Konjac glucomannan microspheres with controllable particle size and preparation method and application thereof

Abstract

The invention relates to a preparation method of konjac glucomannan microspheres with controllable size and particle size, which comprises the following steps: dissolving konjac powder in an acid solution, fully stirring to form a jelly shape, placing in a high-pressure environment for high-temperature degradation, taking out a degradation product, and cooling to obtain a solid degradation product, mixing and stirring the solid degradation product and an alkali solution until the solid degradation product is completely dissolved, filtering to obtain a konjac glucomannan clarified solution, and taking the konjac glucomannan clarified solution as a water phase; providing an oil phase containing a surfactant; and adding the water phase into the oil phase, stirring the water phase at a constant speed by adopting a stirring device to crush the water phase into drops, and dropwise adding a cross-linking agent to cross-link and cure the drops to generate the konjac glucomannan microspheres with consistent particle sizes.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of bioengineering, in particular to a konjac glucomannan microsphere with controllable size and particle diameter, a preparation method and application thereof. BACKGROUND

[0002] In the field of medical cosmetic filling, microsphere materials are widely used to provide structural support and stimulate autologous collagen regeneration to achieve the effect of long-acting shaping; at present, agarose microspheres are widely used in this field, agarose is a linear polysaccharide extracted from red algae, which has stable source and relatively mature preparation process, however, the existing agarose microspheres are difficult to directly form microspheres with uniform particle diameter in the preparation process, and must rely on subsequent complex screening process to screen out agarose microspheres with target particle diameter, which increases the production cost and reduces the production efficiency, and the cost of extracting agarose from red algae is high. SUMMARY

[0003] Therefore, the present application provides a preparation method of a konjac glucomannan microsphere with controllable size and particle diameter, which comprises the following steps: The konjac powder is dissolved in an acid solution and stirred to form a jelly, and then subjected to high-temperature degradation in a high-pressure environment, and the degraded product is taken out and cooled to obtain a solid degraded product, which is mixed with an alkali solution and stirred until completely dissolved, and then filtered to obtain a konjac glucomannan clear liquid, which is used as an aqueous phase; An oil phase containing a surfactant is provided; The aqueous phase is added to the oil phase, and a stirring device is used to stir at a constant speed to break the aqueous phase into droplets, and a crosslinking agent is added dropwise to crosslink and solidify the droplets to form konjac glucomannan microspheres with uniform particle diameter.

[0004] According to an aspect of the present application, a konjac glucomannan microsphere is provided, which is prepared by the method of any one of the above claims, and has an average particle diameter of 10-300 pm.

[0005] According to another aspect of the present application, a medical cosmetic filling composition is provided, which comprises the konjac glucomannan microsphere prepared by the method of any one of the above claims and water for injection.

[0006] Advantages of the present application The application adjusts the amount of acid solution to control the viscosity of the water phase (i.e., the konjac glucomannan clarified liquid), avoids the difficulty of dispersion of the water phase due to too high viscosity or the breakage of droplets due to too low viscosity; at the same time, the stirring device provides stable shear force to ensure that the water phase with suitable viscosity is uniformly dispersed into droplets with uniform size in the oil phase, and the droplets can directly obtain konjac glucomannan microspheres after cross-linking and solidification. The konjac glucomannan microspheres prepared by the application can directly obtain konjac glucomannan microspheres with uniform particle size without a complex screening process, greatly improving the production efficiency and reducing the production cost. At the same time, the application adjusts the rotating speed of the stirring device to change the size of the droplets, and then flexibly controls the particle size of the konjac glucomannan microspheres, so as to meet the demand for different particle sizes of konjac glucomannan microspheres in the medical and beauty filling field. The application uses konjac glucomannan instead of traditional agarose as raw material. Konjac is a high-yield crop and has a relatively low price. The extraction cost of konjac glucomannan is lower than that of agarose extracted from red algae, which greatly reduces the cost of using microsphere materials in the medical and beauty filling field, and improves the market competitiveness of the product.

[0007] Other features and aspects of the present application will become apparent from the following detailed description of exemplary embodiments with reference to the accompanying drawings. BRIEF DESCRIPTION OF DRAWINGS

[0008] The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments, features, and aspects of the application and serve to explain the principles of the present application.

[0009] Figure 1 Representative images showing the observation of rat muscle tissue after injection for 1 week, 4 weeks, 8 weeks, and 16 weeks; Figure 2 Representative images showing the observation of rat heart tissue after injection for 1 week, 4 weeks, 8 weeks, and 16 weeks; Figure 3 Representative images showing the observation of rat lung tissue after injection for 1 week, 4 weeks, 8 weeks, and 16 weeks; Figure 4 Representative images showing the observation of rat kidney tissue after injection for 1 week, 4 weeks, 8 weeks, and 16 weeks; Figure 5 Optical microscope photos of konjac glucomannan microspheres prepared in Example 1 are shown; Figure 6 Optical microscope photos of konjac glucomannan microspheres prepared in Example 2 are shown; Figure 7 Optical microscope photos of konjac glucomannan microspheres prepared in Example 3 are shown; Figure 8 Optical microscope photos of konjac glucomannan microspheres prepared in Example 4 are shown; Figure 9 An optical microscope photo of konjac glucomannan microspheres prepared in Example 5 is shown. Figure 10 An optical microscope photo of konjac glucomannan microspheres prepared in Example 6 is shown. DETAILED DESCRIPTION

[0010] Various exemplary embodiments, features, and aspects of the present application will be described below in detail with reference to accompanying drawings. The same reference numbers in different drawings represent the same or similar elements. Although various aspects of embodiments are illustrated in the drawings, the drawings are not necessarily drawn to scale unless specifically noted, and the embodiments are not necessarily limited to the drawings.

[0011] It should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", and the like indicate relative or positional relationships based on the orientation or position shown in the drawings, and are used only for convenience of description or simplification of description, and do not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be construed as limiting the present application.

[0012] In addition, the terms "first", "second", are used only for descriptive purposes and should not be construed as indicating or implying relative importance or an indicated number of technical features. Therefore, features defined as "first", "second" can explicitly or implicitly include one or more of the features. In the description of the present application, the meaning of "a plurality of" is two or more, unless otherwise specifically limited.

[0013] The word "exemplary" is used herein to mean "serving as an example, instance, or illustration." Any implementation described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other implementations.

[0014] In addition, in order to better illustrate the present application, numerous specific details are given in the following detailed description. Those skilled in the art will understand that the present application can also be implemented without certain specific details. In some examples, methods, means, elements and circuits well known to those skilled in the art are not described in detail in order to highlight the main idea of the present application.

[0015] The present application proposes a method for preparing konjac glucomannan microspheres with controllable size and particle size, comprising the following steps: S1, the konjac powder is dissolved in an acid solution, and after being fully stirred to form a jelly, it is placed in an environment for high-temperature degradation. The degraded product is taken out and cooled to obtain a solid degraded product. The solid degraded product is mixed with an alkali solution and stirred until completely dissolved, and then filtered to obtain a konjac glucomannan clear liquid, which is used as the water phase; S2, an oil phase containing a surfactant is provided; S3, the water phase is added to the oil phase, and a stirring device is used to stir it at a uniform speed to break the water phase into droplets. A crosslinking agent is added dropwise to crosslink and solidify the droplets to generate konjac glucomannan microspheres with consistent particle sizes.

[0016] The konjac glucomannan microspheres prepared in the application can be used as a filling material in medical and cosmetic applications. The konjac glucomannan in the application is a natural complex polysaccharide with a molecular weight of about 80-100 million. Typically, 1% of the glucomannan in water is jelly-like and difficult to stir, with no flowability. Such viscous substances cannot be dispersed in oily substances, so the glucomannan must be degraded to obtain a liquid that is easy to flow. In the application, a certain amount of konjac powder is dissolved in an acid solution and fully stirred to form a jelly-like konjac glucomannan solution. In this stirring operation, the stirring speed of the konjac powder and the acid solution can be adjusted according to the actual situation, which is not limited in the application. The stirred konjac glucomannan solution is placed in a high-temperature, high-pressure sterilization environment for degradation for 50-60 minutes. When the stirred konjac glucomannan solution is placed in a high-temperature, high-pressure environment for degradation, higher pressure can promote more intense intermolecular reactions and accelerate the degradation process. Higher temperatures can provide enough energy for the degradation reaction, causing the konjac glucomannan molecular chain to break, achieving the degradation effect. The degraded product is taken out and cooled to room temperature (where room temperature conventionally refers to a temperature of 10-30°C) to obtain a solid degraded product. In the application, the cooling process can be performed by placing the solid degraded product in a room temperature environment for natural cooling. Then, an alkali solution and the solid degraded product are placed in a container in a certain proportion and fully stirred to completely dissolve the solid degraded product, and then filtered to obtain a golden yellow konjac glucomannan clear liquid. In this stirring operation, the stirring speed of the alkali solution and the solid degraded product can be adjusted according to the actual situation, which is not limited in the application. The viscosity of the konjac glucomannan clear liquid obtained in the application is 10-60 mPa·s.

[0017] Further, the mass ratio of konjac fine powder to acid solution is 1:0.6; the value range of high-temperature environment is 115-121°C, and the value range of high-pressure environment is 0.13Mpa-0.21Mpa (wherein, at standard atmospheric pressure, 100°C corresponds to 0.1 MPa, 115°C corresponds to about 0.13 MPa, 121°C corresponds to about 0.21 MPa, and the high-pressure environment of 0.21MPa is commonly used for high-pressure sterilization); preferably, the value of the high-pressure environment is 0.21Mpa; and the mass ratio of solid-state degradation product to alkali solution is 3:1.

[0018] By adjusting the amount of acid solution added, the degradation degree of konjac glucomannan molecular chain in the konjac glucomannan solution and the viscosity of the konjac glucomannan solution are controlled, so that the viscosity of the water phase (i.e., the konjac glucomannan clear liquid) is controlled within the range of 10 mPa·s-60 mPa·s, avoiding the difficulty of uniform dispersion of droplets due to too high viscosity of the water phase and the oil phase in the emulsification process; the alkali solution is suitable for neutralizing the residual acid solution in the solid-state degradation product, and at the same time, the solid-state degradation product is re-dissolved into a liquid state; the oil phase of the surfactant is suitable for promoting the dispersion of the water phase in the oil phase; the introduction of the surfactant effectively reduces the interfacial tension between the water phase and the oil phase; uniform stirring is provided by the stirring device, thereby providing stable shear force for the dispersion of the water phase in the oil phase to form droplets, avoiding the dispersion of the water phase into droplets of different sizes due to fluctuation of the stirring rate, ensuring that the water phase forms droplets of uniform size in the oil phase during the emulsification process, and further realizing uniform particle size of the konjac glucomannan microspheres formed by solidification and cross-linking; the cross-linking agent reacts with the konjac glucomannan molecules in the droplets to form a three-dimensional network structure, fixing the droplet form into konjac glucomannan microspheres, and avoiding changes in the size of the konjac glucomannan microspheres in subsequent processing.

[0019] It should be noted that the water phase, as a dispersed phase, needs to be broken into small droplets by external shear force and uniformly dispersed in the oil phase; by adjusting the stirring speed of the stirring device, the shear force on the water phase is changed, thereby realizing control of the droplet size, i.e., control of the particle size of the konjac glucomannan microspheres; the faster the stirring speed of the stirring device, the larger the droplet size, i.e., the larger the particle size of the konjac glucomannan microspheres, and vice versa; the smaller the droplet size, i.e., the smaller the particle size of the konjac glucomannan microspheres; by adjusting the stirring speed of the stirring device, the average particle size of the konjac glucomannan microspheres is controlled within the range of 10μm-300μm.

[0020] The application adjusts the amount of acid solution to control the viscosity of the water phase (i.e. konjac glucomannan clarified liquid), avoids the difficulty of dispersion of the water phase due to too high viscosity or the breakage of droplets due to too low viscosity; at the same time, the stirring device provides stable shear force to ensure that the water phase with suitable viscosity is uniformly dispersed into droplets with uniform size in the oil phase, and the droplets can directly obtain konjac glucomannan microspheres after crosslinking and solidification. The konjac glucomannan microspheres prepared by the application can directly obtain konjac glucomannan microspheres with uniform particle size without complex screening process, greatly improving the production efficiency and reducing the production cost. At the same time, the application changes the droplet size by adjusting the rotating speed of the stirring device, and then realizes flexible control of the particle size of the konjac glucomannan microspheres, so as to meet the demand of different particle sizes of the konjac glucomannan microspheres in the medical and beauty filling field. The application uses konjac glucomannan instead of traditional agarose as raw material. Konjac is a high-yield crop and has relatively low price. The extraction cost of konjac glucomannan is lower than that of agarose extracted from red algae, so that the cost of using microsphere material in the medical and beauty filling field is greatly reduced, and the market competitiveness of the product is improved.

[0021] In a possible implementation manner, the acid solution is hydrochloric acid with a concentration of 0.5 mol / L; the alkali solution is a sodium hydroxide solution, and further, the concentration of the sodium hydroxide solution ranges from 35% to 40%.

[0022] In a possible implementation manner, the specific steps of step S3 are as follows: S301, placing the water phase and the oil phase into a reaction kettle and performing water bath heating on the reaction kettle; S302, uniformly stirring the water phase and the oil phase in the reaction kettle by using a stirring device to disperse the water phase into uniform droplets; S303, gradually adding the crosslinking agent into the reaction kettle to make the konjac glucomannan molecular chains in the droplets crosslink and solidify with the crosslinking agent, thereby generating solid konjac glucomannan microspheres.

[0023] It should be noted that the water phase and the oil phase are transferred to the reaction kettle at a ratio of 1:3, and the reaction kettle is heated; the water bath heating specifically refers to: heating the outside of the reaction kettle through a constant temperature water bath equipment, so as to provide stable temperature conditions for the emulsification and crosslinking of the water phase, and avoid uneven emulsification caused by temperature fluctuations; wherein the reaction kettle can be directly heated, the reaction kettle is heated to 40-60℃, heating the reaction kettle to the appropriate temperature can significantly improve the fluidity of the konjac glucomannan slurry, while avoiding the degradation of the molecular chain in the konjac glucomannan slurry caused by high temperature, thereby laying a foundation for subsequent droplet formation; the stirring device is suitable for providing continuous and stable shear force generated by uniform stirring, so as to uniformly break the water phase into small droplets and disperse them in the oil phase; during the stirring process, the droplets will have a tendency to agglomerate due to Brownian motion, and uniform stirring can prevent the droplets from colliding and merging through continuous fluid disturbance, ensuring that the droplets always maintain a dispersed and uniform size before crosslinking and solidification; wherein the stirring speed of the stirring device is 30-300rpm, and the stirring device can use an electric stirring rod with adjustable speed in the prior art; after the water phase and the oil phase are stirred for 1 hour, the crosslinking agent is added dropwise, the adding process needs to be added uniformly under stirring, and it is ensured that the adding process is completed within 1-1.5 hours; the gradual and uniform addition can make the crosslinking agent fully react with the glucomannan molecular chain in the droplet, the surface of the droplet first undergoes slight crosslinking to form a protective layer, and then gradually penetrates into the interior of the droplet to react, thereby forming a dense and uniform three-dimensional network structure, so that the droplet is converted from liquid to solid konjac glucomannan microspheres, and the insufficient solidification is avoided to cause the konjac glucomannan microspheres to dissolve or deform during subsequent washing and storage; wherein the dropwise adding speed of the crosslinking agent is 0.56-0.83g / min, and the mass of each drop of the crosslinking agent is 0.05g; the crosslinking reaction time between the glucomannan molecular chain in the droplet and the crosslinking agent is 8-12 hours.

[0024] In one possible implementation, the oil phase is selected from liquid paraffin, and the surface active agent is sorbitan monooleate, wherein the ratio of liquid paraffin to sorbitan monooleate is 20:1.

[0025] In one possible implementation, the crosslinking agent is glutaraldehyde.

[0026] In one possible implementation, the method further comprises washing the konjac glucomannan microspheres, and the specific steps are as follows: S401, transferring the konjac glucomannan microspheres containing the oil phase to a filtering device to separate the oil phase from the konjac glucomannan microspheres; S402, adding petroleum ether to the filtering device to wash the separated konjac glucomannan microspheres; S403, adding ethanol again to the konjac glucomannan microspheres for the second washing; S404, adding purified water again to the konjac glucomannan microspheres for the third washing.

[0027] It should be noted that after the cross-linking in step S3, petroleum ether, ethanol and purified water are sequentially added to the konjac glucomannan microspheres to remove the residual oil phase, surfactant, unreacted cross-linking agent and other impurities on the surface of the konjac glucomannan microspheres, so as to avoid the interference of the residual substances on the performance of the konjac glucomannan microspheres. Specifically, first, petroleum ether is added to the konjac glucomannan microspheres, wherein the volume ratio of the konjac glucomannan microspheres to the petroleum ether is 1:2. The petroleum ether can efficiently dissolve the residual oil phase components on the surface and in the pores of the konjac glucomannan microspheres. Through two rounds of petroleum ether washing, the oil phase components adsorbed on the surface and in the pores of the konjac glucomannan microspheres are removed to the maximum extent. Then, ethanol is added to the microspheres washed by the petroleum ether, wherein the volume ratio of the konjac glucomannan microspheres to the ethanol is 1:1. The ethanol is used to fully dissolve the residual petroleum ether in the pores and on the surface of the microspheres. Through two rounds of ethanol washing, the petroleum ether is completely removed. Then, purified water is added to the microspheres washed by the ethanol, wherein the volume ratio of the konjac glucomannan microspheres to the purified water is 1:3. The purified water is used to dissolve the residual ethanol, unreacted cross-linking agent and polar impurities such as emulsifiers on the surface of the microspheres. The residual reagents on the surface and in the pores of the microspheres are thoroughly washed to obtain konjac glucomannan microspheres with high purity.

[0028] The filtering device can be a sand core funnel and a suction filter pump in the prior art.

[0029] In the following examples, the konjac glucomannan clarified solution with a viscosity of 10 mPa·-60 mPa·s is used as the water phase. 86.98 g of konjac powder is dissolved in 1111.02 g of acid solution, and stirred thoroughly to form a jelly-like konjac glucomannan solution. Then, the solution is placed in a high-pressure environment at 115°C for degradation for 55 minutes. The degraded product is taken out and cooled to room temperature to obtain a solid degraded product. Then, 400 g of alkali solution is added and stirred thoroughly to dissolve the solid degraded product. After filtration, a golden yellow konjac glucomannan clarified solution is obtained, and the viscosity of the konjac glucomannan clarified solution is 10 mPa·-60 mPa·s. The specific preparation steps of the acid solution are as follows: 45 ml of 0.5 mol / L hydrochloric acid is transferred to 1000 ml of purified water, and the acid solution is obtained after shaking. The specific preparation steps of the alkali solution are as follows: 140 g of solid sodium hydroxide with a purity of 35% is weighed and added to 260 g of purified water, and the alkali solution is obtained after stirring until completely dissolved.

[0030] In the following embodiments, a Haixinrui HL3100 wet laser particle size analyzer was used to measure the particle size of konjac glucomannan microspheres. 0.5g to 5g of konjac glucomannan microspheres were placed in 500mL of purified water and then positioned at the detection location of the wet laser particle size analyzer. The analyzer measured the particle size of the konjac glucomannan microspheres, and the operator read the values ​​from the analyzer. The specific measurement procedure is common knowledge in the field and will not be described in detail here.

[0031] Example 1: Preparation of konjac glucomannan microspheres with a particle size of approximately 10 μm Under the condition that the water bath of the reactor is heated to 60℃, 200g of the prepared konjac glucomannan clarified solution and 600g of oil phase containing surfactant are transferred into the reactor. The stirring device is started and the stirring speed is set to 280rpm-300rpm / min. Stirring is continued at this stirring speed. After stirring for 1 hour, 50g of glutaraldehyde solution is added dropwise. The dropwise addition process must be carried out under stirring and must be completed uniformly within 1 to 1.5 hours. After the glutaraldehyde solution is added, the reaction is continued under stirring for 8 hours to allow the aqueous phase droplets to fully cross-link and solidify to form konjac glucomannan microspheres. After the reaction, the konjac glucomannan microspheres are washed to remove impurities, thereby obtaining konjac glucomannan microspheres with uniform particle size.

[0032] The particle size of the konjac glucomannan microspheres prepared in this embodiment was determined to be 10 μm. Figure 5 As shown, under a microscope, the konjac glucomannan microspheres exhibit good sphericity and a smooth surface.

[0033] Example 2: Preparation of konjac glucomannan microspheres with a particle size of approximately 100 μm Under the condition that the water bath of the reactor is heated to 60℃, 200g of the prepared konjac glucomannan clarified solution and 600g of oil phase containing surfactant are transferred into the reactor. The stirring device is started and the stirring speed is set to 200rpm-210rpm / min. Stirring is continued at this stirring speed. After stirring for 1 hour, 50g of glutaraldehyde solution is added dropwise. The dropwise addition process must be carried out under stirring and must be completed uniformly within 1 to 1.5 hours. After the glutaraldehyde solution is added, the reaction is continued under stirring for 8 hours to allow the aqueous phase droplets to fully cross-link and solidify to form konjac glucomannan microspheres. After the reaction, the konjac glucomannan microspheres are washed to remove impurities, thereby obtaining konjac glucomannan microspheres with uniform particle size.

[0034] The particle size of the konjac glucomannan microspheres prepared in this embodiment was determined to be 100 μm. Figure 6As shown, under a microscope, the konjac glucomannan microspheres exhibit good sphericity and a smooth surface.

[0035] Example 3: Preparation of konjac glucomannan microspheres with a particle size of approximately 150 μm Under the condition that the water bath of the reactor is heated to 60℃, 200g of the prepared konjac glucomannan clarified solution and 600g of oil phase containing surfactant are transferred into the reactor. The stirring device is started and the stirring speed is set to 130rpm-140rpm / min. Stirring is continued at this stirring speed. After stirring for 1 hour, 50g of glutaraldehyde solution is added dropwise. The dropwise addition process must be carried out under stirring and must be completed uniformly within 1 to 1.5 hours. After the glutaraldehyde solution is added, the reaction is continued under stirring for 8 hours to allow the aqueous phase droplets to fully cross-link and solidify to form konjac glucomannan microspheres. After the reaction, the konjac glucomannan microspheres are washed to remove impurities, thereby obtaining konjac glucomannan microspheres with uniform particle size.

[0036] The particle size of the konjac glucomannan microspheres prepared in this embodiment was determined to be 150 μm. Figure 7 As shown, under a microscope, the konjac glucomannan microspheres exhibit good sphericity and a smooth surface.

[0037] Example 4: Preparation of konjac glucomannan microspheres with a particle size of approximately 200 μm Under the condition that the water bath of the reactor is heated to 60℃, 200g of the prepared konjac glucomannan clarified solution and 600g of oil phase containing surfactant are transferred into the reactor. The stirring device is started and the stirring speed is set to 110rpm-120rpm / min. Stirring is continued at this stirring speed. After stirring for 1 hour, 50g of glutaraldehyde solution is added dropwise. The dropwise addition process must be carried out under stirring and must be completed uniformly within 1 to 1.5 hours. After the glutaraldehyde solution is added, the reaction is continued under stirring for 8 hours to allow the aqueous phase droplets to fully cross-link and solidify to form konjac glucomannan microspheres. After the reaction, the konjac glucomannan microspheres are washed to remove impurities, thereby obtaining konjac glucomannan microspheres with uniform particle size.

[0038] The particle size of the konjac glucomannan microspheres prepared in this embodiment was determined to be 200 μm. Figure 8 As shown, under a microscope, the konjac glucomannan microspheres exhibit good sphericity and a smooth surface.

[0039] Example 5: Preparation of konjac glucomannan microspheres with a particle size of approximately 250 μm Under the condition that the water bath of the reactor is heated to 60℃, 200g of the prepared konjac glucomannan clarified solution and 600g of oil phase containing surfactant are transferred into the reactor. The stirring device is started and the stirring speed is set to 85rpm-95rpm / min. Stirring is continued at this stirring speed. After stirring for 1 hour, 50g of glutaraldehyde solution is added dropwise. The dropwise addition process must be carried out under stirring and must be ensured to be completed uniformly within 1 to 1.5 hours. After the glutaraldehyde solution is added, the reaction is continued under stirring for 8 hours to allow the aqueous phase droplets to fully cross-link and solidify to form konjac glucomannan microspheres. After the reaction, the konjac glucomannan microspheres are washed to remove impurities, thereby obtaining konjac glucomannan microspheres with uniform particle size.

[0040] The particle size of the konjac glucomannan microspheres prepared in this embodiment was determined to be 250 μm. Figure 9 As shown, under a microscope, the konjac glucomannan microspheres exhibit good sphericity and a smooth surface.

[0041] Example 6: Preparation of konjac glucomannan microspheres with a particle size of approximately 300 μm Under the condition that the water bath of the reactor is heated to 60℃, 200g of the prepared konjac glucomannan clarified solution and 600g of oil phase containing surfactant are transferred into the reactor. The stirring device is started and the stirring speed is set to 30rpm-45rpm / min. Stirring is continued at this stirring speed. After stirring for 1 hour, 50g of glutaraldehyde solution is added dropwise. The dropwise addition process must be carried out under stirring and must be completed uniformly within 1 to 1.5 hours. After the glutaraldehyde solution is added, the reaction is continued under stirring for 8 hours to allow the aqueous phase droplets to fully cross-link and solidify to form konjac glucomannan microspheres. After the reaction, the konjac glucomannan microspheres are washed to remove impurities, thereby obtaining konjac glucomannan microspheres with uniform particle size.

[0042] The particle size of the konjac glucomannan microspheres prepared in this embodiment was determined to be 300 μm. Figure 10 As shown, under a microscope, the konjac glucomannan microspheres exhibit good sphericity and a smooth surface.

[0043] Example 7: A type of konjac glucomannan microsphere was prepared by mixing and diluting 70 μm konjac glucomannan microspheres (in the dry state after filtration) with 1 ml of water for injection, and then injecting 500 μL into the hind leg muscle tissue of rats using a 1 ml syringe; the muscle tissue at the injection site was then observed.

[0044] Figure 1No obvious inflammation was observed in the muscle tissue images 1 week, 4 weeks, 8 weeks and 16 weeks after injection, nor was there any fusion of konjac glucomannan microspheres with surrounding tissue; and no konjac glucomannan microsphere residue was found in the muscle tissue 16 weeks after injection, indicating that the konjac glucomannan microspheres were basically completely degraded.

[0045] Example 8: A type of konjac glucomannan microsphere is prepared by mixing and diluting 70 μm konjac glucomannan microspheres (in the filtered state) with 1 ml of water for injection, and then injecting 100 μL into the heart tissue of a rat using a 1 ml syringe. The heart tissue at the injection site is then observed.

[0046] Figure 2 Images of heart tissue observed 1 week, 4 weeks, 8 weeks, and 16 weeks after injection show that the rat myocardial fibers are distributed in a normal short cylindrical shape, with branches and interconnected in a network, which is a normal myocardial tissue structure, and no tissue abnormalities were observed.

[0047] Example 9: A type of konjac glucomannan microsphere is prepared by mixing and diluting 70 μm konjac glucomannan microspheres (in the filtered state) with 1 ml of water for injection, and then injecting 100 μL into the lung tissue of rats using a 1 ml syringe. The lung tissue at the injection site is then observed.

[0048] Figure 3 Images of lung tissue observed 1 week, 4 weeks, 8 weeks and 16 weeks after injection. The lung histological morphology of rats was normal, with no abnormal pathological features. The single-layer epithelial structure in the small bronchi was intact, the pulmonary blood vessels were not ruptured, and the vesicular alveoli did not collapse.

[0049] Example 10: A type of konjac glucomannan microsphere is prepared by mixing and diluting 70 μm konjac glucomannan microspheres (in the filtered state) with 1 ml of water for injection, and then injecting 100 μL into the kidney tissue of rats using a 1 ml syringe. The kidney tissue at the injection site is then observed.

[0050] Figure 4 Images of kidney tissue observed 1 week, 4 weeks, 8 weeks, and 16 weeks after injection. The kidney histology was normal, with no abnormal pathological features. The glomeruli, which are important structural and functional components of the kidney, were normal in morphology, with no glomerular sclerosis or hyalinization.

[0051] As can be seen from Examples 7-10, the konjac glucomannan microspheres of this application are non-toxic and have good biocompatibility and low foreign body reaction. The raw material cost of the konjac glucomannan microspheres of this application is lower than that of agarose, and the preparation method of this application can achieve konjac glucomannan microspheres with uniform particle size without screening, which improves production efficiency and reduces preparation cost. Therefore, the konjac glucomannan microspheres of this application can be used to replace agarose microspheres.

[0052] The various embodiments of this application have been described above. These descriptions are exemplary and not exhaustive, nor are they limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen to best explain the principles, practical application, or improvement of the technology in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. A method for preparing konjac glucomannan microspheres with controllable particle size, characterized in that, Includes the following steps: Konjac flour is dissolved in an acidic solution and stirred thoroughly to form a gel. Then, it is placed in a high-pressure environment for high-temperature degradation. The degradation product is taken out and cooled to obtain a solid degradation product. The solid degradation product is mixed with an alkaline solution and stirred until completely dissolved. Then, it is filtered to obtain a clear konjac glucomannan solution. The clear konjac glucomannan solution is used as the aqueous phase. Provide an oil phase containing surfactants; The aqueous phase is added to the oil phase, and the aqueous phase is stirred at a constant speed using a stirring device to break it into droplets. A crosslinking agent is then added to crosslink and solidify the droplets to generate konjac glucomannan microspheres with uniform particle size.

2. The method for preparing konjac glucomannan microspheres with controllable particle size according to claim 1, characterized in that... ； The aqueous phase and the oil phase are placed in a reaction vessel, and the reaction vessel is heated in a water bath. The stirring device is used to stir the aqueous phase and the oil phase in the reaction vessel at a constant speed so that the aqueous phase is dispersed into uniform droplets; The crosslinking agent is gradually added to the reaction vessel so that the konjac glucomannan molecular chains in the droplets undergo a crosslinking and solidification reaction with the crosslinking agent, thereby generating solid konjac glucomannan microspheres.

3. The method for preparing konjac glucomannan microspheres with controllable particle size according to claim 2, characterized in that, The reactor is heated to 40℃-60℃, and the stirring speed of the stirring device in step S302 is 30rpm-300rpm; the cross-linking and curing reaction time between the konjac glucomannan molecular chains in the droplets and the cross-linking agent in step S303 is 8-12 hours.

4. The method for preparing konjac glucomannan microspheres with controllable particle size according to claim 1, characterized in that, The viscosity of the clarified konjac glucomannan solution is 10 mPa·s-60 mPa·s.

5. The method for preparing konjac glucomannan microspheres with controllable particle size according to claim 1, characterized in that, It also includes washing the konjac glucomannan microspheres, with the specific steps as follows: The konjac glucomannan microspheres containing the oil phase are transferred to a filtration device to separate the oil phase from the konjac glucomannan microspheres; Petroleum ether was added to the filtration device to wash the separated konjac glucomannan microspheres. The konjac glucomannan microspheres were washed a second time with ethanol. The konjac glucomannan microspheres were washed a third time with purified water.

6. The method for preparing konjac glucomannan microspheres with controllable particle size according to claim 1, characterized in that, The oil phase is selected from liquid paraffin, and the surfactant is sorbitan monooleate.

7. The method for preparing konjac glucomannan microspheres with controllable particle size according to claim 1, characterized in that, The crosslinking agent is glutaraldehyde.

8. The method for preparing konjac glucomannan microspheres with controllable particle size according to claim 1, characterized in that, The acid solution is hydrochloric acid with a concentration of 0.5 mol / L.

9. A type of konjac glucomannan microsphere, characterized in that, The konjac glucomannan microspheres are prepared by the method described in any one of claims 1 to 8, and their average particle size is between 10 μm and 300 μm.

10. A composition for medical aesthetic filling, characterized in that, include: The konjac glucomannan microspheres prepared by the method according to any one of claims 1 to 8 and water for injection.

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