KR102227720B1 - Organic and inorganic mixture granules and manufacturing method thereof - Google Patents

Abstract

It is an object of the present invention to provide an organic-inorganic composite granule having a uniform size and a method for producing the same. To this end, the present invention includes an organic member and an inorganic member, and the weight ratio of the inorganic member to the organic member is 1 to 10, the size is 100 to 2000 μµm, and provides an organic-inorganic composite granule characterized in that it is in the form of a hydrogel. do. In addition, the present invention provides a method for producing organic-inorganic composite granules. The organic-inorganic composite granules of the present invention have an effect of having a uniform size, and also have an effect of having a sustained release property when a functional member is supported. In addition, there is an advantage in that cell culture is easy. Further, the manufacturing method of the present invention has the advantage of being able to produce a large amount of organic-inorganic composite granules having a uniform size in a short time, and to produce granules with a high yield. Accordingly, the organic-inorganic composite granules and their manufacturing method according to the present invention have advantages that can be applied to various fields such as pharmaceutical field, medical field, cosmetic field, and food field.

Description

Organic-inorganic complex granules and manufacturing method thereof {ORGANIC AND INORGANIC MIXTURE GRANULES AND MANUFACTURING METHOD THEREOF}

The present invention relates to an organic-inorganic composite granule and a method for producing the same.

The bioceramic representative of calcium phosphate is a bioceramic alone or a ceramic-biopolymer organic-inorganic complex, 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 the granule form, since it can be easily applied alone or in the form of a paste to the irregularly shaped defect, it is widely applied in the fields of dentistry and orthopedics. In addition, in order to increase the biofunctionality of the granules, various drugs are adsorbed onto ceramic or organic-inorganic complex granules.

In order to manufacture such ceramic or organic-inorganic composite granules, spray drying method and chemical reaction method (emulsion, sol-gel method, etc.) are generally used. The spray drying method has the advantage of being able to produce a large amount of granules in a short time, but has a disadvantage in that the production yield at 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 being able to produce granules of relatively uniform size, but it has the disadvantages that mass production is difficult and the manufacturing process is complicated.

Regarding bone fillers carrying sustained-release osteoporosis treatments, Korean Patent Laid-Open No. 10-2010-0026910 is a bone used in osteoporosis-caused fracture patients by adding calcium phosphate microspheres loaded with alendronate, an osteoporosis treatment, to human demineralized bone. The filler was started. However, this has a disadvantage in that it is difficult to mass-produce within a short time by obtaining microspheres through a sol-gel process, and it is not easy to control the size of the produced microspheres.

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

Electrostatic spraying is a method of atomizing liquid by electric force (electric field). Since droplets formed by electrospray have high charging properties, they are attracting attention as a useful nanotechnology recently because they have the advantage of preventing agglomeration by self-dispersing. In particular, the electrospray technology is expected to be applied in various fields because it is possible to deposit fine and complex structures with inexpensive equipment and simple operation in an atmospheric environment.

As described above, an electrospray device using an electrostatic charge method may be a fine granule coating machine. The microgranule coating machine is mainly applied to organic matter. In other words, by making granules using polymers and hydrogels and forming core-shell granules containing oil or various drugs inside, they are applied in fields such as pharmaceuticals, chemistry, cosmetics, foodstuffs, and agriculture for the purpose of delivery of active ingredients. Has become. In particular, by providing a microgranule coating machine that can be encapsulated, it is applied to the food industry by implementing effects such as improved aging, storage stability, and blocking harmful substances. In addition, it is applied to the pharmaceutical industry by implementing effects such as release control, solubility and osmotic improvement. In addition, it realizes various effects in in vivo tests and is applied to the bio-pharmaceutical industry. However, in the case of manufacturing granules by a fine granule coating machine, a problem such as clogging of a nozzle may occur in the case of a raw material having a high viscosity or a material that is well agglomerated.

Accordingly, the inventors of the present invention can produce granules of uniform size in a short time and the content of inorganic members (for example, ceramic members) is increased, so that the organic-inorganic composite granules and methods of manufacturing the same are highly likely to be utilized as bone graft materials and bone fillers. The present invention was completed by studying.

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 granule having a uniform size and containing an inorganic substance, and a method for producing the same.

For this

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 organic-inorganic composite granules, the average size of the organic-inorganic composite granules is 100 to 500 μm, It provides an organic-inorganic composite granule, characterized in that.

In addition, the present invention comprises the steps of preparing an organic member solution; Forming an organic-inorganic composite solution by uniformly dispersing 1 to 10 inorganic members relative to the organic member in a weight ratio in the organic member solution; Spraying the organic-inorganic complex solution in an electrostatically charged manner; And polymerizing the sprayed organic-inorganic composite solution to form a hydrogel phase.

The organic-inorganic composite granules of the present invention have an effect of having a uniform size, and also have an effect of having a sustained release property when a functional member is supported. In addition, there is an advantage in that cell culture is easy. Further, the manufacturing method of the present invention has the advantage of being able to produce a large amount of organic-inorganic composite granules having a uniform size in a short time, and to produce granules with a high yield. Accordingly, the organic-inorganic composite granules and their manufacturing method according to the present invention have advantages that can be applied to various fields such as pharmaceutical field, medical field, cosmetic field, and food field.

1 is a view showing a photograph and size of a composite granule manufactured according to embodiments of the present invention,
2 is a view showing the drug release characteristics of the composite granules prepared according to the embodiments of the present invention,
3 is a graph showing the cell proliferation behavior by the composite granules prepared according to the examples of the present invention,
Figure 4 is a photograph showing the cell delivery ability of the composite granules prepared according to 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
6 is a photograph and a graph showing the shape and size of the organic-inorganic composite granules prepared according to embodiments of the present invention.

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

Objects and effects of the present invention may be naturally understood or more clearly understood by 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 a detailed description of a known technology related to the present invention may unnecessarily obscure the subject matter of the present invention, a detailed description thereof will be omitted.

In the present invention,'size' means the diameter when the object is spherical, and means the length of the major axis when the object is elliptical.

The present invention

Including an organic member and an inorganic member,

The weight ratio of the inorganic member to the organic member is an organic-inorganic composite granule of 1 to 10,

The average size of the organic-inorganic composite granules is 100 to 500 μm,

delete

It provides an organic-inorganic composite granule characterized in that it is in the form of a hydrogel. Hereinafter, the organic-inorganic composite granules according to the present invention will be described in detail for each configuration.

The organic-inorganic composite granule according to the present invention includes an organic member and an inorganic member. At this time, 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 an appropriate combination of these. I can. In addition, the inorganic member of the present invention may be a ceramic member, and specifically, may be any one of calcium phosphate, biological glass, alumina, zirconia, and composites thereof. The calcium phosphate-based is hydroxyapatite (HA), dicalcium phosphate (DCP), tricalcium phosphate (TCP), tetracalcium phosphate (TTCP), and octane calcium phosphate (TTCP). OCP: octacalcium phosphate), but is not limited thereto.

The organic-inorganic composite granule according to the present invention has a structure in which the inorganic member is uniformly dispersed in the organic member, so that the size of the composite granule can be uniform, and the presence or absence of the added functional member sustained release, effective cultivation of cells, etc. There is an effect of greatly improving the utility of the composite granules.

Specifically, in the case of manufacturing granules with only an organic member or an inorganic member, there is a problem in that it is difficult to perform the function as a carrier of the functional member due to the immediate release of the granules. In addition, in the case of manufacturing granules with only an inorganic member, cells cannot be cultured or transferred on the granules in addition to this, and there is a problem in that, for example, it is limited in application to the medical field. The organic-inorganic composite granules according to the present invention have a characteristic of sustained release when a functional member is supported, and also have the advantage of being able to greatly expand the field of application by enabling the culture and delivery of cells.

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

The organic-inorganic composite granules according to the present invention have an average size of 100 to 500 μm. The organic-inorganic composite granules according to the present invention have the same size range as described above, so that the standard stability of the granules is excellent, and thus the ease of application to various fields of application is very excellent.

In addition, the organic-inorganic composite granules according to the present invention are in the form of a hydrogel. Accordingly, the composite granules of the present invention are advantageous in delivering physiologically active substances such as cells, drugs, and proteins compared to simple solid granules. In addition, it is advantageous to maintain the viability of cells and deliver them into the human body, and higher loading efficiency of drugs, etc. than simple solid granules can be expected. In addition, the organic-inorganic composite granules of the present invention, which are hydrogels, have an advantage in controlling/controlling the drug release behavior with sustained release compared to the simple solid granules. Simple solid granules are delivered by adsorption on the surface of solid granules to support drugs or proteins. This is because the surface on which the drug or protein is loaded is directly exposed to the portion to be delivered after delivery, so that an initial excess is released, and it is difficult to control the release behavior for a long period of time thereafter. On the other hand, in the case of complex granules on a hydrogel, since an inorganic member carrying a drug or the like is supported and delivered in the hydrogel, the initial excessive release phenomenon can be reduced, and the release behavior can be controlled by using the physical/chemical properties of the hydrogel. In the case of complex granules on a hydrogel, since cells or drugs can be delivered by being contained in the hydrogel, two functional drugs or proteins such as growth factors can be simultaneously delivered and their sequential release behavior can be implemented. In addition, in order to deliver solid granules into the human body, they must be delivered through a surgical operation that exposes the area to be delivered to the outside or mixed with other carriers on a hydrogel and delivered by a syringe. There is an advantage that can be delivered through a non-invasive method.

The organic-inorganic composite granules according to the present invention may further include a functional member or cells. At this time, the functional member may be a bisphosphonate drug or a polyphenol-based naturally derived material as a target supported and delivered on the organic-inorganic composite granules.

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

In addition, the functional member is a polyphenol-based natural substance such as Quercetein, Genistein, Curcumin, Saurolactam, Sauchinone, Baicalin, etc. Daizein, Rutin, Anthocyanidin, Fisetin, Icariin, Kaempferol, E. Koreanum Nakei and Ikuol It may contain one or more substances selected from the group consisting of (Equol).

The functional member is not limited to the above material, and may be selected and applied according to the required functionality.

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

The case where the organic-inorganic composite granules according to the present invention include cells can be applied, for example, when stem cells are cultured on the organic-inorganic composite granules of the present invention and then delivered to the body.

The organic-inorganic composite granules according to the present invention may not contain a dispersant. The organic-inorganic composite granules according to the present invention may be used in the pharmaceutical field, medical field, cosmetic field, food field, etc. In this case, when a chemical substance such as a dispersant is included, there may be restrictions on its application. In consideration of this, the organic-inorganic composite granules according to the present invention may not contain a dispersant, especially when used in the above fields. Specific examples of application of the organic-inorganic composite granules according to the present invention include bone fillers, bone graft materials, and fillers. Bone fillers that are supported in spongy bone pores to induce bone formation promotion, filler materials for cosmetic and plastic surgery, and microplastic materials can be replaced and used in the field.

In addition, the present invention

Preparing an organic member solution;

Forming an organic-inorganic composite solution by uniformly dispersing 1 to 10 inorganic members relative to the organic member in a weight ratio in the organic member solution;

Spraying the organic-inorganic complex solution in an electrostatically charged manner; And

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

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

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

The manufacturing method of the present invention includes the step of preparing an organic member solution. At this time, 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 an appropriate combination of these. I can. The organic member is dissolved in, for example, phosphate buffer saline to prepare an organic member solution. Alternatively, it can be dissolved and dispersed in water, glycerin, or lipid oil.

At this time, it is preferable not to use an organic solvent in the step of preparing the organic member solution in the manufacturing method of the present invention. When an organic solvent is used in the manufacturing process, there may be limitations 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 when applying the composite granules according to the present invention to the fields of pharmaceuticals, medical care, food, or cosmetics can be eliminated.

Next, the manufacturing method of the present invention includes the step of uniformly dispersing 1 to 10 inorganic members relative 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, biological glass, alumina, zirconia, and composites thereof. The calcium phosphate-based is hydroxyapatite (HA), dicalcium phosphate (DCP), tricalcium phosphate (TCP), tetracalcium phosphate (TTCP), and octane calcium phosphate (TTCP). OCP: octacalcium phosphate), but is not limited thereto.

In this case, 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 drug or a polyphenol-based natural material, as described above.

In the manufacturing method of the present invention, it is preferable to introduce 1 to 10 inorganic members to the organic member solution in a weight ratio to the organic member, and to introduce it in a weight ratio of 5 to 10. The present invention has the advantage of further improving the sustained-release properties of the granules by including a large amount of the inorganic member in the composite solution as described above. In addition, when the granules are used as a bone filler, a relatively large amount of bone-forming components is included. There is an advantage to be doing it. For example, when the weight ratio of the inorganic member is less than 1, the effect of improving the sustained-release properties of the granules may be insufficient, and when used as a bone filler, there may be a problem in that bone forming components are 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 problems such as clogging of the agglomeration nozzle of the inorganic member, and thus it may be difficult to form granules having uniform properties.

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

As described below, the manufacturing method of the present invention forms granules by spraying in an electrostatically charged manner. Although the electrostatic charge method has the advantage of being able to manufacture granules of a relatively uniform size, there may be a problem that the nozzle is clogged during the process, making it impossible to manufacture granules. In particular, when the organic-inorganic composite solution containing an inorganic member having very high cohesiveness as in the present invention is used as a raw material, the nozzle is very easily clogged by the aggregation of the inorganic member among the raw material, so that the raw material containing the inorganic member is used. It is not easy to consider making granules by spraying in an electrostatically charged manner.

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

However, when using a co-rotating mixer and an ultrasonic mixer, when used for an excessively long time, the temperature of the complex solution itself increases significantly, which may be a problem when a functional member or cells are included in the complex solution. That is, when the temperature of the complex solution is too high, there is a problem in that the properties of drugs that may be included therein may change or cells may be killed. In consideration of these points, mixing using a co-rotating mixer and an ultrasonic mixer is preferably carried out within a range where the temperature of the complex solution does not exceed 40°C. For example, the co-rotating mixer is within 6 minutes, and the ultrasonic mixer is It can be done in less than 30 minutes.

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

Next, the manufacturing method of the present invention includes the step of spraying the organic-inorganic composite solution in an electrostatic charge method, for example, the electrostatic charge method spraying may be performed with a fine granule coating machine. In the case of producing granules by electrostatic charge method, most of the introduced raw materials can be converted into granules, so the yield is excellent, a large amount of granules can be produced in a short time, and the granules can be produced in a relatively uniform size. There is this. However, when the raw material has a high viscosity or a high degree of cohesion, there is a problem in that the nozzle may be clogged. The organic member included in the raw material used in the manufacturing method of the present invention has a high viscosity, and the inorganic member has a very high degree of aggregation, so it is not generally suitable for making granules by using an electrostatic charge method, but when forming an organic-inorganic complex solution By uniformly dispersing the inorganic member in the member solution and, if necessary, further dispersing the inorganic member with a post co-rotation mixer and stirring with an ultrasonic mixer, granules may be prepared in an electrostatically charged manner.

When the above step is performed with a microgranule coating machine, the size of the spray nozzle of the microgranule coating machine is preferably 50 μm to 1000 μm. If the size is less than 50 μm, there is a problem that it is difficult to spray the organic-inorganic composite solution including the inorganic member through the nozzle due to problems such as clogging of the nozzle, and if it exceeds 1000 μm, it is difficult to spray the organic-inorganic composite solution through the nozzle. There is a problem in that it is difficult to manufacture micro-sized particles. Further, the voltage is preferably 500 V to 2,500 V. When the voltage is less than 500 V, it is difficult to uniformly spray the sprayed solution, making it difficult to make spherical particles, and when it exceeds 2,500 V, the solution formed into a sphere after spraying is sprayed in an excessively spread form, resulting in high yield. There is a problem in that it is difficult to manufacture granules. In addition, the pressure is preferably between 100 mbar and 1500 mbar. When the pressure is less than 100 mbar, frequent nozzle clogging is caused, and the solution passing through the nozzle is excessively spread out, and when it exceeds 1500 mbar, the solution formed in a spherical shape is a polymerization inducing solution (e.g., CaCl ₂ ) There is a problem in that it is difficult to form spherical granules because a larger force than necessary is applied when dropped into the solution). Furthermore, it is preferable that the vibration frequency is 100 Hz to 6,000 Hz. When the vibration frequency exceeds 6,000 Hz, there is a problem in that spherical granules in the shape of a tail are formed.

The manufacturing method according to the present invention includes the step of polymerizing the organic-inorganic composite 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. Among these, ion crosslinking may be performed using at least one polymerization inducing material among _{calcium chloride (CaCl 2} ), calcium sulfate (CaSO ₄ ), and calcium carbonate (CaCO _{3 ).} It can be carried out by dropping the complex solution into a CaCl _{2 polymerization inducing solution.} According to the manufacturing method of the present invention, since the granules are formed in the form of a hydrogel rather than a simple solid, it is advantageous for delivering physiologically active substances such as cells, drugs, and proteins as compared to the simple solid granules. In addition, it is advantageous to maintain the viability of cells and deliver them into the human body, and higher loading efficiency of drugs, etc. than simple solid granules can be expected. In addition, the organic-inorganic composite granules of the present invention, which are hydrogels, have an advantage in controlling/controlling the drug release behavior with sustained release compared to the simple solid granules. Simple solid granules are delivered by adsorption on the surface of solid granules to support drugs or proteins. This is because the surface on which the drug or protein is loaded is directly exposed to the portion to be delivered after delivery, so that an initial excess is released, and it is difficult to control the release behavior for a long period of time thereafter. On the other hand, in the case of complex granules on a hydrogel, since an inorganic member carrying a drug or the like is supported and delivered in the hydrogel, the initial excessive release phenomenon can be reduced, and the release behavior can be controlled using the physical/chemical properties of the hydrogel. In the case of complex granules on a hydrogel, since cells or drugs can be delivered by being contained in the hydrogel, two functional drugs or proteins such as growth factors can be simultaneously delivered and their sequential release behavior can be implemented. In addition, in order to deliver solid granules into the human body, they must be delivered through a surgical operation that exposes the area to be delivered to the outside, or mixed with other carriers on a hydrogel and delivered through a syringe. There is an advantage that can be delivered through a non-invasive method.

It is preferable that the concentration of the organic member solution used in the manufacturing method of the present invention is 0.5 to 2.0% by 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, and thus it is difficult to maintain sufficient mechanical properties of the prepared granules, and if it exceeds 2.0% by weight, the concentration is too high. There is a problem in that it is difficult to uniformly disperse the complex solution due to the formation of a high viscosity solution, and it is difficult to form uniform spherical droplets using a spraying device.

In addition, the size of the inorganic member used in the manufacturing 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 (eg, ceramic powder) increases, and it may be difficult to evenly disperse in the organic member (eg, alginate solution). There is a possibility of discharge, and in addition, when cells are supported, endocytosis of cells cannot be neglected. When it exceeds 50 μm, it is difficult to pass through micro-sized nozzles, making particle production difficult. There is a difficult problem.

In the production method of the present invention, it is preferable not to use a dispersant for dispersing the inorganic member. In general, a dispersant is used for uniform dispersion due to the cohesiveness of the inorganic member. However, when a dispersant is included in the produced granules, a problem may arise in application to, for example, medical, pharmaceutical, food, and cosmetic fields. Therefore, the manufacturing method of the present invention does not use a dispersant, and thus has the advantage of greatly expanding the field of application of the granules to be produced. In the manufacturing method of the present invention, instead of using a dispersant, agitation and dispersion are performed through a co-rotating mixer or an ultrasonic mixer for uniform dispersion of the inorganic member.

According to the manufacturing method of the present invention, in manufacturing the organic-inorganic complex granules having sustained release and cell transfer properties, the yield is improved by manufacturing the granules in a static charge method, and the advantage of producing a large amount of granules in a short time is the advantage. have. In addition, since it is possible to manufacture granules of a uniform size, there is an advantage in that it is easy to apply to various fields. In addition, functional members or cells can be easily supported, and since there is no need to use organic solvents or dispersants during the manufacturing process, there is an advantage that the application field can be greatly expanded to various fields such as medical, pharmaceutical, food, and cosmetic fields. .

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 only for describing the present invention, and the scope of the present invention is limited by the following contents and is not construed.

In the following examples, when manufacturing organic-inorganic composite granules using a micro-sized nozzle using an electrostatic charge method, the process of uniformly dispersing the inorganic member in the organic solution is the most important. 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 degree of dispersion was not significantly improved, and there was a concern for denaturation due to heat as an organic substance was used, so it was fixed to less than 6 minutes. Then, it mixed using an ultrasonic mixer. It was shown that the degree of dispersion greatly increased with the mixing time, but the temperature in the solution reached about 40°C after 30 minutes of ultrasonic mixer treatment time, so it is a high temperature to deliver physiologically active substances such as cells and proteins after treatment. May be a limitation, so the processing time of the ultrasonic mixer was limited to within 30 minutes.

<Example 1>

A 1.0 wt% alginate solution was prepared by dissolving alginate, an organic member, in tertiary distilled water, and nanoapatite, an inorganic member, was used in a ratio of 1:1 (organic member: inorganic member) to the organic member by using a co-rotation mixer and an ultrasonic mixer. And mixed to prepare an organic-inorganic complex solution. At this time, a Paste mixer was used as a co-rotation mixer, and a rotational sonicator was used as an ultrasonic mixer, and each was performed for 6 minutes and 15 minutes.

The prepared organic-inorganic composite solution was introduced into a microgranule coating machine (Buchi, B-395 pro), sprayed with a nozzle having a diameter of 150 μm, and _{added dropwise to the CaCl 2} solution to prepare an organic-inorganic composite granule on a hydrogel. After dropping, it was cross-linked in _{CaCl 2 solution for 30 minutes and washed twice with PBS.} In order to measure the size, a certain amount of organic/inorganic composite granules were transferred to a petri dish and images were obtained through an optical microscope. The size of the particles was analyzed using the ImageJ program and the average size of the particles was calculated.

<Example 2>

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

<Example 3>

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

<Example 4>

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

<Example 5>

A 1.0 wt% alginate solution was prepared by dissolving alginate as an organic member in tertiary distilled water, and nanoapatite as an inorganic member was used in a weight ratio of 1:4 (organic member: inorganic member) to the organic member using a co-rotation mixer and an ultrasonic mixer. And mixed to prepare an organic-inorganic complex solution. At this time, a Paste mixer was used as a co-rotation mixer, and a rotational sonicator was used as an ultrasonic mixer, and each was performed for 6 minutes and 15 minutes.

The prepared organic-inorganic composite solution was introduced into a microgranule coating machine (Buchi Co., B-395 pro), sprayed with a nozzle having a diameter of 150 µm, _{and dropped into a CaCl 2} solution to prepare an organic-inorganic composite granule on a hydrogel. . After dropping, it was cross-linked in _{CaCl 2 solution for 30 minutes and washed twice with PBS.} In order to measure the size, a certain amount of organic/inorganic composite granules were transferred to a petri dish and images were obtained through an optical microscope. The size of the particles was analyzed using the ImageJ program and the average size of the particles was calculated.

<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: Organic-inorganic composite granules were prepared in the same manner as in Example 5, except that the inorganic member was 1:8.

<Example 8>

Organic member: Organic-inorganic composite granules were 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 performed in order to prepare an organic-inorganic composite granule in the same manner as in Example 1, except that 1:20 (organic member: inorganic member) was mixed with the inorganic member nano-apatite in a weight ratio to the organic member.

However, it was confirmed that the nozzle clogging of the microgranule coating machine occurred seriously during the process, and the ceramic content in the produced granules was significantly lower than the theoretical content, making it impossible to produce uniform granules, and accordingly, the production yield was significantly lowered.

<Experimental Example 1>

Confirmation of composite granule size

The following experiment was performed to confirm the size of the organic-inorganic composite granules prepared according to the present invention.

The organic-inorganic composite granules prepared in Examples 1 to 4 were checked for the shape of the granules through an optical microscope, and the sizes of each were checked using ImageJ software, and the results are shown in FIG. 1.

According to FIG. 1, it can be seen that the organic-inorganic composite granules according to the present invention are spherical and have an average size range of about 250 to about 270 μm.

<Experimental Example 2>

Checking the amount of inorganic members supported

The following experiment was performed to confirm the loading amount of the inorganic member of the prepared organic-inorganic composite granules.

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, except that the contents of the organic member and the inorganic member were adjusted as shown in Table 1 below. It is shown in Table 1 below.

Organic Absence: Inorganic Absence 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 above, the content of the inorganic member in the organic-inorganic composite granules actually produced substantially matches the amount of the inorganic member introduced as a raw material, through which precipitation of the inorganic member or clogging of the nozzle does not occur during the manufacturing process, and a high yield It can be seen that the composite granules can be prepared.

<Experimental Example 3>

Confirmation of drug sustained release

The following experiment was performed to confirm the drug sustained release of the organic-inorganic composite granules of the present invention.

On MC3T3 osteoblasts, the organic-inorganic composite granules prepared in Examples 1 to 4 of the present invention were placed as shown in FIG. 2B, and the amount of drug released over time and the degree of proliferation of cells over time were measured, respectively. It is shown in 2a and 2c.

The organic-inorganic composite granules prepared in Examples 1 to 4 were put in a phosphate buffer solution (PBS), and the PBS solution was taken for each time using the total replacement method, and the released concentration of the drug through absorbance measurement using a spectroscopic analysis method. By calculating the release behavior was confirmed.

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

According to FIG. 2A, it can be confirmed that the drug is slowly released over time, and according to FIG. 2C, it can be confirmed that the degree of cell proliferation is increased by the released drug. Through this, it can be seen that the organic-inorganic complex granules of the present invention can be used for drug delivery, and specifically, for example, can be used for treating osteoporosis in the body.

<Experimental Example 4>

Confirmation of drug sustained release through cell culture

The following experiment was performed to confirm the cell culture properties of the organic-inorganic composite granules of the present invention.

Method for delivering organic-inorganic complex granules containing quercetin at each concentration prepared in Examples 1 to 4 of the present invention using a transwell, and analyzing cell proliferation and osteodifferentiation behavior of quercetin released from the complex granules By culturing osteoblast (MC3T3) cells, and through the Cell Lysis method, the degree of bone differentiation of the cultured cells was confirmed by its representative factor, ALP activity, and DNA quantification was also performed, and the results are shown in FIGS. 3A and 3A, respectively. It is shown in 3b.

According to FIG. 3, in the case of week 1, the degree of cell proliferation is almost the same, but in the second week, when the quercetin content is high, cell proliferation occurs more. I can confirm.

<Experimental Example 5>

Confirmation of cell loading of organic/inorganic granules

After preparing the organic-inorganic complex solution in the same manner as in Example 5, osteoblasts (MC3T3) were 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, spraying was performed in the same manner as in Example 5, and this was _{added dropwise to a CaCl 2} solution to prepare a hydrogel-like organic-inorganic composite granule. Thereafter, in order to check whether the cells are supported in the complex granules, staining was performed using a DAPI solution capable of discriminating the cell nucleus, and confirmed through a fluorescence microscope, which is shown in FIG. 4, it can be seen that cells are uniformly supported in the organic-inorganic composite granules of the present invention.

<Experimental Example 6>

Confirmation of uniform dispersion according to the mixing method

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

An organic-inorganic composite solution was prepared in the same manner as in Example 5 above. The prepared organic-inorganic composite solution was mixed for 6 minutes and 21 minutes using a magnetic stirrer, and on the other hand, the prepared organic-inorganic composite solution was mixed with a co-rotating mixer for 6 minutes, followed by an ultrasonic mixer for 15 minutes. Stirring was performed with, and then dyed using an Alizarin red solution, and the result was confirmed through an optical microscope. The results are shown in FIG. 5. 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 severely aggregated with each other. In addition, in the case of stirring with a general magnetic stirrer, it can be seen that the aggregation phenomenon becomes more severe as time passes. On the other hand, referring to FIG. 5B, in the case of mixing using a co-rotation mixer and an 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 composite granule size

The following experiment was performed to confirm the size of the organic-inorganic composite granules prepared according to the present invention.

The organic-inorganic composite granules prepared in Examples 5 to 16 were checked for the shape of the granules through an optical microscope, and the sizes of each were checked using ImageJ software, and the results are shown in FIG. 6.

According to FIG. 6, it can be seen that the organic-inorganic composite granules according to the present invention are spherical and have an average size range of about 200 to about 500 μm.

Claims (19)

Including an organic member and an inorganic member,
The weight ratio of the inorganic member to the organic member is 1 to 10 organic-inorganic composite granules,
The average size of the organic-inorganic composite granules is 100 to 457 μm,
Organic-inorganic complex granules, characterized in that the hydrogel phase.
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.
The method of claim 1.
The organic-inorganic composite granules further comprise a functional member or cells.
The method of claim 1,
Organic-inorganic composite granules, characterized in that the organic member is a natural biopolymer or a synthetic biopolymer.
The method of claim 1,
Organic-inorganic composite granules, characterized in that the inorganic member is a ceramic member.
The method of claim 3,
The functional member is an organic-inorganic composite granule comprising a bisphosphonate-based drug, a polyphenol-based natural substance, and a combination thereof.
The method of claim 1,
The organic-inorganic composite granules are used in one field selected from the group consisting of a pharmaceutical field, a medical field, a cosmetic field, and a food field.
The method of claim 1,
Organic-inorganic composite granules, characterized in that it does not contain a dispersant.
Preparing an organic absent solution;
Forming an organic-inorganic composite solution by uniformly dispersing 1 to 10 inorganic members relative to the organic member in a weight ratio in the organic member solution;
After forming the organic-inorganic composite solution, dispersing the inorganic member with a co-rotation mixer and stirring with an ultrasonic mixer;
Spraying the organic-inorganic complex solution in an electrostatically charged manner; And
Polymerizing the sprayed organic-inorganic composite solution to form a hydrogel phase;
Method for producing an organic-inorganic composite granule comprising a.
The method of claim 9,
The method of manufacturing an organic-inorganic composite granule, wherein the inorganic member dispersed in the organic member solution is 5 to 10 in weight ratio compared to the organic member.
delete The method of claim 9,
The step of polymerizing the organic-inorganic composite solution is performed by dropping the sprayed granules into a polymerization inducing solution.
The method of claim 9,
The method of manufacturing an organic-inorganic composite granule, wherein the inorganic member comprises a functional member.
The method of claim 9,
The method of manufacturing an organic-inorganic composite granule, further comprising the step of supporting a functional member or cells in the formed organic-inorganic composite solution.
The method of claim 9,
The organic-inorganic composite granule production method, characterized in that the concentration of the organic member solution is 0.5 to 5% by weight.
The method of claim 9,
The method of manufacturing an organic-inorganic composite granule, characterized in that the size of the inorganic member is 50 nm to 50 μm.
The method of claim 9,
The step of dispersing the inorganic member with the co-rotating mixer and stirring with the ultrasonic mixer is performed in a range in which the temperature of the organic-inorganic composite solution does not exceed 40°C.
The method of claim 9,
The step of spraying the organic-inorganic composite solution in an electrostatic charge method is performed by a fine granule coating machine,
The size of the spray nozzle of the microgranule 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. Method for producing organic-inorganic composite granules as described above.
The method of claim 9,
The organic-inorganic composite granules manufacturing method of the organic-inorganic composite granules, characterized in that it does not contain a dispersing agent.

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