WO2018190688A1 - Organic and inorganic composite granules and production method therefor - Google Patents

Abstract

The purpose of the present invention is to provide organic and inorganic composite granules of a uniform size and a production method therefor. To this end, the present invention provides inorganic and organic composite granules which are in the form of hydrogel and comprise organic members and inorganic members, wherein the weight ratio of the inorganic members to the organic members is 1 to 10, the sizes of the organic members and the inorganic members are 100 to 2000 μμ㎛, and the size distribution ranges from -20% to +20% with respect to the size of the granules. Also, the present invention provides a method for producing organic and inorganic composite granules. The organic and inorganic composite granules of the present invention have the effect of having a uniform size, and also have the effect of sustained release when a functional member is contained. In addition, there is an advantage in that cell culturing is easy. Furthermore, the production method of the present invention has advantages in that a large number of organic and inorganic composite granules of a uniform size can be produced in a short time, and the granules can be produced in a high yield. Therefore, the organic and inorganic composite granules and the production method therefor according to the present invention can be applied to various fields such as the pharmaceutical field, medical field, cosmetics field, food field, etc.

Description

Organic-inorganic complex granules and preparation method thereof

The present invention relates to an organic-inorganic composite granules and a preparation method thereof.

Bioceramic, which is representative of calcium phosphate, is a bioceramic alone or a ceramic-biopolymer organic-inorganic composite, and is used as a bone graft material or a bone filler in various forms such as powder, granule, paste, and support. In particular, in the case of granular form, since it can be easily applied to the irregular defects alone or in the form of paste, it has been variously applied in the fields of dentistry and orthopedics. In addition, in order to increase the biofunctionality of the granules it is also used to adsorb various drugs to ceramic or organic-inorganic composite granules.

In order to manufacture such ceramic or organic-inorganic composite granules, spray drying and chemical reaction methods (emulsion, sol-gel, etc.) are generally used. Spray drying method has the advantage of producing a large amount of granules in a short time, but has a disadvantage in that the production yield in the required size is very low due to the large size distribution of the granules. On the other hand, the chemical reaction method has the advantage of producing a relatively uniform size granules, but the mass production is difficult and the manufacturing process has a disadvantage.

In connection with a bone filling material carrying a sustained release osteoporosis treatment, Korean Laid-Open Patent Publication No. 10-2010-0026910 adds calcium phosphate microspheres carrying alendronate, a osteoporosis treatment, to bone fractures caused by osteoporosis. Filling materials are disclosed. However, this is difficult to mass production in a short time by obtaining a microsphere through the sol-gel process, there is a disadvantage that the size control of the produced microsphere is not easy.

In addition, Korean Patent Publication No. 10-2012-0021899 discloses a method for preparing a porous organic-inorganic hybrid, specifically, a crystal comprising a step of supporting an ionic compound or a polar compound on the porous organic-inorganic hybrid. Disclosed is a method for producing a porous, organic-inorganic hybrid. However, the above technique cannot obtain granules having a uniform particle size, and there is a problem in mass production of granules.

Electrostatic spraying is a method of atomizing a liquid by an electric force (electric field). Liquid droplets formed by electrospray have attracted attention as a useful nanotechnology in recent years because they have a high chargeability and have the advantage of preventing aggregation by their own dispersion. In particular, the electrospray technology is expected to be applied in a variety of fields because it is possible to deposit a fine and complex structure with low-cost equipment and easy operation in the air environment.

As described above, the microspray coater may be exemplified as the electrospray apparatus using the electrostatic charge method. Microgranular coating machine is mainly applied to organic matter. That is, it is used in the fields of pharmaceuticals, chemistry, cosmetics, foodstuffs, agriculture, etc. for the purpose of delivering active ingredients by forming granules using polymers and hydrogels and forming core-shell granules containing oils and various drugs therein. It is becoming. In particular, by providing an encapsulated microgranular coating machine has been applied to the food industry by realizing the effects of aging, storage stability, blocking harmful substances. In addition, it is being applied to the pharmaceutical industry by realizing effects such as release control, solubility and osmoticity improvement. In addition, it has been applied to the bio-pharmaceutical industry by implementing various effects in the in vivo test. However, in the case of producing granules with a fine granule coating machine, a raw material may have a high viscosity, or in the case of a material having good aggregation, problems such as clogging of the nozzle may occur.

Therefore, the inventors of the present invention can produce granules of uniform size in a short time and have high content of an inorganic member (for example, a ceramic member), which is highly applicable to bone graft materials and bone fillers, and a method for preparing organic-inorganic composite granules. The present invention was completed by studying.

<Preceding technical literature>

Korean Patent Publication No. 10-2010-0026910

Korean Patent Publication No. 10-2012-0021899

It is an object of the present invention to provide an organic-inorganic composite granules having a uniform size and containing an inorganic material and a method for preparing the same.

For this purpose

The present invention includes an organic member and an inorganic member, the weight ratio of the inorganic member to the organic member is 1 to 10, the size is 100 to 2000 μm, and the distribution range of the size is -20% to +20 relative to the size of the granules. It provides the organic-inorganic complex granules, characterized in that the range of%, hydrogel phase.

In addition, the present invention comprises the steps of preparing an organic member solution; Uniformly dispersing an inorganic member having a weight ratio of 1 to 10 with respect to the organic member in the organic member solution to form an organic-inorganic composite solution; Spraying the organic-inorganic composite solution in an electrostatic charge manner; It provides a method for producing an organic-inorganic composite granules comprising a; and forming a hydrogel phase by polymerizing the sprayed organic-inorganic composite solution.

The organic-inorganic composite granules of the present invention have an effect of having a uniform size, and also have an effect of sustained release when supporting a functional member. In addition, there is an advantage of easy cell culture. Furthermore, the production method of the present invention can produce a large amount of organic-inorganic composite granules of uniform size in a short time, there is an advantage that can be produced in a high yield granules. Therefore, the organic-inorganic composite granules and the manufacturing method thereof according to the present invention has an advantage that can be applied to various fields, such as pharmaceutical, medical, cosmetics, food.

1 is a view showing a photograph and the size and size distribution of the composite granules prepared by the embodiments of the present invention,

2 is a view showing the drug release properties of the multi-granules prepared by the embodiments of the present invention,

Figure 3 is a graph showing the cell proliferation behavior by the composite granules prepared by the embodiments of the present invention,

Figure 4 is a photograph showing the cell delivery capacity of the composite granules prepared by the embodiments of the present invention,

5 is a photograph showing the degree of dispersion of the inorganic member in the organic member according to the mixing method, and

Figure 6 is a photograph and graph showing the shape, size and size distribution of the organic-inorganic composite granules prepared by the embodiments of the present invention.

Hereinafter, with reference to the contents described in the accompanying drawings will be described in detail the present invention. However, the present invention is not limited or limited by the exemplary embodiments. Like reference numerals in the drawings denote members that perform substantially the same function.

The objects and effects of the present invention may be naturally understood or more apparent from the following description, and the objects and effects of the present invention are not limited only by the following description. In addition, in describing the present invention, when it is determined that the detailed description of the known technology related to the present invention may unnecessarily obscure the subject matter of the present invention, the detailed description thereof will be omitted.

In the present invention, 'size' means a diameter when the object is a spherical shape, the length of the long axis in the case of an elliptical shape.

The present invention

Including an organic member and an inorganic member,

The weight ratio of the inorganic member to the organic member is 1 to 10,

The size is from 100 to 2000 μm,

The distribution range of the size ranges from-20% to + 20% with respect to the size of the granules,

It provides an organic-inorganic complex granules characterized in that the hydrogel phase. Hereinafter, the organic-inorganic composite granules according to the present invention will be described in detail for each component.

The organic-inorganic composite granules according to the present invention include an organic member and an inorganic member. In this case, the organic member may be a natural biopolymer or a synthetic biopolymer, and specifically, may be at least one of alginate, collagen, gelatin, chitosan, cellulose, hyaluronate, and biopolymer, and a combination of these may be appropriate. Can be. In addition, the inorganic member of the present invention may be a ceramic member, and specifically, may be any one of calcium phosphate, bioglass, alumina, zirconia, and a composite thereof. The calcium phosphate is hydroxyapatite (HA), dicalcium phosphate (DCP: dicalcium phosphate), tricalcium phosphate (TCP: tricalcium phosphate), calcium tetraphosphate (TTCP: tetracalcium phosphate) and octacalcium phosphate ( OCP: may be any one of octacalcium phosphate, but is not limited thereto.

The organic-inorganic composite granules according to the present invention have a structure in which the inorganic member is uniformly dispersed in the organic member, so that the size of the composite granules can be uniform, and the presence or absence of additional sustained release of the functional member, effective culture of cells, etc. There is an effect to greatly improve the effectiveness of the multi-granular granules.

Specifically, when the granules are manufactured using only the organic member or the inorganic member, there is a problem in that it is difficult to perform a function as a carrier of the functional member due to the rapid release of the granules. In addition, in the case of preparing the granules by using only the inorganic member, in addition, it is not possible to culture or deliver the cells on the granules, for example, there is a problem that there is a limit in application in the medical field. The organic-inorganic composite granules according to the present invention have a sustained release property when carrying a functional member, and also have an advantage of greatly expanding the field of application by enabling the culture and delivery of cells.

At this time, the organic-inorganic composite granules of the present invention preferably have a weight ratio of the inorganic member to the organic member of 1 to 10, and 5 to 10. The present invention has the advantage of further improving the sustained release of the granules by including a large amount of the inorganic member in the granules, and, when the granules are used as a bone filler, etc. There is an advantage to being there. For example, when the weight ratio of the inorganic member is less than 1, the effect of improving the sustained release of the granules may be inadequate, and when used as a bone filler, there may be a problem that the bone formation component is insufficient. In addition, when the weight ratio of the inorganic member exceeds 10, it may be difficult to form granules having uniform characteristics due to problems such as aggregation of the inorganic member.

Organic-inorganic complex granules according to the present invention has a size of 100 to 2000 μm, the size distribution ranges from -20% to + 20% with respect to the size of the granules. Organic-inorganic composite granules according to the present invention has the size range and the distribution range of the size as described above, excellent stability of the specification of the granules has the advantage of very easy to apply to various applications.

In addition, the organic-inorganic composite granules according to the present invention are hydrogel phase. Accordingly, the composite granules of the present invention are advantageous for delivering bioactive substances such as cells, drugs, and proteins as compared with simple solid granules. In addition, it is advantageous to maintain the viability of the cells and to deliver them to the human body, and higher loading efficiency of drugs and the like than simple solid granules can be expected. In addition, the organic-inorganic composite granules of the present invention, which are hydrogels, are advantageous in controlling / controlling the drug release behavior in a sustained release form as compared with the granules of the solid solid phase. Simple solid granules are adsorbed and delivered to the surface of the solid granules to support the drug or protein. This is because the surface of the drug or protein-carrying surface is directly exposed to the portion to be delivered after delivery, and thus an initial excess is released, and it is difficult to control the release behavior for a long time thereafter. On the other hand, in the case of the composite granules on the hydrogel, since the inorganic member carrying the drug and the like is carried in the hydrogel, the initial excessive release phenomenon can be lowered and the release behavior can be controlled by using the physical / chemical properties of the hydrogel. In the case of the composite granules on the hydrogel, the cells or drugs can be delivered in the hydrogel so that two functional drugs or proteins such as growth factors can be delivered at the same time and their sequential release behavior can be realized. In addition, the solid granules must be delivered through a surgical operation that exposes the area to be delivered to the outside to be delivered to the human body, or mixed with other carriers on the hydrogel, and then delivered by syringe, but the granules on the hydrogel are themselves There is an advantage that can be delivered through non-invasive methods.

Organic-inorganic complex granules according to the present invention may further comprise a functional member or cells. In this case, the functional member may be a bisphosphonate-based drug, a polyphenol-based natural material, and the like, which is supported and delivered to the organic-inorganic composite granules.

More specifically, the functional member is a bisphosphonate-based drug alendronate, risedronate, etidronate, clodronate, neridronate, which are bisphosphonate-based drugs used as bone resorption inhibitors. It may include one or more materials selected from the group consisting of ibandronate, zoleronate and olpadronate.

In addition, the functional member is a polyphenol-based natural material, quercetein, genistein, curcumin, saurolactam, sauchinone, baicalin, and many others. Daidzein, Rutin, Anthocyanidin, Fisetin, Icariin, Kaempferol, E. Koreanum Nakei and Ikuol It may include one or more substances selected from the group consisting of (Equol).

The functional member is not limited to the above materials and can be selected and applied according to the required functionality.

The functional member may be an organic material or an inorganic material. In particular, growth factors such as BMP, which can induce the production of bone tissue as cells, tissues, hormones, and bone formation promoters, are included in the functional member, thereby promoting bone formation as the release of the functional member occurs to target cells in the body. can do.

When the organic-inorganic complex granules according to the present invention include cells, for example, after culturing stem cells on the organic-inorganic complex granules of the present invention, it can be applied to deliver them into the body.

The organic-inorganic composite granules according to the present invention may not include a dispersant. Organic-inorganic composite granules according to the present invention can be used in the pharmaceutical, medical, cosmetics or food, and the like, in this case, there may be a limit to the application in the case of containing a chemical such as a dispersant. In consideration of this, the organic-inorganic composite granules according to the present invention may not include a dispersant, especially when used in the above fields and the like. Examples of the application of the organic-inorganic complex granules according to the present invention include bone fillers, bone grafts and fillers. The bone filling material, which is supported on the sponge bone pores and induces bone formation promotion, fillers for cosmetic and molding, and fine plastic materials may be replaced and used.

In addition, the present invention

Preparing an organic member solution;

Uniformly dispersing an inorganic member having a weight ratio of 1 to 10 with respect to the organic member in the organic member solution to form an organic-inorganic composite solution;

Spraying the organic-inorganic composite solution in an electrostatic charge manner; And

Polymerizing the sprayed organic-inorganic complex solution to form a hydrogel phase;

It provides a method for producing an organic-inorganic composite granules comprising a.

Hereinafter, the manufacturing method of the present invention will be described in detail for each step.

The preparation method of the present invention includes the step of preparing an organic member solution. In this case, the organic member may be a natural biopolymer or a synthetic biopolymer, and specifically, may be at least one of alginate, collagen, gelatin, chitosan, cellulose, hyaluronate, and biopolymer, and a combination of these may be appropriate. Can be. The organic member is dissolved in, for example, phosphate buffer saline to prepare an organic member solution. Alternatively, it may be dissolved in water, glycerin, lipid oil, or the like and dispersed.

At this time, it is preferable not to use an organic solvent in the step of preparing an organic member solution of the production method of the present invention. In the case of using an organic solvent in the manufacturing process, there may be a limit in using the resultant in the medical field due to problems such as cytotoxicity. Therefore, in the manufacturing method of the present invention, by not using an organic solvent, problems that may occur in applying the composite granules according to the present invention to the pharmaceutical, medical, food, or cosmetic fields can be eliminated.

Next, the manufacturing method of the present invention comprises the step of uniformly dispersing the inorganic member of 1 to 10 compared to the organic member in a weight ratio in the organic member solution to form an organic-inorganic composite solution.

The inorganic member dispersed in the above step may be a ceramic member, and specifically, may be any one of calcium phosphate, bioglass, alumina, zirconia, and a composite thereof. The calcium phosphate is hydroxyapatite (HA), dicalcium phosphate (DCP: dicalcium phosphate), tricalcium phosphate (TCP: tricalcium phosphate), calcium tetraphosphate (TTCP: tetracalcium phosphate) and octacalcium phosphate ( OCP: may be any one of octacalcium phosphate, but is not limited thereto.

At this time, the inorganic member dispersed in the manufacturing method of the present invention may include a functional member. The functional member may be included in the inorganic member by various methods such as adsorption, and the specific functional member may be a bisphosphonate-based drug, a polyphenol-based natural material, or the like as described above.

In the production method of the present invention, it is preferable to introduce an inorganic member of 1 to 10 relative to the organic member in a weight ratio of the organic member solution, and to introduce a weight ratio of 5 to 10. The present invention has the advantage of further improving the sustained release of the granules by including a large amount of the inorganic member in the composite solution, and also, when the granules are used as a bone filler or the like, it contains a relatively large amount of bone forming components There is an advantage to doing this. For example, when the weight ratio of the inorganic member is less than 1, the effect of improving the sustained release of the granules may be inadequate, and when used as a bone filler, there may be a problem that the bone formation component is insufficient. In addition, when the weight ratio of the inorganic member exceeds 10, a problem may occur in the step of forming the granules due to clogging of the agglomeration nozzle of the inorganic member, thereby making it difficult to form granules having uniform characteristics.

On the other hand, since the inorganic member has strong cohesiveness and very low dispersibility, it is necessary to introduce the inorganic member while uniformly dispersing the inorganic member in the organic member solution. For example, the manufacturing method of the present invention preferably further comprises the step of forming an organic-inorganic complex solution as described above for the dispersion, and then dispersing the inorganic member with a co-rotating mixer, and stirring with an ultrasonic mixer. .

As described below, the manufacturing method of the present invention forms a granule by spraying in an electrostatic manner. The electrostatic charge method has an advantage of producing granules having a relatively uniform size, but may cause a problem in that the nozzle is clogged during the process to make granules impossible. In particular, when the organic-inorganic composite solution containing the highly cohesive inorganic member as in the present invention is used as a raw material, since the nozzle is clogged very easily by the aggregation of the inorganic member in the raw material, the raw material containing the inorganic member is used. It is not easy to consider making granules by spraying with electrostatic charge.

In the present invention, in order to solve such a problem, it is to further include the step of dispersing the inorganic member with a co-rotating mixer, the inorganic-inorganic composite solution formed of the raw material of the electrostatic charge injection, and stirring with an ultrasonic mixer as described above desirable. Through this process, it is possible to prevent the nozzle from clogging and to prepare granules having uniform physical properties.

However, when using a co-rotation mixer and an ultrasonic mixer, the temperature of the complex solution itself is greatly increased when used for too long, which may be a problem when the functional solution or cells in the complex solution. That is, if the temperature of the complex solution is too high, there is a problem that the properties of the drug that may be included therein, or the cells may die. In consideration of the above, the mixing using the co-rotating mixer and the ultrasonic mixer is preferably performed within a range in which the temperature of the complex solution does not exceed 40 ° C. For example, the co-rotating mixer is within 6 minutes, It can be performed within 30 minutes.

On the other hand, the organic-inorganic complex solution formed in the manufacturing method of the present invention may further comprise the step of supporting a functional member or cells. As described above, the functional member may be mixed with the organic member solution in the state contained in the inorganic member, or may be included after preparing the organic-inorganic composite solution. As described above, when the organic-inorganic complex solution supports the functional member or the cell, it is necessary to adjust the process conditions so as not to adversely affect the supported functional member or the cell when using the co-rotating mixer or the ultrasonic mixer as described above.

Next, the manufacturing method of the present invention includes the step of spraying the organic-inorganic complex solution in the electrostatic charge method, for example, the electrostatic charge injection may be performed with a microgranular coating. In the case of producing granules by the electrostatic charge method, most of the raw materials to be introduced can be converted into granules so that the yield is excellent, a large amount of granules can be prepared in a short time, and granules can be manufactured in a relatively uniform size. There is this. However, there is a problem that the nozzle may be clogged if the raw material has a high viscosity or a high degree of aggregation. Although the organic member included in the raw material used in the production method of the present invention has a high viscosity and the inorganic member has a very high cohesiveness, it is generally not suitable for making granules by an electrostatic charge method. The granules can be prepared in an electrostatic manner by adding a step of uniformly dispersing the inorganic member in the member solution, and further, if necessary, further dispersing the inorganic member with a post-coagulation mixer and stirring with an ultrasonic mixer.

When the step is performed with a microgranular coater, the size of the spray nozzle of the microgranular coater is preferably 50 μm to 1000 μm. When the size is less than 50 μm, it is difficult to spray the organic-inorganic composite solution including the inorganic member through the nozzle due to nozzle clogging, etc., and when the size is more than 1000 μm, the size is easily applicable to the current clinical practice. There is a problem that the production of micro-sized particles is difficult. In addition, the voltage is preferably 500V to 2,500V. If the voltage is less than 500 V, uniform spraying of the sprayed solution is difficult, making it difficult to form spherical particles. If the voltage is higher than 2,500, the spherical solution is sprayed in an unfolded form after spraying, thus yielding high yield. There is a problem that is difficult to manufacture granules. In addition, the pressure is preferably 100 mbar to 1500 mbar. If the pressure is less than 100 mbar causes frequent nozzle clogging and the solution passed through the nozzle is excessively wide spread, if the pressure exceeds 1500 mbar spherical solution is a polymerization-induced solution (for example, CaCl ₂ solution) is difficult to form spherical granules by applying a greater force than necessary when dropping. Furthermore, the vibration frequency is preferably 100 Hz to 6,000 Hz. If the vibration frequency exceeds 6,000 Hz, there is a problem in that spherical granules with a tail are formed.

The production method according to the present invention includes the step of polymerizing the organic-inorganic complex solution sprayed by the above method to form a hydrogel phase. The polymerization method may be performed by any one of ion crosslinking, chemical crosslinking, and photocrosslinking. Double ion crosslinking may be carried out using at least one polymerization inducing substance of calcium chloride (CaCl ₂ ), calcium sulfate (CaSO ₄ ), calcium carbonate (CaCO ₃ ), and specifically, this step may be carried out, for example The combined solution can be carried out by dropping into a CaCl ₂ polymerization induction solution. According to the production method of the present invention, since the granules are formed as a hydrogel rather than a simple solid, it is advantageous to deliver bioactive substances such as cells, drugs, and proteins as compared with the granules of the simple solid phase. In addition, it is advantageous to maintain the viability of the cells and to deliver them to the human body, and higher loading efficiency of drugs and the like than simple solid granules can be expected. In addition, the organic-inorganic composite granules of the present invention, which are hydrogels, are advantageous in controlling / controlling the drug release behavior in a sustained release form as compared with the granules of the solid solid phase. Simple solid granules are adsorbed and delivered to the surface of the solid granules to support the drug or protein. This is because the surface of the drug or protein-carrying surface is directly exposed to the portion to be delivered after delivery, and thus an initial excess is released, and it is difficult to control the release behavior for a long time thereafter. On the other hand, in the case of the composite granules on the hydrogel, since the inorganic member carrying the drug and the like is carried in the hydrogel, the initial excessive release phenomenon can be lowered and the release behavior can be controlled by using the physical / chemical properties of the hydrogel. In the case of the composite granules on the hydrogel, the cells or drugs can be delivered in the hydrogel so that two functional drugs or proteins such as growth factors can be delivered at the same time and their sequential release behavior can be realized. In addition, the solid granules must be delivered through a surgical operation that exposes the area to be delivered to the outside to be delivered to the human body, or mixed with other carriers on the hydrogel, and then delivered by syringe, but the granules on the hydrogel are themselves There is an advantage that can be delivered through non-invasive methods.

It is preferable that the density | concentration of the organic member solution used by the manufacturing method of this invention is 0.5 to 2.0 weight%. If the concentration is less than 0.5% by weight, the concentration of the organic member is too low to achieve a sufficient crosslinking density, so that it is difficult to maintain sufficient mechanical properties of the manufactured granules. When the concentration is higher than 2.0% by weight, the concentration is too high. It is difficult to uniformly disperse the complex solution due to the high viscosity of the solution is formed, there is a problem that it is difficult to form a uniform spherical droplets using the injection equipment.

In addition, the size of the inorganic member used in the production method of the present invention is preferably in the range of 50 nm to 50 μm. If the size is less than 50 nm, the surface area of the inorganic member (e.g., ceramic powder) may increase, making it difficult to disperse evenly in the organic member (e.g., alginate solution), and through the pores inside the hydrogel to the outside There is a possibility to be discharged, and also, if the cells are supported, the endocytosis of the cells also can not be ignored, and if it exceeds 50 μm, it is difficult to pass through the micro-sized nozzle, the particle production is There is a difficult problem.

In the manufacturing method of this invention, it is preferable not to use a dispersing agent for the dispersion of an inorganic member. In general, dispersants are used for uniform dispersion due to the cohesiveness of the inorganic member. If the dispersant is included in the granules to be produced, problems may arise, for example, in the medical, pharmaceutical, food, and cosmetic fields. Therefore, in the production method of the present invention, there is an advantage that can greatly extend the field of application of the granules to be produced, without using a dispersant. Instead of using a dispersant in the production method of the present invention, for uniform dispersion of the inorganic member, stirring and dispersion are performed through a co-rotating mixer or an ultrasonic mixer.

According to the production method of the present invention, in the preparation of organic-inorganic composite granules having sustained release and cell transferability, by producing the granules by the electrostatic charge method, the yield is improved, and the advantage of producing a large amount of granules in a short time have. In addition, it is possible to produce a granule of uniform size has the advantage that it is easy to apply to various fields. In addition, the functional member or the cell can be easily supported, there is no need to use an organic solvent or dispersant in the manufacturing process, there is an advantage that can be greatly extended to the field of application, such as medical, pharmaceutical, food, cosmetics .

Hereinafter, the present invention will be described in more detail based on Examples, Comparative Examples, and Experimental Examples. The following Examples, Comparative Examples, and Experimental Examples are merely illustrative of the present invention, and the scope of the present invention is not limited to the following description.

In the following examples, when manufacturing the organic-inorganic composite granules by using a micro-sized nozzle using an electrostatic charge method, the process of uniformly dispersing the inorganic member in the organic solution is most important. The organic-inorganic composite granules were prepared by dispersing as much as possible using an ultrasonic mixer. Even if the time of the co-rotating mixer was increased, the dispersibility did not improve significantly, and there was a fear of denaturation by heat as much as the organic material was used. Thereafter, the mixture was mixed using an ultrasonic mixer. Dispersibility increased significantly with the mixing time, but after 30 minutes of ultrasonic mixer treatment, the temperature in solution reached about 40 ℃, which was high for delivering bioactive substances such as cells and proteins after treatment. May be a constraint, the treatment time of the ultrasonic mixer was limited to 30 minutes.

<Example 1>

Dissolve the alginate, an organic member, in distilled water in a tertiary distilled water to prepare a 1.0 wt% alginate solution, and use a co-rotating mixer and an ultrasonic mixer in a ratio of 1: 1 (organic member: inorganic member) to inorganic apatite in a weight ratio of organic member. And mixed to prepare an organic-inorganic complex solution. At this time, Paste mixer was used as co-rotation mixer, Rotational sonicator was used as ultrasonic mixer, and each was performed for 6 minutes and 15 minutes.

The organic-inorganic composite solution prepared above was introduced into a microgranular coating machine (Buchi, B-395 pro), sprayed with a nozzle having a diameter of 150 μm, and dropped into a CaCl ₂ solution to prepare organic-inorganic composite granules on a hydrogel. After dropping, the mixture was crosslinked in CaCl ₂ solution for 30 minutes, and then washed twice with PBS. In order to measure the size, a certain amount of organic / inorganic composite granules were transferred to a petri dish to obtain an image through an optical microscope, and the size of the particles was calculated using an ImageJ program to calculate the average size of the particles.

<Example 2>

An organic-inorganic composite granule on a hydrogel was prepared in the same manner as in Example 1, except that quercetin, which is a functional member, was mixed with the organic-inorganic composite solution in an amount of 1 wt% based on the weight of the organic-inorganic composite solution.

<Example 3>

The organic-inorganic composite granules on the hydrogel were prepared in the same manner as in Example 1 except that the quoscetin, which is a functional member, was mixed with the organic-inorganic composite solution in an amount of 2.5 wt% based on the weight of the organic-inorganic composite solution.

<Example 4>

The organic-inorganic composite granules on the hydrogel were prepared in the same manner as in Example 1, except that quercetin, which is a functional member, was mixed with the organic-inorganic composite solution in an amount of 5 wt% based on the weight of the organic-inorganic composite solution.

Example 5

Alginate, an organic member, was dissolved in tertiary distilled water to prepare a 1.0 wt% alginate solution, and a nano-apatite, which is an inorganic member, was used at a weight ratio of 1: 4 (organic member: inorganic member) at a weight ratio of the organic member, using a co-rotating mixer and an ultrasonic mixer. And mixed to prepare an organic-inorganic complex solution. At this time, Paste mixer was used as co-rotation mixer, Rotational sonicator was used as ultrasonic mixer, and each was performed for 6 minutes and 15 minutes.

The organic-inorganic composite solution prepared above was introduced into a microgranular coating machine (Buchi, B-395 pro), sprayed with a nozzle having a diameter of 150 μm, and dropped into a CaCl ₂ solution to prepare organic-inorganic composite granules on a hydrogel. . After dropping, the mixture was crosslinked in CaCl ₂ solution for 30 minutes, and then washed twice with PBS. In order to measure the size, a certain amount of organic / inorganic composite granules were transferred to a petri dish to obtain an image through an optical microscope, and the size of the particles was calculated using an ImageJ program to calculate the average size of the particles.

<Example 6>

Organic member: An organic-inorganic composite granule was prepared in the same manner as in Example 5 except that the inorganic member was 1: 6.

<Example 7>

Organic member: An organic-inorganic composite granule was prepared in the same manner as in Example 5 except that the inorganic member was 1: 8.

<Example 8>

Organic member: An organic-inorganic composite granule was prepared in the same manner as in Example 5 except that the inorganic member was 1:10.

<Examples 9 to 12>

Organic-inorganic composite granules were prepared in the same manner as in Examples 5 to 8 except that a nozzle having a diameter of 120 μm was used.

<Examples 13 to 16>

Organic-inorganic composite granules were prepared in the same manner as in Examples 5 to 8 except that a nozzle having a diameter of 200 μm was used.

Comparative Example 1

An experiment was conducted to prepare organic-inorganic composite granules in the same manner as in Example 1 except that 1:20 (organic member: inorganic member) was mixed in a weight ratio of the inorganic apatite to the organic member.

However, the clogging of the nozzle of the microgranular coating machine occurred seriously during the process, and the ceramic content in the manufactured granules was significantly lower than the theoretical content, so that it was impossible to produce uniform granules, and thus the production yield was significantly reduced.

Experimental Example 1

Confirmation of Compound Granule Size

In order to confirm the size and size distribution of the organic-inorganic composite granules prepared according to the present invention, the following experiment was performed.

The organic-inorganic composite granules prepared in Examples 1 to 4 were confirmed the form of the granules through an optical microscope, and the size and distribution of the size were confirmed using ImageJ software, and the results are shown in FIG. 1.

According to Figure 1, it can be seen that the organic-inorganic composite granules according to the present invention are spherical, having a size range of about 250 to about 270 μm, and having a uniform size that does not deviate from ± 20%.

Experimental Example 2

Check the amount of supporting weapon

In order to confirm the inorganic member loading of the prepared organic-inorganic composite granules, the following experiment was performed.

Except for adjusting the content of the organic member and the inorganic member as shown in Table 1 below, the weight ratio of the actual organic member and the inorganic member was measured for the organic-inorganic composite granules prepared in the same manner as in Example 1. It is shown in Table 1 below.

Organic member: Inorganic member	Theoretical weight ratio	Actual weight ratio
1: 0.1	0.1	0.124
1: 0.25	0.25	0.271
1: 1	One	0.962
1: 2.5	2.5	2.796
1:10	10	7.760

According to Table 1, the inorganic member content of the actually prepared organic-inorganic composite granules is almost equal to the amount of the inorganic member introduced into the raw material, and thus, no precipitation or clogging of the inorganic member occurs in the manufacturing process, and thus a high yield. It can be seen that the composite granules can be prepared.

Experimental Example 3

Drug Sustained Release Check

In order to confirm drug sustained release of the organic-inorganic complex granules of the present invention, the following experiment was performed.

Place the organic-inorganic complex granules prepared in Examples 1 to 4 of the present invention on MC3T3 osteoblasts as shown in Figure 2b, and measure the amount of release of the drug over time and the degree of proliferation of the cells over time, respectively 2a and 2c.

The organic-inorganic complex granules prepared in Examples 1 to 4 were placed in a phosphate buffer solution (PBS), and the PBS solution was taken using the total substitution method for each time, and then the concentration of the drug was released by measuring the absorbance using the spectroscopic analysis method. The release behavior was confirmed by calculating.

In addition, the organic-inorganic complex granules prepared in Examples 1 to 4 were placed on the transwell, and the cells were attached to the well plate surface, and then cultured together in the culture medium. In order to quantify the proliferation of the cells, the quercetin-sensing organic-inorganic complex granules were removed, and the culture solution was also removed. After washing with PBS, the culture solution containing the MTS assay was applied and then left in a cell culture incubator for 2 hours. The cultures were then taken and measured for absorbance at 495 nm using a plater reader to analyze cell proliferation trends.

According to Figure 2a, it can be seen that the drug is gradually released over time, and according to Figure 2c, it can be seen that the degree of cell proliferation is increased by the released drug. Through this, in fact, the organic-inorganic complex granules of the present invention can be used for drug delivery, and specifically, for example, it can be seen that it can be used for treating osteoporosis and the like in the body.

Experimental Example 4

Confirmation of drug sustained release through cell culture

In order to confirm the cell culture of the organic-inorganic complex granules of the present invention, the following experiment was performed.

Method for delivering the organic-inorganic complex granules containing quercetin at each concentration prepared in Examples 1 to 4 of the present invention using a transwell, and analyzing the cell proliferation and bone differentiation behavior of the quercetin released from the composite granules Osteoblasts (MC3T3) cells were cultivated, and through the Cell Lysis method, the degree of bone differentiation of the cultured cells was confirmed by ALP activity, which is a representative factor thereof, and DNA quantification was performed, respectively. Shown in 3b.

According to Figure 3, the degree of cell proliferation in the first week is almost the same, but in the second week through the fact that the cell proliferation occurs more if the quercetin content, the sustained-release characteristics of the organic-inorganic complex granules according to the present invention You can check it.

Experimental Example 5

Confirmation of Cell Support of Organic-Inorganic Granules

After preparing the organic-inorganic complex solution in the same manner as described in Example 5, osteoblasts (MC3T3) is introduced into the complex solution at a concentration of 1.0 X 106 cells / ml and 5.0 X 106 cells / ml, and then slowly Stirred. Thereafter, the method was sprayed in the same manner as described in Example 5, and this was added dropwise to a CaCl ₂ solution to prepare an organic-inorganic composite granule on a hydrogel. Thereafter, in order to confirm whether the cells were supported in the composite granules, staining was performed using a DAPI solution capable of discriminating cell nuclei, and confirmed through a fluorescence microscope, which is shown in FIG. 4. According to Figure 4, it can be seen that cells are uniformly supported in the organic-inorganic complex granules of the present invention.

Experimental Example 6

Check uniform dispersion by mixing method

In the mixing of the organic member and the inorganic member, the following experiment was performed to confirm the uniform dispersion degree of the organic-inorganic composite solution according to the mixing method.

Inorganic-inorganic composite solution was prepared by the same method as described in Example 5. The prepared organic-inorganic composite solution was mixed for 6 minutes using a magnetic stirrer and 21 minutes of mixing. On the other hand, the prepared organic-inorganic composite solution was mixed for 6 minutes with a co-rotating mixer, followed by an ultrasonic mixer for 15 minutes. Agitation was carried out, and then stained using an Alizarin red solution, and the result was confirmed through an optical microscope. The results are shown in FIG. 5A, it can be seen that when the stirring is performed with a general magnetic stirrer, the organic member and the inorganic member are not sufficiently mixed, and the inorganic members are heavily aggregated with each other. In addition, in the case of stirring with a general magnetic stirrer, it can be confirmed that as time passes, the aggregation phenomenon becomes more severe. On the other hand, in FIG. 5B, when mixing using the co-rotation mixer and the ultrasonic mixer, it can be seen that the organic member and the inorganic member are uniformly mixed and the inorganic member is uniformly dispersed on the organic member.

Experimental Example 7

Confirmation of Compound Granule Size

In order to confirm the size and size distribution of the organic-inorganic composite granules prepared according to the present invention, the following experiment was performed.

The organic-inorganic composite granules prepared in Examples 5 to 16 were confirmed the form of the granules through an optical microscope, and the size and distribution of size were confirmed using ImageJ software, and the results are shown in FIG. 6.

According to Figure 6, it can be seen that the organic-inorganic composite granules according to the present invention have a spherical shape, have a size range of about 200 to about 500 μm, and the size distribution has a uniform size that does not deviate from ± 15%.

Claims (19)

1. Including an organic member and an inorganic member,

   The weight ratio of the inorganic member to the organic member is 1 to 10,

   The size is from 100 to 2000 μm,

   The distribution range of the size ranges from-20% to + 20% with respect to the size of the granules,

   Organic-inorganic complex granules, characterized in that the hydrogel phase.
2. The method of claim 1,

   Organic-inorganic composite granules, characterized in that the weight ratio of the inorganic member to the organic member is 5 to 10.
3. The method of claim 1.

   The organic-inorganic complex granules are characterized in that the organic-inorganic complex granules further comprises a functional member or cells.
4. The method of claim 1,

   The organic member is an organic-inorganic composite granules, characterized in that the natural biopolymer or synthetic biopolymer.
5. The method of claim 1,

   The inorganic member is an organic-inorganic composite granules, characterized in that the ceramic member.
6. The method of claim 3,

   The functional member is an organic-inorganic composite granules, characterized in that it comprises a bisponate-based drug, a polyphenol-based natural material and a combination thereof.
7. The method of claim 1,

   The organic-inorganic composite granules are used in one of the fields selected from the group consisting of pharmaceutical, medical, cosmetics and food.
8. The method of claim 1,

   An organic-inorganic composite granule, characterized in that it does not contain a dispersant.
9. Preparing an organic member solution;

   Uniformly dispersing an inorganic member having a weight ratio of 1 to 10 with respect to the organic member in the organic member solution to form an organic-inorganic composite solution;

   Spraying the organic-inorganic composite solution in an electrostatic charge manner; And

   Polymerizing the sprayed organic-inorganic complex solution to form a hydrogel phase;

   Organic-inorganic composite granules manufacturing method comprising a.
10. The method of claim 9,

    The inorganic member dispersed in the organic member solution is a method for producing an organic-inorganic composite granules, characterized in that 5 to 10 compared to the organic member in weight ratio.
11. The method of claim 9,

    After forming the organic-inorganic complex solution, dispersing the inorganic member with a co-rotating mixer, the method of manufacturing an organic-inorganic composite granules, characterized in that further comprising stirring with an ultrasonic mixer.
12. The method of claim 9,

    The step of polymerizing the organic-inorganic composite solution is a method for producing organic-inorganic composite granules, characterized in that the granules are sprayed dropwise into the polymerization induction solution.
13. The method of claim 9,

    The inorganic member is a method for producing an organic-inorganic composite granules, characterized in that it comprises a functional member.
14. The method of claim 9,

    Method for producing an organic-inorganic composite granules further comprising the step of supporting a functional member or cells in the formed organic-inorganic composite solution.
15. The method of claim 9,

    The concentration of the organic member solution is a method for producing an organic-inorganic composite granules, characterized in that 0.5 to 5% by weight.
16. The method of claim 9,

    The inorganic member has a size of 20 nm to 10 μm.
17. The method of claim 11,

    Dispersing the inorganic member with the co-rotating mixer, and stirring with an ultrasonic mixer is a method for producing an organic-inorganic composite granules, characterized in that the temperature of the organic-inorganic composite solution does not exceed 40 ℃.
18. The method of claim 9,

    The step of spraying the organic-inorganic composite solution in the electrostatic charge method may be performed by a fine granule coating machine,

    The size of the injection nozzle of the microgranular coating machine is in the range of 50 to 1,000 μm, the voltage is in the range of 500 to 2,500 V, the pressure is in the range of 100 to 1,500 mbar, and the vibration frequency is in the range of 100 to 6,000 Hz. Process for producing organic-inorganic composite granules.
19. The method of claim 9,

    The organic-inorganic composite gel production method of the organic-inorganic composite granules, characterized in that it does not contain a dispersant.

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