CN115300482A - Bovine serum albumin coated chlorhexidine loaded nickel phosphide nano-capsule, preparation method and antibacterial application thereof - Google Patents
Bovine serum albumin coated chlorhexidine loaded nickel phosphide nano-capsule, preparation method and antibacterial application thereof Download PDFInfo
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- CN115300482A CN115300482A CN202210987189.2A CN202210987189A CN115300482A CN 115300482 A CN115300482 A CN 115300482A CN 202210987189 A CN202210987189 A CN 202210987189A CN 115300482 A CN115300482 A CN 115300482A
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- chlorhexidine
- serum albumin
- bovine serum
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- GHXZTYHSJHQHIJ-UHFFFAOYSA-N Chlorhexidine Chemical compound C=1C=C(Cl)C=CC=1NC(N)=NC(N)=NCCCCCCN=C(N)N=C(N)NC1=CC=C(Cl)C=C1 GHXZTYHSJHQHIJ-UHFFFAOYSA-N 0.000 title claims abstract description 72
- 229960003260 chlorhexidine Drugs 0.000 title claims abstract description 72
- 239000002088 nanocapsule Substances 0.000 title claims abstract description 58
- 108091003079 Bovine Serum Albumin Proteins 0.000 title claims abstract description 55
- 229940098773 bovine serum albumin Drugs 0.000 title claims abstract description 55
- FBMUYWXYWIZLNE-UHFFFAOYSA-N nickel phosphide Chemical compound [Ni]=P#[Ni] FBMUYWXYWIZLNE-UHFFFAOYSA-N 0.000 title claims abstract description 45
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- 239000002091 nanocage Substances 0.000 claims abstract description 34
- 239000000463 material Substances 0.000 claims abstract description 11
- 238000000034 method Methods 0.000 claims description 19
- PXHVJJICTQNCMI-UHFFFAOYSA-N nickel Substances [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 claims description 19
- 229910000159 nickel phosphate Inorganic materials 0.000 claims description 19
- JOCJYBPHESYFOK-UHFFFAOYSA-K nickel(3+);phosphate Chemical compound [Ni+3].[O-]P([O-])([O-])=O JOCJYBPHESYFOK-UHFFFAOYSA-K 0.000 claims description 19
- 239000013154 zeolitic imidazolate framework-8 Substances 0.000 claims description 16
- MFLKDEMTKSVIBK-UHFFFAOYSA-N zinc;2-methylimidazol-3-ide Chemical compound [Zn+2].CC1=NC=C[N-]1.CC1=NC=C[N-]1 MFLKDEMTKSVIBK-UHFFFAOYSA-N 0.000 claims description 16
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- 238000002156 mixing Methods 0.000 claims description 13
- 238000000576 coating method Methods 0.000 claims description 7
- 238000011068 loading method Methods 0.000 claims description 7
- 239000011248 coating agent Substances 0.000 claims description 6
- 238000001027 hydrothermal synthesis Methods 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000008367 deionised water Substances 0.000 claims description 5
- 229910021641 deionized water Inorganic materials 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- 239000002904 solvent Substances 0.000 claims description 4
- 239000011261 inert gas Substances 0.000 claims description 3
- 239000012621 metal-organic framework Substances 0.000 claims description 3
- 238000011144 upstream manufacturing Methods 0.000 claims description 3
- 238000005229 chemical vapour deposition Methods 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 239000002114 nanocomposite Substances 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims description 2
- 229910019142 PO4 Inorganic materials 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- 239000002184 metal Substances 0.000 claims 1
- 229910052751 metal Inorganic materials 0.000 claims 1
- 239000010452 phosphate Substances 0.000 claims 1
- 238000000926 separation method Methods 0.000 claims 1
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 abstract description 4
- 241000589517 Pseudomonas aeruginosa Species 0.000 abstract description 4
- 241000191967 Staphylococcus aureus Species 0.000 abstract description 4
- 229960003085 meticillin Drugs 0.000 abstract description 4
- 239000005416 organic matter Substances 0.000 abstract description 4
- 239000002131 composite material Substances 0.000 abstract description 2
- 239000003242 anti bacterial agent Substances 0.000 description 18
- 238000005530 etching Methods 0.000 description 8
- 241000894006 Bacteria Species 0.000 description 5
- 208000015181 infectious disease Diseases 0.000 description 5
- 230000000813 microbial effect Effects 0.000 description 5
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 239000002243 precursor Substances 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
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- 238000002441 X-ray diffraction Methods 0.000 description 3
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- LXBGSDVWAMZHDD-UHFFFAOYSA-N 2-methyl-1h-imidazole Chemical compound CC1=NC=CN1 LXBGSDVWAMZHDD-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000000862 absorption spectrum Methods 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 229910052786 argon Inorganic materials 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
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- 238000009210 therapy by ultrasound Methods 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
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- 208000035473 Communicable disease Diseases 0.000 description 1
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- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 201000009906 Meningitis Diseases 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
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- 206010034674 peritonitis Diseases 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 239000012221 photothermal agent Substances 0.000 description 1
- 238000007626 photothermal therapy Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000001878 scanning electron micrograph Methods 0.000 description 1
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- 238000000992 sputter etching Methods 0.000 description 1
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5107—Excipients; Inactive ingredients
- A61K9/513—Organic macromolecular compounds; Dendrimers
- A61K9/5169—Proteins, e.g. albumin, gelatin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/13—Amines
- A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0052—Thermotherapy; Hyperthermia; Magnetic induction; Induction heating therapy
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
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- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/51—Nanocapsules; Nanoparticles
- A61K9/5192—Processes
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
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- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
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Abstract
The invention belongs to the technical field of antibacterial materials, and relates to a bovine serum albumin coated chlorhexidine loaded nickel phosphide nano-capsule, and a preparation method and an antibacterial application thereof. The material is a composite material of bovine serum albumin, chlorhexidine and nickel phosphide nanocages. The material is presented as a nano-capsule structure encapsulated by bovine serum albumin, soThe carrier of the nano capsule structure is a nickel phosphide nanocage, the size of the nanocage is about 400nm, and the nanocage structure is hollow. The organic matter package can be clearly seen from the outside. The material is at 1W/cm 2 Under the irradiation of near infrared light of 808nm, 99.4 percent of methicillin-resistant staphylococcus aureus and 99.9 percent of pseudomonas aeruginosa can be killed quickly (15 min) at a mild temperature.
Description
Technical Field
The invention belongs to the technical field of antibacterial materials, and particularly relates to a bovine serum albumin coated chlorhexidine loaded nickel phosphide nano-capsule, and a preparation method and an antibacterial application thereof.
Background
The information disclosed in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
Diseases related to microbial infection are one of important public health problems, and can cause a series of diseases such as meningitis, urinary tract infection, inflammation, septicemia, peritonitis and the like. In the world, public health problems such as epidemics and infectious diseases caused by microbial infections put the human society into a predicament, resulting in economic stagnation and social confusion. In addition, the lack of effective antibacterial treatment modalities has hindered the development of modern medical technology. A large number of patients are still struggling with microbial-related infections, seriously affecting their quality of life. However, our weapons against microbial infections (antibiotics) are weakening due to the increased resistance of bacteria to traditional antibiotics. Therefore, the development of highly effective antibacterial agents to eliminate the risk of microbial infections is becoming more and more urgent.
Photothermal therapy is a novel antibacterial treatment mode, and photothermal antibacterial agents are gathered at affected parts to cause local high temperature, so that the cell membranes of bacteria can be depolarized, and intracellular substances are leaked. The chlorhexidine is a broad-spectrum antibacterial agent, has high cost performance, and is not easy to cause anaphylaxis and drug resistance. Currently, in the antibacterial field, antibacterial materials combining photothermal and antibacterial treatments are a research focus, but there is still a lack of bactericides that can rapidly inactivate bacteria under mild conditions.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a bovine serum albumin coated chlorhexidine loaded nickel phosphide nano-capsule, a preparation method and an antibacterial application thereof. The invention combines an organic matter coating method, an ion etching method and a tubular furnace phosphating method to prepare a chlorhexidine loaded nickel phosphide nanocapsule coated by bovine serum albumin, and applies the nanocapsule to the antibacterial field.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect of the invention, a preparation method of a bovine serum albumin coated chlorhexidine loaded nickel phosphide nano-capsule is provided, which comprises the following steps:
mixing the metal organic framework ZIF-8 and NiCl 2 ·6H 2 Mixing O in solvent, hydrothermal reacting to obtain Ni (OH) 2 A nanocage;
mixing the Ni (OH) 2 Phosphorizing the nano cage to obtain Ni 2 A P nanocage;
adding the Ni 2 And (3) loading chlorhexidine on the P nano cage, and then coating by adopting bovine serum albumin to obtain the nano-composite material.
The invention designs a chlorhexidine loaded nickel phosphide nano capsule based on bovine serum albumin coating, which can realize rapid sterilization under mild conditions.
In a second aspect of the invention, the bovine serum albumin coated chlorhexidine loaded nickel phosphate nanocapsule prepared by the method is provided.
The third aspect of the invention provides the application of the above-mentioned bovine serum albumin coated chlorhexidine loaded nickel phosphate nano capsule in the antibacterial field.
The invention has the advantages of
(1) The invention provides a novel light-driven nano antibacterial agent: the chlorhexidine coated by the bovine serum albumin loads the nickel phosphide nano-capsule. The drug carrier of the antibacterial system is a nickel phosphide nanocage, and the antibacterial agent chlorhexidine is loaded on the nanocage and coated with bovine serum albumin. The sterilization performance of the antibacterial agent is effectively improved by utilizing the synergistic effect of photo-heat and the antibacterial agent;
(2) The chlorhexidine loaded nickel phosphide nano capsule coated by the bovine serum albumin can quickly kill bacteria under mild conditions. The methicillin-resistant staphylococcus aureus of more than 99.4 percent and the pseudomonas aeruginosa of 99.9 percent can be killed by only lighting for 15 min;
(3) The invention provides a preparation method of a nickel phosphide nanocage drug carrier with excellent photo-thermal performance and drug loading capacity, and the preparation method is simple, strong in practicability and easy to popularize;
(4) The invention provides a preparation method of a bovine serum albumin coated chlorhexidine loaded nickel phosphide nano-capsule, and the antibacterial agent has good biocompatibility and good practical application prospect.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are incorporated in and constitute a part of this specification, illustrate exemplary embodiments of the invention and together with the description serve to explain the invention and not to limit the invention.
FIG. 1 is a Scanning Electron Microscope (SEM) photograph of bovine serum albumin coated chlorhexidine loaded nickel phosphate nanocapsules prepared in example 1 of the present invention;
FIG. 2 is an X-ray diffraction (XRD) pattern of bovine serum albumin coated chlorhexidine loaded nickel phosphate nanocapsules prepared in example 1 of the present invention;
FIG. 3 is an infrared absorption spectrum (FT-IR) spectrum of a bovine serum albumin coated chlorhexidine supported nickel phosphate nanocapsule prepared in example 1 of the present invention;
FIG. 4 is a spectrum of ultraviolet-visible light absorption spectrum (UV-Vis) of bovine serum albumin coated chlorhexidine supported nickel phosphate nanocapsule prepared in example 1 of the present invention;
fig. 5 is a demonstration of the antibacterial properties of the bovine serum albumin coated chlorhexidine loaded nickel phosphate nanocapsule prepared in example 1 of the present invention.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
In a first aspect, the bovine serum albumin coated chlorhexidine loaded nickel phosphide nanocapsule is a composite material of bovine serum albumin, chlorhexidine and nickel phosphide nanocages. The material is presented as a nano-capsule structure encapsulated by bovine serum albumin. The carrier of the nano capsule structure is a nickel phosphide nano cage, the size is about 400nm, and the nano capsule structure is a hollow porous structure. The organic matter package can be obviously seen from the outside.
Compared with the existing chlorhexidine antibacterial agent, the bovine serum albumin-coated chlorhexidine-loaded nickel phosphide nanocapsule provided by the invention is characterized in that a nickel phosphide nanocage material is firstly used for antibiosis and is used as a drug carrier of chlorhexidine, and in addition, the three components are organically combined for the first time, so that the property of the nickel phosphide nanocage capsule is more excellent photo-thermal and antibacterial agent synergistic antibacterial performance.
In a second aspect, the specific preparation steps of the above-mentioned bovine serum albumin-coated chlorhexidine-loaded nickel phosphate nanocapsule are: adding Zn (NO) 3 ) 2 ·6H 2 Dissolving O and 2-methylimidazole in absolute methanol to obtain a solution, and then standing the solution after ultrasonic treatment to obtain a product, namely a metal organic framework ZIF-8.
Carrying out hydrothermal reaction on ZIF-8 serving as an etching template to obtain Ni (OH) 2 The nano cage is used as a precursor.
Mixing Ni (OH) 2 Phosphorization of nano cage to obtain Ni 2 P nanometer cage.
By stirring to realize Ni 2 And coating the chlorhexidine load on the P nano cage with bovine serum albumin to obtain the chlorhexidine load nickel phosphide nano capsule coated with the bovine serum albumin.
In some embodiments of the present invention, the etch template is ZIF-8.
In some embodiments of the invention, the temperature of the hydrothermal reaction to prepare the precursor is in the range of 100-120 ℃. The reaction time is 1-2h.
The solvent of the hydrothermal reaction is absolute methanol or deionized water.
In some embodiments of the inventionIn one embodiment, the etchant NiCl 2 ·6H 2 The molar ratio of O to ZIF-8 is 1.5-10; preferably 2 to 3.
In a third aspect, the nickel phosphide nanocage has good photo-thermal performance.
In some embodiments of the present invention, the phosphating is performed by chemical vapor deposition, and the specific steps of phosphating are as follows: mixing Ni (OH) 2 Nanocage and NaH 2 PO 2 ·H 2 Placing O at the downstream and the upstream of the tube furnace respectively, raising the temperature to 300-350 ℃ under the protection of inert gas, and then preserving the heat for 3-3.5 h; cooling, washing and drying to obtain the product.
In some embodiments of the invention, the molar ratio of precursor to phosphorus source during the tube furnace phosphating is from 1.
In some embodiments of the present invention, the carrier gas is an inert gas such as nitrogen, argon, etc. during the tube furnace phosphating process.
In some embodiments of the invention, the rate of temperature increase during the tube furnace phosphating is from 2 to 5 ℃/min.
In the invention, the nickel phosphide nanocage with a thin shell layer has better photo-thermal property.
In a fourth aspect, the nickel phosphide nanocages are applied to drug loading, and the nickel phosphide nanocages can effectively load an antibacterial agent chlorhexidine.
In some embodiments of the invention, the mass ratio of chlorhexidine to nickel phosphide nanocages during drug loading is 0.1-1.
In the invention, the load capacity of the nickel phosphide nano cage on the chlorhexidine is far stronger than that of ZIF-8 nano particles and Ni (OH) 2 A nanocage.
In the fifth aspect, the application of the chlorhexidine loaded nickel phosphate nanocapsule coated with the bovine serum albumin in the field of light-driven antibiosis is provided.
In the invention, pseudomonas aeruginosa and methicillin-resistant staphylococcus aureus are used as standard strain samples. 808nm near infrared laser is used as a light source, and the optical power density is set at 1W/cm 2 。
In the sixth aspect, the nickel phosphide nanocapsule is loaded on the chlorhexidine coated by the bovine serum albumin, and the photo-thermal effect of the nickel phosphide improves the sterilization effect of the chlorhexidine.
The antibacterial agent provided by the invention has the advantages of rapid sterilization under mild conditions, low biological toxicity and the like.
The present invention is described in further detail below with reference to specific examples, which are intended to be illustrative of the invention and not limiting.
Example 1
A preparation method of a bovine serum albumin coated chlorhexidine loaded nickel phosphide nanocapsule comprises the following steps:
1) 0.892g of Zn (NO) 3 ) 2 ·6H 2 O and 0.985g of 2-methylimidazole were dissolved in 25mL of anhydrous methanol, respectively.
2) Mixing the solution obtained in the step 1), performing ultrasonic treatment for 20min, and standing for 24h to obtain ZIF-8 powder.
3) 0.75mmol of ZIF-8 powder and 1.5mmol of NiCl 2 ·6H 2 O is dispersed in 15mL of anhydrous methanol, then stirred for 20min, transferred to a 25mL stainless steel autoclave and hydrothermally treated at 120 ℃ for 1h.
4) Mixing the precursor powder obtained in the step 3) with NaH 2 PO 2 ·H 2 O is 1:10 into two separate crucibles, which are placed downstream and upstream of the tube furnace, respectively, and then N 2 Raising the temperature (the temperature raising speed is 2 ℃/min) to 300 ℃ in the atmosphere, and then preserving the heat for 3h.
5) Dispersing the black powder obtained in the step 4) in 10mL of deionized water (the concentration is 1 mg/mL). 3mL of a chlorhexidine solution (1 mg/mL) was mixed with the above solution, followed by stirring for 24h. Followed by encapsulation by the addition of 5mL BSA (1 mg/mL) and stirring was continued for 12h.
Example 2
A method for preparing a bovine serum albumin coated chlorhexidine loaded nickel phosphide nanocapsule, which is different from the first embodiment in that ZIF-67 is used as a template in the step 2).
Example 3
A method for preparing a bovine serum albumin coated chlorhexidine supported nickel phosphate nano capsule, which is the same as the first embodiment except that the dispersing agent in the step 3) is deionized water.
Example 4
A preparation method of a bovine serum albumin coated chlorhexidine loaded nickel phosphide nanocapsule is different from the first embodiment in that the etching agent NiCl in the step 3) 2 ·6H 2 The molar ratio of O to ZIF-8 is 3:1.
example 5
A preparation method of a bovine serum albumin coated chlorhexidine loaded nickel phosphide nano-capsule is different from the first embodiment in that the etching agent NiCl in the step 3) 2 ·6H 2 The molar ratio of O to ZIF-8 is 5:1.
example 6
A method for preparing a bovine serum albumin coated chlorhexidine supported nickel phosphide nano capsule, which is the same as the first embodiment except that the hydrothermal time in the step 3) is 2 hours.
Example 7
A method for preparing a bovine serum albumin coated chlorhexidine supported nickel phosphide nano capsule, which is the same as the first embodiment except that the hydrothermal temperature in the step 3) is 100 ℃.
Example 8
A method for preparing a bovine serum albumin coated chlorhexidine supported nickel phosphide nano-capsule, which is the same as the first embodiment except that the phosphating gas in the step 4) is argon.
Example 9
A method for preparing a bovine serum albumin coated chlorhexidine supported nickel phosphide nano capsule, which is the same as the first embodiment, and is different from the first embodiment in that the addition amount of the chlorhexidine in the step 5) is 5mL.
Different from the etching template of the embodiment 1 and the etching template of the embodiment 2, through a comparative experiment, the ZIF-8 can be obtained as the template to obtain a nano cage structure, and the ZIF-67 can be used as the template to grow a flower-shaped structure under the same subsequent treatment. Different from the dispersant used in the etching in the embodiment 1 and the embodiment 3, the nano cage-like structure cannot be obtained by successfully etching the ZIF-8 by using deionized water as the dispersant. The molar ratio of the etchant of the embodiment 1, the embodiment 4 and the embodiment 5 to the ZIF-8 is different, and the molar ratio of the etchant to the ZIF-8 can be obtained by a comparative experiment when the molar ratio of Ni: the Zn molar ratio is 5:1, the thickness of the nanometer cage shell layer is obviously increased, and the photo-thermal performance is deteriorated. The hydrothermal processes of example 1, example 6 and example 7 are different, and the hydrothermal temperature plays a decisive role in the etching process, while the hydrothermal time has little influence. The atmosphere of phosphating in example 1 and example 8 was different and had a small influence on the material. The different amounts of chlorhexidine addition for example 1 and example 9 mainly affected the drug loading in the final nanocapsule. The addition amount is large, and the content of chlorhexidine in the nano capsule is increased.
Experimental example 1
And (3) testing the application of the chlorhexidine loaded nickel phosphide nano-capsule coated by the bovine serum albumin as an antibacterial agent.
Fig. 1 is an SEM image of the bovine serum albumin-coated chlorhexidine-loaded nickel phosphate nanocapsule prepared in example 1. It can be seen that the antimicrobial agent was prepared with an organic coating on the outer layer.
Fig. 2 is an XRD image of the bovine serum albumin-coated chlorhexidine-loaded nickel phosphide nanocapsule prepared in example 1. As can be seen from FIG. 2, the prepared antibacterial agent has X-ray diffraction peaks corresponding to those of standard PDF cards one-to-one, and has an envelope at a low angle, indicating the presence of organic matter.
Fig. 3 is the FT-IR spectrum of the bovine serum albumin coated chlorhexidine loaded nickel phosphate nanocapsule prepared in example 1. As can be seen from FIG. 3, the prepared antibacterial agent had a peak of chlorhexidine, of which 2700-3400cm -1 Broad peaks in the range, due to N-H stretching vibrations. And at 1600 and 1100cm -1 The band in between is the characteristic peak of CHX, related to the stretching vibration of C = N and C-N-C bonds.
FIG. 4 is a UV-Vis spectrum of the BSA-loaded nickel phosphide nanocapsule prepared in example 1. It can be seen that a peak appears at the 255nm absorption peak, corresponding to the presence of chlorhexidine.
Fig. 5 is a graph comparing the antibacterial performance of the bovine serum albumin coated chlorhexidine loaded nickel phosphate nanocapsule prepared in example 1. The process uses pseudomonas aeruginosa (CMCC (B) 10104) and methicillin resistant staphylococcus aureus (ATCC 43300) as typical bacterial models to study the antibacterial effect. By applying a coating plateThe antibacterial performance of the samples was evaluated by counting. Specifically, the bacteria were cultured in Luria-Bertani (LB) and brain-heart infusion (BHI) culture solutions at 37 ℃ for 24 hours, respectively. The activated bacterial suspension (1.0X 10) 8 CFU/mL) was diluted ten-fold for use. Adding 50 μ L of the prepared antibacterial agent solution into 950 μ L of the bacterial suspension, and then using 808nm laser (1.0W cm) -2 ) Standing and culturing for 15min with or without illumination. Finally, 250. Mu.L of the treated suspension was added to 5mL of PBS buffer, 50. Mu.L of the suspension was spread on an agar plate, incubated at 37 ℃ for 24 hours, and the number of colonies was quantitatively counted. It can be seen that the antibacterial performance of the nickel phosphide-loaded chlorhexidine nano-capsule coated by the bovine serum albumin is superior to that of the single chlorhexidine and Ni 2 P nanometer cage.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A preparation method of a bovine serum albumin coated chlorhexidine loaded nickel phosphide nanocapsule is characterized by comprising the following steps:
metal organic frame ZIF-8 and NiCl 2 ·6H 2 Mixing O in solvent, hydrothermal reacting to obtain Ni (OH) 2 A nanocage;
mixing the Ni (OH) 2 Phosphorizing the nano cage to obtain Ni 2 A P nanocage;
adding the Ni 2 And (3) loading chlorhexidine on the P nano cage, and then coating by adopting bovine serum albumin to obtain the nano-composite material.
2. The method of claim 1, wherein the NiCl is in the form of NiCl-coated chlorhexidine loaded nickel phosphate nanocapsules 2 ·6H 2 The molar ratio of O to the metal organic framework ZIF-8 is 1.5-10; preferably 2 to 3.
3. The method for preparing the bovine serum albumin coated chlorhexidine supported nickel phosphide nano-capsule according to claim 1, wherein the hydrothermal reaction is carried out for 1-2h at 100-120 ℃.
4. The method for preparing the bovine serum albumin-coated chlorhexidine-loaded nickel phosphate nanocapsule of claim 1, wherein the solvent of the hydrothermal reaction is absolute methanol or deionized water.
5. The preparation method of the bovine serum albumin coated chlorhexidine loaded nickel phosphide nano-capsule of claim 1, wherein the phosphorization adopts a chemical vapor deposition method, and preferably, the specific steps of the phosphorization are as follows: mixing Ni (OH) 2 Nanocage and NaH 2 PO 2 ·H 2 Placing O at the downstream and the upstream of the tube furnace respectively, raising the temperature to 300-350 ℃ under the protection of inert gas, and then preserving the heat for 3-3.5 h; cooling, washing and drying to obtain the product.
6. The method for preparing the bovine serum albumin coated chlorhexidine loaded nickel phosphate nano-capsule according to claim 1, wherein the specific steps of the chlorhexidine loading are as follows: mixing Ni 2 And uniformly mixing the water solution of the P nano cage with the chlorhexidine solution, and stirring for 24-32 hours to obtain the compound.
7. The method of claim 1, wherein chlorhexidine-loaded Ni-phosphate nanocapsules are loaded with chlorhexidine-loaded Ni 2 Adding BSA into the P nano cage solution, uniformly mixing, stirring for 12-16 h, and carrying out solid-liquid separation to obtain the product.
8. The bovine serum albumin-coated chlorhexidine-loaded nickel phosphate nanocapsule prepared by the method of any one of claims 1-7.
9. The use of the bovine serum albumin coated chlorhexidine loaded nickel phosphate nanocapsule of claim 8 in the antibacterial field.
10. The use of claim 9, wherein the bovine serum albumin-coated chlorhexidine loaded nickel phosphate nanocapsule is sterilized under near infrared light.
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