KR20220067109A - HUMIDITY AND pH SENSITIVE SELF-HEALING POROUS NANOCAUSULES AND MANUFACTURING METHOD THEREOF - Google Patents

Abstract

One aspect of the present invention provides a core comprising an antimicrobial agent, a ligand brush formed on at least a part of the surface of the core, and a porous nanocapsule, in which the antimicrobial agent is released under conditions of humidity of 45 to 100 %, and pH of 5 to 7. The present invention can selectively release antibacterial substances under environmental conditions in which bacteria, Escherichia coli, etc. are actively proliferating, thereby accurately and effectively improving antibacterial activity.

Description

Humidity and pH-sensitive porous nanocapsules and manufacturing method thereof

The present invention relates to a porous nanocapsule capable of self-healing because it has excellent antibacterial properties and is sensitive to humidity and pH, and a method for manufacturing the same.

Antibacterial agents are used for the purpose of inactivating microorganisms that cause not only bad odors but also infectious diseases. Numerous antibacterial agents have been developed for inactivation of these microorganisms, and the form of the antibacterial agent is variously used as a spray type, a patch type, and a film type. However, since this type of antibacterial agent has only a single or one-time effect, there is a problem in that the continuous antibacterial effect is not maintained.

In order to solve this problem, an antibacterial agent in the form of a nanocapsule carrying an antibacterial substance in the capsule has been proposed and applied to special textile products such as protective products and protective clothing, medical supplies, and special packaging materials. It is a constant release system using the form of beads, and the release period of the capsule cannot be controlled, so once the release begins, it is impossible to control. These existing nanocapsules have a problem in that the antibacterial material is wasted heavily and the lifespan thereof is consumed within a short period of about 6 months.

On the other hand, many types of bacteria, Escherichia coli, and fungi can survive in the conditions of pH 1-9, but they proliferate the most at pH 6-7. can be lowered

Accordingly, there is a demand for the development of a nanocapsule technology capable of controlling the release of antibacterial substances by selectively reacting under specific conditions, particularly, in an environment in which the proliferation of bacteria, E. coli, etc. is active.

The present invention is to solve the problems of the prior art, and an object of the present invention is to control the release and suppression of the release of antibacterial substances under specific conditions, so that the lifespan characteristics are excellent, and the porous nanocapsules having excellent antibacterial properties and a method for manufacturing the same is to provide

One aspect of the present invention is a core comprising an antibacterial agent; a ligand brush formed on at least a portion of the surface of the core; And the antibacterial agent provides a porous nanocapsule that is released under the conditions of 45-100% humidity and pH 5-7.

In one embodiment, the core includes one or more pores, and the pores are closed by the ligand brush under a condition of less than pH 5, and open by the ligand brush at a condition of pH 5-7 ( can be open).

In an embodiment, the ligand brush may react the acrylic compound after bonding the silane compound to the core.

In one embodiment, the silane-based compound is 3-(trimethoxysilyl)propylmethacrylate (MPS), 3-(glycidoxy)trimethoxysilane (3-(Glycidyloxypropyl) ) may be one selected from the group consisting of trimethoxysilane, GPS), 3-(mercaptopropyl) trimethoxysilane (3-(mercaptopropyl) trimethoxysilane, MCPS), and a mixture of two or more thereof.

In one embodiment, the acrylic compound is acrylic acid, methacrylic acid, methyl acrylate, methyl methacrylate, ethyl acrylate, ethyl methacrylate, propyl acrylate, propyl methacrylate, butyl acrylate, butyl methacrylic It may be one selected from the group consisting of rate, pentyl acrylate, pentyl methacrylate, polymethyl acrylate, polymethyl methacrylate, polyethyl acrylate, polyethyl methacrylate, and a mixture of two or more thereof.

In one embodiment, the antibacterial agent may be triclosan (Triclosan) or an antibacterial substance in a solution state.

In one embodiment, the average particle diameter of the porous nanocapsules may be 200 ~ 800㎚.

In addition, in order to achieve the above object, another aspect of the present invention, (a) preparing a silica core support by reacting an aqueous solution containing a silica (SiO ₂ ) precursor and an antibacterial agent; (b) reacting by adding a silane-based compound to the silica core support; and (c) adding and reacting an acrylic compound to the product of step (b) to form a ligand brush; wherein the antibacterial agent is a porous nano-particle released under conditions of 45-100% humidity and pH 5-7. A method for manufacturing a capsule is provided.

In one embodiment, the silica core support may include one or more pores.

In one embodiment, the average particle diameter of the porous nanocapsules may be 200 ~ 800㎚.

A porous nanocapsule capable of self-healing according to an aspect of the present invention and a method for manufacturing the same, by forming a core supporting an anionic antibacterial agent and a ligand brush on the core, are effective in the expansion of the ligand brush in a specific range of humidity and pH conditions. Accordingly, the antibacterial agent in the core is released, and when it is out of the above range, the antibacterial life can be extended by self-healing to its original form, and the antibacterial material can be selectively released in a specific range, that is, under environmental conditions where the proliferation of bacteria, E. coli, etc. is active. Therefore, it is possible to accurately and effectively improve the antibacterial action.

It should be understood that the effects of the present invention are not limited to the above-described effects, and include all effects that can be inferred from the configuration of the invention described in the detailed description or claims of the present invention.

1 is a schematic view of the release mechanism according to the pH of the porous nanocapsule according to an embodiment of the present invention.
FIG. 2 is a schematic diagram illustrating a method for manufacturing a porous nanocapsule according to an embodiment of the present invention and a mechanism for releasing and inhibiting the porous nanocapsule according to humidity/pH conditions.
3 is a result of EDS and XPS analysis of the PNC@SiO ₂ support according to an embodiment of the present invention.
4 is a SEM and TEM image of a PNC@SiO ₂ -g-MAA porous nanocapsule according to an embodiment of the present invention.
5 is a result of FT-IR analysis by performing a release test according to pH conditions of PNC@SiO ₂ -g-MAA porous nanocapsules according to an embodiment of the present invention.
6 is a UV-Vis analysis result by performing a release test according to pH conditions of PNC@SiO ₂ -g-MAA porous nanocapsules according to an embodiment of the present invention.
7 is a result of evaluation of antibacterial properties of PNC@SiO ₂ -g-MAA porous nanocapsules according to an embodiment of the present invention.
8 is a cytotoxicity evaluation result of PNC@SiO ₂ -g-MAA porous nanocapsules according to an embodiment of the present invention.

Hereinafter, the present invention will be described with reference to the accompanying drawings. However, the present invention may be embodied in several different forms, and thus is not limited to the embodiments described herein. And in order to clearly explain the present invention in the drawings, parts irrelevant to the description are omitted, and similar reference numerals are attached to similar parts throughout the specification.

Throughout the specification, when a part is "connected" with another part, this includes not only the case where it is "directly connected" but also the case where it is "indirectly connected" with another member interposed therebetween. . In addition, when a part "includes" a certain component, this means that other components may be further provided without excluding other components unless otherwise stated.

When a range of numerical values is recited herein, the values have the precision of the significant figures provided in accordance with the standard rules in chemistry for significant figures, unless the specific range is otherwise stated. For example, 10 includes the range of 5.0 to 14.9 and the number 10.0 includes the range of 9.50 to 10.49.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings.

One aspect of the present invention, a core comprising an antibacterial agent; a ligand brush formed on at least a portion of the surface of the core; And the antibacterial agent provides a porous nanocapsule that is released under the conditions of 45-100% humidity and pH 5-7.

Conventional antibacterial nanocapsules have a problem in that when the release of the antibacterial material starts, it continues until the time when the release of the antibacterial material is terminated, so that the release of the antibacterial material cannot be controlled, so that lifespan characteristics and antibacterial efficiency are lowered.

In the porous nanocapsule having excellent antibacterial properties of the present invention, the core containing the antimicrobial agent includes one or more pores, the ligand brush may be formed on at least a part of the surface of the core, and a specific range of conditions, specifically, humidity 45~ Under the conditions of 100% and pH 5-7, the pores may be opened according to the expansion of the ligand brush, and accordingly, the antibacterial agent contained in the core may be released. release can be prevented.

The core may be silica (SiO ₂ ), and as the silica precursor, tetraethylorthosilicate (TEOS), tetramethylorthosilicate (TMOS), tetramethoxysilicate (TMS), and 2 of these It may be one selected from the group consisting of the above mixtures, but is not limited thereto.

The core may be loaded with an antimicrobial agent, and the antimicrobial agent may be triclosan or an antimicrobial substance in a solution state. The triclosan has excellent sterilization and antibacterial effects against bacteria, bacteria and fungi, and can be rapidly dispersed within a short time to have excellent deodorization, anti-inflammatory and cleaning effects. The antibacterial material may be, for example, an amide-based compound, a pyridine-based compound, a carboxylic acid-based compound, nano-silver, a halogen compound, or a natural material having antibacterial properties, but is not limited thereto.

Meanwhile, a ligand brush may be formed on the core, and the ligand brush may react with the acrylic compound after bonding the silane compound to the core. Since the core is made of silica and has low binding force with the acrylic compound forming the ligand brush, uniform formation of the ligand brush is difficult. can form.

The silane-based compound is 3- (trimethoxysilyl) propylmethacrylate (3- (Trimethoxysilyl) propylmethacrylate, MPS), 3- (glycidoxy) trimethoxysilane (3- (Glycidyloxypropyl) trimethoxysilane, GPS), It may be one selected from the group consisting of 3-(mercaptopropyl) trimethoxysilane (3-(mercaptopropyl) trimethoxysilane, MCPS) and a mixture of two or more thereof, but is not limited thereto.

In addition, the acrylic compound is acrylic acid, methacrylic acid, methyl acrylate, methyl methacrylate, ethyl acrylate, ethyl methacrylate, propyl acrylate, propyl methacrylate, butyl acrylate, butyl methacrylate, pentyl acryl rate, pentyl methacrylate, polymethyl acrylate, polymethyl methacrylate, polyethyl acrylate, polyethyl methacrylate, and may be one selected from the group consisting of mixtures of two or more thereof, but is not limited thereto

On the other hand, the core may include one or more pores, and the pores may be closed by the ligand brush under a condition of less than pH 5, and open by the ligand brush under a condition of pH 5-7. have.

1 is a schematic view of the release mechanism according to the pH of the porous nanocapsule according to an embodiment of the present invention.

Referring to FIG. 1( a ), the core includes pores, and the silica support, which is the core, may be a PNC@SiO ₂ -MPS intermediate whose surface is modified by a silane-based compound, and the PNC@SiO ₂ -MPS The intermediate may be prepared as PNC@SiO ₂ -g-MAA porous nanocapsules by combining the acrylic compound. On the surface of the PNC@SiO ₂ -g-MAA porous nanocapsule, a ligand brush formed of a carboxyl group of MAA may be formed, an antibacterial agent is supported inside the capsule, and the pores of the capsule are opened by the ligand brush ( open) or closed (closed).

1(b) and (c) show that the pH conditions satisfy pH 4 and pH 6, respectively. In FIG. 1(b), hydrogen bonds are formed between the carboxyl groups of MAA, that is, between the ligand brushes, and the ligand brush expands. (swelling) does not occur, and the formed pores of the core are closed due to the interaction between MAA and Si—O—Si group molecules, that is, electrostatic attraction and intermolecular hydrogen bonding on the surface of the nanocapsule, so that the core The release of the antimicrobial agent supported on the can be controlled. On the other hand, in FIG. 1(c), as the carboxylic acid group of MAA is anionized to -COO- ^, a repulsive force is generated to repel each other, and accordingly, the ligand brush is expanded, and the pores of the core are opened, and the antibacterial agent can be released. .

On the other hand, the average particle diameter of the porous nanocapsules may be 200 ~ 800㎚. In the porous nanocapsule, the average particle diameter may increase when the antimicrobial agent is released, and the average particle diameter may decrease when the release is suppressed. If the average particle diameter of the porous nanocapsules is less than 200 nm, the antibacterial effect may be reduced, and if it exceeds 800 nm, the uniformity of the porous nanocapsules particles may be reduced and enlarged to reduce workability.

In addition, in order to achieve the above object, another aspect of the present invention, (a) preparing a silica core support by reacting an aqueous solution containing a silica (SiO ₂ ) precursor and an antibacterial agent; (b) reacting by adding a silane-based compound to the silica core support; and (c) adding and reacting an acrylic compound to the product of step (b) to form a ligand brush; wherein the antibacterial agent is a porous nano-particle released under conditions of 45-100% humidity and pH 5-7. A method for manufacturing a capsule is provided.

2 is a schematic diagram illustrating a method for manufacturing a porous nanocapsule according to an embodiment of the present invention and a mechanism for releasing and inhibiting the porous nanocapsule according to humidity/pH conditions.

Referring to FIG. 2, referring to FIGS. 2 (a) and (b), a silica core support (PNC@SiO ₂ ) can be prepared by reacting an aqueous solution containing a silica precursor and an antibacterial agent, and on the surface of the silica core support It may be prepared by including one or more pores. The silica precursor may be one selected from the group consisting of tetraethylorthosilicate (TEOS), tetramethylorthosilicate (TMOS), tetramethoxysilicate (TMS), and mixtures of two or more thereof, but not limited

Referring to FIG. 2( c ), an intermediate (PNC@SiO ₂ -MPS) in which the surface of the silica core support is modified by adding and reacting a silane-based compound to the PNC@SiO ₂ support can be prepared, and the surface of the intermediate PNC@SiO ₂ -g-MAA porous nanocapsules having a ligand brush formed thereon can be prepared by adding and reacting an acryl-based compound thereon. The PNC@SiO ₂ -g-MAA porous nanocapsule may release the antibacterial agent under a specific range of environmental conditions, specifically, the porous nanocapsule has a humidity of 45-100% and a pH of 5-7. In Figure 2(c), the release inhibition of the porous nanocapsules at pH 4 and the release of the antimicrobial agent at the pH 6 were shown. At pH 4, electrostatic attraction and hydrogen bonding between the ligand brushes of the porous nanocapsules. Due to this, the pores of the core are closed and the release of the antibacterial agent can be suppressed, and at pH 6, the carboxylic acid group of MAA forming the ligand brush of the porous nanocapsule is anionized into -COO- ^, and the repulsive force repels each other. As it occurs, the ligand brush swells, which in turn opens the pores of the core so that the antimicrobial agent can be released through the pores.

Hereinafter, an embodiment of the present invention will be described in more detail. However, the following experimental results describe only representative experimental results among the above examples, and the scope and contents of the present invention may not be construed as being reduced or limited by the examples. Each effect of the various embodiments of the present invention not explicitly presented below will be specifically described in the corresponding section. The experimental results below were performed at room temperature (25° C.) and atmospheric pressure (1 atm) unless otherwise specified.

Samples and Analytical Instruments

Pentanol, polyvinylpyrrolidone (Mw=40,000, Polyvinylpyrrolidone), tetraethyl orthosilicate (TEOS), ammonium hydroxide (28-30%, ammonium hydroxide), sodium citrate (0.18M, Sodium Citrate) ), 3-(Trimethoxysilyl)propylmethacrylate (MPS), methyl methacrylate poly (methalic acid, sodium salt) solution (Poly (methalic acid, sodium salt) ) solution, MAA), N-N'-methylenebisacrylamide (N,N'-methylenebisacrylamide, BIS), ammonium persulfate (APS), triclosan (anionic antibacterial substance), sodium laurylbenzene sulfate ( sodium lauryl benzenesulfate), buffer (Buffer, pH 4, 6), and absolute ethanol were used in the manufacturing process of nanocapsules, and all the samples were purchased from Korea-Sigma Aldrich, and deionized water was used in all processes. was used

The surface structure, such as the shape of the porous nanocapsule, was analyzed using a field emission scanning electron microscope (FE-SEM, LEO SUPRA 55, Carl Zeiss, Germany) and a transmission electron microscopy (TEM).

The element composition constituting the surface of the porous nanocapsule was analyzed by X-ray photoelectron spectroscopy (X-ray photoelectron spectroscopy, XPS, JEM-2100F (JEOL), USA).

Particle size was measured using particle size analysis (Otsuka (ELSZ-2000ZS), Japan).

Chemical composition and component changes were analyzed using a Fourier transform infrared spectrometer (FT-IR, Spectrum One System, PerkinElmer) in the range of 4,000-500 cm ^-1 according to the shift of the absorption peak.

The emission behavior test was analyzed using UV-Vis spectrometry (CARY 300 Bio, Varian).

In the cytotoxicity test, A549 cells were cultured using a cell viability assay kit (EZ-Cytox, Dogen, Seoul Korea), and viable cells were observed with an optical microscope (Olympus, CKX41).

Example 1: PNC@SiO ₂ support manufacturing

2.5 g of PVP was dispersed in 25 ml of pentanol and stirred at 250 rpm at 60° C. for about 50 minutes to prepare a homogeneous mixture. 80 mg (3.0 mg/mL) of triclosan was added to the prepared mixture, 3.5 ml of water, 250 μl of 0.18 M concentration of sodium citrate solution (243.12 μl of water contains 11.6 mg of sodium citrate), 2.4 ml of absolute ethanol, hydroxide After adding 250 μl of ammonium and stirring for 1 minute and 30 seconds, 1.0 ml of TEOS was added and stirred again for 1 minute and 30 seconds. After stirring was completed, the nanocapsules were separated by centrifugation at 10,000 rpm for 10 minutes after maintaining at room temperature for 12 hours, washing with ethanol (20vt%), and vacuum drying. The separated nanocapsules were dried at 550° C. using a high-temperature furnace to prepare a porous nanocapsule PNC@SiO ₂ support.

Example 2: PNC@SiO ₂ -MPS intermediate preparation

After adding 210 mg of ethanol and 1.05 ml of MPS to the prepared PNC@SiO ₂ support 210 mg, stirring at 250 rpm for 24 hours at 25 ° C., centrifugation again at 10,000 rpm for 10 minutes, and washing with 20vt% ethanol , and vacuum at 40 °C for 6 hours to prepare a PNC@SiO ₂ -MPS intermediate.

Example 3: PNC@SiO ₂ Preparation of -g-MAA porous nanocapsules

The PNC@SiO ₂ - In 100 mg of the MPS intermediate, 50 ml of water and 5 mg of sodium laurylbenzene sulfate were dispersed in a mixed solution, sonicated at 250 rpm for 1 hour, and MMA monomer 200 mg and BIS 15 mg were added. After bubbling in an N ₂ gas atmosphere for 30 minutes, ammonium persulfate solution (2.0 mL, 5.0 mg/mL) was quickly added and sonicated at 75° C. for 4 hours. The sonicated porous nanocapsule was removed Washed with ionized water and dried at room temperature to prepare PNC@SiO ₂ -g-MAA nanocapsules.

Experimental Example 1: PNC@SiO ₂ EDS and XPS analysis of scaffolds

3 is a PNC@SiO ₂ EDS and XPS analysis results of the support according to an embodiment of the present invention. 3(a) to 3(c), the surface of the PNC@SiO ₂ has a round spherical shape mainly forming a Si-O-Si bond, and in particular, FIGS. 3(b) and 3( c) is to confirm the element composition of the PNC@SiO ₂ by EDS analysis of PNC@SiO ₂ , it is possible to confirm the presence of Si and O elements forming the surface of the PNC@SiO ₂ , the PNC@SiO ₂ support is It can be confirmed that the porous nanocapsules are obtained by bonding Si-O-Si and C=O.

On the other hand, FIGS. 3(a1) to 3(c1) are the results of XPS analysis of the PNC@SiO ₂ support, and each peak represents an element composition corresponding to a chemical bond, and in FIG. 3(d), the element for each peak Atom (%) is shown. 3(a1) to 3(c1) and 3(d), it can be seen that PNC@SiO ₂ is a porous nanocapsule formed of a high molecular polymer and a silanol bond (Si-O-Si bond), It can be seen that the component ratios of Si and O, which are the main components, are 34.87 At% and 65.13 At%, respectively.

Experimental Example 2: PNC@SiO ₂ -SEM and TEM analysis of g-MAA porous nanocapsules

4 is a SEM and TEM image of a PNC@SiO ₂ -g-MAA porous nanocapsule according to an embodiment of the present invention.

4(a) and 4(b) are images of the surface shape by SEM of PNC@SiO ₂ -g-MAA porous nanocapsules, which are formed by the Pickering method, so that very small silica-based particles are gathered and spherical. It can be confirmed that the raspberry-type porous nanocapsules of In particular, the PNC@SiO ₂ -g-MAA porous nanocapsule in FIG. 4(b), which enlarged the surface of the PNC@SiO ₂ -g-MAA porous nanocapsule of FIG. 4(a), was manufactured by the Pickering method and the sintering process. The pores formed between the beads with the porous nanocapsule can be confirmed, and the shape of the ligand brush formed on the surface covering the pores thinly can be confirmed.

In addition, FIGS. 4(c) and 4(d) are analysis images by TEM of PNC@SiO ₂ -g-MAA porous nanocapsules. The black particles on the surface are parts due to Si-O-Si bonding, In particular, referring to FIG. 4( d ), it can be confirmed that, in the open space between the black particles, the pores formed between the particles are formed on the surface of the nanocapsule, and thus the nanocapsule having a porous structure is manufactured.

Specifically, in the preparation of PNC@SiO ₂ -g-MAA porous nanocapsules, PNC@SiO ₂ is formed by the Pickering method, and the PNC@SiO ₂ is very small silica particles on the surface due to the reaction of the polymer polymer. The PNC@SiO ₂ support, a porous nanocapsule of the core-shell type covering the pore surface, can be prepared by forming the shell portion of the capsule while self-assembly on the surface of the material droplets. By modifying the prepared PNC@SiO ₂ support with MPS, a PNC@SiO ₂ -MPS intermediate having a Si-O and (C=O)-OH structure formed on the surface thereof can be prepared, and the PNC@SiO ₂ -MPS intermediate can be reacted with the carboxyl group (-COOH) of MAA to prepare a porous nanocapsule (PNC@SiO ₂ -g-MAA) having a ligand brush grafted onto the surface thereof.

The PNC@SiO ₂ -g-MAA porous nanocapsule can release an antibacterial material when the condition of pH 5 (pKa > 2) or higher is satisfied by the -COO ^- group present on the surface on which the ligand brush is formed.

Experimental Example 3: PNC@SiO ₂ , PNC@SiO ₂ -MPS, PNC@SiO ₂ -Assessment of physical properties of g-MAA

Zeta potential
[mV] Mobility
[cm2/Vs] Diameter
[nm] Std. Dev.
[nm] PNC@SiO ₂ -16.58 -1.06E-04 414.8 105.6 PNC@SiO ₂ -MPS -51.95 -4.05E-04 487.6 88.6 PNC@SiO ₂ -g-MAA -39.09 -3.05E-04 527.8 90.5

Table 1 is a PNC@SiO ₂ support, PNC@SiO ₂ -MPS intermediate, and PNC@SiO ₂ -g-MAA of each manufacturing step according to an embodiment of the present invention. Surface to measure the surface zeta potential value of the porous nanocapsule The zeta potential value due to each chemical unit is shown by confirming the change in energy and anionization.

The surface charge value changes due to the equilibrium of chemical potentials such as Si-O-Si, C=O and COO- on the surface of the porous nanocapsule. Referring to Table 1, in the case of the PNC@SiO ₂ porous support, it can be seen that the surface electron value is anionized (-16.58 mV) due to the Si-O group of TEOS, and the zeta potential value after MPS modification is a C=O bond was decreased (-51.95 mV), and in PNC@SiO ₂ -g-MAA, MAA was graft-bonded, and thus, due to anionization of the carboxyl group, the electron density of negative charges on the surface of PNC@SiO ₂ -g-MAA increased. As a result, it can be confirmed that the surface potential value rose again (-39.09mV).

In addition, in the case of the PNC@SiO ₂ support in the average diameter, 414.8 (±105.6) nm, and the PNC@SiO ₂ -MPS intermediate, the average diameter was measured to be 487.6 (± 88.6) nm, PNC@SiO ₂ -g-MAA It can be seen that the average diameter of the porous nanocapsules increased to 527.8 (±90.5) nm. The average diameter of the porous nanocapsules increased as the surface modification progressed, and it can be seen that the size of the PNC@SiO ₂ -g-MAA porous nanocapsules on which MAA is grafted and ligand brushes are formed. Accordingly, it can be confirmed that the average diameter of the porous nanocapsules increases according to the surface modification process, and the standard deviation of the porous nanocapsule particles of the present invention is 90.5 nm, so that the particle size of the porous nanocapsules and It can be seen that the uniformity of the form is excellent, and accordingly, it can be predicted that excellent and uniform antibacterial and sterilization effects can be obtained when applied to coatings, spray forms, packaging materials, and special fabrics for protection.

Experimental Example 4: Evaluation of physical properties of porous nanocapsules 1

5 is an antibacterial agent triclosan, PNC@SiO ₂ -g-MAA containing an antibacterial agent, and PNC@SiO ₂ -g-MAA porous nanocapsule containing an antibacterial agent Chemicals when the antibacterial agent is released at pH 4 and pH 6, respectively It is the result of FT-IR analysis to confirm the composition and component change, and the FR-IR analysis of triclosan was performed together to confirm the presence or absence of the antibacterial agent.

Referring to Figure 5 (a), in the FT-IR spectrum of triclosan, an antibacterial agent, a specific peak was observed at 3,312 cm ^{-1 due to a hydroxyl group (-OH), and a C-Cl absorption band was observed at 722-570 cm -1 .} , a strong CH vibration band for CH ₂ -Cl was observed at 1,300-1,150 cm ^-1 . Referring to FIG. 5(b), in the case of PNC@SiO ₂ -g-MAA containing an antibacterial agent, a peak of 1,115 cm ^-1 by -COO ^- was observed by the MAA-grafted ligand brush on the surface of the nanocapsule. In addition, the characteristic peak of C=O, the MAA absorption band, was observed at 1,709 cm ^-1 , the C=OC group was observed by stretching vibration absorption in the band at 1,500 cm ^-1 , and a small amount of C-CI characteristic peak was 722 . cm ⁻¹ .

On the other hand, in order to confirm the release behavior of the antimicrobial agent according to the pH conditions of the porous nanocapsules of the present invention, chemical composition and component changes were analyzed when the release test was performed using buffers of pH 4 and pH 6.

Referring to FIG. 5(c), looking at the peak measured after conducting a release test of PNC@SiO ₂ -g-MAA containing an antibacterial agent at pH 4, PNC@SiO ₂ -g-MAA in the core of the porous nanocapsule. A small amount of the supported antibacterial agent was released, and the triclosan characteristic peak, C-CI absorption band, was observed at 722-570 cm ^-1 , and a strong CH oscillation band for CH ₂ -Cl was observed at 1,300-1,150 cm ^-1 , whereas Referring to FIG. 5(d), in the case of the porous nanocapsules subjected to the release experiment at pH 6, a large amount of the antibacterial agent supported on the core was released, and the -COO ^- group of MAA ^, which is the main component of the porous nanocapsule surface, was 1,150 cm- ¹ was observed as a main characteristic peak, and a small amount of C-CI characteristic peak remaining after triclosan, an antibacterial agent, was released was also observed at 722 cm ^-1 .

Experimental Example 5: Evaluation of physical properties of porous nanocapsules 2

6 is a graph showing the results of UV-Vis measurement of PNC@SiO ₂ -g-MAA porous nanocapsules according to an embodiment of the present invention to evaluate the release behavior of antibacterial agents at pH 4 and pH 6, respectively. To compare the release or not, UV-Vis measurement of PNC@SiO ₂ -g-MAA porous nanocapsules without antibacterial agent triclosan and antibacterial agent was also performed.

Referring to FIG. 6 , it can be seen that the PNC@SiO ₂ -g-MAA porous nanocapsules of the antibacterial agent triclosan and a buffer of pH 6 exhibit the triclosan release behavior at 240 nm. In contrast, it can be seen that the characteristic peak of triclosan does not appear in the PNC@SiO ₂ -g-MAA porous nanocapsules that do not contain an antibacterial agent and the PNC@SiO ₂ -g-MAA porous nanocapsules in the pH 4 buffer. Therefore, it can be expected that the release behavior of the antimicrobial agent, which is the core material of the PNC@SiO ₂ -g-MAA porous nanocapsule, can be controlled according to the pH condition.

In detail, the entangling ligand brush causes electrostatic interaction with the anionic antibacterial agent while swelling occurs only under specific humidity and pH conditions, and the PNC@SiO ₂ -g-MAA porous nanometer of the present invention In the case of capsules, the surface of the PNC@SiO ₂ -g-MAA porous nanocapsule is formed due to the electrostatic repulsion between negatively charged groups by ionization of the carboxyl group in an environment of pH 5 or higher, and the ligand brushes covering the pores are rapidly removed. It can be confirmed that the antibacterial agent is released as it swells due to the strong electrostatic repulsion with triclosan, which is an antibacterial agent having an anion value.

On the other hand, in the PNC@SiO ₂ -g-MAA porous nanocapsule at pH 4, the release of the antibacterial agent did not occur, in detail, the formation of hydrogen bonds between the carboxyl groups of MAA and the Si-O-Si groups present on the surface. Swelling of the ligand brushes hardly occurs due to the presence of electrostatic attraction, but at pH 6, the carboxyl group of MAA is ionized and electrically negatively charged, causing electrostatic repulsion between the groups to rapidly expand the ligand brush. Accordingly, it can be confirmed that the antimicrobial agent, which is the core material, is released by electrostatic repulsion.

Experimental Example 6: Evaluation of physical properties of nanocapsules 3

7 and Table 2 show the evaluation of the antibacterial properties of PNC@SiO ₂ -g-MAA porous nanocapsules according to an embodiment of the present invention.

7 shows (a1, b1, c1) control samples (Control sample, CS), (a2, b2, c2) PNC@ that does not contain an antibacterial agent in the medium pH 4, pH 6 and pH 7 for the evaluation of the antibacterial properties. SiO ₂ -g-MAA, (a3, b3, c3) Formation of a clear zone after leaving a sample of PNC@SiO ₂ -g-MAA containing an antibacterial agent in the core at regular intervals and at 100% humidity for 8 hours is an observed image.

On the other hand, in the evaluation of antibacterial properties, the density change of E. coli was measured for 8 hours using 55 mg/10 mL of porous nanocapsules, and the results are shown in Table 2. The release of the antibacterial agent from the support of the capsule and the formation of a clear zone through the antibacterial action were observed.

Referring to Figure 7, it can be confirmed that the clear zone of E. coli was not observed because the antibacterial agent was not released in all samples (a1, a2, a3) in the medium of pH 4, which is a porous nanometer under the condition of pH 4 As a result of strong hydrogen bonding between the ligand brushes formed in the capsule, it can be seen that the ligand brush does not have a swelling phenomenon. In contrast, it can be seen that clear zones of 12 mm and 13 mm were formed in the medium of pH 6 and 7, respectively. This is because the electrostatic repulsive force between the anionized ligand brushes on the surface of the porous nanocapsule containing the antibacterial agent is strengthened, and the ligand brush expands. Accordingly, the pores of the porous nanocapsule are opened and the antibacterial agent supported in the core is released to release the antibacterial agent. It can be confirmed according to the action effect.

pH 4 pH 6 pH 7 Early
OD value after 8h
OD value Early
OD value after 8h
OD value Early
OD value after 8h
OD value control sample
(CS) 0.028 0.080 0.034 0.100 0.037 2.350 PNC@SiO ₂ -g-MAA
(including antibacterial x) 0.028 9.390 0.034 2.490 0.037 3.400 PNC@SiO ₂ -g-MAA
(including antibacterial) 0.028 11.46 0.034 6.020 0.037 8.300

On the other hand, referring to Table 2 showing the results of changes in E. coli density, the initial density of E. coli in all cases at pH 4 was 0.028 (OD), and in CS, the density of E. coli increased to 0.080 (OD) after 8 hours, and antibacterial agents In the case of PNC@SiO 2 -g-MAA porous nanocapsules without PNC@SiO ₂ -g-MAA, it increased to 9.390 (OD), and in the case of PNC@SiO ₂ -g-MAA porous nanocapsules containing antibacterial agent, it increased the most to 11.46 (OD). did

At pH 6, the initial density of E. coli in all cases was 0.034 (OD), and after 8 hours, the density of E. coli was 0.100 (OD) at CS, PNC@SiO ₂ -g-MAA porous nano The capsule increased to 2.490 (OD), and 6.020 (OD) for the PNC@SiO ₂ -g-MAA porous nanocapsule containing the antibacterial agent. At pH 7, the initial density of E. coli was 0.037 (OD) in all cases, and the density of E. coli after 8 hours was 2.350 (OD), and PNC@SiO ₂ -g-MAA porous nanocapsules without antibacterial agents were 3.400 (OD), and increased to 8.300 (OD) in the case of PNC@SiO ₂ -g-MAA porous nanocapsules containing the antibacterial agent.

Accordingly, it can be confirmed that the PNC@SiO ₂ -g-MAA porous nanocapsule containing the antibacterial agent has the best effect of inhibiting the proliferation of E. coli by the antibacterial agent under the pH 6 condition, and in particular, the E. coli at pH 6 than at the pH 4 condition It can be confirmed that the density of the exhibits a proliferation inhibitory effect of about 48%. This is because when the PNC@SiO ₂ -g-MAA porous nanocapsule containing the antibacterial agent satisfies specific humidity and pH conditions, the ligand brush on the surface of the PNC@SiO ₂ -g-MAA porous nanocapsule expands, and the closed pores It can be confirmed that this is opened, and the antibacterial agent supported on the core is released, thereby realizing the antibacterial effect.

Experimental Example 7: Evaluation of physical properties of porous nanocapsules 4

8 and Table 3 are cytotoxicity evaluation results of PNC@SiO ₂ -g-MAA porous nanocapsules according to an embodiment of the present invention.

To perform the cytotoxicity test, a control sample (CS), PNC@SiO ₂ -g-MAA porous nanocapsules without antibacterial agent, and PNC@SiO ₂ -g-MAA porous nanocapsules containing antibacterial agent were used. A solution obtained by immersion in a buffer of pH 6 for 24 hours was used as an exposure solution.

The sample was diluted to a specific concentration and 3.12 × 10 ⁵ human lung epithelial cells (A549 cells) were distributed in 10 mL medium, and the distributed cells were cultured for 24 hours, sonicated for 5 minutes, and three exposure solutions After culturing for 24 hours after exposure to , the number of proliferated cells was observed under an optical microscope, the number of cells before exposure and after 24 hours of incubation was measured, and the cell adhesion rate according to the change in the number of adhesions is shown in Table 3.

In addition, the concentration of the exposure solution was performed in the range of 1/10 of the drug toxicity concentration standard, and accordingly, the concentration of the antimicrobial agent extracted from the capsule was 100ug/mL, and the cell adhesion activity was compared with the control sample.

number of cell attachments
(Number of cell adhesion) cell adhesion rate
(Cell adhesion activity, %) 0 h 24 h control sample
(CS) 104 166 160 PNC@SiO ₂ -g-MAA
(including antibacterial x) 85 112 132 PNC@SiO ₂ -g-MAA
(including antibacterial) 113 113 100

Referring to Table 3, it can be seen that the number of viable cells in CS increased from 104 to 166, and after culturing by exposure to the exposure solution for 24 hours, it can be confirmed that the number increased by 160% compared to before culturing. In addition, in the case of the PNC@SiO ₂ -g-MAA porous nanocapsule containing no antibacterial agent, the number of viable cells increased from 85 to 112, and it can be seen that the cell adhesion rate increased to 132%. On the other hand, in the case of the PNC@SiO ₂ -g-MAA porous nanocapsule containing the antibacterial agent, the number of surviving cells did not increase or decrease to 113 both before and after exposure, which is the PNC@SiO ₂ -g-MAA porous nanocapsule. It can be predicted that this is due to the antimicrobial agent released from the capsule.

Specifically, in the PNC@SiO ₂ -g-MAA porous nanocapsule of the present invention, the closed pores are opened by the expansion of the ligand brush on the surface of the porous nanocapsule while the antibacterial agent is supported on the core and specific pH and humidity conditions are met. , it can be predicted that the antimicrobial agent is released through the pores.

On the other hand, Figure 8 is a cytotoxicity test, and is a medium image observed before and after exposure, Figure 8 (a1, a2) is a control sample (Control sample, CS), Figure 8 (b1, b2) is an antibacterial agent PNC@SiO ₂ -g-MAA porous nanocapsules not containing, FIGS. 8 (c1, c2) are images before and after exposure of PNC@SiO ₂ -g-MAA porous nanocapsules containing an antibacterial agent. As shown in FIGS. 8(a1) and (a2), dead cells indicate that they are not attached to the side of the medium, and surviving cells indicate that the cells are attached to the side of the medium.

According to Table 3 and Figure 8, the PNC@SiO ₂ -g-MAA porous nanocapsule of the present invention showed a 60% lower cell adhesion rate than the control CS, but the change in the number of viable cells before and after exposure was It can be seen that the PNC@SiO ₂ -g-MAA porous nanocapsule has no toxicity to A549 cells while the antibacterial effect of the released antibacterial agent is present.

The description of the present invention described above is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a dispersed form, and likewise components described as distributed may also be implemented in a combined form.

The scope of the present invention is indicated by the following claims, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included in the scope of the present invention.

Claims (10)

a core comprising an antimicrobial agent;
a ligand brush formed on at least a portion of the surface of the core; and
The antibacterial agent is a porous nanocapsule that is released under the conditions of 45-100% humidity and pH 5-7. According to claim 1,
The core comprises one or more pores,
The pores are closed (closed) by the ligand brush at a condition of less than pH 5, and open (opened) by the ligand brush at a condition of pH 5 to 7 porous nanocapsules. According to claim 1,
The ligand brush is a porous nanocapsule in which an acryl-based compound is reacted after binding a silane-based compound to the core. 4. The method of claim 3,
The silane-based compound is 3- (trimethoxysilyl) propylmethacrylate (3- (Trimethoxysilyl) propylmethacrylate, MPS), 3- (glycidoxy) trimethoxysilane (3- (Glycidyloxypropyl) trimethoxysilane, GPS), A porous nanocapsule which is one selected from the group consisting of 3-(mercaptopropyl)trimethoxysilane (3-(mercaptopropyl)trimethoxysilane, MCPS) and a mixture of two or more thereof. 4. The method of claim 3,
The acrylic compound is acrylic acid, methacrylic acid, methyl acrylate, methyl methacrylate, ethyl acrylate, ethyl methacrylate, propyl acrylate, propyl methacrylate, butyl acrylate, butyl methacrylate, pentyl acrylate, A porous nanocapsule which is one selected from the group consisting of pentyl methacrylate, polymethyl acrylate, polymethyl methacrylate, polyethyl acrylate, polyethyl methacrylate, and a mixture of two or more thereof. According to claim 1,
The antibacterial agent is triclosan or a porous nanocapsule which is an antibacterial material in a solution state. According to claim 1,
The average particle diameter of the porous nanocapsules is 200 ~ 800nm porous nanocapsules. (a) preparing a silica core support by reacting an aqueous solution containing a silica (SiO ₂ ) precursor and an antibacterial agent;
(b) reacting by adding a silane-based compound to the silica core support; and
(c) adding an acrylic compound to the product of step (b) and reacting to form a ligand brush;
The method for producing a porous nanocapsule, wherein the antibacterial agent is released under the conditions of 45-100% humidity and pH 5-7. 9. The method of claim 8,
The silica core support is a method of manufacturing a porous nanocapsule comprising one or more pores. 9. The method of claim 8,
The average particle diameter of the porous nanocapsules is a method of manufacturing a porous nanocapsule of 200 ~ 800㎚.

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