CN111333774A - Antibacterial hydrogel based on coumarin skeleton and preparation method and application thereof - Google Patents
Antibacterial hydrogel based on coumarin skeleton and preparation method and application thereof Download PDFInfo
- Publication number
- CN111333774A CN111333774A CN202010290097.XA CN202010290097A CN111333774A CN 111333774 A CN111333774 A CN 111333774A CN 202010290097 A CN202010290097 A CN 202010290097A CN 111333774 A CN111333774 A CN 111333774A
- Authority
- CN
- China
- Prior art keywords
- photoinitiator
- solution
- hydrogel
- concentration
- active monomer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000000017 hydrogel Substances 0.000 title claims abstract description 79
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 52
- 125000000332 coumarinyl group Chemical group O1C(=O)C(=CC2=CC=CC=C12)* 0.000 title claims abstract description 46
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 239000000178 monomer Substances 0.000 claims abstract description 88
- 238000006243 chemical reaction Methods 0.000 claims abstract description 43
- 238000005286 illumination Methods 0.000 claims abstract description 33
- IWPNSXBCZANXBC-UHFFFAOYSA-N 7-hydroxy-4-(hydroxymethyl)chromen-2-one Chemical compound C1=C(O)C=CC2=C1OC(=O)C=C2CO IWPNSXBCZANXBC-UHFFFAOYSA-N 0.000 claims abstract description 32
- HFBMWMNUJJDEQZ-UHFFFAOYSA-N acryloyl chloride Chemical compound ClC(=O)C=C HFBMWMNUJJDEQZ-UHFFFAOYSA-N 0.000 claims abstract description 31
- 238000000034 method Methods 0.000 claims abstract description 16
- 238000004132 cross linking Methods 0.000 claims abstract description 15
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 42
- 239000003960 organic solvent Substances 0.000 claims description 42
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 41
- 238000002156 mixing Methods 0.000 claims description 29
- 238000003756 stirring Methods 0.000 claims description 26
- 229910052760 oxygen Inorganic materials 0.000 claims description 22
- 229910052736 halogen Inorganic materials 0.000 claims description 18
- 150000002367 halogens Chemical class 0.000 claims description 18
- 125000000217 alkyl group Chemical group 0.000 claims description 17
- 239000008367 deionised water Substances 0.000 claims description 15
- 229910021641 deionized water Inorganic materials 0.000 claims description 15
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 14
- 239000001301 oxygen Substances 0.000 claims description 14
- 238000001035 drying Methods 0.000 claims description 13
- 230000007935 neutral effect Effects 0.000 claims description 13
- 125000004093 cyano group Chemical group *C#N 0.000 claims description 11
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 claims description 10
- 229910052757 nitrogen Inorganic materials 0.000 claims description 9
- 229910052717 sulfur Inorganic materials 0.000 claims description 9
- 229910052698 phosphorus Inorganic materials 0.000 claims description 8
- 229910052710 silicon Inorganic materials 0.000 claims description 8
- 230000000977 initiatory effect Effects 0.000 claims description 6
- 229910052739 hydrogen Inorganic materials 0.000 claims description 5
- 239000001257 hydrogen Substances 0.000 claims description 5
- 230000001678 irradiating effect Effects 0.000 claims description 5
- 230000002194 synthesizing effect Effects 0.000 claims description 5
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 claims description 4
- 230000000845 anti-microbial effect Effects 0.000 claims description 4
- 230000001580 bacterial effect Effects 0.000 claims description 3
- 150000002431 hydrogen Chemical class 0.000 claims description 3
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 claims description 2
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims description 2
- OMIGHNLMNHATMP-UHFFFAOYSA-N 2-hydroxyethyl prop-2-enoate Chemical compound OCCOC(=O)C=C OMIGHNLMNHATMP-UHFFFAOYSA-N 0.000 claims description 2
- NLHHRLWOUZZQLW-UHFFFAOYSA-N Acrylonitrile Chemical compound C=CC#N NLHHRLWOUZZQLW-UHFFFAOYSA-N 0.000 claims description 2
- BZHJMEDXRYGGRV-UHFFFAOYSA-N Vinyl chloride Chemical compound ClC=C BZHJMEDXRYGGRV-UHFFFAOYSA-N 0.000 claims description 2
- 125000005250 alkyl acrylate group Chemical group 0.000 claims description 2
- QNILTEGFHQSKFF-UHFFFAOYSA-N n-propan-2-ylprop-2-enamide Chemical compound CC(C)NC(=O)C=C QNILTEGFHQSKFF-UHFFFAOYSA-N 0.000 claims description 2
- 239000003999 initiator Substances 0.000 abstract description 26
- 230000015572 biosynthetic process Effects 0.000 abstract description 11
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 abstract description 6
- 238000003786 synthesis reaction Methods 0.000 abstract description 5
- 150000001875 compounds Chemical class 0.000 abstract description 4
- 239000000463 material Substances 0.000 abstract description 4
- 229960000956 coumarin Drugs 0.000 abstract description 3
- 235000001671 coumarin Nutrition 0.000 abstract description 3
- 238000013461 design Methods 0.000 abstract description 3
- 239000003431 cross linking reagent Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 79
- 239000000499 gel Substances 0.000 description 16
- 241000588724 Escherichia coli Species 0.000 description 13
- 239000002609 medium Substances 0.000 description 11
- 239000007787 solid Substances 0.000 description 11
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 9
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 7
- 238000000576 coating method Methods 0.000 description 7
- 238000012986 modification Methods 0.000 description 7
- 230000004048 modification Effects 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 239000011248 coating agent Substances 0.000 description 6
- 238000006116 polymerization reaction Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000001879 gelation Methods 0.000 description 3
- 150000002430 hydrocarbons Chemical group 0.000 description 3
- 239000011148 porous material Substances 0.000 description 3
- 238000012604 3D cell culture Methods 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 239000002274 desiccant Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 125000001183 hydrocarbyl group Chemical group 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 239000012452 mother liquor Substances 0.000 description 2
- 235000019645 odor Nutrition 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 125000003386 piperidinyl group Chemical group 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- 125000000026 trimethylsilyl group Chemical group [H]C([H])([H])[Si]([*])(C([H])([H])[H])C([H])([H])[H] 0.000 description 2
- 239000004215 Carbon black (E152) Substances 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010015866 Extravasation Diseases 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical group [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000009881 electrostatic interaction Effects 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000036251 extravasation Effects 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 description 1
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 230000007505 plaque formation Effects 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
- 230000005641 tunneling Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F220/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical or a salt, anhydride ester, amide, imide or nitrile thereof
- C08F220/02—Monocarboxylic acids having less than ten carbon atoms; Derivatives thereof
- C08F220/52—Amides or imides
- C08F220/54—Amides, e.g. N,N-dimethylacrylamide or N-isopropylacrylamide
- C08F220/56—Acrylamide; Methacrylamide
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F2/00—Processes of polymerisation
- C08F2/46—Polymerisation initiated by wave energy or particle radiation
- C08F2/48—Polymerisation initiated by wave energy or particle radiation by ultraviolet or visible light
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
- C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
- C08J3/075—Macromolecular gels
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention discloses an antibacterial hydrogel based on a coumarin skeleton and a preparation method and application thereof, and belongs to the technical field of hydrogels. The invention uses 4-hydroxymethyl-7-hydroxycoumarin or derivatives thereof and acryloyl chloride to synthesize a photoinitiator containing a coumarin skeleton, and then uses the photoinitiator to initiate active monomer crosslinking reaction under the illumination condition to prepare the antibacterial hydrogel. The coumarin skeleton-based photo-crosslinking initiator synthesized by the method can efficiently and quickly initiate active monomers to form hydrogel with high transparency and certain mechanical strength, effectively expands the application of a photoresponse coumarin micromolecule compound in the field of antibacterial materials, and provides a new idea for design synthesis and application expansion of a functional photo-crosslinking agent.
Description
Technical Field
The invention relates to the technical field of hydrogel, in particular to antibacterial hydrogel based on coumarin skeleton and a preparation method and application thereof.
Background
The hydrogel is a high molecular polymer with a three-dimensional cross-linked network structure, has wide application in the fields of chemical industry, agriculture, medicine and the like due to good hygroscopicity and high water retention, and particularly becomes a research hotspot in the aspects of tissue engineering, wound dressing, drug wrapping release, 3D cell culture and the like. At present, the literature reports various methods for the preparation and formation of hydrogels, including supramolecular interaction gelation, electrostatic interaction gelation and copolymeric cross-linking gelation. The hydrogel formation method based on non-covalent interaction generally has higher requirements on the structure of the reactive monomer, the temperature, the pH value and the like of a system. The hydrogel preparation method based on covalent crosslinking can rapidly crosslink active monomers in aqueous solution so as to realize the conversion from small molecular monomers to a three-dimensional network, the preparation method can realize the high-efficiency forming of high-performance hydrogel, and can realize the regulation and control of various performances of the hydrogel by adjusting the structure of an initiator, the structure of the active monomers or the ratio of the initiator and the active monomers, so the application is very wide.
At present, the widely used small molecule photoinitiator still has some performance defects, for example, the photoinitiator has a certain color, and the transparency of the hydrogel obtained by initiation is limited; in addition, most photoinitiators release undesirable odors during initiation, which also limits their use; more importantly, to impart more functionality to the formed hydrogel, further modification of the reactive monomer, or further modification of the formed hydrogel, is required; however, the modification of the reactive monomer has higher requirements on the universality of the initiator, and the modification of the hydrogel is likely to influence the properties of the hydrogel.
Coumarin and derivatives thereof are a series of micromolecular compounds which are simple and stable in structure and have various biological activities. The skeleton has strong fluorescence emission and can endow the material with the required fluorescence characteristic. Fluorescence is used as an important visual analysis means, has the advantages of short response time, simple operation, naked eye observation and the like, and is sequentially and widely applied to detection of biological targets such as active oxygen, active sulfur, enzyme and the like in living cells and even in animals. In particular, organic small molecule fluorophores are favored by a plurality of scientific researchers because of the characteristics of easy modification, low cost and the like.
At present, researchers modify a fluorophore on an active monomer or physically wrap a compound with fluorescence in a crosslinking process, so as to prepare a series of hydrogels with a luminescence property, and the hydrogels are applied to aspects of information encryption, 3D cell culture and the like. However, the modification process of the active monomer is usually complicated, and the strategy of physical entrapment also has the disadvantages of gel network and fluorophore size mismatch, which leads to fluorophore extravasation and the like.
Disclosure of Invention
The invention aims to provide an antibacterial hydrogel based on a coumarin skeleton and a preparation method and application thereof, and aims to solve the problems that the existing hydrogel is single in function and complicated in preparation process.
The technical scheme for solving the technical problems is as follows:
4-hydroxymethyl-7-hydroxycoumarin or derivatives thereof and acryloyl chloride are used for synthesizing a photoinitiator containing a coumarin skeleton, and then the photoinitiator is used for initiating an active monomer crosslinking reaction under the illumination condition to prepare the antibacterial hydrogel.
The photoinitiator containing the coumarin skeleton is synthesized by using the 4-hydroxymethyl-7-hydroxycoumarin and the derivatives thereof and acryloyl chloride, the fluorescent characteristic required by the hydrogel material is endowed by utilizing the stronger fluorescence emission performance of the coumarin skeleton, and meanwhile, the photoinitiator synthesized by the invention uses double bonds at two ends of the molecular structure thereof to initiate active monomers to be crosslinked to form hydrogel.
Further, in a preferred embodiment of the present invention, the derivative of 4-hydroxymethyl-7-hydroxycoumarin has the following structural formula:
wherein R is0Is one of cyano, halogen, alkyl and O, N, halogen, P, S or Si-containing alkyl.
The structural formula of the 4-hydroxymethyl-7-hydroxycoumarin is as follows:
further, in a preferred embodiment of the present invention, the photoinitiator has the following structural formula:
wherein R is1Is one of hydrogen, cyano, halogen, alkyl and O, N, halogen, P, S or Si-containing alkyl.
The photoinitiator is synthesized by the following reaction process:
corresponding to R in the above structural formula1Is hydrogen.
Or:
wherein R is0Is one of cyano, halogen, alkyl and O, N, halogen, P, S or Si-containing alkyl; r1Is one of cyano, halogen, alkyl and O, N, halogen, P, S or Si-containing alkyl.
Furthermore, in the preferred embodiment of the present invention, the reaction concentration a of the photoinitiator is more than 0 and less than or equal to 1mmol/L, and the reaction concentration b of the reactive monomer is 20 wt% to 30 wt%; the illumination conditions are as follows: irradiating for 10 min-24 h under the illumination with the wavelength of 200 nm-405 nm.
Further, in a preferred embodiment of the present invention, the reaction concentration a of the photoinitiator is 0.125 mmol/L. ltoreq. a.ltoreq.1 mmol/L. More preferably, the reaction concentration a of the initiator is 0.125 mmol/L. ltoreq. a.ltoreq.0.25 mmol/L. Most preferably, the reaction concentration a of the initiator is 0.25 mmol/L.
Further, in a preferred embodiment of the present invention, the illumination conditions are: irradiating for 30-120 min under the illumination with the wavelength of 270-405 nm. More preferably, the wavelength is 270nm to 365nm, most preferably 365 nm. More preferably, the irradiation time is from 30min to 90min, most preferably 60 min.
Further, in the preferred embodiment of the present invention, the reaction concentration a of the photoinitiator is 0.25mmol/L, and the reaction concentration b of the reactive monomer is 25 wt%; the illumination conditions are as follows: irradiating under 365nm light for 60 min.
Further, in a preferred embodiment of the present invention, the reactive monomer is one or more selected from the group consisting of acrylamide, acrylic acid, N-isopropylacrylamide, 2-hydroxyethyl acrylate, vinyl chloride, styrene, acrylonitrile, and alkyl acrylates. In the case of a plurality of combinations, the mixing ratio is arbitrary.
Further, in a preferred embodiment of the present invention, the preparation method comprises the following specific steps:
(1) dissolving 4-hydroxymethyl-7-hydroxycoumarin or a derivative thereof in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, then dropwise adding the acryloyl chloride into the system, reacting for 1.5-2.5h under the ice bath condition, then stirring for 8-15 h at room temperature, adjusting the pH value of the system to be neutral after the reaction is completed, drying, separating and purifying to obtain a photoinitiator containing a coumarin skeleton;
(2) mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 10min to 24h under illumination with the wavelength of 200nm to 405nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is more than 0 and less than or equal to 1mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Further, in a preferred embodiment of the present invention, the organic solvent is one or more of dichloromethane, tetrahydrofuran, chloroform, methanol and dimethylsulfoxide. When it is a combination of plural kinds, it is an arbitrary mixing ratio.
Triethylamine is added in the step (1) and has the function of serving as an acid-binding agent to inhibit the generated hydrogen chloride.
The antibacterial hydrogel based on the coumarin skeleton is prepared by the preparation method.
The application of the antibacterial hydrogel based on the coumarin skeleton in bacterial culture.
Such bacteria include, but are not limited to: gram-negative bacteria and gram-positive bacteria such as escherichia coli, staphylococcus aureus, pseudomonas aeruginosa and the like.
The invention has the following beneficial effects:
the invention firstly synthesizes the photoinitiator based on the coumarin skeleton, and initiates the active monomer to realize the formation of the photo-crosslinking polymerized hydrogel. Because both ends of the photoinitiator contain double bonds and the skeleton has photosensitive characteristics, the photoinitiator can quickly initiate the reactive monomer to form gel under the irradiation of light with specific wavelength. Further, after the synthesis of the photo-crosslinking polymeric hydrogel was successful, the preparation of the hydrogel coating and the culture of E.coli were achieved in combination with the determination of the IC50 value of the initiator.
The antibacterial hydrogel prepared by utilizing the photo-crosslinking polymerization reaction has high transparency, and can be conveniently and rapidly prepared into a uniform antibacterial coating on a substrate. In addition, the hydrogel formed by the initiation of the low-concentration photoinitiator prepared by the invention has a very good inhibition effect on the growth of escherichia coli and the formation of colonies. The preferred photo-crosslinking polymeric hydrogel effectively makes up the defects of color or certain odor of common micromolecular photoinitiators.
The invention has mild reaction conditions, high efficiency and high speed, effectively expands the application of the photoinitiated hydrogel in the biological field and provides a new idea for the design and synthesis of the functional photoinitiator.
Drawings
FIG. 1 is an acrylamide standard curve.
FIG. 2 shows the conversion of acrylamide monomer at various initiator concentrations.
FIG. 3 shows the conversion of acrylamide monomer under different illumination times.
Figure 4 is a scanning tunneling microscope (SEM) image of photoinitiated hydrogel formation.
FIG. 5 shows HeLa cytotoxicity experiments (A) and calculation of IC50 values (B) for synthetic photoinitiators of the invention.
FIG. 6 is a schematic diagram of the synthesis of photoinitiator to initiate gelling of acrylamide as an antibacterial coating for culturing Escherichia coli.
FIG. 7 shows the growth of E.coli on the antibacterial gel coat in solid medium and different initiator concentrations.
FIG. 8 is a SEM of the morphology of E.coli grown on solid LB medium and antimicrobial gel coating; (A, D) blank-LB solid medium; (B, E) initiator concentration of 0.125 mmol/L; the initiator concentration of (C, F) was 0.5 mmol/L.
Detailed Description
The principles and features of this invention are described below in conjunction with the following drawings, which are set forth by way of illustration only and are not intended to limit the scope of the invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
In the following examples of the present invention, in the step (1) of synthesizing the photoinitiator, the organic solvent used is dichloromethane, and the organic solution used in the step (2) of dissolving the photoinitiator is dimethyl sulfoxide. Other examples of the present invention may be selected from dichloromethane, tetrahydrofuran, chloroform, methanol and dimethylsulfoxide, and the organic solvent that may be used is not limited to those mentioned in the following examples.
In the following embodiments of the present invention, the drying agent is anhydrous sodium sulfate, and the specific process of separation and purification is as follows: and evaporating the solvent from the dried sample under reduced pressure to obtain an oily white solid, separating by 300-400-mesh silica gel column chromatography, and taking ethyl acetate/petroleum ether-1/5 as an eluent to finally obtain a white solid powdery substance, namely the photoinitiator. Of course, those skilled in the art can use other known drying agents and separation and purification methods for processing, which is not limited by the invention.
The coumarin skeleton-based antibacterial hydrogel mainly comprises the following two steps: step one, synthesizing a photoinitiator containing a coumarin skeleton by using 4-hydroxymethyl-7-hydroxycoumarin and derivatives thereof and acryloyl chloride; and secondly, initiating a crosslinking reaction of the active monomer by using a photoinitiator under the illumination condition to prepare the antibacterial hydrogel.
The structural formula of the 4-hydroxymethyl-7-hydroxycoumarin is as follows:
the structural formula of the derivative of the 4-hydroxymethyl-7-hydroxycoumarin is as follows:
wherein R is0Is one of cyano, halogen, alkyl and O, N, halogen, P, S or Si-containing alkyl.
The structural formula of the photoinitiator synthesized by the 4-hydroxymethyl-7-hydroxycoumarin and the derivative thereof is as follows:
wherein R is1Is one of hydrogen, cyano, halogen, alkyl and O, N, halogen, P, S or Si-containing alkyl. Namely: r1Is hydrogen; or R1Is cyano; or R1Is halogen; or R1Is an alkyl group; or R1Is an O-containing hydrocarbon group; or R1Is an N-containing hydrocarbon group; or R1Being a halogen-containing hydrocarbon radical(ii) a Or R1Is a P-containing hydrocarbyl group; or R1Is an S-containing hydrocarbyl group; or R1Is a Si-containing hydrocarbon group.
The reaction processes of synthesizing the photoinitiator by the 4-hydroxymethyl-7-hydroxycoumarin and the derivatives thereof are respectively as follows:
the present invention will be described in detail with reference to examples.
Example 1:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, dropwise adding the acryloyl chloride into the system, reacting for 1.5h under the condition of ice bath, then stirring overnight at room temperature, after complete reaction, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 180min under the illumination with the wavelength of 200nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is 0.1mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Example 2:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, dropwise adding the acryloyl chloride into the system, reacting for 2.5h under the condition of ice bath, then stirring overnight at room temperature, after complete reaction, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 15min under the illumination with the wavelength of 405nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is 0.125mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Example 3:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, dropwise adding the acryloyl chloride into the system, reacting for 2 hours under the ice bath condition, stirring overnight at room temperature, after complete reaction, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 120min under illumination with a wavelength of 270 to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is 0.13mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Example 4:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving a derivative of 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, then dropwise adding the acryloyl chloride into the system, reacting for 1.5-2.5h under the ice bath condition, then stirring overnight at room temperature, after complete reaction, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
The structural formula of the derivative of 4-hydroxymethyl-7-hydroxycoumarin:
R0is cyano. Correspondingly, the formula of the photoinitiator in this example is:
R1is cyano.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating under the illumination of 350nm wavelength for 100min to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is more than 0 and less than or equal to 1mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Example 5:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving a derivative of 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, then dropwise adding the acryloyl chloride into the system, reacting for 2 hours under the ice bath condition, then stirring overnight at room temperature, after the reaction is completed, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
The structural formula of the derivative of 4-hydroxymethyl-7-hydroxycoumarin:
R0is Br. Correspondingly, the formula of the photoinitiator in this example is:
R1is Br.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 150min under the illumination with the wavelength of 300nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is 0.15mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Example 6:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving a derivative of 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, then dropwise adding the acryloyl chloride into the system, reacting for 2 hours under the ice bath condition, then stirring overnight at room temperature, after the reaction is completed, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
The structural formula of the derivative of 4-hydroxymethyl-7-hydroxycoumarin:
R0is methyl. Correspondingly, the formula of the photoinitiator in this example is:
R1is methyl.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 90min under the illumination with the wavelength of 320nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is 0.2mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Example 7:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving a derivative of 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, then dropwise adding the acryloyl chloride into the system, reacting for 2 hours under the ice bath condition, then stirring overnight at room temperature, after the reaction is completed, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
The structural formula of the derivative of 4-hydroxymethyl-7-hydroxycoumarin:
R0is a hydroxyl group. Correspondingly, the formula of the photoinitiator in this example is:
R1is a hydroxyl group.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 30min under the illumination with the wavelength of 390nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is 0.25mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Example 8:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving a derivative of 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, then dropwise adding the acryloyl chloride into the system, reacting for 2 hours under the ice bath condition, then stirring overnight at room temperature, after the reaction is completed, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
The structural formula of the derivative of 4-hydroxymethyl-7-hydroxycoumarin:
R0is piperidinyl. Correspondingly, the formula of the photoinitiator in this example is:
R1is a piperidine group-containing group.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 70min under the illumination with the wavelength of 350nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is 0.18mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Example 9:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving a derivative of 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, then dropwise adding the acryloyl chloride into the system, reacting for 2 hours under the ice bath condition, then stirring overnight at room temperature, after the reaction is completed, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
The structural formula of the derivative of 4-hydroxymethyl-7-hydroxycoumarin:
R0is a phosphoric acid triester group. Correspondingly, the formula of the photoinitiator in this example is:
R1is a phosphoric acid triester group.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 90min under the illumination with the wavelength of 300nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is 0.2mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Example 10:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving a derivative of 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, then dropwise adding the acryloyl chloride into the system, reacting for 2 hours under the ice bath condition, then stirring overnight at room temperature, after the reaction is completed, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
The structural formula of the derivative of 4-hydroxymethyl-7-hydroxycoumarin:
R0is a trimethylsilyl group. Correspondingly, the formula of the photoinitiator in this example is:
R1is a trimethylsilyl group.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 60min under the illumination with the wavelength of 365nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is 0.25mmol/L, and the concentration b of the active monomer solution is 25 wt%.
Example 11:
the preparation method of the coumarin skeleton-based antibacterial hydrogel comprises the following specific steps:
(1) dissolving 4-hydroxymethyl-7-hydroxycoumarin in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, dropwise adding the acryloyl chloride into the system, reacting for 2 hours under the ice bath condition, stirring overnight at room temperature, after complete reaction, adjusting the pH value of the system to be neutral, drying, separating and purifying to obtain the photoinitiator containing the coumarin skeleton.
(2) Mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 60min under the illumination with the wavelength of 365nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is 0.25mmol/L, and the concentration b of the active monomer solution is 25 wt%.
The following will specifically explain the performance and application of the antibacterial hydrogel prepared by the present invention with reference to the test examples.
Test example 1: determination of optimal initiator concentration and optimal illumination conditions
1. The experimental method comprises the following steps:
preparing a solution of acrylamide with the mass concentration of 25 wt% by using deionized water, adding dimethyl sulfoxide mother liquor (the mother liquor concentration is 20mmol/L) of initiators with different volumes until the concentration of a final photoinitiator is 0, 0.0625, 0.125, 0.25, 0.5 and 1mmol/L, then uniformly mixing the photoinitiator solution and an active monomer acrylamide solution, vertically applying a light source with the wavelength of 365nm, and observing the gelling condition of the system, wherein the illumination time is 90 min. After the reaction is completed and gel is formed in part of the system, deionized water is added into the system, the gel is washed for multiple times to ensure that the residual monomers in the gel are completely washed out, then the washed monomers are lyophilized to prepare the concentration and the volume required by ultraviolet measurement, the residual amount of the monomers under different initiator equivalent weights is measured, and then the conversion rate of the monomers under different initiator concentrations is calculated according to the standard curve of acrylamide in the figure 1, so that the optimal initiator concentration is determined.
2. The experimental results are as follows:
as shown in fig. 2, the conversion of the reactive monomer-acrylamide was plotted against the concentration of the photoinitiator. As can be seen from FIG. 2, the conversion of the monomer is unfavorable when the initiator concentration is too large or too small, and the conversion can reach 95% or more when the initiator concentration is 0.125mmol/L or 0.25mmol/L, so that the concentration range of 0.125mmol/L to 0.25mmol/L is the optimum concentration range of the photoinitiator. Wherein, the optimal concentration of the photoinitiator is 0.125mmol/L, so that the invention can initiate the cross-linking reaction of the active monomer at a rather low concentration to form the hydrogel. It should be noted that the concentration range of the photoinitiator determined by the present invention is determined by the reaction concentration of the corresponding reactive monomer being 25 wt%, and when the concentration of the reactive monomer changes, the corresponding photoinitiator also changes, and the skilled person can adjust accordingly.
As shown in FIG. 3, after the initiator concentration is determined to be 0.125mmol/L, the illumination time is subsequently screened, the conversion efficiency of the acrylamide monomer under different illumination times is determined, and the monomer can be almost completely converted after the illumination time reaches 30min, the conversion rate is still kept at a higher level after the illumination time is continuously prolonged, which indicates that the photoinitiation rate of the polymerization reaction is fast. Therefore, the present invention determines 30min to 60min as the optimum illumination time period in consideration of the sufficient reaction progress and the time cost.
Test example 2: morphology study of antibacterial hydrogel
The hydrogel was prepared according to the experimental method in experimental example 1, and the prepared hydrogel was lyophilized, brittle-broken in liquid nitrogen, and SEM was used to observe the internal morphology of the gel.
As shown in fig. 4(a) and (b), an oriented network structure can be observed from the gel fracture, indicating that the photoinitiator gradually initiates the polymerization of the monomer. Meanwhile, the pore diameter of the hydrogel is measured to be about 500nm, which shows that the synthesized photoinitiator containing the coumarin skeleton successfully initiates the polymerization of the monomer to form the hydrogel with small pores. The photoinitiation reagent can efficiently initiate monomers to form hydrogel, and the hydrogel has good transmittance and compact pores and can be used for wrapping and releasing medicines, cells and the like.
Test example 3: antimicrobial hydrogel antimicrobial Performance testing
1. Biocompatibility investigation
The in vitro cytotoxicity test of the photoinitiators of the invention was determined by the MTS method. The specific process is as follows: HeLa cells were seeded into 96-well culture plates at 7000 cells/well in 37 ℃ with 5% CO2Is cultured in the incubator for 24h to a cell density of about 70-80%. Is replaced again100 μ L of DMEM medium containing FBS at different initiator concentrations was added. After culturing was continued in an incubator for 48 hours, the stock culture was removed, and 5mg mL of the stock culture was added to each 100. mu.L well-1The MTS solution of (2) was diluted with a PBS solution at a ratio of 20%. The incubation is carried out at 37 ℃ for 30 min. The absorbance I at 490nm was measured for each well using a microplate reader (Bio-Rad 680).
The relative survival rate of the cells is calculated by the formula:
cell viability (%) - (I sample/I control) × 100%,
wherein the measurement values of I are the values measured for 3 independent replicates of which the cell viability is expressed as mean ± Standard Deviation (SD).
The control group was a blank without addition of the photoinitiator solution of the invention, only HeLa cells.
As shown in fig. 5, the initiator was highly toxic to HeLa cells, and the IC50 value of the initiator was calculated to be 60 micrograms per ml.
2. Antibacterial experiments
As shown in FIG. 6, hydrogel coatings were prepared in LB-solid medium with the photoinitiators of the invention, while a blank set was provided (left medium in the figure). The specific process is as follows: firstly, selecting an LB solid culture medium as a blank control group to carry out escherichia coli culture; subsequently, the experimental group prepared the antibacterial hydrogel of the present invention on the surface of the LB solid medium under the illumination of 365nm wavelength; coli was inoculated on the experimental group and the blank group, respectively, and cultured under the same conditions, and growth of E.coli was observed on the blank medium and the gel surface. The culture conditions were 37 ℃ and 200r.m.p., 24 hours.
As shown in FIG. 7, it was clearly found that the blank group of E.coli was not restricted in growth, and that there was a large number of macroscopic E.coli colonies growing on the surface of the medium; when the initiator concentration was 0.125mmol/L, plaque formation was observed only on the surface of a very small area; when the initiator concentration is continuously increased to 0.5mmol/L, the formation of bacterial plaque is hardly observed on the surface of the gel coating, which shows that the gel formed by the photo-initiation has good antibacterial effect, and the antibacterial coating is more obvious when the initiator concentration is increased.
As shown in FIG. 8, from the SEM results, it can be seen that the upper surface of the LB-solid medium had the appearance of a torn chip (FIG. 8A) on which Escherichia coli had densely grown (FIG. 8D); in the experimental group, the surface of the LB-solid medium was covered with a gel layer, which is in the form of a porous structure of gel (FIG. 8B, FIG. 8C), the amount of E.coli grown was significantly less than in the blank group (FIG. 8E, FIG. 8F), and the deformation of E.coli was clearly observed in the embedded images in FIGS. 8E, 8F. The result shows that the photoinitiator prepared by the invention can successfully use light to initiate cross-linking polymerization, antibacterial gel is prepared on the surface of LB-solid culture medium, bacteria are respectively cultured in blank groups and experimental groups, the antibacterial property of the gel can be observed by naked eyes, and the result of SEM also confirms the point.
The coumarin skeleton-based photo-crosslinking initiator synthesized by the method can efficiently and quickly initiate active monomers to form hydrogel with high transparency and certain mechanical strength, effectively expands the application of a photoresponse coumarin micromolecule compound in the field of antibacterial materials, and provides a new idea for design synthesis and application expansion of a functional photo-crosslinking agent.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A preparation method of an antibacterial hydrogel based on a coumarin skeleton is characterized in that 4-hydroxymethyl-7-hydroxycoumarin or derivatives thereof and acryloyl chloride are used for synthesizing a photoinitiator containing the coumarin skeleton, and then the photoinitiator is used for initiating a crosslinking reaction of an active monomer under a lighting condition to prepare the antibacterial hydrogel.
2. The method of claim 1, wherein the derivative of 4-hydroxymethyl-7-hydroxycoumarin has the following formula:
wherein R is0Is one of cyano, halogen, alkyl and O, N, halogen, P, S or Si-containing alkyl.
3. The method of claim 2, wherein the photoinitiator has the formula:
wherein R is1Is one of hydrogen, cyano, halogen, alkyl and O, N, halogen, P, S or Si-containing alkyl.
4. The preparation method of claim 1, wherein the reaction concentration a of the photoinitiator is more than 0 and less than or equal to 1mmol/L, and the reaction concentration b of the active monomer is 20-30 wt%; the illumination conditions are as follows: irradiating for 10 min-24 h under the illumination with the wavelength of 200 nm-405 nm. .
5. The method of claim 3, wherein the reaction concentration a of the photoinitiator is 0.125 mmol/L.ltoreq.a.ltoreq.1 mmol/L.
6. The method according to claim 3, wherein the lighting conditions are: irradiating for 30-120 min under the illumination with the wavelength of 270-405 nm.
7. The method of claim 1, wherein the reactive monomer is one or more of acrylamide, acrylic acid, N-isopropylacrylamide, 2-hydroxyethyl acrylate, vinyl chloride, styrene, acrylonitrile, and alkyl acrylates.
8. The preparation method based on the coumarin skeleton according to any one of claims 1 to 7, characterized by comprising the following specific steps:
(1) dissolving 4-hydroxymethyl-7-hydroxycoumarin or a derivative thereof in an organic solvent under the conditions of no water and no oxygen and at the temperature of 0 ℃, then adding triethylamine, stirring uniformly at room temperature, dissolving acryloyl chloride in the organic solvent, then dropwise adding the acryloyl chloride into the system, reacting for 1.5-2.5h under the ice bath condition, then stirring for 8-15 h at room temperature, adjusting the pH value of the system to be neutral after the reaction is completed, drying, separating and purifying to obtain a photoinitiator containing a coumarin skeleton;
(2) mixing an active monomer with deionized water to obtain an active monomer solution, dissolving a photoinitiator in an organic solvent to obtain a photoinitiator solution, adding the photoinitiator solution into the active monomer solution, uniformly mixing, and radiating for 10min to 24h under illumination with the wavelength of 200nm to 405nm to obtain the antibacterial hydrogel; wherein the concentration a of the photoinitiator solution is more than 0 and less than or equal to 1mmol/L, and the concentration b of the active monomer solution is 25 wt%.
9. The coumarin skeleton-based antibacterial hydrogel prepared by the preparation method according to any one of claims 1 to 8.
10. Use of the coumarin scaffold-based antimicrobial hydrogel according to claim 9 in bacterial culture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010290097.XA CN111333774B (en) | 2020-04-14 | 2020-04-14 | Antibacterial hydrogel based on coumarin skeleton and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010290097.XA CN111333774B (en) | 2020-04-14 | 2020-04-14 | Antibacterial hydrogel based on coumarin skeleton and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111333774A true CN111333774A (en) | 2020-06-26 |
| CN111333774B CN111333774B (en) | 2020-12-08 |
Family
ID=71178935
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010290097.XA Active CN111333774B (en) | 2020-04-14 | 2020-04-14 | Antibacterial hydrogel based on coumarin skeleton and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111333774B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4015512A1 (en) | 2020-12-16 | 2022-06-22 | AMO Ireland | Optically active devices |
| CN114848653A (en) * | 2021-02-03 | 2022-08-05 | 四川大学 | Application of RORa protein and agonist thereof in preparation of acute DNA injury repair agent |
| CN116102671A (en) * | 2022-12-22 | 2023-05-12 | 浙江理工大学嵊州创新研究院有限公司 | Preparation method and antibacterial application of 7-hydroxy-4-methylcoumarin-based polymer |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090318582A1 (en) * | 2006-07-19 | 2009-12-24 | Basf Se A German Corporation | Method for Producing Water-Absorbing Polymer Particles by Polymerizing Droplets of a Monomer Solution |
| CN101914175A (en) * | 2010-08-05 | 2010-12-15 | 江南大学 | Method for preparing self-initiation ultraviolet-curing amphiphilic macromolecular resin |
| KR20120132145A (en) * | 2011-05-27 | 2012-12-05 | 강원대학교산학협력단 | Photo-responsive hydrogels and preparation method thereof |
| CN103113600A (en) * | 2012-10-27 | 2013-05-22 | 盐城工学院 | Photoresponsive reversible chitosan hydrogel and preparation method thereof |
| US20140054497A1 (en) * | 2011-04-20 | 2014-02-27 | Evonik Degussa Gmbh | Water-absorbing polymer having a high absorption rate |
| CN106083791B (en) * | 2016-06-08 | 2017-10-24 | 深圳市前海金卓生物技术有限公司 | A kind of coumarin derivative, its preparation method and by its obtained hydrogel |
| CN107987209A (en) * | 2018-01-08 | 2018-05-04 | 深圳市佶达德科技有限公司 | A kind of Coumarins polymer semiconductor laser material and its application |
| CN109970696A (en) * | 2019-04-11 | 2019-07-05 | 江南大学 | A kind of cumarin oxime ester lightlike initiating agent |
| CN110511400A (en) * | 2019-08-08 | 2019-11-29 | 天津大学 | Copolymerization object light reversible hydrogel and preparation method based on zwitterionic compound and coumarin derivative |
-
2020
- 2020-04-14 CN CN202010290097.XA patent/CN111333774B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090318582A1 (en) * | 2006-07-19 | 2009-12-24 | Basf Se A German Corporation | Method for Producing Water-Absorbing Polymer Particles by Polymerizing Droplets of a Monomer Solution |
| CN101914175A (en) * | 2010-08-05 | 2010-12-15 | 江南大学 | Method for preparing self-initiation ultraviolet-curing amphiphilic macromolecular resin |
| US20140054497A1 (en) * | 2011-04-20 | 2014-02-27 | Evonik Degussa Gmbh | Water-absorbing polymer having a high absorption rate |
| KR20120132145A (en) * | 2011-05-27 | 2012-12-05 | 강원대학교산학협력단 | Photo-responsive hydrogels and preparation method thereof |
| CN103113600A (en) * | 2012-10-27 | 2013-05-22 | 盐城工学院 | Photoresponsive reversible chitosan hydrogel and preparation method thereof |
| CN106083791B (en) * | 2016-06-08 | 2017-10-24 | 深圳市前海金卓生物技术有限公司 | A kind of coumarin derivative, its preparation method and by its obtained hydrogel |
| CN107987209A (en) * | 2018-01-08 | 2018-05-04 | 深圳市佶达德科技有限公司 | A kind of Coumarins polymer semiconductor laser material and its application |
| CN109970696A (en) * | 2019-04-11 | 2019-07-05 | 江南大学 | A kind of cumarin oxime ester lightlike initiating agent |
| CN110511400A (en) * | 2019-08-08 | 2019-11-29 | 天津大学 | Copolymerization object light reversible hydrogel and preparation method based on zwitterionic compound and coumarin derivative |
Non-Patent Citations (3)
| Title |
|---|
| CAROLINE DE GRACIA LUX，ET.AL: "Short Soluble Coumarin Crosslinkers for Light-Controlled Release ofCells and Proteins from Hydrogels"", 《BIOMACROMOLECULES》 * |
| S.VENKATESAN, ET.AL: "Synthesis, characterization, thermal stability and antibacterial activity of coumarin based methacrylate copolymers:", 《CHINESE JOURNAL OF POLYMER SCIENCE》 * |
| 李梦琦等: ""香豆素肟酯类双光子引发剂的合成及性能研究"", 《影响科学与光化学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP4015512A1 (en) | 2020-12-16 | 2022-06-22 | AMO Ireland | Optically active devices |
| WO2022128832A1 (en) | 2020-12-16 | 2022-06-23 | Merck Patent Gmbh | Optically active devices |
| CN114848653A (en) * | 2021-02-03 | 2022-08-05 | 四川大学 | Application of RORa protein and agonist thereof in preparation of acute DNA injury repair agent |
| CN114848653B (en) * | 2021-02-03 | 2023-05-30 | 四川大学 | Application of RORa protein and agonist thereof in preparation of acute DNA damage repair agent |
| CN116102671A (en) * | 2022-12-22 | 2023-05-12 | 浙江理工大学嵊州创新研究院有限公司 | Preparation method and antibacterial application of 7-hydroxy-4-methylcoumarin-based polymer |
| CN116102671B (en) * | 2022-12-22 | 2024-03-29 | 浙江理工大学嵊州创新研究院有限公司 | Preparation method and antibacterial application of 7-hydroxy-4-methylcoumarin-based polymer |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111333774B (en) | 2020-12-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111333774B (en) | Antibacterial hydrogel based on coumarin skeleton and preparation method and application thereof | |
| US20130142763A1 (en) | Crosslinked cellulosic polymers | |
| US4279795A (en) | Hydrophilic-hydrophobic graft copolymers for self-reinforcing hydrogels | |
| Han et al. | Photopolymerization of methacrylated chitosan/PNIPAAm hybrid dual-sensitive hydrogels as carrier for drug delivery | |
| US20040266992A1 (en) | Method for the preparation of silk fibron hydrogels | |
| CN110804193B (en) | 3D printing method of hydrogel support | |
| CN104356319A (en) | Porous biological material using modified gelatin as crosslinking agent and preparation method of porous biological material | |
| CN112375191B (en) | Block copolymer, preparation method and application thereof | |
| CN109232885B (en) | Modified dendritic PAMAM polymer and preparation method and application thereof | |
| CN111214695A (en) | Novel 3D structure biological high molecular material prepared by covalent reaction and synthetic method thereof | |
| CN108641092B (en) | Preparation method of supramolecular polymer composite micelle based on hydrogen bond | |
| Ahmadi et al. | Photo-and thermo-responsive extracellular matrix mimicking nano-coatings prepared from poly (N-isopropylacrylamide)-spiropyran copolymer for effective cell sheet harvesting | |
| CN112543791B (en) | Polymers based on difunctional modified biopolymers and hydrogels obtainable from such polymers based on difunctional modified biopolymers | |
| CN115636884B (en) | Preparation method of sodium hyaluronate derivative, cross-linked sodium hyaluronate and application | |
| CN109810265B (en) | DNA-polysaccharide hybrid hydrogel with volume change driven by solvent and preparation method thereof | |
| CN115260337A (en) | Arginine grafted carboxylated pullulan polysaccharide and preparation method and application thereof | |
| CN110105508B (en) | Preparation method of aqueous photoresponse nanogel, prepared nanogel and application of nanogel | |
| KR20230081755A (en) | Preparation of non-covalent interpenetrating network hydrogel and method for manufacturing product of it | |
| CN113754564A (en) | Antibacterial amidine oligomer with anti-drug resistance and preparation method and application thereof | |
| CN112592446A (en) | Three-dimensional macroporous frozen gel scaffold and preparation method and application thereof | |
| Bonartsev et al. | BSA adsorption on porous scaffolds prepared from bioPEGylated poly (3-hydroxybutyrate) | |
| JP6718610B2 (en) | Cell culture substrate, method for producing the same, and method for controlling physical properties of cell culture substrate | |
| CN115501378B (en) | Modified polyvinyl alcohol embolism microsphere and preparation method thereof | |
| CN113527602B (en) | Photo-responsive block polymer synthesized by ATRP method and preparation method and application thereof | |
| KR20240030783A (en) | Method for producing nanofibrils bound to hyaluronic acid and nanofibrils produced thereby |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |