio of the emulsifier to the oil phase is 0.08:1g/L.
The preparation method of the porous polymer microsphere comprises the following steps:
mixing hyaluronic acid and polyethylene glycol (stirring speed is 90rpm, stirring time is 24 h), heating in water bath at 60deg.C, and reacting to obtain oil phase; preparing sodium alginate solution by using water as a solvent to prepare an inner water phase; mixing the oil phase with an emulsifier, mixing with an internal water phase to prepare emulsion, and carrying out ultrasonic treatment on the emulsion for 3 hours under the condition of 50 kHZ; preparing a polyvinyl alcohol solution by taking water as a solvent to prepare a continuous phase;
pumping the emulsion and the continuous phase into a microfluidic device (two microfluidic injection pump devices are started), wherein the flow rate of the emulsion is 0.06mL/min, the flow rate of the continuous phase is 2.5mL/min, microspheres are prepared, and the microspheres are soaked in alkali liquor (alkali liquor is 0.01mmoL/L sodium bicarbonate, and the pH value of the alkali liquor is 8.5), so that the porous microspheres are prepared. And then the porous microspheres are washed by ultrapure water until the pH value of the washing liquid is 6.0-6.5. After washing with water, the microspheres were placed in a freeze dryer for freeze drying under the following conditions: the temperature is minus 20 ℃ and the time is 48 hours.
Effect example 1
The porous polymer microspheres prepared in examples 1 to 5 were taken and designated as examples 1 to 5, the number of microspheres in each group was 300, the particle size and pore size of the microspheres were measured, the particle size measurements were shown in FIGS. 2 to 6, the particle size distribution bar charts of the microspheres in examples 1 to 5 were shown in FIGS. 2 to 6, and the pore size measurements were shown in Table 1.
TABLE 1 microsphere pore size measurement results
As can be seen from fig. 2 to 5, the particle size of the microspheres prepared in example 1 is larger than that of the microspheres prepared in the other four groups, and is more concentrated in 420 μm to 460 μm, while the particle size of the microspheres in example 2 is more concentrated in 400 μm to 420 μm due to the better emulsion state of the droplets after the emulsifier is added, and the emulsion is further subjected to ultrasonic treatment while the emulsifier is added in the groups of example 3, example 4 and example 5, so that the particle size of the microspheres is more concentrated in 320 μm to 360 μm. It can be seen that the size of the microspheres can be controlled by changing the manufacturing process, and the particle size of the microspheres can be further controlled by changing the ratio of the emulsion flow rate to the continuous phase flow rate in the group 3-5, and the particle sizes of the microspheres in the group 4 and 5 are smaller.
In addition, in the present application, since the size of the microspheres prepared in the example 3 group to the example 5 group is smaller, the pore size is smaller.
Effect example 2
Microspheres (25 mg) prepared in example 4 and example 5 were immersed in phosphate buffer, and the volume of phosphate buffer was 20ml and ph was 6.8. The soaking time was 40 days, and samples were taken after 5 days, 10 days, 20 days and 40 days, and the quality of the microspheres was measured, and the results are shown in table 2. The soaking conditions were 37℃and the rotation speed was 40rpm, and the number of phosphate buffer changes was 10 days/time.
The weight loss rate calculation mode comprises the following steps: weight loss = (mass before microsphere soaking-mass after microsphere soaking) ×100%/mass before microsphere soaking.
Buffer pH test method: phosphate buffer solutions before and after soaking are tested by a Metrele-tolith pH tester, and the test results are obtained as three average values.
TABLE 2 Mass loss results
As shown in the table above, the microspheres prepared in examples 1 to 5 had good degradability after 40 days of soaking. Further, after soaking, the pH of the phosphate buffer tends to be acidic, mainly because the molecular structure of the microspheres is decomposed when the microspheres are affected by the phosphate buffer, and free carboxylate ions in the soaking solution are increased at this time, so that the pH of the solution is lowered.
Effect example 3
The porous microspheres prepared in example 5 were taken for cytotoxicity study, and the following are concrete:
cell selection: rat bone marrow mesenchymal stem cells (BMSCs);
cell digestion: resuscitated BMSCs were grown to a cell concentration of 1.0X10 9 Inoculating to 25cm 2 CO of (c) 2 In culture flask and at 37℃and CO 2 Culturing in a saturated humidity incubator. Changing culture medium at a frequency of 3 d/time in the culture process, adding 0.25% pancreatin when the cells are paved on the bottle bottom of the culture bottle until the cells are fused into a single layer and the density is 75%, and putting the mixture into an incubator for digestion for 2min;
cell passage: uniformly mixing the digested cell suspension, sucking the mixture into a 10mL centrifuge tube, centrifuging the mixture for 2min at 1000rpm, removing supernatant, and performing subculture in a DMEM culture medium, wherein the sugar content of the culture medium is 6mmol/L;
cell growth curve: taking CO 2 BMSCs grown well in culture flasks were treated with 0.25% pancreatin and then cell concentration was adjusted to 2X 10 4 1mL of each well was inoculated onto a 12-well plate, and then counted at 24h intervals to obtain a cell growth curve, as shown in FIG. 7, wherein the ordinate represents the number of cells, i.e. the value of the ordinate is 10 5 individual/mL;
as can be seen from FIG. 7, cells were in the adaptation phase during 12h-84h, while during 84h-132h, cells entered the logarithmic phase, were actively metabolized, and then entered the plateau phase. Thus, the optimal period of cell activity is 84h-132h.
Cell proliferation assay:
1) Sample group: the microspheres prepared in example 5 were prepared into suspensions at a concentration of 0.5mg/L, and then the microspheres were broken by ultrasound at an ultrasonic frequencyThe rate is 55kHZ, the mixture is centrifuged for 3min under the condition of 2000rpm after ultrasonic treatment, and the supernatant is removed to prepare broken microspheres; sterilizing the crushed microspheres, and mixing the sterilized crushed microspheres with a DMEM culture medium to prepare a microsphere-DMEM culture medium; then the cell concentration was adjusted to 2X 10 4 Culturing in microsphere-DMEM medium at a rate of one mL/mL, and counting at intervals of 24 h;
2) The control group is identical to the above conditions except that the culture medium is replaced by DMEM culture medium without microspheres;
3) Blank group: DMEM medium was placed in the same environment as the sample group and the control group for incubation, and counted at 24h intervals.
Cell relative proliferation rate (RGR) calculation mode: RGR= (sample group-blank group)/(control group-blank group)
The results of the above calculation and analysis of the cell proliferation rate are shown in FIG. 8.
As is clear from FIG. 8, the medium containing microspheres has little effect on cells, i.e., the microspheres have no toxic effect on cells, and the use value is high. Specifically, the growth of cells in the sample group was still at an active level, and the number of cell proliferation was hardly reduced, compared with the control group. However, as the culture time increases, the cell proliferation rate decreases, and this phenomenon occurs because the cell growth gradually progresses toward the plateau.
In conclusion, the microsphere is prepared by taking the microfluidic chip with the homodromous structure as a main microfluidic device, the microsphere size can be unified by the method, the microsphere size can be controlled by adjusting the flow rate ratio (emulsion/continuous phase), the applicability is higher, and meanwhile, the microsphere preparation effect is better. The microsphere prepared by the method takes polyethylene glycol as a framework, hyaluronic acid as a grafting monomer, macromolecular substances are formed together, emulsion formed by the macromolecular substances is taken as an oil phase, sodium alginate solution is taken as an internal water phase, polyvinyl alcohol solution is taken as a continuous phase, and the porous polymer microsphere with higher biocompatibility and cell compatibility is formed by matching, and the porous polymer microsphere has no toxicity on cell activity, so that the porous polymer microsphere can be used as a filling material in biomedical engineering, has higher use value and achieves the expected effect.
The embodiments described above are some, but not all embodiments of the invention. The detailed description of the embodiments of the invention is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Claims (9)
1. The preparation method of the porous polymer microsphere is characterized by comprising the following steps of:
mixing hyaluronic acid and salts thereof with polyethylene glycol according to the mass ratio of (1.5-2.3): 2, and heating in water bath to obtain an oil phase; preparing a sodium alginate solution with the concentration of 6-9 w/v% by taking water as a solvent to prepare an inner water phase; mixing the oil phase with the inner water phase to prepare emulsion; preparing a polyvinyl alcohol solution with the concentration of 0.5-3 w/v% by taking water as a solvent to prepare a continuous phase;
pumping the emulsion and the continuous phase into a microfluidic device respectively, assembling a microfluidic chip with a homodromous structure in the microfluidic device to prepare microspheres, and then placing the microspheres in alkali liquor for soaking to prepare the porous microspheres.
2. The method of claim 1, wherein the emulsion has a flow rate of 0.03mL/min to 0.08mL/min and the continuous phase has a flow rate of 1.5mL/min to 3mL/min when the emulsion and the continuous phase are pumped into the microfluidic device, respectively.
3. The method according to claim 1 or 2, wherein the emulsion is subjected to ultrasound with a frequency of 50kHZ-55kHZ for a time of 1h-4h.
4. The method according to claim 3, wherein the stirring is performed during the mixing of hyaluronic acid and polyethylene glycol at a speed of 50rpm to 70rpm for 24 hours to 48 hours.
5. The method of claim 1, wherein the water bath heating is at a temperature of 50 ℃ to 60 ℃.
6. The process according to claim 1, wherein the alkaline solution has a pH of 8.0-8.5.
7. The method according to claim 1, wherein the emulsion is obtained by mixing the oil phase with the emulsifier and then with the internal water phase, and the ratio of the emulsifier to the oil phase is (0.05-0.08): 1g/L.
8. The method of claim 7, wherein the emulsifier is Span 80 or Tween 80.
9. A porous polymeric microsphere prepared by the method for preparing a porous polymeric microsphere according to any one of claims 1 to 8.
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005097292A (en) * | 2003-09-01 | 2005-04-14 | Taisho Pharmaceut Co Ltd | W/o/w type double emulsion |
KR20120080267A (en) * | 2011-01-07 | 2012-07-17 | 공주대학교 산학협력단 | Preparation of biodegradable microparticles with structural complexity on the surface and inside by using a microfluidic device |
CN103371974A (en) * | 2012-04-25 | 2013-10-30 | 中国科学院大连化学物理研究所 | Drug sustained release polymeric microspheres prepared based on micro-fluidic technology and application |
CN113634208A (en) * | 2021-08-20 | 2021-11-12 | 西南交通大学 | Method for preparing porous calcium alginate microspheres by using microfluidic double-aqueous-phase emulsion as template |
WO2023284107A1 (en) * | 2021-07-16 | 2023-01-19 | 江南大学 | Hydrogel microsphere for adsorbing growth factor in stem cell supernatant, and preparation thereof |
-
2023
- 2023-05-09 CN CN202310516758.XA patent/CN116535728B/en active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005097292A (en) * | 2003-09-01 | 2005-04-14 | Taisho Pharmaceut Co Ltd | W/o/w type double emulsion |
KR20120080267A (en) * | 2011-01-07 | 2012-07-17 | 공주대학교 산학협력단 | Preparation of biodegradable microparticles with structural complexity on the surface and inside by using a microfluidic device |
CN103371974A (en) * | 2012-04-25 | 2013-10-30 | 中国科学院大连化学物理研究所 | Drug sustained release polymeric microspheres prepared based on micro-fluidic technology and application |
WO2023284107A1 (en) * | 2021-07-16 | 2023-01-19 | 江南大学 | Hydrogel microsphere for adsorbing growth factor in stem cell supernatant, and preparation thereof |
CN113634208A (en) * | 2021-08-20 | 2021-11-12 | 西南交通大学 | Method for preparing porous calcium alginate microspheres by using microfluidic double-aqueous-phase emulsion as template |
Non-Patent Citations (1)
Title |
---|
基于微流控液滴技术的载药缓释微球研究进展;杨兴远;夏曾子露;温维佳;高兴华;;自然杂志(第02期);第115-119页 * |
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