WO2012060620A2

WO2012060620A2 - Anti-biofouling ssq/peg network and preparation method thereof

Info

Publication number: WO2012060620A2
Application number: PCT/KR2011/008278
Authority: WO
Inventors: 정봉현; 이봉국
Original assignee: 한국생명공학연구원
Priority date: 2010-11-02
Filing date: 2011-11-02
Publication date: 2012-05-10
Also published as: WO2012060620A3; KR20120046532A

Abstract

The present invention relates to an anti-biofouling SSQ/PEG network and a preparation method thereof. More particularly, the present invention relates to an SSQ/PEG network, and a preparation method thereof, wherein the network is prepared by curing a mixture of polyethylene glycol (PEG) and photocurable silsesquioxane (SSQ) with UV rays, has high resistance to non-specific adsorption that is used for coating a solid substrate, and can be directly nano-patterned. The anti-biofouling SSQ/PEG network according to the present invention not only has an anti-biofouling property but also has low viscosity, high optical transparency, high hydrophilicity, an anti-swelling property in organic/aqueous solution, high stabilities against biological, chemical and thermal stresses, and high mechanical strength, and can also be finely patterned into sub-25 nm-scale directly to be easily used for high-tech performance in a wide range of biomedical applications.

Description

Antibiotic Adhesion SV / PEB Network and Manufacturing Method Thereof

The present invention relates to an anti-bioadhesive SSQ / PEG network and a method for manufacturing the same, and more specifically, polyethylene glycol (PEG) and photocurable silsesquioxane (SSQ) are mixed and prepared by UV curing, and a solid substrate. The present invention relates to an SSQ / PEG network having high resistance to non-specific adsorption used for coating a film and capable of direct nano-patterning and a method of manufacturing the same.

In the manufacture of biomedical devices, biosenseors, diagnostic arrays, implants and drug delivery systems, the prevention of nonspecific adsorption of biopolymers on solid surfaces Of great importance (Senaratne W. et al., Biomacromolecules , 6: 2427, 2005; Cooper S. L. et al., Biomaterials. Interfacial phenomenaand applications, 1982).

The problem of nonspecific adsorption can be prevented by precoating the surface with a material that is resistant to the adsorption of biomaterials, and can be avoided by poly (vinylalcohol), polyethylene glycol (PEG, poly (ethyleneglycol)), poly Various polymeric materials, such as acrylamide, dextran, and methacrylated phosphatidylcholine, have been used as coatings for the prevention or minimization of nonspecific adsorption (Amanda A. etal. Mallapragada, Biotechnol. Prog. , 17: 917, 2001; Harris J. M. et al., Poly (ethyleneglycol) chemistry. Biotechnicaland Biomedical Applications, 1992; Park S. et al., J. Biomed. Mater. Res., 53: 568,2000 Masson J. F. et al., Taralan , 67: 918 , 2005; Ishihara K., Sci . Technol . Adv . Mater ., 1: 131, 2000).

The coating material was used in the form of self-assembled monolayer (SAM), polymer brush, hydrogel, and immobilized on the solid surface by covalent bonds such as physical adsorption and chemical coupling. (Amanda A. et al., Mallapragada, Biotechnol. Prog., 17: 917 , 2001; Harris J. M. et al., Poly (ethyleneglycol) chemistry. Biotechnicaland Biomedical Applications, 1992; Park S. etal., J. Biomed. Mater. Res. 53: 568 , 2000; Masson J. F. et al., Talanta , 67: 918 , 2005; Ishihara K., Sci. Technol. Adv. Mater. , 1: 131, 2000; Dulcey C. S. et al., Science , 252 : 551,1991; Milner S. T., Science , 251: 905,1991; Spargo B. J. et al., Proc . Natl . Acad . Sci . USA., 91: 11070, 1994; Ruhe J. et al., J. Biomater Sci., Polym. Ed. , 10: 859, 1999; Ratner B. D. etal., Annu. Rev. Biomed. Eng., 6:41, 2004; Sibarani J. etal., Colloids Surf ., B , 54:88. (2007), these materials are used for deep-ultraviolet lith to fabricate miniaturized biomedical devices and biosensors. ography, Dulcey CS et al., Science , 252: 551,1991), standard photolithography, Revzin A et al., Langmuir , 19: 9855,2003), dip-penlithography, Lee KB etal. , Nano Lett ., 4: 1869, 2004), electron-beam lithography, Smith JC et al., Nano Lett . , 3: 883, 2003), soft lithography, Whitesides GM et al., Annu. Rev. Biomed. Eng. , 3: 335, 2001; Mendes P. M. et al., Nanoscale Res. Lett . , 2: 373, 2007), and nanoimprint lithography, Falconnet D. et al., Nano Lett . , 4: 1909, 2004; Lee B. K. et al., Small , 4: 342, 2008; Lee B. K. et al., Lab Chip , 9: 132,2009), etc., were fabricated directly / indirectly in the form of micro / nanoscale on a solid substrate.

However, the use of precoatings such as antimicrobial self-assembled monolayers, polymer brushes, or hydrogels on the solid surface is well established to prevent nonspecific adsorption of biomaterials, but such precoatings There is a disadvantage to stability. PEG based SAMs are unstable and have been reported to oxidize rapidly, especially in the presence of oxygen and transition metal ions (Ostuni et al., Langmuir , 17: 5605,2001; Chen S. et al., J. Am. Chem. Soc. , 127: 14473,2005; Crouzet C. et al., Makromol. Chem . , 177: 145,1976), PEG brushes have been reported to gradually lose their repulsive properties to their proteins above 35 ° C (Leckband D. et al. , J. Phys. Chem. B, 10: 1125, 1998). Hydrogels are also reported to be too unstable in aqueous solutions (Priola A. et al., Polymer , 34: 3653,1993), and their three-dimensional structures collapse due to significant swelling by aqueous solutions (Kim P. etal). , LabChip , 6: 1432,2006).

Therefore, the fabrication of biomedical devices, biosensors, and lab-on-a-chips is important because the instability of antimicrobial materials is directly related to accuracy, sensitivity, and reproducibility in biosensing. This requires the development of stable antimicrobial materials.

On the other hand, stable anti-bioadhesive materials should be made of nanostructures by appropriate patterning methods with high productivity, low cost and high reproducibility for the production and production of biomedical devices, nanobiosensors, and lab-on-a-chips. In addition, to simplify their fabrication process, direct nanopatterning of antimicrobial materials is much more efficient than indirect patterning methods (Revzin A et al., Langmuir , 19: 9855,2003; Lee B.K. etal., Small , 4: 342, 2008; Lee B. K. et al., Lab Chip , 9: 132 , 2009; Kim P. et al., Adv. Mater. , 20:31 , 2008). To produce a pattern, a sufficiently hard material with a tensile modulus of 100 MPa or more is required (Palmieri F. et al., ACSNano , 1: 307, 2007).

In this respect, the ideal anti-bioadhesive material for advanced performance in a wide range of biomedical applications is not only high anti-biofouling property (Revzin A et al., Langmuir , 19: 9855,2003; Lee B.K. et al., Small , 4: 342, 2008; Lee B. K. etal., LabChip , 9: 132, 2009; Kim P. et al., Adv. Mater. , 20:31, 2008), low viscosity, high optical transparency , High hydrophilicity (Jeong HE et al., Small , 3: 778,2007), high resistance to swelling in organic / aqueous solutions (Kim P. etal., LabChip , 6: 1432,2006), biological, chemical, thermal stress Various properties are required such as high stability, high mechanical strength, and direct patterning possibility. However, antimicrobial materials that satisfy all of the above-mentioned various properties have not been developed yet.

Accordingly, the present inventors have made intensive efforts to develop an ideal anti-bioadhesive material having various properties for the above-mentioned wide range of biomedical applications, and thus photocurable silsesquioxane in photocurable PEG that can be directly patterned into an anti-bioadhesive material. (SSQ) was added to prepare a low viscosity photocurable SSQ / PEG mixture with improved PEG's unstable thermal stability, mechanical strength, insulation, swelling, and the like, followed by addition of 2 wt% UV initiator to the mixture, followed by simple UV Irradiation crosslinks the low viscosity photocurable SSQ / PEG mixture to produce an SSQ / PEG network, wherein the prepared SSQ / PEG network has a high anti-biofouling of less than 4.6%. Low viscosity of 20.8 to 175 cP, high optical transparency of 90% or more, high hydrophilicity (water contact angle = 42.2 to 54.5 °), high resistance to swelling in organic / aqueous solution (1.3 to 20.5 wt%), In addition to confirming that it has high stability against physical, chemical and thermal stress, and high mechanical strength of 1.898 to 2.815 GPa Young's modulus, UV embossing enables the fabrication of direct micropatterns of 25 nm or less. To complete.

Summary of the Invention

It is an object of the present invention to provide an anti-bioadhesive material for a wide range of biomedical applications and methods of making the same.

In order to achieve the above object, the present invention comprises the steps of (a) mixing polyethylene glycol (PEG) and photocurable silsesquioxane (SSQ); And (b) curing the mixture (a) by irradiating UV in the presence of a UV initiator to obtain an SSQ / PEG network.

The present invention also provides an SSQ / PEG network produced by the above method.

The present invention also provides a method of manufacturing a nanopattern, characterized in that using the SSQ / PEG network.

The present invention also provides nanodevices having high resistance to nonspecific adsorption including nanopatterns prepared by the above method.

Figure 1 shows the anti-bioadhesive SSQ / PEG network components and reaction schemes capable of free radical polymerization.

2 graphically shows the viscosity measurements of the SSQ / PEG mixture.

FIG. 3 shows UV-Vis transmission spectra of SSQ / PEG networks polymerized by free radical polymerization on a quartz substrate.

4 shows the initial static contact angle of the SSQ / PEG network and the equilibrium static contact angle after 20 minutes.

Figure 5 graphically shows the expansion rate of SSQ / PEG network for toluene and PBS.

6 graphically shows the Young's modulus for an SSQ / PEG network.

7 is a schematic diagram of a nanostructure fabrication process of SSQ / PEG mixture using UV embossing (a), Si master (b) with 50 nm features, Si master (c) with 25 nm features, 50 nm FE-SEM images of 50SSQMA / 50PEGDMA330 (d) nano patterned with features and 50SSQMA / 50PEGDMA330 (e) nano patterned with 25 nm features are shown.

Figure 8 shows the AFM height images and shrinkage of NIM-80L master mold (a), UV-embossed 50SSQMA / 50PEGDMA330 (b), UV-embossed 50SSQMA / 50PEGDMA550 (c) and UV-embossed 50SSQMA / 50PEGDMA750 (d). .

Figure 9 shows the optical image of the glass (a), 50SSQMA / 50PEGDMA330 network (b) and 50SSQOG / PEGGDG526 network (c) and component (d) of the 50SSQOG / 50PEGDG526 network capable of cationic polymerization.

FIG. 10 shows liposomes (ac) and EGFP (optionally adsorbed on 50SSQMA / 50PEGDMA330 (b, e) and 50SSQOG / 50PEGDG526 (c, f) prepared on glass (a, d), PET film) df) fluorescence image.

FIG. 11 shows a non-adhesive SSQ / PEG network prepared by free radical polymerization for biomedical applications up to 25 nm in size.

Detailed Description of the Invention and Specific Embodiments

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.

In one aspect, the present invention, in order to increase the crosslinking density of the PEG network, a photocurable SSQ was mixed and a photoinitiator was added to prepare an SSQ / PEG network using free radical polymerization. As a result of measuring the viscosity of the SSQ / PEG mixture mixed with the PEG and SSQ, it showed a low viscosity in the range of 20.8 ~ 175 cP.

The present invention in one aspect, (a) mixing polyethylene glycol (PEG) and photocurable silsesquioxane (SSQ); And (b) curing the mixture (a) by irradiating UV in the presence of a UV initiator to obtain an SSQ / PEG network.

In the present invention, the polyethylene glycol (PEG) may be selected from the group consisting of polyethylene glycol dimethacrylate (PEGDMA) and polyethylene glycol diacrylate (PEGDA), the photocurable silsesquioxane (SSQ) can be characterized as functionalized with methacrylate or acrylate.

In the present invention, the UV initiator is 2,2'-dimethoxy-2-phenylacetophenone (DMPA, 2,2'-dimethoxy-2-phenylacetophenone), 2-hydroxy-2-methyl-1-phenyl- Propane-1-one (HMPP, 2-hydroxy-2-methyl-1-phenyl-propane-1-one), 2,4,6-trimethylbenzoyl diphenylphosphine oxide (2,4,6-Trimethylbenzoyl-diphenylphosphine Oxide) and diphenyl 2,4,6-trimethylbenzoyl phosphine oxide (Diphenyl 2,4,6-trimethylbenzoyl phosphine oxide) may be selected from the group consisting of.

In another aspect, the present invention confirmed the UV-Vis permeability, surface hydrophilicity, swelling and mechanical properties of the SSQ / PEG network. As a result, the SSQ / PEG network showed high UV transmission (> 90% at 365 nm), high hydrophilicity (water contact angle = 42.2-54.5 °), and non-swelling or very small expansion rate in both toluene and PBS solutions, and excellent mechanical properties (Young's modulus = 1.898 ~ 2.815GPa).

In another aspect, the present invention relates to an SSQ / PEG network produced by the above method.

In the present invention, the SSQ / PEG network is (a) at least 90% UV transmission at a wavelength of 365nm or more; (b) a positive contact angle of 42.2-54.5 °; (c) mechanical strength of 1.898 to 2.815GPa; (d) an expansion ratio of 1.3 to 20.5 wt% based on the organic solvent and the aqueous solution; (e) shrinkage of 3% or less; And (f) less than 4.6% antimicrobial adhesion.

In the present invention, the SSQ / PEG network may be characterized by direct nanopatterning, direct nanopatterning is UV nanoimprint (UV nanoimprint), UV embossing (UV embossing) and UV replica molding (UV replica molding) It may be characterized in that it is performed by any one method selected from the group consisting of.

In another aspect, the present invention provides an SSQ / PEG mixture using a UV embossing method from a master mold to a high line-to-space density (1: 1), high aspect ratio (4: 1) and low shrinkage (< SSQ / PEG networks were patterned with relief nanostructures of 25-nm or smaller features with 3%).

In another aspect, the present invention confirmed the anti-bioadhesion of the nano patterned SSQ / PEG network using a fluorescence method. As a result, the SSQ / PEG network strongly prevented the adsorption of negatively charged liposomes and confirmed that they have long-term stability against chemical stress, thermal stress and biological stress.

In still another aspect, the present invention relates to a method of manufacturing a nanopattern, which uses an SSQ / PEG network.

In the present invention, the size of the nanopattern may be characterized in that less than 25nm.

In still another aspect, the present invention relates to a nanodevice having high resistance to nonspecific adsorption including a nanopattern produced by the above method.

In the present invention, the nanodevice may be selected from the group consisting of biomedical devices, biosensors, diagnostic arrays, implantation and delivery systems, and lab-on-a-chips.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only to illustrate the invention, it will be apparent to those of ordinary skill in the art that the scope of the present invention is not to be construed as limited by these examples.

Example 1 SSQ / PEG Network Fabrication

SSQ multifunctionalized with methacrylate, a mixture of various SSQMAs [(C ₇ H ₁₁ O ₂ ) _n (SiO _1.5 ) _n ], where n is 8, 10 or 12: the weight ratio of PEG is 2: 8 and 5: 5 In order to fabricate 20SSQ / 80PEG network and 50SSQ / 50PEG network, SSQ (SSQMA, Methacrylate multi-functionalized SSQ; Hybrid Plastics) was used for PEGDMA330 (Sigma Aldrich), PEGDMA550 (Sigma Aldrich), PEGDMA750 (Sigma Aldrich) and PEGDA575 (Sigma Aldrich). ) And the weight ratio was mixed for 5 minutes each to obtain a clear liquid mixture for all weight ratios.

2% (w / w) of UV initiator DMPA (2,2'-dimethoxy-2-phenylacetophenone; Sigma Aldrich) was added to the clear liquid mixture obtained as a mixture of SSQ / PEG, and 2% (w / w) of SSPA / PEG mixtures containing DMPA were spin coated onto quartz substrates modified with TMSPM (3- (trimethoxysilyl) propyl methacrylate; Sigma Aldrich) in the form of a 500 nm thick film. Then, the SSQ / PEG network was prepared by curing the SSQ / PEG mixture by irradiating 365 nm of UV (ultraviolet dose: 1000 mJ / cm ² ) for 30 minutes using a UV lamp (Toscure251; Toshiba) in a vacuum state ( 1).

In addition, the viscosity of the SSQ / PEG mixture was measured using a Brookfield viscometer Model DV-II Pro (Brookfield Engineering Labs Inc.) at 25 ° C., as shown in FIG. 2, depending on the SSQ / PEG mixture, ranging from 20.8 to 175 cP. Low viscosity.

Example 2: Characteristics of SSQ / PEG Network

(1) UV-Vis transmittance

In order to confirm UV-Vis transmittance of the SSQ / PEG network prepared in Example 1, UV-Vis transmittance of the 50SSQ / 50PEG network was measured in a wavelength range of 200 to 800 nm using a spectrophotometer (UVmini-1420; Shimadzu). Measured.

As a result, as shown in Figure 3, it was confirmed that the UV transmittance of more than 90% at a wavelength larger than 365nm. Such highly transmissive networks can be used as components in optical applications.

In addition, as a result of measuring the weight of SSQ / PEG network heat treated at 170 ° C. and SSQ / PEG not heat treated for 1 hour, there was no difference in weight before and after heat treatment. This means that the SSQ / PEG network has high thermal stability, and this result indicates that the first decomposition temperature of SSQ (OMPS) functionalized with PEG-8 methacrylate is 180 ° C or higher ( Pielichowski K. et al., Adv. Polym. Sci., 201: 225, 2006).

(2) surface hydrophilicity

Surface hydrophilicity is a desirable property for preventing nonspecific adsorption of biomolecules, especially in micro / nanofluid channel applications, which allows for the introduction of biological reagents without a pump (Jeong HE et al., Small , 3: 778, 2007).

Therefore, the surface hydrophilic static contact angle of SSQ / PEG network was measured and confirmed. To produce a flat SSQ / PEG network, the SSQ / PEG mixture was dispensed into small droplets on a Si wafer modified with PFOS (trichloro (1H, 1H, 2H, 2H-perfluorooctyl) silane; Sigma Aldrich). The transparent support made of 188 μm thick PET (poly (ethylene terephthalate) film was placed on the surface and a 1 μm thick SSQ / PEG mixture was placed under vacuum using a UV lamp (Toscure251; Toshiba) for 30 minutes. UV (ultraviolet dose: 1000 mJ / cm ² ) was irradiated to cure the SSQ / PEG mixture to prepare a flat SSQ / PEG network A 5 μl drop of distilled water was placed on a flat SSQ / PEG network using a contact angle analyzer (Mouse-X; SurfaceTech Co.). , Dispensing automatically and measuring the static contact angle to determine the initial static contact angle (SCA) of PEG homopolymer and SSQ / PEG networks after 20 minutes. The equilibrium static contact angle was compared Jung contact angle is the average value for at least 6 times.

As a result, as shown in FIG. 4, the equilibrium static contact angle of all networks was measured from 42.2 to 54.5 °, and the equilibrium static contact angle of all networks was slightly smaller than the initial static contact angle by surface hydration by continuous exposure to water. Lowered. The positive contact angle of PEGDMA330 decreased as the ratio of SSQMA increased, whereas the positive contact angle of PEGDMA550, PEGDMA575 and PEGDMA750 increased as the ratio of SSQMA increased. However, the static contact angle values of all kinds of SSQ / PEG networks were smaller than the static contact angle values of PEGDMA330 homopolymers, which means that all kinds of SSQ / PEG networks remain hydrophilic.

(3) swellability

Toluene and PBS were used to confirm the effect of SSQMA on the expansion of the PEG homopolymer.

Independent SSQ / PEG networks isolated from Si wafers modified with PFOS were immersed in 10 mM PBS (phosphate-buffered saline) at toluene and pH 7.4, and the SSQ / PEG networks were taken out at specific intervals to filter the remaining toluene and PBS on the surface. It was dried using paper. The expansion mass (Ws) was measured at a constant weight for 30 seconds, dried for 48 hours in a dryer at room temperature, and then again measured for the dry weight (Wd). The expansion rate (Qr) of SSQ / PEG was calculated by the following equation.

Equation 1

As a result, as shown in Figure 5, the expansion ratio of the PEG / SSQ network in toluene and PBS was measured to be 1.3 ~ 20.5 wt%, the expansion ratio of the PEG homopolymer increased with increasing the molecular weight of the PEG, PEG expansion rate of the homopolymer It was confirmed that the relative concentration of the SSQ increases, and decreases as the molecular weight of the PEG homopolymer decreases. This result is due to the increased crosslinking density of the network by SSQMA. In particular, the 50SSQMA / 50PEGDMA330 network exhibited negligible water expansion (1.3 wt .-%). Non-swellable 50SSQMA / 50PEGDMA330 networks can be used as insulating films for electrochemical biosensors because redox agents, such as ferricyanide compounds dissolved in buffer solutions, cannot pass through non-swellable films.

In addition, it was confirmed that the expansion rate for all 50SSQ / 50PEG networks was maintained for more than 1 month in toluene and PBS solution, which is stable in hydrophobic and hydrophilic environments.

Thus, for the development of non-swellable and stable hydrophobic films in hydrophobic and hydrophilic environments, a network of low molecular weight PEGs with high ratios of SSQ and relatively low expansion rates can be used.

(4) mechanical strength

Nanopatterning up to 25 nm with high line-to-space ratio (1: 1) and high aspect ratio (4: 1) is possible when the polymer network has sufficient mechanical strength to maintain the structure. In general, the Young's modulus of the crosslinked PEGDMA330 is 1GPa, the Young's modulus of the PEGDMA550 is 40.3 MPa and the PEGDMA750 Young's modulus is 16.5 MPa. LabChip , 6: 1432,2006; HuZ. Et al., J. Am. Chem. Soc. , 130: 14244, 2008) . To demonstrate that the mechanical strength of PEG networks is enhanced by the addition of SSQ, 50SSQ / 50PEG Each Young's modulus of the network was measured using a conventional Nanoindentation System (Nanoindenter XP; MTS Nano Instruments) at room temperature. In order to eliminate the influence of the substrate, a 50SSQ / 50PEG network film of approximately 5 μm thickness was prepared on a SiO ₂ wafer, and the Young's modulus was calculated at least 10 times to calculate the statistical mean value to minimize the position-dependent factor. The Posson's ratio of all 50SSQ / 50PEG networks was 0.35.

As a result, as shown in FIG. 6, the smallest Young's modulus for the 50SSQMA / 50PEGDMA330, 50SSQMA / 50PEGDMA550 and 50SSQMA / 50PEGDMA750 networks were measured to be 2.815, 2.038 and 1.898 GPa, respectively. These results are significantly higher than the Young's modulus of PEG homopolymers and higher than the Young's modulus values of previously studied polymer materials used as anti-bioadhesive materials (Amanda A. etal., Biotechnol. Prog. , 17: 917,2001; Kim A. et al., Lab Chip , 6: 1432,2006; HuZ. Et al., J. Am. Chem. Soc. , 130: 14244, 2008).

Thus, SSQ / PEG networks with high Young's modulus are capable of direct nanopatterning up to 25nm.

Example 3: Direct Nanopatterning of SSQ / PEG Mixtures

UV embossing method with high productivity, low cost and high reproducibility was used for direct nanopatterning of the 50SSQ / 50PEG mixture (FIG. 7A).

About 1 μl of SSQ / PEG mixture was dispensed drop by drop onto a silicone master modified with PFOS as a release agent. The silicon master mold was used by purchasing NIM25L / 100, NIM-80L, and NIM-100H having a line-to-space ratio of 1: 1 and a height of 100 nm in a size of 25-200 nm in NTT-AT Coporation. A transparent support such as glass or PET film modified with TMSPM was carefully transferred onto the surface and in vacuum the support was compressed at an imprinting pressure of 0.1 MPa for 10 seconds. The 50SSQ / 50PEG mixture was UV irradiated for 3 minutes from the top of the transparent substrate while the pressure was maintained by a nanoimprinter system (NM-401; Meisyo Kiko Co., Ltd) equipped with a UV lamp (Toscure251; Toshiba) under vacuum. UV dose: 200 mJ / cm ² ), and then the mold was separated from the substrate.

In order to confirm the surface morphology of the silicon master mold and the nanopatterned 50SSQ / 50PEG network, FE-SEM (Field emission scanning electron microscopy, S-4300 type microscope; Hitachi Co.) was used. To prevent charging, the 50SSQ / 50PEG network was coated with a 10 nm gold layer before analysis using the Quick Coater SC-701HMC (Sanyu Electron Co., Ltd.), and the patterned nanostructures were vibrated at ambient temperature (tapping mode). Photographs were taken with a Digital Instruments NanoScope III atomic force microscope (Veeco Instruments). Data was processed using SPIP V3.3.7.0 software.

As a result, as shown in FIG. 7, the low viscosity 50SSQMA / 50PEGDMA330 mixture was replicated from the silicon master to the nanostructures using UV embossing at a relatively short time of 190 seconds at room temperature and low pressure of 0.1 MPa. The pattern of uniformly replicated parallel lines with half-pitches of 50 nm and 25 nm did not differ from the silicon master and no defects were found at smaller sizes. In addition, the same results as for the 50SSMQA / 50PEGDMA330 were confirmed for nanopatterning of other 50SSQ / 50PEG mixtures.

In addition, the shrinkage of the nano-patterned 50SSQ / 50PEG network showed a low shrinkage of less than 3% as shown in FIG. 8, which improves the accuracy of the nanopattern and extends the life of the master mold. .

Example 4: Antibiotic Adhesion of SSQ / PEG Network

In order to confirm that the SSQ / PEG network prevents non-specific adsorption of biomolecules, SSQMA / PEGDMA having an 800 nm dot pattern and SSQOG / PEGDG having an 800 nm dot pattern were prepared as a comparative group (FIG. 9). SSQOG / PEGDG having the 800 nm dot pattern is a 50:50 wt .-% glycidyl ether octa-functionalized SSQ (Toagosei Co., Ltd.) / PEGDG (PEG) which is a cationic polymerizable mixture for UV embossing. 3 wt .-% Irgacure 250 (Ciba Special Chemicals) acting as a cationic photoinitiator and 1.5 wt .-% Darocur ITX (Ciba Special) acting as a cationic photoinitiator in a diglycidyl ether (Sigma Aldrich) mixture Chemicals) was prepared by mixing.

To measure the adsorption of biomolecules using a fluorescence method, liposomes containing texas-red 1,2-dihexadecanoyl-sn-glycero-phosphoethanolamine (molecular probes) and enhanced green fluorescence protein (EGFP) are modeled. Used as a biomolecule. POPC (phospholipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine; Avanti Polar Lipids Inc.) / SA (stearic acid; Sigma Aldrich) / with a molar ratio of 80: 20: 1 of 50 nm diameter 10 mM liposome suspensions of TR-DHPE were prepared using an extrusion method in PBS at pH 7.4. 3 mM EGFP purified from E. coli was suspended in 10 mM PBS. The liposomes and protein solution were dropped on a glass, SSQMA / PEGDMA having an 800 nm dot pattern and an SSQOG / PEGDG substrate having an 800 nm dot pattern, incubated at room temperature for 1 hour, and washed with PBS, followed by fluorescence addition apparatus (IX-FLA; Fluorescence images for each substrate were obtained using an Olympus BX51 inverted research microscope with Olympus and a high resolution digital camera (DP70; Olympus) for image acquisition. Red luminescence of 590 nm and more and green light emission of 480-550 nm were filtered using U-MWG and U-MSWG Olympus filter cubes, respectively.

As a result, as shown in FIG. 10, SSQMA / PEGDMA having an 800 nm dot pattern strongly prevented nonspecific adsorption of liposomes compared to SSQOG / PEGDG having a glass and 800 nm dot pattern. In order to measure the adsorption amount of liposomes and EGFP on each surface, the fluorescence intensity of TR-DHPE and EGFP on the surface was systematically measured, and when compared with the glass at 100%, all SSQMA / PEGDMA networks were liposome and It was confirmed to prevent the adsorption of EGFP to less than 4.6%. The prevention of biomolecule adsorption is due to the heavy hydration of PEG, good structural flexibility and high chain mobility. In this regard, hydrated PEGs with flexibility and mobility in SSQ / PEG networks can prevent nonspecific adsorption of organisms, whereas SSQOG / PEGDG networks prepared by cationic polymerization react with liposomes containing negatively charged SAs and Electrostatic interactions between the EGFP and the positively charged SSQOG / PEGDG surface resulted in uniform and high adsorption of biomolecules. This means that the free radical polymerization step is more appropriate than the cationic polymerization step. In addition, the pattern of the 50SSQMA / 50PEGDMA network did not degrade even after long standing for 12 hours at a high concentration of 10 mM liposomes in 10 mM PBS at pH 7.4. These results indicate that the SSQMA / PEGDMA network polymerized by free radicals not only has good stability in the biological environment, but also has strong resistance to nonspecific adsorption of biomolecules (FIG. 11).

Thus, SSQ / PEG networks with high optical transmission, low viscosity, non-swellability, hydrophilicity, high mechanical strength and high stability against chemical stress, thermal stress and biological stress can be free radical polymerized and directly nanopatterned. It can be used in a variety of biomedical applications such as nanobiodevices, nanobiosensors and lab-on-a-chips.

As described above in detail specific parts of the present invention, it is apparent to those skilled in the art that such specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. something to do. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

The anti-bioadhesive SSQ / PEG network according to the present invention not only has high anti-biofouling, but also low viscosity, high optical transparency, high hydrophilicity, high resistance to swelling in organic / aqueous solutions, biological and chemical And it has high stability against thermal stress and high mechanical strength, it is possible to manufacture direct micropattern up to 25nm can be easily used for advanced performance of a wide range of biomedical applications.

Claims

Method for preparing an anti-bioadhesive SSQ / PEG network comprising the following steps:

(a) mixing polyethylene glycol (PEG) and photocurable silsesquioxane (SSQ); And

(b) curing the mixture (a) by irradiation with UV in the presence of a UV initiator to obtain an SSQ / PEG network.
The method of claim 1, wherein the polyethylene glycol (PEG) is selected from the group consisting of polyethylene glycol dimethacrylate (PEGDMA) and polyethylene glycol diacrylate (PEGDA) of the anti-bioadhesive SSQ / PEG network Manufacturing method.
The method of claim 1, wherein the photocurable silsesquioxane (SSQ) is functionalized with methacrylate or acrylate.
The method of claim 1, wherein the UV initiator is 2,2'-dimethoxy-2-phenylacetophenone (DMPA, 2,2'-dimethoxy-2-phenylacetophenone), 2-hydroxy-2-methyl-1-phenyl Propane-1-one (HMPP, 2-hydroxy-2-methyl-1-phenyl-propane-1-one), 2,4,6-trimethylbenzoyl diphenylphosphine oxide (2,4,6-Trimethylbenzoyl- Diphenylphosphine Oxide) and diphenyl 2,4,6-trimethylbenzoyl phosphine oxide (Diphenyl 2,4,6-trimethylbenzoyl phosphine oxide) method for producing an anti-bioadhesive SSQ / PEG network characterized in that selected from the group consisting of .
SSQ / PEG network prepared by the method of any one of claims 1 to 4.
6. The SSQ / PEG network of claim 5, wherein the SSQ / PEG network is capable of direct nanopatterning.
The method of claim 6, wherein the direct nanopatterning is performed by any one method selected from the group consisting of UV nanoimprint, UV embossing, and UV replica molding. SSQ / PEG network.
6. The SSQ / PEG network of claim 5, wherein the SSQ / PEG network has the following physical properties:

(a) at least 90% UV transmission at wavelengths of at least 365 nm;

(b) a positive contact angle of 42.2-54.5 °;

(c) mechanical strength of 1.898 to 2.815GPa;

(d) an expansion ratio of 1.3 to 20.5 wt% based on the organic solvent and the aqueous solution;

(e) shrinkage of 3% or less; And

(f) less than 4.6% antimicrobial adhesion.
The method of manufacturing a nanopattern characterized by using the SSQ / PEG network of claim 5.
10. The method of claim 9, wherein the size of the nanopattern is 25nm or less.
Nanodevice having a high resistance to non-specific adsorption including the nanopattern produced by the method of claim 9.
The nanodevice of claim 11, wherein the nanodevice is selected from the group consisting of a biomedical device, a biosensor, a diagnostic array, an implantation and delivery system, and a lab-on-a-chip.