KR20160136345A - Nano-scaffold containing functional factor and method for producing same - Google Patents

Abstract

The present invention relates to a nano-scaffolds and methods for their preparation containing functional factors, but more specifically, by using the SiO ₂ beads for each unit size as a template to prepare a capsule of the nanoscale, TiO ₂ on SiO ₂ beads template , HA (Hydroxyapatite), TCP (Tri Calcium Phosphate), and then SiO ₂ beads are removed to prepare hollow capsules. Hollow capsules of each unit diameter are mixed in various combinations to form aggregates a method for producing a synthetic bone material containing a functional factor by loading a functional agent exhibiting osteogenesis promotion or anti-inflammation, antibacterial action or the like into a hollow capsule after producing aggregates, and a method for producing a functional factor ≪ / RTI >

Description

Nano scaffold containing functional factor and method for producing same

The present invention relates to a nano-scaffolds and methods for their preparation containing functional factors, but more specifically, by using the SiO ₂ beads for each unit size as a template to prepare a capsule of the nanoscale, TiO ₂ on SiO ₂ beads template , HA (Hydroxyapatite), TCP (Tri Calcium Phosphate), and then SiO ₂ beads are removed to prepare hollow capsules. Hollow capsules of each unit diameter are mixed in various combinations to form aggregates a method for producing a synthetic bone material containing a functional factor by loading a functional agent exhibiting osteogenesis promotion or anti-inflammation, antibacterial action or the like into a hollow capsule after producing aggregates, and a method for producing a functional factor ≪ / RTI >

It is very important to have bone loss, slow down of bone growth, improve bone growth, and have anatomical structure due to diseases such as trauma, surgery, inflammation, osteoporosis or periodontitis. A scaffold can be used as a replacement for the lost bone structure or as a carrier for delivery of a drug or cell therapy agent involved in the bone regeneration process.

Artificial bones and techniques for their manufacture (Korean Patent No. 1,031,121). The present invention relates to a method for manufacturing an artificial bone which is formed by a composite structure of a porous body having a plurality of holes and a dense body having no hole, Preparing a slurry by adding a slurry, and gelling the slurry prepared in the step, and then pouring the slurry having different pore-forming agent contents into the prepared slurry, gelling the slurry, drying and sintering the slurry to produce artificial bones. However, in this case, a pore-forming agent is used to make a porous article, and a portion where the pore-forming agent is not contained is dense. Since PMMA, polymer beads, and naphthalene are used as a pore-forming agent, ~ 700μm, which is relatively large and diverse.

There is a description of a bone prosthesis having a protective coating for enhancing bone cross-growth (US Application No. 08 / 521,111). This technique includes a boundary surface that is in contact with and attached to a living bone, a plurality of shallow indentations extending inwardly with respect to the indentation floor, and a bone growth promoting coating that is over the floor and is shielded by the bath- The present invention relates to an implantable bone prosthesis having a body attached to a biological bone for replacing or repairing a part of the biological bone. Hydroxyapatite is used as a bone growth promoting coating.

There is a technique for manufacturing a porous implant (Korean Patent Laid-Open No. 10-2004-0064794). This technique involves mixing pure titanium (Ti) powder and hydroxyapatite (HA) powder to form a mixed powder; Mixing the mixed powder with a binder, extruding the mixed powder through an extruder, and injecting the mixed powder to produce a molded body; Removing the binder from the injection-molded body; And sintering the injection-molded article from which the binder has been removed.

There is a technique relating to a porous bone-cartilage composite support for tissue engineering (Korean Patent Publication No. 2006-0054236). This technique comprises the steps of (i) a porous bone regeneration layer comprising a biocompatible polymer and a physiologically active ceramic, wherein the ceramic is exposed to the surface, and (ii) a porous cartilage regeneration layer comprising a biocompatible elastomeric polymer. TECHNICAL FIELD The present invention relates to a porous bone-cartilage composite support.

There is a description of a living bone-induced artificial bone (PCT / JP2004 / 00042). This technique consists of a single mass of titanium or titanium alloy and has a through hole in the shape of a three-dimensional network with a diameter of 100 to 3,000 μm and a hole with a diameter of 50 μm or less on the inner surface of the hole and a porosity of 30 to 80% And a coating film formed on at least a part of the surface of the hole and the hole in the porous body, the film being made of a porous material, an amorphous titanium oxide phase, an amorphous alkaline titanium carbonate phase or the like, .

There is also a description of a ZrO ₂ scaffold having a hydroxyapatite (HA) layer. In the case of the scaffold, however, the HA layer may be peeled off or worn during implantation, and cracking, tearing and slacking may occur from the surface of the implant due to differences in mechanical properties between ZrO ₂ and HA. HA fragments can form small body bodies (sequestered bodies), which can interfere with osteointegration.

Other known scaffolds are made of degradable material, which means that they are degraded after implantation in the subject. Although it is advantageous in that it can be disassembled, it may be a disadvantage because it is not desirable from the standpoint of stabilization of the implant itself in other respects. In addition, some of the scaffolds known in the prior art may trigger inflammatory reactions or cause infections. For example, human or animal-derived bone implant scaffolds may cause allergic reactions when implanted in another animal.

On the other hand, a layer of ceramic material was coated on the surface of the implant to stimulate the regeneration of the bone tissue. However, the ceramic coating is fragile and may peel off or fall off the implant surface, which may prevent the implant from functioning properly.

The present invention provides synthetic bone in the form of aggregates of various sizes of nano-capsules having various sizes of pores, and by mounting a functional factor in a hollow capsule, it can be used for proliferation of capillary blood vessels and attachment, differentiation and proliferation of osteogenic cells The present invention also provides a nano scaffold which can be used as a carrier for delivering a nano drug or a cell therapeutic agent, and a method for producing the same.

The invention nano scaffolds and methods for their preparation containing functional factors, but more specifically, by using the SiO ₂ beads for each unit size as a template to prepare a capsule of the nanoscale, TiO _2, on the SiO ₂ beads template HA ( Hydroxy apatite), TCP (Tri Calcium Phosphate) coating a bio-compatible material, and the like thereafter (agglomerates by mixing the hollow capsules of removing SiO ₂ beads were prepared hollow capsule, and each unit of the diameter in different combinations aggregates) A method for producing a synthetic bone material containing a functional factor by loading a functional factor showing bone formation promotion or anti-inflammation, antibacterial action or the like into a hollow capsule, and a method for producing a synthetic bone material containing a functional factor The above problem is solved by providing a nanoscale scaffold.

The synthetic bone material containing the functional factor according to the present invention provides a nano-unit scaffold proved to have a comparative advantage in settlement of cells such as osteoblasts involved in bone formation, Because it is equipped with a functional factor, it is advantageous that a customized therapy can be performed by applying an appropriate functional factor according to the application target.

In addition, the manufacturing cost of the synthetic bone material according to the present invention is significantly lower than that of the conventional synthetic bone material. In the case of the synthetic bone material according to the present invention, the drug effect on the traumatic bone defect, By inducing the release by time difference by the drug delivery system, the existing wound healing process can be safely and reliably performed, the treatment period can be shortened, and the bone formation process can be promptly induced.

By controlling the selection of raw materials such as TiO ₂ , HA or Tcp, capsule pore size, capsule thickness, etc., it is possible to control the release rate of the factors loaded in each capsule, thereby providing various drug delivery systems (DDS) have.

1 is a photograph of a SiO ₂ template.
2 is a photograph of a TiO ₂ nanocapsule.
3 is an electron microscopic view of the MBCP third-party synthetic bone.
4 is an electron microscopic view of TIO ₂ according to the present invention.
Fig. 5 shows the appearance of HA hollow capsule aggregates according to the present invention attached to an implant surface. Fig.
Figure 6 is an illustration of an example of a hollow capsule aggregate according to the present invention.

Best Mode for Carrying Out the Invention

The present invention provides nanoscrafolds containing functional agents and methods of making the same.

In vivo The present invention as more specifically, by using the SiO ₂ beads for each unit size as a template prepared in the capsules of the _{nano-scale, TiO 2, HA (Hydroxy apatite} ), TCP (Tri Calcium Phosphate) on SiO ₂ beads template The hollow capsules are prepared by coating the affinity material, then the SiO ₂ beads are removed, and the hollow capsules of each unit diameter are mixed in various combinations to produce aggregates. And the like, into a hollow capsule to provide a synthetic bone material containing a functional factor and a nanoscale containing the functional agent prepared according to such a production method.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a process for preparing (a) an SiO ₂ bead; (b) using a SiO ₂ bead as a template to produce a nanoscale capsule, wherein the biocompatible material is coated on the SiO ₂ template; (c) removing the SiO ₂ beads to prepare a hollow capsule; And (d) mixing the hollow capsules to produce aggregates, and then loading the functional agents into the hollow capsules, or loading the functional agents into the hollow capsules, followed by mixing the hollow capsules to form agglomerates. Lt; RTI ID = 0.0 > nanoscale < / RTI >

In one embodiment according to the invention, in step (a), the diameter of the SiO ₂ beads is in the range of 500 nm to 1 μm, specifically 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, Beads.

In one aspect, SiO ₂ beads in accordance with the present invention can be produced through the following procedure.

(1) Preparation of dispersion of silica nanoparticles

The Stober method can be used to produce silica nanoparticles of various sizes. In the sol-gel reaction, the type of solvent, the amount of catalyst, the amount of water, and the amount of tetraethylorthosilicate (TEOS), the precursor, affect particle size. First, anhydrous ethanol is quantified in a reaction vessel to dissolve tetraethylorthosilicate, which is a precursor of a certain concentration. Tetraethylorthosilicate is then added with water to remove tetraethyl ether through hydrolysis and substitution of the hydroxyl groups of the water. At this time, since the spherical nanoparticles are uniformly formed in the basic state, the reaction is carried out by adding ammonia water. Particularly, the tetraethyl orthosilicate solution dissolved in anhydrous ethanol produces a silica nanoparticle dispersion while water is added uniformly through a separating funnel by a dropwise addition while uniformly forming a silica precursor.

(2) Preparation of core-shell silica particles having a single mesopore

The silica nanoparticle dispersion is added to distilled water containing ammonia water and the like and then stirred to prepare solution A. The surfactant solution is stirred and then added to the solution A and stirred. A silica precursor is added while stirring, and a silica cast body can be produced by applying heat.

(3) Production of encapsulated silica-titania composite

The silica mold formed in the above step is put into a mixed solution of an organic solvent and water and dispersed. Titanium butoxide or the like is added to a solvent such as ethylene glycol and the mixture is stirred to uniformly disperse the TiO ₂ precursor in the solution. When stirring vigorously was added dropwise to the dispersed state of silica polymorphs dropwise to the TiO ₂ precursor and the reaction proceeds TiO ₂ precursor to the surface of the silica primary molded product is deposited through hydrolysis, and the coating is started. After addition and stirring for more than 30 minutes, centrifuge the coated capsule-type silica-titania complex using a centrifuge. At this time, impurities adhered to the surfaces of these particles are washed with an organic solvent and air dried. At this time, a pure capsule-type silica composite in which the solvent and the impurities are completely removed can be produced through a heat treatment drying process of 60 degrees or more.

(4) Production of porous hollow capsule

The prepared capsule-type silica composite is uniformly dispersed in a solvent mixed with water and an organic solvent, and the silica is appropriately diluted with an aqueous solution in an aqueous solution, followed by heating and stirring for 30 minutes or more. At this time, the silica cast body in the core reacts with the base ion of the permeated aqueous solution so that the silica is gradually eluted to prepare the porous hollow capsule from which the core is removed.

In one embodiment according to the present invention, bio-compatible material in step (b) is a _{TiO 2, Ti 3 O, Ti} 2 O, Ti 3 O 2, TiO, Ti 2 O 3, Ti 3 O 5 , and titanium butoxide Titanium oxide selected from the group consisting of; Tricalcium phosphate, calcium phosphate; Apatite selected from the group consisting of hydroxy apatite, silicon and magnesium substituted hydroxy apatite; Calcium sulfate; Zirconium dioxide; Silicon dioxide; And combinations thereof.

In an embodiment according to the present invention, the biocompatible material in step (b) may be any one selected from the group consisting of TiO ₂ , hydroxide apatite and tricalcium phosphate more specifically.

By using the material as a material, it can be implanted in a subject without causing side effects such as immunological problems, cross-infection, or allergic reaction that may occur in allograft (human bone) or heterogeneous bone (bones, horses, etc.).

In one embodiment according to the present invention, in step (d), the functional agent may be a functional factor selected from the group consisting of factors that promote the promotion of bone formation, antibacterial, antiinflammatory and acidity. At this time, the scaffold may be loaded with an osteogenesis factor. As the osteophyte, an osteogenic protein such as a recombinant human bone morphogenetic protein may be used. Biomolecules to be mounted on the scaffold according to the present invention include natural biomolecules, synthetic biomolecules, and recombinant biomolecules. Specifically, it is a cell attachment factor, enzyme, blood protein, antibody, growth factor, hormone, DNA, RNA, RNAi, drug, protein drug, peptide, inorganic vitamin,

In an embodiment according to the present invention, in step (d), the functional agent may be loaded into the hollow capsule using one or more methods selected from the group consisting of dipping, centrifugation and sonication.

In one aspect of the present invention, there is provided a nanocascaphrode containing a functional agent produced according to the above production method.

In one embodiment of the present invention, the nanoscale may be used as a medical implant, a drug delivery carrier or a cell delivery carrier, and in particular, the scaffold may be a bone- Play, value

Replacement, and recovery; Or for cells or drug delivery bodies.

Hereinafter, the present invention will be described in more detail with reference to Examples of the present invention and Comparative Examples which are not based on the present invention, but the scope of the present invention is not limited by the following Examples.

Example 1. Preparation of synthetic bone graft material

(1) SiO ₂ beads having a size of 500 nm or more and 1 μm or less were prepared. The SiO ₂ beads were prepared according to the following procedure.

1) Preparation of dispersion of silica nanoparticles

Stober method was used to prepare silica nanoparticles having various sizes. In the sol-gel reaction, the type of solvent, the amount of catalyst, the amount of water, and the amount of tetraethylorthosilicate (TEOS), the precursor, affect particle size. A more specific manufacturing process is as follows:

The reaction vessel was charged with 1,000 mL of ethanol, and 0.14 mol of tetraethylorthosilicate, which is a precursor, was added thereto to sufficiently disperse it. At this time, tetraethylorthosilicate is very weak to moisture and is not exposed to the atmosphere for a long time. On the other hand, in the tetraethyl orthosilicate solution dispersed in anhydrous ethanol, 1 mol of 28 wt% ammonia water as a catalyst was added to 10 mL of tertiary distilled water, and 1 drop was added dropwise to the organic solution of tetraethyl orthosilicate through a separating funnel. The tetraethyl ether of tetraethyl orthosilicate is hydrolyzed by water to separate into a solution and the hydroxyl group of water is substituted for it to form silica. At this time, these reaction solutions were evenly stirred at room temperature for 1 hour to form uniform spherical silica nanoparticles. The resulting silica particles were found to have a diameter of about 500 nm.

2) Preparation of core-shell silica particles having a single mesopore

To the reaction vessel, 10 mL of the silica nanoparticle dispersion prepared in 1) was added to 20 mL of distilled water containing ammonia water (28 wt%, 0.1 mL) and stirred for 30 minutes to prepare Solution A. And the molar ratio of cetyltrimethylammonium bromide: 1,3,5-trimethylbenzene: decane: distilled water: ethanol was 1: 1: 1: 113.99: 17.77 6.24 mL of the prepared surfactant solution was stirred at room temperature for 30 minutes, then added to Solution A, and stirred at room temperature for 30 minutes. Then 0.43 mL of TEOS was added with stirring and stirred for 10 minutes. After the stirring, hydrothermal reaction was carried out in an oven set at 70 DEG C for 15 hours. The recovered sample is dried at 70 ° C and then heated to a temperature of 500 ° C from room temperature while blowing oxygen using a tube furnace. The temperature was gradually raised for 40 minutes, maintained at 500 < 0 > C for 5 hours, and then cooled to room temperature to remove organic matter.

3) Preparation of capsule-type silica-titania composite

0.1 g of the silica precipitate prepared in step 2) was added to a mixed solution of 50 mL of acetone and 0.1 mL of tertiary distilled water and dispersed evenly using an ultrasonic machine. To 60 mL of ethylene glycol, 0.4 mL of titanium butoxide was added, followed by stirring for 12 hours to prepare a TiO ₂ precursor. 10 mL of the TiO ₂ precursor was added to the dispersed silica mold, stirred for 3 hours, washed with ethanol, and dried at 70 ° C for 12 hours. Then, a capsule type silica composite was formed by forming a metal or metal oxide layer by heat treatment at a temperature of 450 ° C. for 5 hours using a saccharification furnace. The thickness of the oxide layer was 25 nm.

4) Manufacture of Porous hollow capsule

0.1 g of the prepared capsule-shaped silica composite was dispersed in 3 mL of ethanol, and the resulting mixture was placed in 5 mL of 1M NaOH aqueous solution and reacted in a reaction oven at 70 ° C for 3 to 5 hours to remove the silica cast body to prepare a porous hollow capsule. The prepared porous hollow capsules were separated by centrifugation, washed with ethanol, and then dried at 70 ° C for 12 hours.

(2) TiO ₂ , HA, or Tcp were coated on the SiO ₂ bead template using SiO ₂ beads of each unit diameter as a template, and the SiO ₂ bead template was removed to dissolve TiO ₂ , HA or Tcp hollow capsules capsule) was prepared. In detail, first, the silica mold formed by the Stober method was dispersed in a mixed solution of an organic solvent and water. Titanium butoxide or the like is added to a solvent such as ethylene glycol and the mixture is stirred to uniformly disperse the TiO ₂ precursor in a solution. When stirring vigorously was added dropwise to the dispersed state of silica polymorphs dropwise to the TiO ₂ precursor and the reaction proceeds TiO ₂ precursor to the surface of the silica primary molded product is deposited through hydrolysis, and the coating is started. After the addition and stirring for more than 30 minutes, the coated capsule-type silica-titania complex was centrifuged using a centrifuge. At this time, impurities adhering to the surfaces of these particles were washed with an organic solvent and air dried. At this time, a pure capsule-type silica composite in which the solvent and the impurities are completely removed can be produced through a heat treatment drying process of 60 degrees or more.

Preparation of Porous Hollow Capsule: The prepared capsule-type silica composite was uniformly dispersed in a mixture of water and an organic solvent, and the silica was appropriately diluted in an aqueous solution, followed by heating and stirring for 30 minutes or more. At this time, the silica template in the core reacted with the base ion of the infiltrated aqueous solution, and the silica was gradually eluted to prepare a porous hollow capsule having the core removed.

(3) Hollow capsules of each unit diameter, 100 nm, 200 nm, 300 nm and 500 nm were classified using each unit of a unit, and these capsules were mixed in various combinations to obtain a nano-unit synthetic bone material .

(4) SEM photographs of the hollow capsules prepared in (2) revealed that spherical capsules having irregular pore structures of various sizes were produced, not closed capsules (see FIG. 1). The capsule has a pore size of 1/100 to 1/10 of the capsule diameter. (3), and bone formation promoting factors (peptide, antibiotic, antiinflammatory agent, etc.) having a size of 20 탆 to 100 탆 are loaded into hollow capsules by dipping, centrifugation, To produce synthetic bone containing functional factors. At this time, the functional drug was diluted to a sufficient concentration in an aqueous solution or an organic solvent and physically mixed with a hollow capsule, and then loaded into a hollow capsule through capillary and osmotic phenomenon. In particular, since the hollow capsule is porous, a nano unit synthetic bone material having a function of gradually releasing or gradually releasing the drug through the porous capsule membrane can be produced.

Example 2: Preparation of synthetic bone graft material

Prepared as in Example 1, in step (4), citric acid was loaded into hollow capsules to produce synthetic bone containing factors that increased acidity.

Example 3: Preparation of synthetic bone graft material

Prepared as in Example 1, in step (4), ascorbic acid was loaded into hollow capsules to produce synthetic bone containing factors that increased acidity.

Example 4. Preparation of synthetic bone graft material

Prepared as in Example 1, in step (4), the antibiotic was loaded into a hollow capsule to produce a synthetic bone containing a functional factor with antimicrobial activity.

Example 5: Preparation of synthetic bone graft material

The preparation was carried out as in Example 1, and the anti-inflammatory agent was loaded into the hollow capsules in step (4) to produce a synthetic bone having an anti-inflammatory effect.

Since the synthetic bone material containing the functional factor according to the present invention can induce faster bone formation and is equipped with a functional factor, it is possible to provide a customized therapy by applying an appropriate functional factor according to the application object. In the case of the synthetic bone material, the drug delivery system for traumatic bone defect, surgical bone defect, and osteogenesis promoting factor induces release by time lag, thereby safely and securely performing the existing wound healing process And the treatment period can be shortened, and the bone formation process can be promptly induced, and more economical manufacturing is possible, which is industrially applicable.

Claims (10)

(a) preparing a SiO ₂ beads;
(b) using a SiO ₂ bead as a template to produce a nanoscale capsule, wherein the biocompatible material is coated on the SiO ₂ template;
(c) removing the SiO ₂ beads to prepare a hollow capsule; And
(d) mixing the hollow capsules to produce aggregates, then loading the functional agents into the hollow capsules, or loading the functional agents into the hollow capsules, followed by mixing the hollow capsules to form aggregates. The method comprising the steps of: The method according to claim 1,
A process for producing a nanocascade comprising a functional factor, wherein in step (a), the diameter of the SiO ₂ bead is 500 nm or more and 1 μm or less. The method according to claim 1,
Biocompatible materials are _{TiO 2, Ti 3 O, Ti} 2 O, Ti 3 O 2, TiO, Ti 2 O 3, Ti 3 O 5 and titanium oxide that is selected from the group consisting of titanium butoxide, in step (b); Tricalcium phosphate, calcium phosphate; Apatite selected from the group consisting of hydroxy apatite, silicon and magnesium substituted hydroxy apatite; Calcium sulfate; Zirconium dioxide; Silicon dioxide; And combinations thereof. ≪ RTI ID = 0.0 > 21. < / RTI > The method according to claim 1,
Wherein the biocompatible material in step (b) is any one selected from the group consisting of TiO ₂ , hydroxide apatite, and tricalcium phosphate. The method according to claim 1,
Wherein the functional factor in step (d) is a functional factor selected from the group consisting of factors that promote osteogenesis promotion, antibacterial, antiinflammatory, and acidity. / RTI > The method according to claim 1,
Characterized in that in step (d) the functional agent is loaded into a hollow capsule using one or more methods selected from the group consisting of dipping, centrifugation and sonication methods. Gt; 7. A nanocascade containing functional agents, prepared according to the method of any one of claims 1 to 6. A medical synthetic bone prepared using the nano scaffold of claim 7. A drug delivery carrier prepared using the nanoscale of claim 7. A cell delivery carrier prepared using the nanoscale of claim 7.

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