CN114601964A - Injectable PHA microsphere and preparation method and application thereof - Google Patents
Injectable PHA microsphere and preparation method and application thereof Download PDFInfo
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- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/06—Flowable or injectable implant compositions
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Abstract
The invention discloses an injectable PHA microsphere and a preparation method and application thereof, comprising the following steps: s1, preparing an oil phase: dissolving PHA in organic solvent to prepare 0.1-500mg/mL PHA/organic solvent solution; s2, preparing a water phase: preparing 1.8-5% polyvinyl alcohol aqueous solution by mass; s3, controlling the volume ratio of the oil phase to the water phase, mixing the oil phase and the water phase, and preparing microspheres to obtain the injectable PHA microspheres. The method provided by the invention can prepare the injectable PHA microspheres with good protein adsorption and ZETA potential characteristics, and can be used in the fields of tissue filling and the like.
Description
Technical Field
The invention belongs to the technical field of biochemical engineering, and relates to an injectable PHA microsphere as well as a preparation method and application thereof.
Background
As people age or are affected by certain diseases, muscle and collagen tissues in human bodies can generate functional degeneration with different degrees, which causes problems of skin depression, gastric reflux and the like, so that people invent various fillers to fill the depressed skin or stimulate the regrowth of muscle and collagen through foreign matters, such as hyaluronic acid, bovine collagen and the like. However, these substances as fillers have a short retention time of the filling effect, and thus the filling effect thereof needs to be maintained by frequent injection. In order to achieve the long-term filling effect, people try to use biodegradable materials to prepare microspheres as fillers, such as polyvinyl alcohol (PVA), polymethyl methacrylate (PMMA) and the like. Although the filling effect of these fillers is maintained for a significantly longer time, these materials remain in the body for too long, releasing harmful substances and thus causing a series of side reactions that are harmful to human health.
In recent years, biodegradable high polymer materials come into the visual field of people, the material has no toxicity or rejection reaction to human bodies, can be gradually degraded along with human metabolism and then discharged out of the human bodies, and can make the degradation time vary from one week to several years by adjusting parameters such as the molecular weight of the material.
Polyhydroxyalkanoates, which is called polyhydroxyakanoates in English, called PHA for short, is a natural high-molecular biological material, and is an intracellular polyester synthesized by microorganisms. PHA is one of the most desirable biomedical materials due to its good biocompatibility and biodegradability. PHA has good cell compatibility with cells in vivo, cells can grow well on the scaffold, and the scaffold can be degraded into CO2And H2And O. To facilitate injection, they are usually made as microspheres, which can be injected through a needle. The microspheres are usually over 20 microns in diameter due to the presence of human phagocytes, but too large a microsphere can clog needles and even cause skin breakdown. The microspheres used for injection are therefore typically below 60 microns in size.
In the prior art, when PHA material is prepared into injectable microspheres, the prepared microspheres are easy to agglomerate and adhere to form large massive substances, which brings inconvenience to subsequent injection, and the prepared microspheres have weak capability of adhering cells and can not be well compatible with tissues, thereby greatly hindering the application of biodegradable material microspheres as a filling agent. Therefore, a novel microsphere preparation scheme is urgently needed, and the problems of low cell adhesion, easy agglomeration and adhesion of the microspheres are solved.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides the injectable PHA microspheres and the preparation method and application thereof.
The method is realized by the following technical scheme:
a method for preparing injectable PHA microspheres, comprising the steps of:
s1, preparing an oil phase: dissolving PHA in organic solvent to prepare 0.1-500mg/mL PHA/organic solvent solution;
s2, preparing a water phase: preparing 1.8-5% polyvinyl alcohol aqueous solution by mass; the polyvinyl alcohol is not used as a filler main body, so that the situation that when the polyvinyl alcohol is used as the filler, harmful substances are released after the polyvinyl alcohol is remained in a body for a long time, and a series of side reactions are caused to harm human health can be avoided;
s3, controlling the volume ratio of the oil phase to the water phase, mixing the oil phase and the water phase, and preparing microspheres to obtain the injectable PHA microspheres.
Further, in step S1, it is preferable to prepare 5 to 90mg/mL of PHA/organic solvent solution. When the concentration of the PHA/organic solvent solution is not in the range of 0.1-500mg/mL, it is not miscible in the proper ratio and microspheres cannot be formed. When the concentration of the PHA/organic solvent solution is in the range of 5-90mg/mL, the prepared PHA microspheres have good dispersibility and are not easy to adhere.
Specifically, in step S1, the PHA is one or more of poly (3-hydroxybutyrate-co-3-hydroxypentanoic acid-co-3-hydroxyhexanoic acid), poly (3-hydroxybutyrate-4-hydroxybutyrate) or poly (3-hydroxybutyrate-co-3-hydroxyvalerate), and is preferably poly (3-hydroxybutyrate-4-hydroxybutyrate). Wherein, poly (3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) is referred to as PHBHHX for short, poly (3-hydroxybutyrate-3-hydroxyhexanoate) is referred to as PHBHHx for short, poly (3-hydroxybutyrate-co-3-hydroxyvalerate) is referred to as PHBV for short, and poly (3-hydroxybutyrate-4-hydroxybutyrate) is referred to as P34HB for short.
Specifically, in the polyvinyl alcohol aqueous solution, the polyvinyl alcohol can be polyvinyl alcohol with ultrahigh polymerization degree (molecular weight is 25-30 ten thousand), polyvinyl alcohol with high polymerization degree (molecular weight is 17-22 ten thousand), polyvinyl alcohol with medium polymerization degree (molecular weight is 12-15 ten thousand) and polyvinyl alcohol with low polymerization degree (molecular weight is 2.5-3.5 ten thousand), and the alcoholysis degree of the polyvinyl alcohol is 78% -98%.
Further, in step S3, the volume ratio of the oil phase to the water phase is 1/15-1/3. When the volume ratio of the oil phase to the aqueous phase is less than 1/15 or more than 1/3, the obtained PHA microspheres have poor cell adhesion and dispersibility, and the PHA microspheres cannot be formed.
Specifically, in step S3, the method of making microspheres includes: emulsifying the oil phase into the water phase through a membrane emulsifier, wherein the emulsifying pressure is 0.01-0.1MPA, the length of a membrane tube is 0.8-50um, the length of the membrane tube can influence the particle size of the prepared PHA microspheres, the emulsifying time is 20-500min, the emulsifying stirring speed is 150-1000r/min, after the emulsification is finished, water is added for solidification, the stirring speed is 50-500r/min after the water is added, and the solidification time is 1-48 h. The emulsification pressure, the emulsification time, the stirring manner and the curing time are critical to make the PHA microspheres have proper particle size, uniform distribution and good spherical shape.
Specifically, in step S3, on the basis of steps S1 and S2, a conventional emulsion evaporation method, a phase separation method, a spray drying method, a hot melt extrusion method, a microfluidic technique, or a stirring technique may be used as the method for preparing the microspheres.
Further, in step S1, the organic solvent is one or more of N-methylpyrrolidone, dichloromethane, chloroform, or acetonitrile. By selecting a suitable solvent, the PHA can be dissolved therein.
Further, in step S1, the PHA has a weight-average molecular weight of 2 to 100 ten thousand.
The invention also provides injectable PHA microspheres prepared by the method.
Furthermore, the injectable PHA microspheres prepared by the invention can be applied to the fields of tissue filling, medical and cosmetic injection and the like.
Compared with the prior art, the invention has the following advantages:
when the biomaterial is in contact with the physiological environment, the biomaterial is not rejected in vivo, and has no serious side effect related to the filler, so that the filler is required to have good cell adhesion, and an important point in the cell adhesion is that a large amount of protein is on the cell surface, and protein adsorption is an important driving force for cell adhesion. The PHA microspheres which have high protein adhesion, good dispersibility and no mutual adhesion are successfully prepared by configuring the oil phase and the water phase with specific concentrations and controlling the volume ratio of the oil phase to the water phase, and the PHA microspheres prepared by the method have good protein adhesion without depending on adding hyaluronic acid in PHA or coating hyaluronic acid on the surface of PHA to improve the cell adhesion of the microspheres. The prepared PHA microspheres have proper particle size, and degradation products of PHA are beneficial to cell regeneration and pain relief, and can promote collagen regeneration, so that the PHA microspheres can be used for medical injection, tissue filling and the like.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.
FIG. 1 is a schematic flow chart of the production process of the present invention;
FIG. 2 is an electron micrograph of injectable PHA microspheres prepared in example 1 of the present invention;
FIG. 3 is a graph showing the particle size distribution of injectable PHA microspheres obtained in example 1 of the present invention;
FIG. 4 is a ZETA potential measured in 20% ethanol of injectable PHA microspheres prepared in example 1 of the present invention;
FIG. 5 shows the ZETA potential measured in pure water of injectable PHA microspheres prepared in example 1 of the present invention;
FIG. 6 is a standard curve of BSA adsorption experiment in example 1 of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
S1, preparing an oil phase: dissolving 0.8g of P34HB in 20mL of dichloromethane to prepare 40mg/mL of PHA/organic solvent solution, wherein the weight average molecular weight of P34HB is 15 ten thousand;
s2, preparing a water phase: preparing a polyvinyl alcohol aqueous solution with the mass fraction of 3%;
s3: emulsifying the oil phase into the rotating water phase through a membrane emulsifier, controlling the volume ratio of the oil phase to the water phase to be 1/5, the emulsifying pressure to be 0.02MPA, the length of a membrane tube to be 15 mu m, and the emulsifying time to be 108 min; the length of the magneton is 5cm, and the emulsifying and stirring speed is 400 r/min. And after emulsification is finished, starting curing, wherein an overhead stirrer and a 10cm stirring paddle are used for curing, 280mL of water is additionally added, the stirring speed is 120r/min, and the curing time is 24h, so that the injectable PHA microspheres are obtained.
Fig. 1 is a schematic flow chart of the preparation method of the present invention, and the results of observing the prepared injectable PHA microspheres by SEM scanning electron microscopy are shown in fig. 2, and it can be seen from the electron microscopy of the injectable PHA microspheres prepared in example 1 of the present invention that the obtained microspheres have good dispersibility, no adhesion between them, uniform morphology, and are spherical. The particle size and span values were measured by a laser particle sizer, fig. 3 is a particle size distribution graph of the injectable PHA microspheres obtained in example 1, it can be seen from fig. 3 that the average particle size of the microspheres obtained in example 1 is 27.2 μm, and table 1 shows the span value (the span value is the degree of dispersion of the particle size distribution) of example 1 measured by a laser particle sizer, and the measurement result shows that the span value is 0.776, which can indicate that the injectable PHA microspheres of this example have uniform particle size.
Table 1.
The injectable PHA microspheres prepared in example 1 were subjected to the ZETA potential test in 20% ethanol and pure water, respectively, and the results are shown in fig. 4 and 5, respectively. As seen in FIGS. 4 and 5, the injectable microspheres had a ZETA potential of-30.3 mV in 20% ethanol and a ZETA potential of-30.2 mV in purified water. The important significance of the ZETA potential is that its value is related to the stability of the colloidal dispersion. The ZETA potential is a measure of the strength of the mutual repulsion or attraction between particles. The smaller the molecule or dispersed particle, the higher the absolute value (positive or negative) of the ZETA potential, the more stable the system, i.e., the dissolution or dispersion can resist aggregation. Conversely, the lower the ZETA potential (positive or negative), the more prone to coagulation or agglomeration, i.e. the attractive force exceeds the repulsive force, and the dispersion is broken and coagulation or agglomeration occurs (note that the absolute value of the ZETA potential represents its magnitude of stability, and positive or negative represents what charge the particles are charged with). The ZETA potential of the injectable PHA microspheres in 20% ethanol is-30.3 mV, and the ZETA potential in pure water is-30.2 mV, which can show that the injectable PHA microspheres have good stability, also show that the surfaces of the microspheres are negatively charged and can be adsorbed and combined with sugar or protein with positive charges on the surfaces of cells, and indirectly prove that the injectable PHA microspheres provided by the invention have good cell adhesion. In addition, considering that the subsequent storage of PHA microspheres was in 20% ethanol, the results of the test also indicate that the injectable PHA microspheres produced by the present invention can be stably stored.
To further examine the protein adsorption of injectable PHA microspheres, the injectable PHA microspheres prepared in this example were used to: (1) drawing a protein growth curve: BSA was used to prepare a 3mg/ml solution, which was scanned at full wavelength using a spectrophotometer to detect the maximum absorption wavelength of BSA albumin at 278.58 nm. Then, a series of concentration gradients of 0.5, 1, 1.5, 3, 4, 5mg/ml solutions were prepared, and the absorbance of each concentration of solution was measured at the maximum absorption wavelength to prepare a standard curve, which is shown in FIG. 6, where the abscissa of FIG. 6 is represented by x, the ordinate is represented by y, x is the BSA concentration, and y is the absorbance. (2) Preparing 5mg/ml BSA solution, incubating injectable PHA microspheres, measuring the light absorption value of the BSA solution to be 2.941 before incubation, centrifuging and taking supernatant after incubation for 5 hours, detecting the light absorption value of the BSA solution (supernatant solution), and calculating the specific adsorption concentration, wherein the result is as follows: after 5 hours of incubation, the supernatant had a BSA concentration of 4.8mg/mL and an absorbance of 2.877, and the difference between the concentrations before and after incubation indicated that BSA protein was adsorbed by the injectable PHA microspheres. (3) Then, the value of protein adsorbed by each gram of microspheres is calculated (the higher the value is, the better the protein adsorption is), the value of protein adsorbed by the injectable PHA microspheres in the embodiment is calculated to be 6700ug/g, and compared with the adsorption value of 600ug/g of a CT1 charge sphere (a CT1 charge sphere is a commonly used cell culture microcarrier, namely Cytodex 1, and the main component of CT1 is GE Healthcare, DEAE-cross-linked dextran), the injectable PHA microspheres provided by the invention can be well adsorbed on the cell surface, and the adsorption effect is better than that of the traditional CT1, which is about 10 times of the adsorption capacity of the CT1 sphere.
Example 2
S1, preparing an oil phase: dissolving 100mg of P34HB in 20mL of dichloromethane to prepare 5mg/mL PHA/organic solvent solution, wherein the weight average molecular weight of P34HB is 3 ten thousand;
s2, preparing a water phase: preparing a polyvinyl alcohol aqueous solution with the mass fraction of 1.8%;
s3: emulsifying the oil phase into the rotating water phase through a membrane emulsifier, controlling the volume ratio of the oil phase to the water phase to be 2/15, the emulsifying pressure to be 0.05MPA, the length of a membrane tube to be 10 mu m, and the emulsifying time to be 100 min; the length of the magneton is 5cm, and the emulsifying and stirring speed is 450 r/min. And after emulsification is finished, starting curing, wherein an overhead stirrer and a 10cm stirring paddle are used for curing, and 300mL of water is additionally added, the stirring speed is 100r/min, and the curing time is 24h, so that the injectable PHA microspheres are obtained.
Example 3
S1, preparing an oil phase: dissolving 1.8g of P34HB in 20mL of dichloromethane to prepare 90mg/mL of PHA/organic solvent solution, wherein the weight average molecular weight of P34HB is 28 ten thousand;
s2, preparing a water phase: preparing a polyvinyl alcohol aqueous solution with the mass fraction of 5%;
s3: emulsifying the oil phase into the rotating water phase through a membrane emulsifier, controlling the volume ratio of the oil phase to the water phase to be 1/15, the emulsifying pressure to be 0.1MPA, the length of a membrane tube to be 30 mu m, and the emulsifying time to be 120 min; the length of the magneton is 5cm, and the emulsifying and stirring speed is 350 r/min. And after emulsification is finished, starting curing, wherein an overhead stirrer and a 10cm stirring paddle are used for curing, and 350mL of water is additionally added, the stirring speed is 180r/min, and the curing time is 48h, so that the injectable PHA microspheres are obtained.
Example 4
S1, preparing an oil phase: dissolving 2mg of P34HB in 20mL of N-methylpyrrolidone to prepare a PHA/organic solvent solution of 0.1mg/mL, wherein the weight average molecular weight of P34HB is 23 ten thousand;
s2, preparing a water phase: preparing a polyvinyl alcohol aqueous solution with the mass fraction of 5%;
s3: controlling the volume ratio of the oil phase to the water phase to be 4/15, mixing the oil phase and the water phase, and preparing the microspheres by a conventional spray drying method, wherein the specific steps are as follows: firstly, introducing hot air into the top of a drying tower, pumping the oil-water mixed solution to the top of the tower, spraying the oil-water mixed solution into atomized liquid drops through an atomizer, contacting with high-temperature hot air, quickly evaporating the organic reagent and water, and discharging a dried product from the bottom of the tower to obtain the injectable PHA microspheres. The temperature of the hot air is significantly lowered and the humidity thereof is increased after the hot air contacts the droplets, and the organic reagent and the moisture entrained in the exhaust gas are extracted by the exhaust fan and recovered by the separation device as the exhaust gas.
Example 5
S1, preparing an oil phase: dissolving 100mg of P34HB in 20mL of chloroform to prepare 500mg/mL PHA/organic solvent solution, wherein the weight average molecular weight of P34HB is 10 ten thousand;
s2, preparing a water phase: preparing a polyvinyl alcohol aqueous solution with the mass fraction of 4%;
s3: controlling the volume ratio of the oil phase to the water phase to be 1/3, mixing the oil phase and the water phase, and preparing the microspheres by a conventional emulsification and volatilization method, wherein the method specifically comprises the following steps: firstly, stirring the oil-water mixed solution by magnetic force at the rotation speed of 200r/min for 5min, then reducing the rotation speed to 100r/min, stirring for 24h to volatilize the organic solvent trichloromethane, finally centrifuging the mixed solution, collecting and cleaning microspheres, and freeze-drying to obtain the injectable PHBV microspheres.
Comparative example 1
S1, preparing an oil phase: dissolving 0.8g of P34HB in 20mL of dichloromethane to prepare 40mg/mL of PHA/organic solvent solution, wherein the weight average molecular weight of P34HB is 15 ten thousand;
s2, preparing a water phase: preparing a polyvinyl alcohol aqueous solution with the mass fraction of 3%;
s3: emulsifying the oil phase into the rotating water phase through a membrane emulsifier, controlling the volume ratio of the oil phase to the water phase to be 1/20, the emulsifying pressure to be 0.02MPA, the length of a membrane tube to be 15 mu m, and the emulsifying time to be 108 min; the length of the magneton is 5cm, and the emulsifying and stirring speed is 400 r/min. And after the emulsification is finished, starting curing, wherein an overhead stirrer and a 10cm stirring paddle are used for curing, 280mL of water is additionally added, the stirring speed is 120r/min, and the curing time is 24 h.
Comparative example 1 compared with example 1, the oil-water ratio of comparative example 1 was less than 1/15, and the microspheres obtained in this way had severe spherical shrinkage, 90% had irregular flake shapes, and could not be made into microspheres.
Comparative example 2
S1, preparing an oil phase: dissolving 0.8g of P34HB in 20mL of dichloromethane to prepare 40mg/mL of PHA/organic solvent solution, wherein the weight average molecular weight of P34HB is 15 ten thousand;
s2, preparing a water phase: preparing a polyvinyl alcohol aqueous solution with the mass fraction of 3%;
s3: emulsifying the oil phase into the rotating water phase through a membrane emulsifier, controlling the volume ratio of the oil phase to the water phase to be 1/2, the emulsifying pressure to be 0.02MPA, the length of a membrane tube to be 15 mu m, and the emulsifying time to be 108 min; the length of the magneton is 5cm, and the emulsifying and stirring speed is 400 r/min. And after the emulsification is finished, starting curing, wherein an overhead stirrer and a 10cm stirring paddle are used for curing, 280mL of water is additionally added, the stirring speed is 120r/min, and the curing time is 24 h.
Comparative example 2 compared to example 1, the oil-water ratio of comparative example 2 was higher than 1/3, and the microspheres prepared by the method of comparative example 2 did not sphere and were in the form of block P34 HB.
The above examples are exemplified by P34HB, but the basic structure of all PHAs is shown by the following general structural formula,
in the structural general formula, x and n represent positive integers, carboxyl of PHA monomer and hydroxyl of adjacent PHA monomer form ester bond, and the monomers are all in R-configuration. Similar in structure and properties, are soluble in forming microspheres, and thus one skilled in the art would be able to deduce that all PHAs are suitable for use in the present invention.
In conclusion, the injectable PHA microspheres prepared by the preparation method provided by the invention have good protein adsorption and ZETA potential characteristics, and can be used in the fields of tissue filling and the like.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A method for preparing injectable PHA microspheres, comprising the steps of:
s1, preparing an oil phase: dissolving PHA in organic solvent to prepare 0.1-500mg/mL PHA/organic solvent solution;
s2, preparing a water phase: preparing 1.8-5% polyvinyl alcohol aqueous solution by mass;
s3, controlling the volume ratio of the oil phase to the water phase, mixing the oil phase and the water phase, and preparing microspheres to obtain the injectable PHA microspheres.
2. The method for preparing injectable PHA microspheres of claim 1, wherein in step S1, the concentration of PHA/organic solvent solution is 5-90 mg/mL.
3. The method of claim 1, wherein the PHA is one or more of poly (3-hydroxybutyrate-co-3-hydroxypentanoic acid-co-3-hydroxyhexanoic acid), poly (3-hydroxybutyrate-4-hydroxybutyrate), or poly (3-hydroxybutyrate-co-3-hydroxyvalerate) in step S1.
4. The method for preparing injectable PHA microspheres of claim 1, wherein in step S3, the volume ratio of oil phase to aqueous phase is 1/15-1/3.
5. The method for preparing injectable PHA microspheres of claim 1, wherein in step S3, the method for making microspheres comprises: emulsifying the oil phase into the water phase through a membrane emulsifier, wherein the emulsifying pressure is 0.01-0.1MPA, the pore diameter of a membrane tube is 0.8-50 mu m, the emulsifying time is 20-500min, the emulsifying stirring speed is 150-1000r/min, after the emulsification is finished, adding water for solidification, and after the water is added, the stirring speed is 50-500r/min, and the solidification time is 1-48 h.
6. The method for preparing injectable PHA microspheres of claim 1, wherein in step S3, the method for making microspheres comprises: emulsion evaporation, phase separation, spray drying, hot melt extrusion, microfluidic technology or stirring technology.
7. The method for preparing injectable PHA microspheres of claim 1, wherein in step S1, the organic solvent is one or more of N-methylpyrrolidone, dichloromethane, trichloromethane or acetonitrile.
8. The method for preparing injectable PHA microspheres of claim 1, wherein in step S1, the weight average molecular weight of PHA is 2-100 ten thousand.
9. Injectable PHA microspheres obtainable by the process of any one of claims 1 to 8.
10. Use of the injectable PHA microspheres of claim 9 in the field of tissue augmentation.
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| CN101708175A (en) * | 2009-12-24 | 2010-05-19 | 辽宁大学 | Sustained release microsphere medicinal preparation using PHBHHx as carrier and method for preparing same |
| US20130323306A1 (en) * | 2012-05-30 | 2013-12-05 | Boston Scientific Scimed, Inc. | Injectable biodegradable particles for controlled therapeutic agent release |
| CN113617305A (en) * | 2021-10-13 | 2021-11-09 | 北京蓝晶微生物科技有限公司 | Polyhydroxyalkanoate microsphere with narrow particle size distribution and preparation method thereof |
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| CN101708175A (en) * | 2009-12-24 | 2010-05-19 | 辽宁大学 | Sustained release microsphere medicinal preparation using PHBHHx as carrier and method for preparing same |
| US20130323306A1 (en) * | 2012-05-30 | 2013-12-05 | Boston Scientific Scimed, Inc. | Injectable biodegradable particles for controlled therapeutic agent release |
| CN113617305A (en) * | 2021-10-13 | 2021-11-09 | 北京蓝晶微生物科技有限公司 | Polyhydroxyalkanoate microsphere with narrow particle size distribution and preparation method thereof |
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