CN116173294A

CN116173294A - Microsphere for injection filling and preparation method thereof

Info

Publication number: CN116173294A
Application number: CN202310141858.9A
Authority: CN
Inventors: 张琳雅; 张红斌; 王佳冕; 宋晓杰
Original assignee: Shenzhen National Research Institute of High Performance Medical Devices Co Ltd
Current assignee: Shenzhen National Research Institute of High Performance Medical Devices Co Ltd
Priority date: 2023-02-13
Filing date: 2023-02-13
Publication date: 2023-05-30

Abstract

The invention discloses a microsphere for injection filling and a preparation method thereof. The microsphere for injection filling comprises a core formed by the aliphatic polyester microsphere and a shell formed by a sodium alginate layer coated outside the aliphatic polyester microsphere, wherein the hydrophilic shell formed by the sodium alginate layer effectively improves the dispersibility of the aliphatic polyester microsphere in aqueous solution and reduces side reactions such as inflammation and nodule caused or aggravated by the dispersibility of the aliphatic polyester microsphere. The hydrophilic shell formed by the sodium alginate layer also enables the aliphatic polyester microspheres to be easy to inject, does not need to be redissolved to simplify injection flow, reduces bacterial pollution risk and reduces the probability of agglomeration of microspheres for injection and filling.

Description

Microsphere for injection filling and preparation method thereof

Technical Field

The invention relates to the technical field of tissue filling materials, in particular to microspheres for injection filling and a preparation method thereof.

Background

Skin aging is a natural and complex human process, and in recent years, as the anti-aging consciousness of people increases, injection of fillers is an indispensable part of cosmetic medicine. The regeneration material is a material which can finally achieve the effect of face rejuvenation by promoting the regeneration of human body self-fibroblasts and collagen.

Compared with traditional filling products with shorter aging, the regenerative material is widely paid attention to due to natural, progressive and long-acting filling effect. The aliphatic polyester microspheres are used as a main body of the regeneration filler because of being capable of stimulating collagen proliferation, thereby achieving the effects of plumping the face, reducing wrinkles and filling depressions.

However, aliphatic polyester microspheres are poor in hydrophilicity and tend to agglomerate, thereby causing or aggravating side reactions such as inflammation and nodules.

Disclosure of Invention

Accordingly, there is a need for microspheres for injection filling that can solve the above-described problems.

In addition, it is necessary to provide a method for preparing the microspheres for injection and filling.

The microsphere for injection filling comprises a core formed by aliphatic polyester microspheres and a shell formed by a sodium alginate layer coated outside the aliphatic polyester microspheres, wherein the particle size of the aliphatic polyester microspheres is 10-40 mu m.

In one embodiment, the mass ratio of the sodium alginate to the aliphatic polyester microspheres is 0.05-0.5: 0.05 to 1.

In one embodiment, the aliphatic polyester microsphere is at least one material selected from the group consisting of PCL, PLA, PLLA, PLGA, PHA, PLGA-PEG, PLA-PEG, PLLA-PEG, PCL-PEG, PLGA-PEG-PLGA, PLA-PEG-PLGA, PLLA-PEG-PLGA, PCL-PEG-PLGA, PLGA-PEG-PLA, PLA-PEG-PLA, PLLA-PEG-PLA, PCL-PEG-PLA, PLGA-PEG-PLLA, PLLA-PEG-PLLA, PCL-PEG-PLLA, PLGA-PEG-PCL, PLA-PEG-PCL, PLLA-PEG-PCL, and PCL-PEG-PCL.

The preparation method of the microsphere for injection filling comprises the following steps:

providing aliphatic polyester microspheres, sodium alginate solution, calcium carbonate, oil phase solution of surfactant and organic acid, wherein the concentration of the sodium alginate solution is 5-50 g/L, and the concentration of the oil phase solution of the surfactant is 1-5 wt%;

the proportion is 0.05 g-1 g:0.1 g-1 g:10mL, dispersing the aliphatic polyester microspheres and the calcium carbonate into the sodium alginate solution, and uniformly mixing to obtain a dispersed phase;

uniformly mixing the disperse phase and the oil phase solution of the surfactant with the concentration of 1-5wt% to obtain emulsion, wherein the ratio of the sodium alginate solution to the oil phase solution of the surfactant is 10mL:20 mL-100 mL; and

uniformly mixing the disperse phase and the organic acid, and fully stirring and solidifying to obtain the needed microsphere for injection filling, wherein the proportion of the sodium alginate solution to the organic acid is 10mL: 1-5 mL, wherein the microsphere for injection filling comprises a core formed by the aliphatic polyester microsphere and a shell formed by a sodium alginate layer coated outside the aliphatic polyester microsphere, and the particle size of the aliphatic polyester microsphere is 10-40 mu m.

In one embodiment, the sodium alginate solution is prepared as follows: dispersing sodium alginate into physiological saline, heating and stirring at 60-80 ℃ until the sodium alginate is dissolved, and obtaining the sodium alginate solution.

In one embodiment, the solute of the oil phase solution of the surfactant is selected from at least one of span 80, tween 80, span 60, and tween 60;

the solvent of the oil phase solution of the surfactant is at least one selected from normal hexane, liquid paraffin, vegetable oil, kerosene and cyclohexane.

In one embodiment, the organic acid is selected from at least one of acetic acid, citric acid, and oxalic acid.

In one embodiment, the method further comprises the following steps after the microspheres for injection filling are obtained: after cleaning the microspheres for injection and filling, dispersing the microspheres into a gel matrix, wherein the proportion of the microspheres for injection and filling to the gel matrix is 0.05 g-1 g:10mL.

In one embodiment, the solute of the gel matrix is selected from at least one of cross-linked hyaluronic acid, cellulose derivatives, cross-linked sodium alginate, chitosan derivatives, amino acids and animal-derived collagen;

the concentration of the solute of the gel matrix is 1-5 wt%.

In one embodiment, the step of cleaning the microspheres for injection filling is: and sequentially cleaning the microspheres for injection and filling with an organic solvent and deionized water, and centrifuging the cleaned microspheres for injection and filling, wherein the organic solvent is at least one of absolute ethyl alcohol, ethyl acetate, petroleum ether and acetone, the centrifuging speed is 3000rpm, and the centrifuging time is 5min.

The microsphere for injection filling comprises a core formed by the aliphatic polyester microsphere and a shell formed by a sodium alginate layer coated outside the aliphatic polyester microsphere, wherein the hydrophilic shell formed by the sodium alginate layer effectively improves the dispersibility of the aliphatic polyester microsphere in aqueous solution and reduces side reactions such as inflammation and nodule caused or aggravated by the dispersibility of the aliphatic polyester microsphere. The hydrophilic shell formed by the sodium alginate layer also enables the aliphatic polyester microspheres to be easy to inject, does not need to be redissolved to simplify injection flow, reduces bacterial pollution risk and reduces the probability of agglomeration of microspheres for injection and filling.

In addition, the sodium alginate layer has good biocompatibility, volume expansion efficiency and relatively slow metabolic behavior, and can effectively improve and prolong the instant filling effect.

Meanwhile, the sodium alginate and other raw materials involved in the invention are common and widely available, and have the advantages of low cost, wide sources and the like.

The preparation method of the microsphere for injection filling can be simply and conveniently applied to various microspheres with the requirements of improving dispersibility and prolonging degradation time, and has universality.

In addition, the microspheres for injection and filling are dispersed into the gel matrix after being cleaned, and the gel outer layer with elasticity and biocompatibility can be wrapped outside the microspheres for injection and filling, so that the degradation time of the microspheres for injection and filling in vivo can be effectively prolonged, and the filling time can be prolonged.

Drawings

In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

Wherein:

FIG. 1 is a flowchart of a method for preparing microspheres for injection filling according to an embodiment.

FIG. 2 is a low-magnification optical microscope image of microspheres for injection filling obtained in example 1.

FIG. 3 is a high-magnification optical microscope image of microspheres for injection filling obtained in example 1.

FIG. 4 is a scanning electron microscope image of microspheres for injection filling obtained in example 1.

Detailed Description

The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. 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.

The invention discloses an injection filling microsphere in an embodiment, which comprises a core formed by aliphatic polyester microspheres and a shell formed by a sodium alginate layer coated outside the aliphatic polyester microspheres, wherein the particle size of the aliphatic polyester microspheres is 10-40 mu m.

In general, sodium alginate can be sodium alginate of different viscosity and different M/G ratio.

Preferably, in the present embodiment, the mass ratio of the sodium alginate to the aliphatic polyester microspheres is 0.05 to 0.5:0.05 to 1.

More preferably, the mass ratio of the sodium alginate to the aliphatic polyester microsphere is 1:1.

preferably, in this embodiment, the aliphatic polyester microspheres are at least one selected from the group consisting of PCL, PLA, PLLA, PLGA, PHA, PLGA-PEG, PLA-PEG, PLLA-PEG, PCL-PEG, PLGA-PEG-PLGA, PLA-PEG-PLGA, PLLA-PEG-PLGA, PCL-PEG-PLGA, PLGA-PEG-PLA, PLA-PEG-PLA, PLLA-PEG-PLA, PCL-PEG-PLA, PLGA-PEG-PLLA, PLA-PEG-PLLA, PLLA-PEG-PLLA, PCL-PEG-PLLA, PLGA-PEG-PCL, PLA-PEG-PCL, PLLA-PEG-PCL and PCL-PEG-PCL.

The aliphatic polyester microspheres can be prepared by an emulsion solvent volatilization method, a membrane emulsification method, a spray drying method, a microfluidic method and an ink-jet printing method or can be directly purchased.

The invention also discloses a preparation method of the microsphere for injection filling according to an embodiment, which comprises the following steps of:

s10, providing aliphatic polyester microspheres, sodium alginate solution with the concentration of 5g/L to 50g/L, calcium carbonate, oil phase solution of surfactant and organic acid.

Calcium carbonate is used to provide calcium ions to ionically crosslink sodium alginate.

Wherein the concentration of the sodium alginate solution is 5 g/L-50 g/L, and the concentration of the oil phase solution of the surfactant is 1wt% -5 wt%.

Preferably, the concentration of the sodium alginate solution is 20g/L, and the concentration of the oil phase solution of the surfactant is 2wt%.

Preferably, in this embodiment, the preparation method of the sodium alginate solution is as follows: dispersing sodium alginate into physiological saline, heating and stirring at 60-80 ℃ until the sodium alginate is dissolved, and obtaining sodium alginate solution.

In other embodiments, the solvent of the sodium alginate solution may also be pure water, deionized water or PBS buffer.

Preferably, in the present embodiment, the solute of the oil phase solution of the surfactant is selected from at least one of span 80, tween 80, span 60 and tween 60.

Preferably, in the present embodiment, the solvent of the oil phase solution of the surfactant is selected from at least one of n-hexane, liquid paraffin, vegetable oil, kerosene and cyclohexane.

Preferably, in the present embodiment, the organic acid is at least one selected from acetic acid, citric acid and oxalic acid.

S20, the proportion is 0.05 g-1 g:0.1 g-1 g:10mL, dispersing aliphatic polyester microspheres and calcium carbonate into a sodium alginate solution, and uniformly mixing to obtain a disperse phase.

Preferably, the ratio of the aliphatic polyester microspheres to the calcium carbonate and sodium alginate solution is 0.2g:0.15g:10mL.

In S20, the mixing may be achieved by stirring or ultrasonic dispersion.

S30, uniformly mixing the disperse phase and the oil phase solution of the surfactant with the concentration of 1-5 wt% to obtain emulsion.

Wherein, the proportion of the sodium alginate solution to the oil phase solution of the surfactant is 10mL:20 mL-100 mL.

Preferably, the ratio of the sodium alginate solution to the oil phase solution of the surfactant is 10mL:300mL.

And S40, uniformly mixing the disperse phase and the organic acid, and fully stirring and solidifying to obtain the required microsphere for injection filling.

Wherein, the proportion of the sodium alginate solution to the organic acid is 10mL:1 mL-5 mL.

Preferably, the ratio of sodium alginate solution to organic acid is 10mL:2mL.

The prepared microsphere for injection filling comprises a core formed by aliphatic polyester microspheres and a shell formed by a sodium alginate layer coated outside the aliphatic polyester microspheres, wherein the particle size of the aliphatic polyester microspheres is 10-40 mu m.

Preferably, in this embodiment, after S40, the following operations are performed: cleaning the microspheres for injection filling, and dispersing the microspheres into a gel matrix, wherein the ratio of the microspheres for injection filling to the gel matrix is 0.05 g-1 g:10mL.

The microspheres for injection and filling are dispersed into a gel matrix after being cleaned, and the gel outer layer with elasticity and biocompatibility can be wrapped outside the microspheres for injection and filling, so that the degradation time of the microspheres for injection and filling in the body can be effectively prolonged, and the filling time can be further prolonged.

Preferably, the solute of the gel matrix is selected from at least one of crosslinked hyaluronic acid, cellulose derivatives, crosslinked sodium alginate, chitosan derivatives, amino acids and animal-derived collagen.

Preferably, the concentration of solute in the gel matrix is 1wt% to 5wt%.

Preferably, the solvent of the gel matrix may be physiological saline, pure water, deionized water or PBS buffer.

Preferably, the operation of cleaning the microspheres for injection filling is as follows: washing and centrifuging the microspheres for injection filling by using an organic solvent, and then washing and centrifuging the microspheres for injection filling by using deionized water, wherein the organic solvent is at least one selected from absolute ethyl alcohol, ethyl acetate, petroleum ether and acetone, the centrifuging speed is 3000rpm, and the centrifuging time is 5min.

The following are specific examples.

Example 1

Preparing sodium alginate solution: 2G of sodium alginate (sodium alginate M/g=1:2, microphone, S875337-100G) was weighed, dispersed in 100mL of physiological saline, heated at 70 ℃ and stirred until dissolved, to obtain sodium alginate solution.

0.2g of PCL microspheres and 0.15g of calcium carbonate are weighed and dispersed in 10mL of sodium alginate solution, and the microspheres are uniformly dispersed by stirring to obtain a dispersed phase. Wherein, PCL microsphere is prepared by solvent volatilization method: dispersing a dichloromethane solution containing 5wt% of PCL in a PVA solution containing 2wt% of PCL, stirring at a high speed for emulsification, stirring at a low speed for solidification, and obtaining PCL microspheres, and washing and drying for later use.

10mL of the dispersed phase was added to 30mL of liquid paraffin containing 2wt% Tween 80, and stirred to obtain an emulsion.

2mL of acetic acid was added to the emulsion, and stirring was continued for 10min to solidify, thereby obtaining the desired microspheres for injection filling.

Washing the microspheres for injection and filling with an organic solvent, centrifuging at 3000rpm for 5min, washing the microspheres for injection and filling with deionized water, centrifuging at 3000rpm for 5min, dispersing 0.1g of the washed microspheres for injection and filling in 10mL of 2wt% carboxymethyl cellulose solution, packaging, and sterilizing by ultraviolet, thus being used for injection.

The microspheres for injection and filling dispersed in a carboxymethyl cellulose solution prepared in example 1 were redispersed in water and then observed by an optical microscope to obtain fig. 2 and 3, and then the aqueous solution of the microspheres for injection and filling was dried and then observed by a scanning electron microscope to obtain fig. 4.

Referring to fig. 2, 3 and 4, it can be seen that the microspheres for injection and filling prepared in example 1 have a remarkable core-shell structure and good dispersibility.

Example 2

Preparing sodium alginate solution: 0.5G sodium alginate (sodium alginate M/g=1:2, microphone, S875337-100G) was weighed, dispersed in 100mL physiological saline, heated at 70 ℃ and stirred until dissolved, to obtain sodium alginate solution.

0.05g of PCL microspheres and 0.1g of calcium carbonate are weighed and dispersed in 10mL of sodium alginate solution, and the microspheres are uniformly dispersed by stirring to obtain a dispersed phase. Wherein, PCL microsphere is prepared by solvent volatilization method: dispersing a dichloromethane solution containing 5wt% of PCL in a PVA solution containing 2wt% of PCL, stirring at a high speed for emulsification, stirring at a low speed for solidification, and obtaining PCL microspheres, and washing and drying for later use.

10mL of the dispersed phase was added to 20mL of liquid paraffin containing 5wt% Tween 80, and stirred to obtain an emulsion.

1mL of acetic acid was added to the emulsion, and stirring was continued for 15min to solidify, thereby obtaining the desired microspheres for injection.

Washing the microspheres for injection and filling with an organic solvent, centrifuging at 3000rpm for 5min, washing the microspheres for injection and filling with deionized water, centrifuging at 3000rpm for 5min, dispersing 1g of the washed microspheres for injection and filling in 10mL of 5wt% carboxymethyl cellulose solution, packaging, and sterilizing by ultraviolet, thus being used for injection.

Example 3

Preparing sodium alginate solution: 5G of sodium alginate (sodium alginate M/g=1:2, microphone, S875337-100G) was weighed, dispersed in 100mL of physiological saline, heated at 70 ℃ and stirred until dissolved, to obtain sodium alginate solution.

1g of PCL microspheres and 1g of calcium carbonate are weighed and dispersed in 10mL of sodium alginate solution, and the microspheres are uniformly dispersed by stirring to obtain a dispersed phase. Wherein, PCL microsphere is prepared by solvent volatilization method: dispersing a dichloromethane solution containing 5wt% of PCL in a PVA solution containing 2wt% of PCL, stirring at a high speed for emulsification, stirring at a low speed for solidification, and obtaining PCL microspheres, and washing and drying for later use.

10mL of the dispersed phase was added to 100mL of liquid paraffin containing 1wt% Tween 80, and stirred to obtain an emulsion.

5mL of acetic acid was added to the emulsion, and stirring was continued for 30min to solidify, thereby obtaining the desired microspheres for injection.

Washing the microspheres for injection and filling with an organic solvent, centrifuging at 3000rpm for 5min, washing the microspheres for injection and filling with deionized water, centrifuging at 3000rpm for 5min, dispersing 0.05g of the washed microspheres for injection and filling in 10mL of 1wt% carboxymethyl cellulose solution, packaging, and sterilizing by ultraviolet, thus being used for injection.

The above examples illustrate only a few embodiments of the invention, which are described in detail and are not to be construed as limiting the scope of the claims. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.

Claims

1. The microsphere for injection filling is characterized by comprising a core formed by aliphatic polyester microspheres and a shell formed by a sodium alginate layer coated outside the aliphatic polyester microspheres, wherein the particle size of the aliphatic polyester microspheres is 10-40 mu m.

2. The microsphere for injection and filling according to claim 1, wherein the mass ratio of the sodium alginate to the aliphatic polyester microsphere is 0.05-0.5: 0.05 to 1.

3. The microspheres for injection filling according to claim 1, wherein the aliphatic polyester microspheres are at least one material selected from the group consisting of PCL, PLA, PLLA, PLGA, PHA, PLGA-PEG, PLA-PEG, PLLA-PEG, PCL-PEG, PLGA-PEG-PLGA, PLA-PEG-PLGA, PLLA-PEG-PLGA, PCL-PEG-PLGA, PLGA-PEG-PLA, PLA-PEG-PLA, PLLA-PEG-PLA, PCL-PEG-PLA, PLGA-PEG-PLLA, PLA-PEG-PLLA, PLLA-PEG-PLLA, PCL-PEG-PLLA, PLGA-PEG-PCL, PLA-PEG-PCL, PLLA-PEG-PCL and PCL-PEG-PCL.

4. A method for producing the microspheres for injection filling according to any one of claims 1 to 3, comprising the steps of:

5. The method for preparing microspheres for injection filling according to claim 4, wherein the sodium alginate solution is prepared by the following steps: dispersing sodium alginate into physiological saline, heating and stirring at 60-80 ℃ until the sodium alginate is dissolved, and obtaining the sodium alginate solution.

6. The method for producing microspheres for injection filling according to claim 4, wherein the solute of the oil phase solution of the surfactant is at least one selected from the group consisting of span 80, tween 80, span 60 and tween 60;

7. The method for producing microspheres for injection filling according to claim 4, wherein the organic acid is at least one selected from acetic acid, citric acid and oxalic acid.

8. The method for producing microspheres for injection and filling according to any one of claims 4 to 7, further comprising, after the microspheres for injection and filling are obtained, performing the following operations: after cleaning the microspheres for injection and filling, dispersing the microspheres into a gel matrix, wherein the proportion of the microspheres for injection and filling to the gel matrix is 0.05 g-1 g:10mL.

9. The method for preparing microspheres for injection filling according to claim 8, wherein the solute of the gel matrix is at least one selected from the group consisting of crosslinked hyaluronic acid, cellulose derivatives, crosslinked sodium alginate, chitosan derivatives, amino acids and animal-derived collagen;

the concentration of the solute of the gel matrix is 1-5 wt%.

10. The method for preparing the microspheres for injection and filling according to claim 8, wherein the step of cleaning the microspheres for injection and filling comprises the following steps: and sequentially cleaning the microspheres for injection and filling with an organic solvent and deionized water, and centrifuging the cleaned microspheres for injection and filling, wherein the organic solvent is at least one of absolute ethyl alcohol, ethyl acetate, petroleum ether and acetone, the centrifuging speed is 3000rpm, and the centrifuging time is 5min.