Chiral multilevel structure antibacterial film as well as preparation method and application thereof

Abstract

The invention belongs to the field of biomedical materials, and provides a chiral multilevel structure antibacterial film and a preparation method and application thereof, and in the preparation process of the film, atomic-level nanosheets are assembled to form a microenvironment with a chiral multilevel structure. The adhesion behavior of bacteria can be selectively regulated and controlled through a chiral multilevel structure, and the growth of a bacterial biofilm is controlled, so that an antibacterial effect is generated. Besides, the nanosheet plays a role of a blade array and also provides a physical tool for destroying a bacterial membrane, the D-CHAPFs with the blade array structure has the effect of inhibiting the growth of the bacterial biomembrane, and the inhibition rate is 90-96%. The antibacterial film is beneficial to repair of infected wounds while resisting bacteria. Based on the above points, construction of a chiral multilevel structure microenvironment, selective regulation and control of bacterial adhesion behaviors and regulation and damage of bacterial membrane growth are realized, and an antibacterial function is realized on the whole.

Description

Technical Field

[0001] The invention belongs to the field of biomedical materials, and in particular relates to a chiral multi-level structure antibacterial film and a preparation method and application thereof. Background Art

[0002] The "Global Antibiotic Resistance Review" report published by the UK Antimicrobial Evaluation Committee in 2016 shows that about 700,000 people die from drug-resistant bacterial infections every year, and the number will gradually increase over time. China is a major antibiotic user, accounting for about half of the world's total. If no monitoring measures are taken, according to the current development trend, it is estimated that by 2050, 1 million people will die from drug-resistant bacterial infections every year, causing a cumulative economic loss of 20 trillion US dollars (J. O'Neill. Nature Reviews Drug Discovery. 2016, 15: 526.). Therefore, with the continuous development of science and technology and economy, people's requirements for hygiene and health in daily life and work are constantly increasing, and antibacterial awareness is also constantly increasing.

[0003] Antibacterial can be divided into two types: antibacterial adhesion and bactericidal. Antibacterial adhesion refers to the process of reducing the initial bacterial adhesion by modifying the surface energy, thereby interfering with the formation of bacterial biofilm. However, due to the lack of effective means to kill bacteria, the antibacterial effect of antibacterial adhesion surfaces cannot achieve long-term effective antibacterial effects. Once a small amount of bacteria can contact this surface, bacterial adhesion, proliferation and biofilm formation will develop rapidly, and it is inevitable that bacteria will be infected again. Sterilization, on the other hand, effectively kills bacteria adhering to the surface of the material through the structure of the material surface or antibacterial agents. For sterilization surfaces that rely on chemical bactericides, the short-term effectiveness of the bactericidal components and the accompanying toxicity to normal human cells are two major problems that need to be solved (TSUI KH, LI X, TSOI JKH, et al. Low-cost, flexible, disinfectant-free and regular-array three-dimensional nanopyramid antibacterial films for clinical applications [J]. Nanoscale, 2018, 10 (22): 10436-10442; WEI T, TANG Z, YU Q, et al. Smart Antibacterial surfaces with switchable bacteria-killing and bacteria-releasing capabilities[J].ACS Applied Materials&Interfaces,2017,9(43):37511-37523.). Therefore, antibacterial materials can be prepared by using antibacterial agents or constructing special structures on the surface of materials. Generally, there are two methods for preparing antibacterial materials: one is to chemically modify the surface by polymer polymerization, surface functionalization and derivatization, coating with bactericidal coatings (antibiotics, metal nanoparticles), etc.; the other is to change the surface morphology by physical methods to achieve antibacterial effect.The antibacterial materials prepared by these two methods have certain bactericidal properties, but chemical antibacterial has significant disadvantages. The development of antibiotics is more difficult and the use of antibiotics will cause bacteria to develop drug resistance. Generally, after a new antibiotic is launched on the market, new drug-resistant bacteria will appear in less than two years, and the bacteria will be stimulated to become more tenacious, making treatment more difficult (ED Brown, GD Wright. Antibacterial drug discovery in the resistance era. Nature. 2016, 529: 336-343.); Physical antibacterial has good physicochemical stability and is not easy to develop drug resistance, but the antibacterial efficiency of physical antibacterial and the types of new antibacterial materials need to be improved.

[0004] Researchers have found that after millions of years of evolution, some animals and plants in nature have some special micro-nano structures on their surfaces, which give them antibacterial functions. For example, there are a large number of micropores on the surface of kapok fibers (Sun Jiaming. Preparation and structural properties of kapok cellulose aerogel-based functional composite materials [D]. Northeast Forestry University, 2024.), ramie fibers have groove-like cavities and porous tube walls. The microporous structures of these natural antibacterial fibers can only kill anaerobic bacteria by forming a polyoxic environment; Hasan et al. first discovered in 2013 that the nanocolumn arrays (diameter: 60nm, height: 200nm) on the wing surface of Clanger cicada can kill bacteria adhering to its surface by contact, and its antibacterial spectrum is broad and can kill a variety of Gram-negative bacteria (Hasan J., Webb HK, Truong VK, Pogodin S., Baulin VA, Watson G.S., Watson JA, Crawford RJ, Ivanova EP Selective Bactericidal Activity ofNanopatterned Superhydrophobic Cicada Psaltoda Claripennis Wing Surfaces[J].Appl.Microbiol.Biotechnol.,2013,97(20):9257-9262.); and the micro-nanostructure of dragonfly wings can destroy the integrity of cell membranes through physical contact to achieve antibacterial effects (Nowlin K., Boseman A., Covell A., LaJeunesse D. Adhesion-Dependent Rupturing of Saccharomyces cerevisiae on Biological Antimicrobial Nanostructured Surfaces[J]. JR Soc. Interface,2015,12(102):20140999.). Inspired by this, researchers have launched antibacterial research focusing on bionic micro-nanostructures.

[0005] Chirality is a basic property of nature. It is ubiquitous in organisms and closely related to life activities, such as various helical structures of DNA, proteins, cells and tissues. It constitutes an important feature of the body's organs, tissues and molecular configurations, and usually exists only in a mirror image form. Multi-level structure is a common phenomenon in nature. Because materials have multi-level organizational structures, they give materials unexpected properties. Living organisms themselves are chiral multi-level structures that have specific selectivity for chiral environments. Natural bone is a chiral multi-level structure organization. For example, chiral drugs play different roles in living organisms, and the chirality of drugs is bound to affect the performance of their pharmacological effects. Antimicrobial peptides have chiral structures. Due to the chiral selectivity and chiral recognition ability of organisms, antimicrobial peptides exhibit excellent antibacterial properties. Based on the excellent antibacterial properties and biocompatibility of antimicrobial peptides, the use of antimicrobial peptides to modify the surface of materials can generate a new type of non-toxic, pollution-free, green and environmentally friendly material. However, the research on the differential biological effects caused by chiral nanomaterials is still in its infancy, and the therapeutic strategies based on chiral nanomaterials have received little attention in microbial infections so far. Compared with antibiotics and small molecule bactericides, metal nanomaterials modified with surfactants or organic molecules have good broad-spectrum bactericidal properties, but they can lead to higher cytotoxicity and cause irreversible damage to normal tissues and organs; while amino acids with chirality and reducing ability can be used as environmentally friendly reducing agents and capping ligands (Wang HX, Qian J, Gu JY, et al. Steric Configuration-enabled selective antimicrobial activity of chiral cysteine[J] Biochemical and Biophysical Research Communications. 2019, 512(3): 505-510.).

[0006] Current research in the field of physical sterilization mainly focuses on the bactericidal properties of bactericidal nanoarrays and their derivative structures. However, with increasingly stringent requirements for the prevention and control of bacterial contamination, it is urgent to further improve the bactericidal efficiency of physical sterilization surfaces and provide a physical antibacterial material with multi-scenario indications. Summary of the invention

[0007] Based on the discovery that the nano-blade array can destroy the bacterial structure and make the chiral multi-level structure have physical antibacterial properties, a chiral multi-level structure antibacterial material is proposed, and the present invention is completed based on this.

[0008] In a first aspect, a chiral multi-level structure antibacterial material is provided, wherein the chiral multi-level structure antibacterial material is composed of a first component, a second component, a chiral inducer, a substrate and / or a nucleation controller to form a microenvironment with a chiral multi-level structure: wherein the first component is a soluble metal source selected from one or more of a soluble calcium source, a soluble gallium source, a soluble molybdenum source, a soluble iron source, a soluble barium source, a soluble bismuth source, a soluble copper source, a soluble strontium source, a soluble titanium source, a soluble silver source and / or a soluble magnesium source; the second component is selected from one or more of a soluble phosphorus source, a soluble sulfur source, a soluble carbon source, a soluble titanium source and / or a soluble bromine source.

[0009] Furthermore, the soluble calcium source is selected from one or more of anhydrous calcium chloride, calcium nitrate tetrahydrate, calcium sulfate dihydrate and calcium oxalate.

[0010] Further, the soluble gallium source is selected from one or more of gallium chloride, gallium bromide, gallium iodide, gallium fluoride, gallium nitride, gallium sulfate, gallium phosphide, gallium oxide and / or gallium selenide.

[0011] Further, the soluble molybdenum source is selected from one or more of ammonium molybdate, sodium molybdate, magnesium molybdate, zinc molybdate, ammonium dimolybdate, iron molybdate, lead molybdate, copper molybdate, sodium paramolybdate, ammonium paramolybdate, ammonium tetramolybdate and / or ammonium dimolybdate.

[0012] Further, the soluble iron source is selected from one or more of ferrous sulfate heptahydrate, ferric chloride, ferric nitrate, ferric oxalate and / or ferric phosphate.

[0013] Furthermore, the soluble barium source is selected from one or more of barium sulfate, barium chloride, barium nitrate, barium carbonate, barium oxalate, barium acetate, barium phosphate, barium tungstate and / or barium titanate.

[0014] Further, the soluble bismuth source is selected from one or more of bismuth nitrate pentahydrate, bismuth sulfate, bismuth chloride, bismuth oxalate and / or bismuth acetate.

[0015] Further, the soluble copper source is selected from one or more of copper chloride, copper sulfate, copper nitrate, copper oxalate, copper tungstate, copper acetate and / or copper phosphate.

[0016] Furthermore, the soluble strontium source is selected from one or more of strontium nitrate, strontium chloride, strontium oxalate, strontium titanate, strontium sulfate and / or strontium carbonate.

[0017] Further, the soluble titanium source is selected from one or more of tetra-n-butyl titanate, titanium acetylacetonate, tetraisopropyl titanate, titanium chloride, titanium sulfate, sodium carbonate, potassium titanate and / or ammonium carbonate.

[0018] Further, the soluble silver source is selected from one or more of silver nitrate, silver chloride, silver sulfate, silver oxide, silver bromide, silver iodide, acetate, oxalate and / or silver citrate.

[0019] Further, the soluble magnesium source is selected from one or more of magnesium chloride, magnesium sulfate, magnesium nitrate, magnesium oxalate, magnesium carbonate, magnesium citrate, magnesium lactate, magnesium phosphate and / or magnesium vanadate.

[0020] Further, the soluble phosphorus source is selected from one or more of diammonium hydrogen phosphate, dipotassium hydrogen phosphate and / or disodium hydrogen phosphate.

[0021] Further, the soluble sulfur source is selected from one or more of thioacetamide, sodium sulfide, sulfuric acid, sodium sulfate, potassium sulfate, ammonium sulfate, thiourea, mercaptan, thioether and / or sulfate ester.

[0022] Further, the soluble carbon source is selected from one or more of ammonium carbonate, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate and / or ammonium bicarbonate.

[0023] Further, the soluble bromine source is selected from one or more of sodium bromide, potassium bromide, hydrogen bromide, ammonium bromide, calcium bromide, zinc bromide, copper bromide and / or ferrous bromide.

[0024] Furthermore, the chiral inducing agent is one or more of chiral amino acids, chiral organic acids, chiral sugars and chiral alcohols.

[0025] Furthermore, the chiral amino acid is selected from one or more of histidine, arginine, lysine, isoleucine, phenylalanine, leucine, tryptophan, alanine, methionine, proline, cysteine, aspartic acid, valine, serine, glutamine, tyrosine, aspartic acid, glutamic acid and threonine; the chiral organic acid is selected from one or more of tartaric acid, malic acid and lactic acid; the chiral sugar is selected from glucose, fructose, galactose, ribose, deoxyribose, furanose, pyranose, maltose, sucrose and lactose; the chiral alcohol includes one or more of mannitol, xylitol, sorbitol, paclitaxel, resveratrol, ginkgo biloba polyprenols, (R)-(+)-1-phenyl-1-propanol, (S)-(-)-1-phenyl-1-propanol and / or phenylpropanol.

[0026] Furthermore, the nucleation control agent is selected from one or more of urea, sodium dodecylbenzene sulfonate, sodium dodecyl sulfate and / or sodium dodecylbenzene sulfonate.

[0027] Furthermore, the support substrate is selected from any one of activated mica sheets, iron sheets, titanium sheets, titanium alloy sheets, magnesium alloy sheets, glass sheets, quartz sheets, FTO sheets, ITO sheets, silicon sheets, copper sheets or nickel sheets, and the thickness of the support substrate is 0.1 to 1 mm.

[0028] Furthermore, the chiral multilevel structure antibacterial material includes a chiral multilevel structure hydroxyapatite antibacterial material, a chiral multilevel structure gallium phosphate antibacterial material, a chiral multilevel structure zinc oxide antibacterial material, a chiral multilevel structure molybdenum disulfide antibacterial material, a chiral multilevel structure iron sulfide antibacterial material, a chiral multilevel structure calcium carbonate antibacterial material, a chiral multilevel structure barium titanate antibacterial material, a chiral multilevel structure titanium dioxide antibacterial material, a chiral multilevel structure bismuth oxygen bromide antibacterial material, a chiral multilevel structure copper oxide antibacterial material, a chiral multilevel structure metallic silver antibacterial material, a chiral multilevel structure copper sulfide antibacterial material, a chiral multilevel structure strontium titanate antibacterial material, a chiral multilevel structure magnesium oxide antibacterial material and a chiral multilevel structure calcium vanadate antibacterial material, etc.

[0029] Furthermore, the chiral multi-level antibacterial material multi-level structure includes D-CHAPFs type, L-CHAPFs type and Racemic type.

[0030] Furthermore, in the crystal of the chiral multi-level antibacterial material, the atomic arrangement is in a spirally twisted state.

[0031] Furthermore, the chiral multi-level antibacterial material is formed by the assembly of atomic-level spirally twisted nanosheets in terms of microstructure.

[0032] Furthermore, the nanosheets act as an array of blades, destroying the structure of bacteria when they adhere.

[0033] Furthermore, the chiral multi-level structure antibacterial material is a chiral multi-level structure antibacterial film.

[0034] In a second aspect, a method for preparing the chiral multi-level antibacterial material according to the first aspect is provided, the method comprising the following steps:

[0035] S1, dissolving the first component in water to form solution A;

[0036] S2, dissolving the chiral inducing agent in water to form solution B;

[0037] S3, dissolving the second component and the nucleation control agent in water to form a solution C;

[0038] S4, rapidly adding solution B into solution A to form a mixed solution, and continuing stirring for 10-60 minutes to obtain an AB mixed solution;

[0039] S5, slowly adding solution C to the mixed solution AB to form a mixed reaction solution, and continuing to stir for 2-30 minutes;

[0040] S6, transferring the mixed reaction liquid into a reaction kettle, adding the pretreated substrate, reacting at 100-200° C. for 2-24 hours, cooling, taking out the substrate, washing and drying, and obtaining a chiral multi-level structure antibacterial material.

[0041] Further, in step S1, the soluble metal source is selected from one or more of a soluble calcium source, a soluble gallium source, a soluble molybdenum source, a soluble iron source, a soluble barium source, a soluble bismuth source, a soluble copper source, a soluble strontium source and / or a soluble titanium source;

[0042] Furthermore, the soluble calcium source is selected from one or more of anhydrous calcium chloride, calcium nitrate tetrahydrate, calcium sulfate dihydrate and / or calcium oxalate; the soluble phosphorus source is selected from one or more of diammonium hydrogen phosphate, dipotassium hydrogen phosphate and / or disodium hydrogen phosphate; the soluble gallium source is selected from one or more of gallium chloride, gallium bromide, gallium iodide, gallium fluoride, gallium nitride, gallium sulfate, gallium phosphide, gallium oxide and / or gallium selenide; the soluble molybdenum source is selected from one or more of ammonium molybdate, sodium molybdate, magnesium molybdate, zinc molybdate, ammonium dimolybdate, iron molybdate, lead molybdate, copper molybdate, sodium paramolybdate, ammonium paramolybdate, ammonium tetramolybdate and / or ammonium dimolybdate; the soluble iron source is selected from one or more of ferrous sulfate heptahydrate, ferric chloride, ferric nitrate, ferric oxalate and / or ferric phosphate; the soluble barium source is selected from one or more of barium sulfate, barium chloride, barium nitrate, barium carbonate, barium oxalate, barium acetate, barium phosphate, barium tungstate and / or barium titanate. The invention relates to a plurality of materials. The invention relates to a plurality of materials. The invention relates to a plurality of materials. The invention relates to a plurality of materials. The invention relates to a plurality of materials. The invention relates to a plurality of materials. The invention relates to a plurality of materials. The invention relates to a plurality of materials. The invention relates to a plurality of materials. The invention relates to a plurality of materials. The invention relates to a plurality of materials.

[0043] Furthermore, the chiral inducing agent in step S2 is one or more of chiral amino acids, chiral organic acids, chiral sugars and chiral alcohols;

[0044] Furthermore, the chiral amino acid is selected from one or more of histidine, arginine, lysine, isoleucine, phenylalanine, leucine, tryptophan, alanine, methionine, proline, cysteine, aspartic acid, valine, serine, glutamine, tyrosine, aspartic acid, glutamic acid and / or threonine; the chiral organic acid is selected from one or more of tartaric acid, malic acid and / or lactic acid; the chiral sugar is selected from glucose, fructose, galactose, ribose, deoxyribose, furanose, pyranose, maltose, sucrose, lactose; the chiral alcohol includes one or more of mannitol, xylitol, sorbitol, paclitaxel, resveratrol, ginkgo biloba polyprenols, (R)-(+)-1-phenyl-1-propanol, (S)-(-)-1-phenyl-1-propanol and / or phenylpropanol.

[0045] Further, in step S3, the second component is selected from one or more of a soluble phosphorus source, a soluble sulfur source, a soluble carbon source, a soluble titanium source and / or a soluble bromine source;

[0046] Furthermore, the soluble phosphorus source is selected from one or more of diammonium hydrogen phosphate, dipotassium hydrogen phosphate and / or disodium hydrogen phosphate; the soluble sulfur source is selected from one or more of thioacetamide, sodium sulfide, sulfuric acid, sodium sulfate, potassium sulfate, ammonium sulfate, thiourea, thiol, thioether and / or sulfate ester; the soluble carbon source is selected from one or more of sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate and / or ammonium bicarbonate; the soluble bromine source is selected from one or more of potassium bromide, hydrogen bromide, ammonium bromide, calcium bromide, zinc bromide, copper bromide and / or ferrous bromide; the soluble titanium source is selected from one or more of tetra-n-butyl titanate, titanium acetylacetonate, tetraisopropyl titanate, titanium chloride, titanium sulfate, sodium carbonate, potassium titanate and / or ammonium carbonate.

[0047] Furthermore, in step S3, the nucleation control agent is selected from one or more of urea, sodium dodecylbenzene sulfonate, sodium dodecyl sulfate and / or sodium dodecylbenzene sulfonate.

[0048] Furthermore, in step S4, the supporting substrate is selected from any one of activated mica sheets, iron sheets, titanium sheets, titanium alloy sheets, magnesium alloy sheets, glass sheets, quartz sheets, FTO sheets, ITO sheets, silicon sheets, copper sheets or nickel sheets, and the thickness of the supporting substrate is 0.1 to 1 mm.

[0049] In a third aspect, a chiral multi-level structure antibacterial film is provided for use in the preparation of a physical antibacterial material, wherein the antibacterial film has the functions of reducing bacterial adhesion, destroying bacterial structure and facilitating wound repair.

[0050] Furthermore, the application scenarios of the physical antibacterial material are any surface or object that needs to be antibacterial.

[0051] Furthermore, the chiral multi-level antibacterial film reduces bacterial adhesion and prevents bacterial adhesion by selectively regulating and influencing the bacterial recognition material surface process.

[0052] Furthermore, the chiral multi-level antibacterial film can selectively regulate the adhesion behavior of bacteria and control the growth of bacterial biofilm, thereby producing an antibacterial effect.

[0053] Furthermore, the chiral multi-level antibacterial film is assembled from atomic-level nanosheets.

[0054] Furthermore, the nanosheets act as a blade array, providing a physical tool for destroying bacterial membranes, achieving an antibacterial effect by destroying the structure of adhered bacteria.

[0055] Furthermore, in the antibacterial application, the inhibited bacteria include autotrophic bacteria and heterotrophic bacteria.

[0056] Furthermore, the inhibited bacteria include Gram-negative bacteria and Gram-positive bacteria.

[0057] Furthermore, the inhibited bacterial forms include cocci, bacilli and spirochetes.

[0058] Furthermore, the inhibited bacteria include Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter, Mycobacterium tuberculosis, Helicobacter pylori, Cholera, Candida albicans, Streptococcus mutans, Actinomycetes, Pseudomonas aeruginosa, Salmonella typhi, Mycobacterium and Bacillus subtilis.

[0059] Beneficial Effects

[0060] The chiral multi-level antibacterial film of the present invention is prepared by hydrothermal reaction. The antibacterial film prepared by this method has the following advantages: 1. The size of the obtained product is controllable, the surface morphology of the antibacterial film is uniform, and it is also easy to scale up and apply to large-scale production;

[0061] 2. During the preparation process, the antibacterial film forms a microenvironment with a chiral multi-level structure through the assembly of nanosheets from the atomic level to the micron level;

[0062] 3. The chiral multi-level structure can selectively regulate the adhesion behavior of bacteria and control the growth of bacterial biofilm, thereby producing an antibacterial effect;

[0063] 4. The nanosheets act as a blade array and provide a physical tool for destroying bacterial membranes. D-CHAPFs with a blade array structure have the effect of inhibiting the growth of bacterial biofilms, with an inhibition rate of 90-96%;

[0064] 5. While being antibacterial, it also helps repair infected wounds. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] Figure 1 A flowchart of a chiral multi-level antibacterial film and a preparation method and application thereof according to an embodiment of the present invention.

[0066] Figure 2 Physical picture of the L-chiral multi-level structure hydroxyapatite antibacterial film and mica substrate of Example 1 of the present invention.

[0067] Figure 3 Scanning electron microscope image of the L-chiral multi-level hydroxyapatite antibacterial film of Example 1 of the present invention.

[0068] Figure 4 XRD spectra of the L-chiral multi-level hydroxyapatite antibacterial film and the corresponding powder of Example 1 of the present invention.

[0069] Figure 5 Circular dichroism spectra of the chiral multi-level structured hydroxyapatite antibacterial film (L-, D- and R-chiral film) of Example 1 of the present invention.

[0070] Figure 6 TEM and corresponding electron diffraction pattern of the L-chiral multi-level hydroxyapatite antibacterial film of Example 1 of the present invention.

[0071] Figure 7 The three-dimensional electron diffraction spectrum of the L-chiral multi-level hydroxyapatite antibacterial film of Example 1 of the present invention.

[0072] Figure 8 Microscopic picture of bacteria growing on the surface of the chiral multi-level hydroxyapatite antibacterial film of Example 1 of the present invention at low concentration.

[0073] Fig. 9 Microscopic picture of bacteria growing on the surface of the chiral multi-level hydroxyapatite antibacterial film of Example 1 of the present invention at high concentration.

[0074] Fig.10 Crystal violet staining of the chiral multi-level hydroxyapatite antibacterial film after bacteria were planted on the surface of Example 1 of the present invention.

[0075] Fig.11 Crystal violet staining OD495 statistical graph after bacteria were planted on the surface of the chiral multi-level hydroxyapatite antibacterial film of Example 1 of the present invention.

[0076] Fig.12 Live-dead staining image of the chiral multi-level hydroxyapatite antibacterial film after bacteria were planted on the surface of the antibacterial film according to Example 1 of the present invention.

[0077] Fig.13 The antibacterial effect diagram of the chiral multi-level hydroxyapatite antibacterial film and bacteria buried under the skin in Example 1 of the present invention.

[0078] Fig.14Comparison of antibacterial effects of chiral multi-level hydroxyapatite antibacterial films.

[0079] Fig.15 The effect diagram of bacterial biofilm formation when the chiral multi-level hydroxyapatite antibacterial film of Example 1 of the present invention and bacteria are buried subcutaneously for 7 days and 15 days.

[0080] Fig.16 Scanning electron microscopy image of the chiral multi-level hydroxyapatite antibacterial film of Example 1 of the present invention after being buried subcutaneously with bacteria for 7 days.

[0081] Fig.17 Statistical chart of logCFU / ml in vivo antibacterial effect of Mica and chiral multi-level hydroxyapatite antibacterial films.

[0082] Fig.18 Antibacterial effect of Escherichia coli on the surface of chiral multi-level structured molybdenum disulfide antibacterial film.

[0083] Fig.19 Antibacterial effect of chiral multi-level structured molybdenum disulfide antibacterial film on Staphylococcus aureus.

[0084] Fig. 20 Comparison of antibacterial effects of chiral multi-level molybdenum disulfide antibacterial films. DETAILED DESCRIPTION

[0085] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in the embodiments described below can be combined with each other as long as they do not conflict with each other.

[0086] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0087] the term

[0088] Chirality: refers to the geometric property that a structure cannot be superimposed on its mirror image through translation and rotation because of its lack of Sn symmetry.

[0089] Hierarchical Structure: It is a structure that is ubiquitous in nature, referring to a structure that is superimposed step by step in scale or dimension from the atomic level to the macroscopic level.

[0090] Chiral Meso-structure: refers to a multi-level structure in which the assembly units of a multi-level structure material are arranged and assembled in a chiral manner during its formation.

[0091] Asymmetric Synthesis: also known as chiral synthesis, stereoselective synthesis, and enantioselective synthesis, is a branch of organic synthesis that studies chemical reactions that introduce one or more chiral elements into reactants (IUPAC (International Union of Pure and Applied Chemistry) Gold Book).

[0092] Chiral Auxiliary Method: Also known as chiral auxiliary induction, this method is the second generation of chiral synthesis method, also known as auxiliary group control method. It refers to the introduction of a chiral auxiliary at a certain stage of the synthesis to form a local induction and produce a new chiral center (developed by the Rhine-Westphalia Institute in Germany, called SAMP, RAMP).

[0093] Antibacterial materials: refers to a new type of functional materials that have the ability to kill or inhibit microorganisms. They have extremely broad application prospects in the medical field, household goods, household appliances, food packaging and other fields.

[0094] Basal membrane (BM): The extracellular matrix widely distributed in the body, found at the bottom of squamous epithelial tissue or glandular epithelium, connected to the dermis or connective tissue, found at the bottom of vascular endothelial cells connected to the vascular wall components, and also found at the bottom of mesothelial cells or synovial cells connected to the surrounding connective tissue. In addition, BM also surrounds nerve sheath cells, smooth muscle cells, striated muscle cells and fat. The composition and function of BM are very complex, and it is the physical support of some cells.

[0095] Nucleating agent: a polymer modification additive that adds crystalline substances to the polymer melt to nucleate the melt at a higher temperature, increase the crystallization rate, and make the polymer crystallize at high temperature, so that it is easy to solidify and demold, thereby shortening the processing cycle of the product. Nucleating agents can further refine the crystal structure of crystalline polypropylene, that is, increase the crystallinity, reduce the diameter of spherulites and control a certain morphology.

[0096] Chemical antibacterial: can be divided into photocatalytic antibacterial and metal ion dissolution antibacterial. Photocatalytic antibacterial means that under the catalytic effect of light, the antibacterial material produces active oxygen or active holes to inhibit or even kill the growth and reproduction of bacteria. Dissolution of metal ion antibacterial means that the antibacterial material dissolves metal ions to produce toxicity while maintaining a high bactericidal concentration in the surrounding environment to destroy the metabolic activities of bacteria.

[0097] Physical antibacterial effect: It utilizes the structure of antibacterial materials to achieve antibacterial effect by restricting the growth of bacteria and destroying the integrity of bacterial cell membranes.

[0098] Antimicrobial peptides (AMPs): They are an important component of the natural immune system of biological organisms and are widely present in nature. Their molecules are usually composed of less than 100 amino acids and have multiple biological activities such as antibacterial and antifungal.

[0099] Example

[0100] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0101] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments may be combined with each other.

[0102] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but they are not intended to limit the present invention.

[0103] Example 1 Preparation and morphology of chiral multi-level hydroxyapatite antibacterial film

[0104] 1.1 Test methods

[0105] Step S1, dissolving a soluble calcium source, calcium nitrate tetrahydrate (Ca(NO3)2·4H2O) in water, and stirring the solution to form a solution A, wherein the content of calcium nitrate tetrahydrate in the solution A is 1.25 mmol;

[0106] Step S2, dissolving a chiral inducing agent, tartaric acid (C4H6O6), in water, and stirring thoroughly to dissolve to form a solution B, wherein the content of tartaric acid in the solution B is 0.625 mmol;

[0107] Step S3, dissolving a soluble phosphorus source diammonium hydrogen phosphate ((NH4)2HPO4) and a nucleation control agent urea (carbonamide: CH4N2O) in water, and stirring and dissolving to form a solution C, wherein the contents of diammonium hydrogen phosphate and urea in the solution C are 0.75 mmol and 1.67 mmol, respectively;

[0108] Step S4, rapidly adding solution B into solution A to form a mixed solution, stirring to obtain an AB mixed solution;

[0109] Step S5, slowly adding solution C to the mixed solution AB while vigorously stirring to form a mixed reaction solution, and then continuing to stir;

[0110] Step S6, transfer the mixed reaction liquid into a reaction kettle, add the pretreated substrate, react at 180°C for 24 hours, cool naturally, take out the substrate, wash it alternately with deionized water and ethanol, and dry it to obtain a chiral mesostructured hydroxyapatite antibacterial film. The substrate used in this embodiment is a mica substrate, which is pretreated in advance to achieve activation, wherein the pretreatment operation is: tear and peel the mica with transparent glue to obtain a fresh exposed surface.

[0111] In the above process, the added amount ratio of each raw material is: the molar ratio of soluble calcium source, soluble phosphorus source, chiral inducer, nucleation control agent and water is 1:0.6:0.5:1.336:1667.

[0112] 1.2 Chiral chiral multi-level structure hydroxyapatite antibacterial film

[0113] In step S2, D-type, L-type and Racemic (i.e., racemic, hereinafter abbreviated as R) tartaric acid are used respectively to synthesize chiral multi-level hydroxyapatite antibacterial films, and the synthesized antibacterial films are hereinafter referred to as D-CHAPFs, L-CHAPFs and R-CHAPFs.

[0114] 1.3 Test results

[0115] The L-CHAPFs chiral multi-level hydroxyapatite antibacterial film is relatively flat with no breakage or fragmentation when observed with the naked eye. Figure 2 ); Scanning electron microscopy showed that the L-CHAPFs chiral multi-level hydroxyapatite antibacterial film had a microstructure in the shape of staggered and neatly arranged thin sheets, with a uniform surface morphology and a multi-level chiral structure ( Figure 3 ); From the XRD pattern, high-magnification transmission electron microscopy and electron diffraction pattern, it can be seen that the crystal structure of the L-CHAPFs chiral multi-level hydroxyapatite antibacterial film is hexagonal, and the space group is Pm / 63, which corresponds to the crystal structure of hydroxyapatite ( Figure 4 and Figure 5 ); The 3D-EDT image shows that the atomic arrangement in the crystal of L-CHAPFs chiral multi-level hydroxyapatite antibacterial film is in a spiral twisted state, which explains the atomic-level chirality of the chiral multi-level hydroxyapatite antibacterial film and proves that the chiral multi-level structure originates from the spiral twisted stacking of atoms ( Figure 6 ).

[0116] From the circular dichroism spectrum of chiral multi-level hydroxyapatite antibacterial film, we can see that ( Figure 7), L-CHAPFs, D-CHAPFs and R-CHAPFs are chiral multi-level structure hydroxyapatite antibacterial films synthesized corresponding to the L-type chiral molecule, D-type chiral molecule and R-type chiral molecule in Example 1 respectively; the chiral multi-level structure hydroxyapatite antibacterial films of Example 1 L-CHAPFs and D-CHAPFs both showed circular dichroism, indicating that both of them formed chiral multi-level structures and had corresponding optical properties, and R-CHAPFs had no circular dichroism signal and did not show chiral multi-level structure.

[0117] Example 2 Preparation of chiral multi-level gallium phosphate antibacterial film

[0118] The preparation method of the chiral multi-level gallium phosphate antibacterial film provided in this embodiment comprises the following steps:

[0119] S1, dissolving a soluble gallium source GaCl3 in water, and stirring thoroughly to dissolve to form a solution A, wherein the content of gallium trichloride in the solution A is 1 mmol.

[0120] S2, dissolving the chiral inducing agent tartaric acid (C4H6O6) in water, stirring thoroughly to dissolve to form solution B, wherein the content of tartaric acid in solution B is 3 mmol.

[0121] S3, dissolving the soluble phosphorus source sodium dihydrogen phosphate (NaH2PO4) and the nucleation control agent urea (carbonamide: CH4N2O) in water, stirring and dissolving to form solution C, wherein the contents of sodium dihydrogen phosphate and urea in solution C are 1 mmol and 3 mmol, respectively.

[0122] S4, at room temperature, solution B is quickly added to solution A to form a mixed solution, and stirring is continued for 30 minutes to obtain an AB mixed solution.

[0123] S5. At room temperature, while vigorously stirring, slowly add solution C dropwise into the mixed solution AB to form a mixed reaction solution, and continue stirring for 30 minutes.

[0124] S6, transfer the mixed reaction liquid into a reaction kettle, add the pretreated substrate, react at 150°C for 24 hours, cool naturally, take out the substrate, wash it alternately with deionized water and ethanol for 3 times, and dry it at 60°C to obtain a chiral multi-level gallium phosphate antibacterial film. The substrate used in this embodiment is a mica substrate, which is pretreated in advance to achieve activation, wherein the pretreatment operation is: tear and peel the mica with transparent glue to obtain a fresh exposed surface.

[0125] In the above process, the ratio of the added amount of each raw material is: the molar ratio of the soluble gallium source, the soluble phosphorus source, the chiral inducer, the nucleation control agent and water is 1:1:3:3:1500.

[0126] In step S2, D-type, L-type and Racemic (i.e., racemic, hereinafter abbreviated as R) tartaric acid are used respectively to synthesize a chiral multi-level gallium phosphate antibacterial film. The synthesized antibacterial films are hereinafter referred to as D-CMS, L-CMS and R-CMS.

[0127] Example 3 Preparation of chiral multi-level structured molybdenum disulfide antibacterial film

[0128] The preparation method of the chiral multi-level structure molybdenum disulfide antibacterial film provided in this embodiment comprises the following steps:

[0129] S1, dissolving soluble molybdenum source ammonium molybdate (H8MoN2O4) in water, stirring and dissolving to form solution A, wherein the content of ammonium molybdate in solution A is 1 mmol.

[0130] S2, the chiral inducing agent phenylalaninol (C9H 13 NO) was dissolved in water, and the solution B was formed by sufficient stirring. The content of phenylalaninol in the solution B was 6 mmol.

[0131] S3, dissolving soluble sulfur source thioacetamide (CH3CSNH2) in water, stirring and dissolving to form solution C, wherein the content of thioacetamide in solution C is 1 mmol.

[0132] S4, at room temperature, solution B is quickly added to solution A to form a mixed solution, and stirring is continued for 60 minutes to obtain an AB mixed solution.

[0133] S5. At room temperature, while vigorously stirring, slowly add solution C dropwise into the mixed solution AB to form a mixed reaction solution, and continue stirring for 10 minutes.

[0134] S6, transfer the mixed reaction liquid into a reactor, add the pretreated substrate, react at 180°C for 8 hours, cool naturally, take out the substrate, wash it alternately with deionized water and ethanol, and dry it for 8 hours to obtain a chiral multi-level structure molybdenum disulfide antibacterial film. The substrate used in this embodiment is a FTO substrate, which is pretreated in advance to achieve activation, wherein the pretreatment operation is: soaking in 1mmol ammonium molybdate solution for 1 hour, taking out and drying for use.

[0135] In the above process, the added amount ratio of each raw material is: the molar ratio of soluble molybdenum source, soluble sulfur source, chiral inducer, and water is 1:1:6:1800.

[0136] In step S2, D-type, L-type and Racemic type (i.e., racemic type, hereinafter abbreviated as R) phenylalaninol are used respectively to synthesize a chiral multi-level structure molybdenum disulfide antibacterial film, and the synthesized antibacterial films are hereinafter respectively referred to as D-CMS, L-CMS and R-CMS.

[0137] Example 4 Preparation of chiral multi-level iron sulfide antibacterial film

[0138] The preparation method of the chiral multi-level iron sulfide antibacterial film provided in this embodiment comprises the following steps:

[0139] S1, dissolving a soluble iron source, ferrous sulfate heptahydrate (FeSO4·7H2O) in water, and stirring thoroughly to dissolve to form a solution A, wherein the content of ferrous sulfate heptahydrate in the solution A is 1 mmol.

[0140] S2, dissolving the chiral inducing agent tartaric acid (C4H6O6) in water, stirring thoroughly to dissolve to form solution B, wherein the content of tartaric acid in solution B is 0.75 mmol.

[0141] S3, a soluble sulfur source sodium sulfide (Na2S) and a nucleation control agent sodium dodecylbenzene sulfonate (C 18 H 29 NaO3S) was dissolved in water and stirred thoroughly to form solution C, in which the contents of sodium sulfide and sodium dodecylbenzene sulfonate were 1 mmol and 1.25 mmol, respectively.

[0142] S4, at room temperature, solution B is quickly added to solution A to form a mixed solution, and stirring is continued for 20 minutes to obtain an AB mixed solution.

[0143] S5. At room temperature, while vigorously stirring, slowly add solution C dropwise into the mixed solution AB to form a mixed reaction solution, and continue stirring for 40 minutes.

[0144] S6, transfer the mixed reaction liquid into a reaction kettle, add the pretreated substrate, react at 180°C for 24 hours, cool naturally, take out the substrate, wash it alternately with deionized water and ethanol, and dry it to obtain a chiral multi-level structure iron sulfide antibacterial film. The substrate used in this embodiment is a substrate, which is pretreated in advance to achieve activation, wherein the pretreatment operation is: tear and peel the mica with transparent glue to obtain a fresh exposed surface.

[0145] In the above process, the added amount ratio of each raw material is: the molar ratio of soluble iron source, soluble sulfur source, chiral inducer, nucleation control agent and water is 1:1:0.75:1.25:1680.

[0146] In step S2, D-type, L-type and Racemic (i.e., racemic, hereinafter abbreviated as R) tartaric acid are used respectively to synthesize a chiral multi-level iron sulfide antibacterial membrane. The synthesized antibacterial membranes are hereinafter referred to as D-CMS, L-CMS and R-CMS.

[0147] Example 5 Preparation of chiral multi-level iron sulfide antibacterial film

[0148] The preparation method of the chiral multi-level structure calcium carbonate antibacterial film provided in this embodiment comprises the following steps:

[0149] S1, dissolving a soluble calcium source, calcium nitrate tetrahydrate (Ca(NO3)2·4H2O) in water, and stirring thoroughly to dissolve to form a solution A, wherein the content of calcium nitrate tetrahydrate in the solution A is 1.2 mmol.

[0150] S2, dissolving the chiral inducing agent aspartic acid (C4H7NO4) in water, stirring thoroughly to dissolve to form solution B, wherein the content of aspartic acid in solution B is 0.6 mmol.

[0151] S3, dissolving the soluble carbon source ammonium carbonate ((NH4)2CO3) in water, stirring thoroughly to dissolve to form solution C, wherein the content of ammonium carbonate in solution C is 1.2 mmol.

[0152] S4, at room temperature, solution B is quickly added to solution A to form a mixed solution, and stirring is continued for 30 minutes to obtain an AB mixed solution.

[0153] S5. At room temperature, while vigorously stirring, slowly add solution C dropwise into the mixed solution AB to form a mixed reaction solution, adjust the pH to 6.7, and continue stirring for 30 minutes.

[0154] S6, the mixed reaction liquid is transferred into a beaker, the pre-treated substrate is added, the reaction is continued for 48 hours, the substrate is cooled naturally, the substrate is taken out, and the substrate is washed alternately with deionized water and ethanol, and dried to obtain a chiral multi-level structure calcium carbonate antibacterial film. The substrate used in this embodiment is a glass substrate, which is cleaned in advance and does not need to be pre-treated.

[0155] In the above process, the ratio of the added amount of each raw material is: the molar ratio of the soluble calcium source, the soluble carbon source, the chiral inducer and water is 1.2:1.2:0.6:5000.

[0156] In step S2, D-type, L-type and Racemic (i.e., racemic, hereinafter abbreviated as R) aspartic acid are used respectively to synthesize a chiral multi-level calcium carbonate antibacterial membrane. The synthesized antibacterial membranes are hereinafter referred to as D-CMS, L-CMS and R-CMS.

[0157] Example 6 Preparation of Chiral Multi-level Barium Titanate Antibacterial Film

[0158] The preparation method of the chiral multi-level barium titanate antibacterial film provided in this embodiment comprises the following steps:

[0159] S1, dissolving soluble barium source barium sulfate (BaSO4) in water, stirring and dissolving to form solution A, wherein the content of barium sulfate in solution A is 0.75 mmol.

[0160] S2, dissolving the chiral inducing agent ascorbic acid (C6H8O6) in water, stirring thoroughly to dissolve to form a solution B, wherein the content of ascorbic acid in the solution B is 1.5 mmol.

[0161] S3, the soluble titanium source acetylacetonate titanium (C 10 H 14 O5Ti) and nucleation control agent urea (carbonamide: CH4N2O) are dissolved in water and fully stirred to dissolve to form solution C, in which the contents of titanium acetylacetonate and urea are 0.75 mmol and 2 mmol, respectively.

[0162] S4, at room temperature, solution B is quickly added to solution A to form a mixed solution, and stirring is continued for 20 minutes to obtain an AB mixed solution.

[0163] S5. At room temperature, while vigorously stirring, slowly add solution C dropwise into the mixed solution AB to form a mixed reaction solution, and continue stirring for 40 minutes.

[0164] S6, transfer the mixed reaction liquid into a reactor, add the pretreated substrate, react at 150°C for 12 hours, cool naturally, take out the substrate, wash it alternately with deionized water and ethanol, and dry it to obtain a chiral multi-level barium titanate antibacterial film. The substrate used in this embodiment is a titanium sheet substrate, which is pretreated in advance to achieve activation, wherein the pretreatment operation is: put the titanium sheet into 1M hydrochloric acid for ultrasonic activation for 30 minutes, and then wash it with deionized water for use.

[0165] In the above process, the added amount ratio of each raw material is: the molar ratio of soluble barium source, soluble titanium source, chiral inducer, nucleation control agent and water is 0.75:0.75:1.5:2:1700.

[0166] In step S2, D-type, L-type and Racemic (i.e., racemic, hereinafter abbreviated as R) ascorbic acid are used respectively to synthesize a chiral multi-level structure barium titanate antibacterial film, and the synthesized antibacterial films are hereinafter referred to as D-CMS, L-CMS and R-CMS.

[0167] Example 7 Preparation of chiral multi-level structure titanium dioxide antibacterial film

[0168] The preparation method of the chiral multi-level structure titanium dioxide antibacterial film provided in this embodiment comprises the following steps:

[0169] S1, tetrabutyl titanate (C 16 H 36 O4Ti) was dissolved in methanol and stirred thoroughly to form solution A, wherein the content of tetra-n-butyl titanate in solution A was 0.5 mmol.

[0170] S2, dissolving the chiral inducing agent tartaric acid (C4H6O6) in water, stirring thoroughly to dissolve to form solution B, wherein the content of tartaric acid in solution B is 1.5 mmol.

[0171] S3, the soluble surfactant sodium dodecyl sulfate (C 12 H 25 OSO3Na) was dissolved in water and stirred thoroughly to form solution C, wherein the content of sodium dodecyl sulfate in solution C was 1.25 mmol.

[0172] S4, at room temperature, solution B is quickly added to solution A to form a mixed solution, and stirring is continued for 10 minutes to obtain an AB mixed solution.

[0173] S5. At room temperature, while vigorously stirring, slowly add solution C dropwise into the mixed solution AB to form a mixed reaction solution, and continue stirring for 10 minutes.

[0174] S6, transfer the mixed reaction liquid into a 60°C water bath, add the pretreated substrate, and continue stirring for 60 minutes. Cool naturally, take out the substrate, wash it alternately with deionized water and ethanol, and dry it to obtain a chiral multi-level structure titanium dioxide antibacterial film. The substrate used in this embodiment is a mica substrate, which is pretreated in advance to achieve activation, wherein the pretreatment operation is: tear and peel the mica with transparent glue to obtain a fresh exposed surface.

[0175] In the above process, the ratio of the added amount of each raw material is: the molar ratio of the soluble surfactant, the soluble titanium source, the chiral inducer, methanol and water is 1.25:0.5:1.5:100:900.

[0176] In step S2, D-type, L-type and Racemic (i.e., racemic, hereinafter abbreviated as R) tartaric acid are used respectively to synthesize a chiral multi-level structure titanium dioxide antibacterial film, and the synthesized antibacterial films are hereinafter referred to as D-CMS, L-CMS and R-CMS.

[0177] Example 8 Preparation of chiral multi-level bismuth oxygen bromide antibacterial film

[0178] The preparation method of the chiral multi-level bismuth oxybromide antibacterial film provided in this embodiment comprises the following steps:

[0179] S1, dissolving a soluble bismuth source, bismuth nitrate pentahydrate (Bi(NO3)3·5H2O) in water, and stirring thoroughly to dissolve to form a solution A, wherein the content of bismuth nitrate pentahydrate in the solution A is 1 mmol.

[0180] S2, the chiral inducing agent mannitol (C6H 14 O6) was dissolved in water and stirred thoroughly to form solution B, wherein the content of mannitol in solution B was 0.5 mmol.

[0181] S3, dissolving a soluble bromine source sodium bromide (NaBr) in water, and stirring thoroughly to dissolve to form a solution C, wherein the content of sodium bromide in the solution C is 1 mmol.

[0182] S4, at room temperature, solution B is quickly added to solution A to form a mixed solution, and stirring is continued for 60 minutes to obtain an AB mixed solution.

[0183] S5. At room temperature, while vigorously stirring, slowly add solution C dropwise into the mixed solution AB to form a mixed reaction solution, and continue stirring for 30 minutes.

[0184] S6, transfer the mixed reaction liquid into a reactor, add the pretreated substrate, react at 200°C for 24 hours, cool naturally, take out the substrate, wash it alternately with deionized water and ethanol, and dry it to obtain a chiral multi-level structure bismuth oxybromide antibacterial film. The substrate used in this embodiment is a FTO substrate, which is pretreated in advance to achieve activation, wherein the pretreatment operation is: immerse the FTO substrate in a 1mmol bismuth nitrate pentahydrate solution for 60min, and then quickly dry it at 100°C to obtain a substrate with seed crystals.

[0185] In the above process, the ratio of the added amount of each raw material is: the molar ratio of the soluble bismuth source, the soluble bromine source, the chiral inducer and water is 1:1:0.5:1580.

[0186] In step S2, D-type, L-type and Racemic type (i.e., racemic type, hereinafter abbreviated as R) mannitol are used respectively to synthesize a chiral multi-level structure bismuth oxybromide antibacterial film, and the synthesized antibacterial films are hereinafter referred to as D-CMS, L-CMS and R-CMS.

[0187] Example 9 Preparation of chiral multi-level copper sulfide antibacterial film

[0188] The preparation method of the chiral multi-level copper sulfide antibacterial film provided in this embodiment comprises the following steps:

[0189] S1, dissolving soluble copper source copper chloride (CuCl2) in water, stirring and dissolving to form solution A, wherein the content of copper chloride in solution A is 1.25 mmol.

[0190] S2, dissolving the chiral inducing agent tartaric acid (C4H6O6) in water, stirring thoroughly to dissolve to form solution B, wherein the content of tartaric acid in solution B is 0.625 mmol.

[0191] S3, a soluble sulfur source sodium sulfide (Na2S) and a nucleation control agent sodium dodecylbenzene sulfonate (C 18 H 29NaO3S) was dissolved in water and stirred thoroughly to form solution C, in which the contents of sodium sulfide and sodium dodecylbenzene sulfonate were 1 mmol and 1.25 mmol, respectively.

[0192] S4, at room temperature, solution B is quickly added to solution A to form a mixed solution, and stirring is continued for 10 minutes to obtain an AB mixed solution.

[0193] S5. At room temperature, while vigorously stirring, slowly add solution C dropwise into the mixed solution AB to form a mixed reaction solution, and continue stirring for 30 minutes.

[0194] S6, transfer the mixed reaction liquid into a reactor, add the pretreated substrate, react at 120°C for 24 hours, cool naturally, take out the substrate, wash it alternately with deionized water and ethanol, and dry it to obtain a chiral multi-level copper sulfide antibacterial film. The substrate used in this embodiment is a FTO substrate, which is pretreated in advance to achieve activation, wherein the pretreatment operation is: immerse the FTO substrate in a 1mmol copper chloride solution for 60min, and then quickly dry it at 100°C to obtain a substrate with seed crystals.

[0195] In the above process, the added amount ratio of each raw material is: the molar ratio of soluble calcium source, soluble phosphorus source, chiral inducer, nucleation control agent and water is 1.25:1.25:0.625:1.5:1500.

[0196] In step S2, D-type, L-type and Racemic (i.e., racemic, hereinafter abbreviated as R) tartaric acid are used respectively to synthesize a chiral multi-level copper sulfide antibacterial film, and the synthesized antibacterial films are hereinafter referred to as D-CMS, L-CMS and R-CMS.

[0197] Example 10 Preparation of chiral multi-level structure strontium titanate antibacterial film

[0198] The preparation method of the chiral multi-level structure strontium titanate antibacterial film provided in this embodiment comprises the following steps:

[0199] S1, dissolving a soluble strontium source strontium nitrate (Sr(NO3)2) in water, stirring and dissolving to form a solution A, wherein the content of strontium nitrate in the solution A is 1 mmol.

[0200] S2, the chiral inducing agent lysine (C6H 14 N2O2) was dissolved in water and stirred thoroughly to form solution B, wherein the content of lysine in solution B was 2 mmol.

[0201] S3, the soluble titanium source acetylacetonate titanium (C 10 H 14O5Ti) and nucleation control agent urea (carbonamide: CH4N2O) are dissolved in water and fully stirred to dissolve to form solution C, in which the contents of titanium acetylacetonate and urea are 1 mmol and 3 mmol, respectively.

[0202] S4, at room temperature, solution B is quickly added to solution A to form a mixed solution, and stirring is continued for 20 minutes to obtain an AB mixed solution.

[0203] S5. At room temperature, while vigorously stirring, slowly add solution C dropwise into the mixed solution AB to form a mixed reaction solution, and continue stirring for 40 minutes.

[0204] S6, transfer the mixed reaction liquid into a reactor, add the pretreated substrate, react at 190°C for 6 hours, cool naturally, take out the substrate, wash it alternately with deionized water and ethanol, and dry it to obtain a chiral multi-level structure strontium titanate antibacterial film. The substrate used in this embodiment is a titanium sheet substrate, which is pretreated in advance to achieve activation, wherein the pretreatment operation is: put the titanium sheet into 1M hydrochloric acid for ultrasonic activation for 30 minutes, and then wash it with deionized water for use.

[0205] In the above process, the added amount ratio of each raw material is: the molar ratio of soluble barium source, soluble titanium source, chiral inducer, nucleation control agent and water is 1:1:2:3:1670.

[0206] In step S2, D-type, L-type and Racemic (i.e., racemic, hereinafter abbreviated as R) lysine are used to synthesize chiral multi-level structure strontium titanate antibacterial films. The synthesized antibacterial films are hereinafter referred to as D-CMS, L-CMS and R-CMS.

[0207] Example 11 In vitro antibacterial test of chiral multi-level hydroxyapatite antibacterial films with different structures

[0208] 11.1 Sample Selection

[0209] Blank control: blank mica basement membrane;

[0210] Test Example: Chiral multi-level structure hydroxyapatite antibacterial film (having a blade array structure) using D-CHAPFs, L-CHAPFs and R-CHAPFs of Example 1;

[0211] Test bacteria: Staphylococcus aureus and Escherichia coli;

[0212] Comparative example: Ach-MHAPFs is a hydroxyapatite antibacterial membrane without a chiral multi-level structure.

[0213] 11.2 Test methods

[0214] 1) Place the substrate on the bottom of the cell culture plate;

[0215] 2) planting dispersed Staphylococcus aureus and Escherichia coli on the chiral substrate respectively for cultivation;

[0216] 3) After a period of time, take out the culture plate, rinse it, and fix it;

[0217] 4) Wash and stain after fixation;

[0218] 5) Observe and take photos under a microscope.

[0219] 11.3 Test results

[0220] like Figure 8 As shown, compared with the blank mica basement membrane in the prior art, the chiral multi-level structure hydroxyapatite antibacterial membrane of Example 1 of the present invention can selectively promote the adhesion and proliferation of bacteria (Escherichia coli), among which D-CHAPFs has obvious anti-bacterial adhesion and anti-proliferation effects, while the antibacterial effect of L-CHAPFs is weaker than that of D-CHAPFs.

[0221] like Fig. 9 As shown, compared with the blank mica basement membrane in the prior art, the chiral multi-level structure hydroxyapatite antibacterial membrane of Example 1 of the present invention can selectively promote the adhesion and proliferation of bacteria (Staphylococcus aureus). The antibacterial adhesion and proliferation effects of D-CHAPFs are obvious, while the antibacterial effect of L-CHAPFs is weaker than that of D-CHAPFs.

[0222] Through the crystal violet staining pictures of the two bacteria on the chiral antibacterial film, it can be seen that the bacterial biofilm formation on the D-CHAPFs surface is less, and the surfaces of the other groups have different degrees of bacterial biofilm formation, and all of them have more D-CHAPFs ( Fig.10 );According to OD of crystal violet staining 495 The statistical value shows that D-CHAPFs has a better resistance effect ( Fig.11 ); At the same time, the live-dead staining of bacteria was used for observation, and the results showed that a large number of bacteria on the surface of D-CHAPFs died, proving its excellent antibacterial effect ( Fig.12 ); Scanning electron microscopy images of bacteria on the chiral antibacterial film show that the number of bacteria on the D-CHAPFs surface is small and their morphology is abnormal, which further proves the excellent antibacterial effect of D-CHAPFs ( Fig.13 ); In summary, D-CHAPFs has excellent antibacterial properties.

[0223] like Fig.14 As shown, compared with the blank mica basement membrane in the prior art, the antibacterial effect of the chiral multi-level hydroxyapatite antibacterial membrane of Example 1 of the present invention is compared as follows:

[0224]

[0225]

[0226] Example 12 Antibacterial test of chiral multi-level hydroxyapatite antibacterial films with different structures in vivo

[0227] 12.1 Sample Selection

[0228] Blank control: blank mica basement membrane;

[0229] Test Example 1: A chiral multi-level structured hydroxyapatite antibacterial film (having a blade array structure) using D-CHAPFs, L-CHAPFs and R-CHAPFs of Example 1;

[0230] Comparative example: Ach-MHAPFs is a hydroxyapatite antibacterial membrane without a chiral multi-level structure.

[0231] Test bacteria: Staphylococcus aureus;

[0232] The experimental animals were nude mice.

[0233] 12.2 Test methods

[0234] 1) Disinfect and create models on the back skin of nude mice;

[0235] 2) Cut the back skin and implant the sterilized chiral antibacterial membrane;

[0236] 2) seeding the dispersed Staphylococcus aureus suspension on the chiral multi-level structure hydroxyapatite antibacterial membrane substrate and culturing in vivo;

[0237] 3) Suture the wound and culture it;

[0238] 4) After a period of incubation, observe the wound status at different time points;

[0239] 5) After the set time point, take out the sample, wash it with PBS, and fix it with 4% PFA; after fixation, wash it several times with PBS, and then stain it;

[0240] 6) Finally, observe and take pictures under a microscope.

[0241] 12.3 Test results

[0242] like Fig.15 As shown, compared with the blank mica basement membrane in the prior art, the chiral multi-level structure hydroxyapatite antibacterial membrane of Example 1 of the present invention can selectively achieve antibacterial. D-CHAPFs helps to repair infected wounds, while other chiral membranes have poor repair effects on infected wounds.

[0243] like Fig.16 As shown, compared with the blank mica basement membrane in the prior art, the chiral multi-level structure hydroxyapatite antibacterial membrane of the present invention can selectively inhibit the formation of bacterial biofilm. The effect of D-CHAPFs in inhibiting the growth of bacterial biofilm is better than that of L-CHAPFs and other groups.

[0244] Such as logCFU / ml (CFU is colony forming unit) Fig.17 As shown, compared with the blank mica basement membrane in the prior art, the chiral multi-level hydroxyapatite antibacterial membrane of Example 1 of the present invention has excellent effect on repairing bacterial infection wounds in vivo.

[0245] In summary, D-CHAPFs with a blade array structure have the effect of inhibiting the growth of bacterial biofilms with an inhibition rate of 90-96%, and are helpful for the repair of infected wounds.

[0246]

[0247] Example 13 In vitro antibacterial test of chiral multi-level molybdenum disulfide antibacterial films with different structures

[0248] 13.1 Sample Selection

[0249] Blank control: blank mica basement membrane;

[0250] Test Example: Chiral multi-level structured molybdenum disulfide antibacterial film (having a blade array structure) of D-CMS, L-CMS and RCMS of Example 3;

[0251] Test bacteria: Staphylococcus aureus and Escherichia coli.

[0252] 13.2 Test methods

[0253] 1) First, place the sterilized substrate on the bottom of the cell culture plate;

[0254] 2) Then, the dispersed Staphylococcus aureus and Escherichia coli are planted on the chiral substrate for cultivation;

[0255] 3) After a period of incubation, the culture plate was removed, washed with PBS, and fixed with 4% PFA;

[0256] 4) After fixation, wash several times with PBS and then stain;

[0257] 5) Finally, observe and take pictures under a microscope.

[0258] 13.3 Test results

[0259] like Fig.18As shown, compared with the blank mica base membrane in the prior art, the chiral multi-level structure molybdenum disulfide antibacterial membrane of Example 3 of the present invention can selectively promote the adhesion and proliferation of bacteria (Escherichia coli), among which D-CMS has obvious anti-bacterial adhesion and anti-proliferation effects, while the antibacterial effect of L-CMS is weaker than that of D-CMS.

[0260] like Fig.19 As shown, compared with the blank mica basement membrane in the prior art, the chiral multi-level structured molybdenum disulfide antibacterial membrane of Example 3 of the present invention can selectively promote the adhesion and proliferation of bacteria (Staphylococcus aureus). The antibacterial adhesion and proliferation effects of D-CMS are obvious, while the antibacterial effect of L-CMS is weaker than that of D-CMS. In summary, D-CMS has excellent antibacterial properties.

[0261] like Fig. 20 As shown, compared with the blank mica base film in the prior art, the antibacterial effect of the chiral multi-level molybdenum disulfide antibacterial film of Example 3 of the present invention is compared as follows:

[0262]

[0263]

Claims

1. A chiral multi-level structure antibacterial material, the chiral multi-level structure antibacterial material is composed of a first component, a second component, a chiral inducing agent, a substrate and / or a nucleation control agent, forming a microenvironment with a chiral multi-level structure: wherein, The first component is a soluble metal source, selected from one or more of a soluble calcium source, a soluble gallium source, a soluble molybdenum source, a soluble iron source, a soluble barium source, a soluble bismuth source, a soluble copper source, a soluble strontium source, a soluble titanium source, a soluble silver source and / or a soluble magnesium source; the second component is selected from one or more of a soluble phosphorus source, a soluble sulfur source, a soluble carbon source, a soluble titanium source and / or a soluble bromine source; the chiral multi-level structure antibacterial material includes a chiral multi-level structure hydroxyapatite antibacterial material, a chiral multi-level structure gallium phosphate antibacterial material, a chiral multi-level structure zinc oxide antibacterial material, a chiral Chiral multilevel structure molybdenum disulfide antibacterial materials, chiral multilevel structure iron sulfide antibacterial materials, chiral multilevel structure calcium carbonate antibacterial materials, chiral multilevel structure barium titanate antibacterial materials, chiral multilevel structure titanium dioxide antibacterial materials, chiral multilevel structure bismuth oxygen bromide antibacterial materials, chiral multilevel structure copper oxide antibacterial materials, chiral multilevel structure metallic silver antibacterial materials, chiral multilevel structure copper sulfide antibacterial materials, chiral multilevel structure strontium titanate antibacterial materials, chiral multilevel structure magnesium oxide antibacterial materials and chiral multilevel structure calcium vanadate antibacterial materials, etc.; the chiral multilevel structure antibacterial material is a chiral multilevel structure antibacterial film.

2. A method for preparing the chiral multi-level antibacterial material according to claim 1, comprising the following steps: S1, dissolving the first component in water to form solution A; S2, dissolving the chiral inducing agent in water to form solution B; S3, dissolving the second component and the nucleation control agent in water to form a solution C; S4, rapidly adding solution B into solution A to form a mixed solution, and continuing stirring for 10-60 minutes to obtain an AB mixed solution; S5, slowly adding solution C to the mixed solution AB to form a mixed reaction solution, and continuing to stir for 2-30 minutes; S6, transferring the mixed reaction liquid into a reaction kettle, adding the pretreated substrate, reacting at 100-200° C. for 2-24 hours, cooling, taking out the substrate, washing and drying, and obtaining a chiral multi-level structure antibacterial material.

3. A use of the chiral multi-level antibacterial film according to claim 1 in the preparation of physical antibacterial materials, wherein the antibacterial film has the effects of reducing bacterial adhesion, destroying bacterial structure and facilitating wound repair; wherein, The inhibited bacteria include autotrophic bacteria and heterotrophic bacteria.

4. The application scenario of the physical antibacterial material as described in claim 3 is any surface or object that needs to be antibacterial; the chiral multi-level structure antibacterial film can selectively regulate the bacterial recognition material surface process to prevent bacterial adhesion; the chiral multi-level structure antibacterial film can selectively regulate the adhesion behavior of bacteria and control the growth of bacterial biofilm, thereby producing an antibacterial effect; the chiral multi-level structure antibacterial film is assembled by atomic-level nanosheets; the nanosheets act as a blade array, providing a physical tool for destroying bacterial membranes, and achieving an antibacterial effect by destroying the structure of adhered bacteria.

5. The first component according to claim 1 or 2, wherein The soluble calcium source is selected from one or more of anhydrous calcium chloride, calcium nitrate tetrahydrate, calcium sulfate dihydrate and calcium oxalate; the soluble gallium source is selected from one or more of gallium chloride, gallium bromide, gallium iodide, gallium fluoride, gallium nitride, gallium sulfate, gallium phosphide, gallium oxide and / or gallium selenide; the soluble molybdenum source is selected from one or more of ammonium molybdate, sodium molybdate, magnesium molybdate, zinc molybdate, ammonium dimolybdate, iron molybdate, lead molybdate, copper molybdate, sodium paramolybdate, ammonium paramolybdate, ammonium tetramolybdate and / or ammonium dimolybdate; the soluble iron source is selected from one or more of ferrous sulfate heptahydrate, ferric chloride, ferric nitrate, ferric oxalate and / or ferric phosphate; the soluble barium source is selected from one or more of barium sulfate, barium chloride, barium nitrate, barium carbonate, barium oxalate, barium acetate, barium phosphate, barium tungstate and / or barium titanate; the soluble bismuth source is selected from one or more of bismuth nitrate pentahydrate, bismuth sulfate , bismuth chloride, bismuth oxalate and / or bismuth acetate; the soluble copper source is selected from one or more of copper chloride, copper sulfate, copper nitrate, copper oxalate, copper tungstate, copper acetate and / or copper phosphate; the soluble strontium source is selected from one or more of strontium nitrate, strontium chloride, strontium oxalate, strontium titanate, strontium sulfate and / or strontium carbonate; the soluble titanium source is selected from one or more of tetra-n-butyl titanate, titanium acetylacetonate, tetraisopropyl titanate, titanium chloride, titanium sulfate, sodium carbonate, potassium titanate and / or ammonium carbonate; the soluble silver source is selected from one or more of silver nitrate, silver chloride, silver sulfate, silver oxide, silver bromide, silver iodide, acetate, oxalate and / or silver citrate; the soluble magnesium source is selected from one or more of magnesium chloride, magnesium sulfate, magnesium nitrate, magnesium oxalate, magnesium carbonate, magnesium citrate, magnesium lactate, magnesium phosphate and / or magnesium vanadate.

6. The second component according to claim 1 or 2, wherein The soluble phosphorus source is selected from one or more of diammonium hydrogen phosphate, dipotassium hydrogen phosphate and / or disodium hydrogen phosphate; the soluble sulfur source is selected from one or more of thioacetamide, sodium sulfide, sulfuric acid, sodium sulfate, potassium sulfate, ammonium sulfate, thiourea, mercaptan, thioether and / or sulfate ester; the soluble carbon source is selected from one or more of ammonium carbonate, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate and / or ammonium bicarbonate; the soluble titanium source is selected from one or more of tetra-n-butyl titanate, titanium acetylacetonate, tetraisopropyl titanate, titanium chloride, titanium sulfate, sodium carbonate, potassium titanate and / or ammonium carbonate; the soluble bromine source is selected from one or more of sodium bromide, potassium bromide, hydrogen bromide, ammonium bromide, calcium bromide, zinc bromide, copper bromide and / or ferrous bromide.

7. The chiral inducing agent according to claim 1 or 2 is one or more of chiral amino acids, chiral organic acids, chiral sugars and chiral alcohols; wherein, The chiral amino acid is selected from one or more of histidine, arginine, lysine, isoleucine, phenylalanine, leucine, tryptophan, alanine, methionine, proline, cysteine, aspartic acid, valine, serine, glutamine, tyrosine, aspartic acid, glutamic acid and threonine; the chiral organic acid is selected from one or more of tartaric acid, malic acid and lactic acid; the chiral sugar is selected from glucose, fructose, galactose, ribose, deoxyribose, furanose, pyranose, maltose, sucrose and lactose; the chiral alcohol includes one or more of mannitol, xylitol, sorbitol, paclitaxel, resveratrol, ginkgo biloba polyprenols, (R)-(+)-1-phenyl-1-propanol, (S)-(-)-1-phenyl-1-propanol and / or phenylpropanol.

8. The nucleation control agent as claimed in claim 1 or 2 is selected from one or more of urea, sodium dodecylbenzene sulfonate, sodium dodecyl sulfate and / or sodium dodecylbenzene sulfonate.

9. The support substrate as claimed in claim 1 or 2 is selected from any one of activated mica sheets, iron sheets, titanium sheets, titanium alloy sheets, magnesium alloy sheets, glass sheets, quartz sheets, FTO sheets, ITO sheets, silicon sheets, copper sheets or nickel sheets, and the thickness of the support substrate is 0.1 to 1 mm.

10. The chiral multi-level antibacterial material according to any one of claims 1 to 3, wherein the multi-level structure comprises D-CHAPFs type, L-CHAPFs type and Racemic type; the chiral multi-level antibacterial material is formed by assembling atomic-level spirally twisted nanosheets in terms of microstructure; the nanosheets act as a blade array, destroying the structure of bacteria when they adhere.

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