CN110732039A - antibacterial peptide bionic calcium phosphate coating and preparation method and application thereof - Google Patents

antibacterial peptide bionic calcium phosphate coating and preparation method and application thereof Download PDF

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CN110732039A
CN110732039A CN201910910453.0A CN201910910453A CN110732039A CN 110732039 A CN110732039 A CN 110732039A CN 201910910453 A CN201910910453 A CN 201910910453A CN 110732039 A CN110732039 A CN 110732039A
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calcium phosphate
phosphate coating
bionic
solution
antibacterial peptide
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刘宁
朱勇
刘昌奎
郭芳
黄硕
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Xian Medical University
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Xian Medical University
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    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/28—Materials for coating prostheses
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    • A61L27/28—Materials for coating prostheses
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    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
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    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
    • A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
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    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
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    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/60—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a special physical form
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    • A61L2400/00—Materials characterised by their function or physical properties
    • A61L2400/18—Modification of implant surfaces in order to improve biocompatibility, cell growth, fixation of biomolecules, e.g. plasma treatment
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    • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
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    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/12—Materials or treatment for tissue regeneration for dental implants or prostheses

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Abstract

The invention discloses antibacterial peptide bionic calcium phosphate coatings, which comprise an amorphous calcium phosphate coating and a crystalline calcium phosphate coating containing antibacterial peptide, wherein the amorphous calcium phosphate coating and the crystalline calcium phosphate coating containing the antibacterial peptide are sequentially coated on the surface layer of a material to be coated from inside to outside.

Description

antibacterial peptide bionic calcium phosphate coating and preparation method and application thereof
Technical Field
The invention belongs to the technical field of medicines, and relates to antibacterial peptide bionic calcium phosphate coatings, a preparation method of the coatings, and application of the antibacterial peptide bionic calcium phosphate coatings.
Background
The method comprises the following steps of carrying out a biomimetic calcium phosphate coating technology to load antibacterial peptide on the surface of the bone substitute material, wherein the antibacterial peptide is which is an important measure for improving the defects, the antibacterial peptide has a mineralization spectrum , has no tissue cell toxicity, is not dependent on the metabolic activity of microorganisms, is not easy to generate drug resistance by bacteria, can still exert antibacterial effect on the surface of a biological material, and the like.
Disclosure of Invention
The invention aims to provide antibacterial peptide bionic calcium phosphate coatings, and solves the problem that a bone substitute material lacks antibacterial performance in the existing oral implant treatment process.
It is another object of the present invention to provide a method for preparing the above coating.
A third object of the invention is to provide the use of the above-mentioned coating.
The technical scheme adopted by the invention is that antibacterial peptide bionic calcium phosphate coatings comprise an amorphous calcium phosphate coating and a crystalline calcium phosphate coating containing antibacterial peptide, wherein the amorphous calcium phosphate coating and the crystalline calcium phosphate coating containing antibacterial peptide are sequentially coated on the surface layer of a material to be coated from inside to outside.
The present invention is also characterized in that,
the amorphous calcium phosphate coating is formed by 650-750 mM NaCl, 15-25 mM NaHCO3，2～7mM Na2HPO4·2H2O，5～10mM MgCl2·2H2O，10～15mM CaCl2·2H2And (4) dipping and coating the solution obtained by the O reaction on the surface layer of the material to be coated.
The crystalline calcium phosphate coating containing antibacterial peptide is made of 1-5 mM Na2HPO4·2H2O，1～5mM HCl，3～7mMCaCl2·2H2And mixing the solution obtained by the O reaction with the antibacterial peptide, and then dipping and coating the surface layer of the material to be coated.
The adopted technical scheme of is that the preparation method of the antibacterial peptide bionic calcium phosphate coating is implemented according to the following steps:
the dosage of each substance in the bionic calcium phosphate coating A liquid is 650-750 mM NaCl and 15-25 mM NaHCO3，2～7mM Na2HPO4·2H2O，5～10mM MgCl2·2H2O，10～15mM CaCl2·2H2O。
The dosage of each substance in the bionic calcium phosphate coating B liquid is 1-5 mM Na2HPO4·2H2O，1～5mM HCl，3～7mM CaCl2·2H2O。
Adjusting the pH value of the primary adjusting solution in the bionic calcium phosphate coating layer A to 2-2.5 by adopting 36-38% HCl and tris (hydroxymethyl) aminomethane; regulating the pH value of the solution again to 6-6.5 by adopting trihydroxymethyl aminomethane;
and (3) regulating the pH value of the primary regulating solution in the bionic calcium phosphate coating B solution to 7-7.5 by adopting trihydroxymethyl aminomethane.
In the step 2, the antibacterial peptide is a cationic antibacterial peptide, and the mass-volume ratio of the antibacterial peptide to the bionic calcium phosphate coating B liquid is more than 10-3mg/mL。
The temperature of the soaking environment in the step 3 and the step 4 is 36.5-37.5 ℃, and the soaking time in the step 3 is 24-48 hours; the soaking time in the step 4 is 48-72 hours;
the specific operation of freeze drying in step 3 and step 4 is as follows: precooling for 2-48 hours at the temperature below-40 ℃ in a freeze dryer, then keeping the temperature, and freeze-drying under the vacuum degree of 5 Pa.
The present invention is also characterized in that,
the third technical scheme adopted by the invention is that antibacterial peptide bionic calcium phosphate coatings are applied to oral implants or bone substitute materials.
The bionic calcium phosphate coating can be deposited under the conditions of physiological temperature and pH, the activity of bioactive protein cannot be damaged, and the formed crystal structure is similar to a bone structure, along with the formation of the calcium phosphate coating crystal, the antibacterial peptide is combined into the network crystal of the coating through codeposition and keeps activity, and the antibacterial peptide is slowly released under the physiological condition, so that the antibacterial material can kill bacteria remained in a bone regeneration area around the implant, does not influence the attachment, survival, growth and differentiation of cells, and has an application prospect of .
Drawings
FIG. 1 is a schematic diagram illustrating the formation of an amorphous calcium phosphate coating according to the present invention;
FIG. 2 is a schematic diagram showing the formation of a crystalline calcium phosphate coating containing an antibacterial peptide according to the present invention.
In the figure, 1, the material to be coated, 2, amorphous calcium phosphate coating, 3, crystalline calcium phosphate coating containing antibacterial peptide, and 4, antibacterial peptide.
Detailed Description
The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.
The invention provides antibacterial peptide bionic calcium phosphate coatings, which comprise an amorphous calcium phosphate coating and a crystalline calcium phosphate coating containing antibacterial peptides, wherein the amorphous calcium phosphate coating and the crystalline calcium phosphate coating containing the antibacterial peptides are sequentially coated on the surface layer of a material to be coated from inside to outside.
The preparation method of the antibacterial peptide bionic calcium phosphate coating is implemented according to the following steps:
step 1, preparation of a bionic calcium phosphate coating solution:
firstly, slowly adding 650-750 mM NaCl into deionized water in sequence; 15-25 mM NaHCO3；2～7mMNa2HPO4·2H2O；5～10mM MgCl2·2H2O, slowly adding HCl (the mass fraction of HCl is 36-38%) and TRIS (hydroxymethyl) aminomethane (TRIS base) into the solution to jointly adjust the pH value of the solution, adjusting the pH value of the solution to 2-2.5, and slowly adding 10-15 mM CaCl2·2H2And O, finally adding TRIS base to adjust the pH value of the solution to 6-6.5, and filtering the solution in a 0.2-micrometer filter membrane ultra-clean workbench after constant volume to prepare the solution.
Step 2, preparation of bionic calcium phosphate coating liquid:
slowly adding 1-5 mM Na into deionized water in sequence2HPO4·2H2O; 1-5 mM HCl (the mass fraction of HCl is 36-38%); 3 to 7mM CaCl2·2H2O; and adding TRIS base to adjust the pH value of the solution to 7-7.5, and filtering the solution in a 0.2-micron filter membrane ultra-clean workbench after constant volume to prepare the solution.
Step 3, preparation of the antibacterial peptide bionic calcium phosphate coating liquid:
according to the mass-volume ratio of the antibacterial peptide to the bionic calcium phosphate coating liquid is more than 10-3And (2) mg/mL, adding the cationic antibacterial peptide into the bionic calcium phosphate coating liquid obtained in the step (2) to obtain the bionic calcium phosphate coating liquid containing the antibacterial peptide.
And 4, forming an amorphous calcium phosphate coating:
as shown in figure 1, a material 1 to be coated is soaked in a bionic calcium phosphate coating liquid for 24-48 hours at the temperature of 36.5-37.5 ℃, an amorphous calcium phosphate coating 2 is formed on the surface layer of the material 1 to be coated, and then freeze drying is carried out, specifically, precooling is carried out for 2-48 hours at the temperature below-40 ℃ in a freeze dryer, then the temperature is kept, and freeze drying is carried out at the vacuum degree below 5 Pa.
And 5, forming a crystalline calcium phosphate coating containing antibacterial peptide:
as shown in fig. 2, under the condition of 36.5-37.5 ℃, the material 1 to be coated with the amorphous calcium phosphate coating 2 is soaked in the antibacterial peptide bionic calcium phosphate coating liquid for 48-72 hours to form a crystalline calcium phosphate coating 3 containing the antibacterial peptide 4, and then freeze-dried, specifically: precooling for 2-48 hours at the temperature below-40 ℃ in a freeze dryer, then keeping the temperature, and freeze-drying under the vacuum degree of 5 Pa.
The antibacterial peptide bionic calcium phosphate coating is prepared and formed under the physiological condition of 35-38 ℃, high-temperature and high-heat treatment is not involved, the energy consumption is low, the activity of bioactive protein is not damaged, the preparation process is simple and easy to implement, secondly, the whole process is carried out in aqueous solution, the surface modification of materials with irregular shapes and porous surfaces is easy to implement, and the bionic coating with uniform thickness is formed.
Example 1
Slowly adding 650mM NaCl into deionized water in sequence; 15mM NaHCO3；2mM Na2HPO4·2H2O；5mMMgCl2·2H2O, slowly adding HCl with the mass fraction of 36-38% and TRIS base into the solution to jointly adjust the pH value of the solution, slowly adjusting the pH value of the solution to 2 by matching the HCl and the TRIS base, and slowly adding 10mM CaCl2·2H2And O, finally adding TRISbase to adjust the pH value of the solution to 6, and filtering the solution in a 0.2-micron filter membrane superclean bench after constant volume to prepare the solution to obtain the bionic calcium phosphate coating solution.
Slowly adding 1mM Na into deionized water in sequence2HPO4·2H2O; 1mM HCl with the mass fraction of 36-38%; 3mM CaCl2·2H2O; adding TRIS base to adjust the pH value of the solution to 7, and filtering the solution in a 0.2-micrometer filter membrane superclean workbench after constant volume to prepare the bionic calcium phosphate coating liquid.
Antibacterial peptide GL13K (amino acid sequence: GKIIKLKASLKLL-CONH)2) Adding the mixture into the bionic calcium phosphate coating liquid to prepare the bionic calcium phosphate coating liquid containing 5mM GL 13K.
Soaking the deproteinized bovine bone inorganic material in the bionic calcium phosphate coating liquid for 24 hours, precooling for 2 hours at the temperature below-40 ℃ in a freeze dryer, then keeping the temperature, freeze-drying at the vacuum degree below 5Pa, then soaking the deproteinized bovine bone inorganic material in the antibacterial peptide bionic calcium phosphate coating liquid for 48 hours, precooling for 2 hours at the temperature below-40 ℃ in the freeze dryer, then keeping the temperature, and freeze-drying at the vacuum degree below 5 Pa.
Example 2
Slowly adding 680mM NaCl into deionized water in sequence; 18mM NaHCO3；3mM Na2HPO4·2H2O；6mMMgCl2·2H2O, slowly adding HCl with the mass fraction of 36% -38% and TRIS base into the solution to jointly adjust the pH value of the solution, slowly adjusting the pH value of the solution to 2.2 by matching the HCl and the TRIS base, and slowly adding 11mM CaCl2·2H2And O, finally adding TRIS base to adjust the pH value of the solution to 6.2, and filtering the solution in a 0.2-micron filter membrane ultra-clean workbench after constant volume to prepare the solution so as to obtain the bionic calcium phosphate coating solution.
Slowly adding 2mM Na into deionized water in sequence2HPO4·2H2O; 2mM HCl with the mass fraction of 36-38%; 4mM CaCl2·2H2O; adding TRIS base to adjust the pH value of the solution to 7.2, and filtering the solution in a 0.2-micron filter membrane ultra-clean workbench after constant volume to prepare the bionic calcium phosphate coating liquid.
Antibacterial peptide RL (amino acid sequence: AcWRKLWRpGLKRWLK-NH)2) Adding into bionic calcium phosphate coating liquid to prepare bionic calcium phosphate coating liquid containing 8mM RL。
Soaking the biological ceramic in the bionic calcium phosphate coating liquid for 30 hours, precooling for 12 hours at the temperature below-40 ℃ in a freeze dryer, then keeping the temperature, freeze-drying at the vacuum degree below 5Pa, then soaking in the antibacterial peptide bionic calcium phosphate coating liquid for 54 hours, precooling for 12 hours at the temperature below-40 ℃ in the freeze dryer, then keeping the temperature, and freeze-drying at the vacuum degree below 5 Pa.
Example 3
Slowly adding 700mM NaCl into deionized water in sequence; 20mM NaHCO3；5mM Na2HPO4·2H2O；7mMMgCl2·2H2O, slowly adding HCl with the mass fraction of 36-38% and TRIS base into the solution to jointly adjust the pH value of the solution, slowly adjusting the pH value of the solution to 2.3 by matching the HCl and the TRIS base, and slowly adding 12mM CaCl2·2H2And O, finally adding TRIS base to adjust the pH value of the solution to 6.3, and filtering the solution in a 0.2-micron filter membrane ultra-clean workbench after constant volume to prepare the solution so as to obtain the bionic calcium phosphate coating solution.
Slowly adding 3mM Na into deionized water in sequence2HPO4·2H2O; 3mM HCl with the mass fraction of 36-38%; 5mM CaCl2·2H2O; adding TRIS base to adjust the pH value of the solution to 7.3, and filtering the solution in a 0.2-micron filter membrane ultra-clean workbench after constant volume to prepare the bionic calcium phosphate coating liquid.
Antibacterial peptide Dhvar-5 (amino acid sequence: LLLFLLKKRKKRKY) is added into the bionic calcium phosphate coating liquid to prepare the bionic calcium phosphate coating liquid containing 7mM Dhvar-5.
Soaking the polylactic acid-glycolic acid copolymer in the bionic calcium phosphate coating liquid for 36 hours, precooling for 24 hours at the temperature below-40 ℃ in a freeze dryer, then keeping the temperature, freeze-drying at the vacuum degree below 5Pa, then soaking in the antibacterial peptide bionic calcium phosphate coating liquid for 60 hours, precooling for 24 hours at the temperature below-40 ℃ in the freeze dryer, then keeping the temperature, and freeze-drying at the vacuum degree below 5 Pa.
Example 4
Slowly adding 730mM NaCl to the deionized water in sequence; 23mM NaHCO3；6mM Na2HPO4·2H2O；8mMMgCl2·2H2O, slowly adding HCl with the mass fraction of 36-38% and TRIS base into the solution to jointly adjust the pH value of the solution, slowly adjusting the pH value of the solution to 2.4 by matching the HCl and the TRIS base, and slowly adding 13mM CaCl2·2H2And O, finally adding TRIS base to adjust the pH value of the solution to 6.4, and filtering the solution in a 0.2-micron filter membrane ultra-clean workbench after constant volume to prepare the solution so as to obtain the bionic calcium phosphate coating solution.
Slowly adding 4mM Na into deionized water in sequence2HPO4·2H2O; 4mM HCl with the mass fraction of 36-38%; 6mM CaCl2·2H2O; adding TRIS base to adjust the pH value of the solution to 7.4, and filtering the solution in a 0.2-micron filter membrane ultra-clean workbench after constant volume to prepare the bionic calcium phosphate coating liquid.
Antibacterial peptide LL37 (amino acid sequence:
LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES) is added into the bionic calcium phosphate coating liquid to prepare the bionic calcium phosphate coating liquid containing 4mM LL 37.
Soaking collagen in the bionic calcium phosphate coating solution for 40 hr, pre-cooling in a freeze drier at-40 deg.c for 36 hr, maintaining the temperature, freeze drying at vacuum degree below 5Pa, soaking in the antibacterial peptide bionic calcium phosphate coating solution for 66 hr, pre-cooling in a freeze drier at-40 deg.c for 36 hr, maintaining the temperature, and freeze drying at vacuum degree below 5 Pa.
Example 5
Slowly adding 750mM NaCl into deionized water in sequence; 25mM NaHCO3；7mM Na2HPO4·2H2O；10mM MgCl2·2H2O, slowly adding HCl with the mass fraction of 36-38% and TRIS base into the solution to jointly adjust the pH value of the solution, slowly adjusting the pH value of the solution to 2.5 by matching the HCl and the TRIS base, and slowly adding 15mM CaCl2·2H2And O, finally adding TRIS base to adjust the pH value of the solution to 6.5, and filtering the solution in a 0.2-micron filter membrane ultra-clean workbench after constant volume to prepare the solution so as to obtain the bionic calcium phosphate coating solution.
Slowly adding 5mM Na into deionized water in sequence2HPO4·2H2O; 5mM HCl with the mass fraction of 36-38%; 7mM CaCl2·2H2O; adding TRIS base to adjust the pH value of the solution to 7.5, and filtering the solution in a 0.2-micron filter membrane ultra-clean workbench after constant volume to prepare the bionic calcium phosphate coating liquid.
The antibacterial peptide KSL (amino acid sequence: KKVVFKVKFK-NH)2) Adding the bionic calcium phosphate coating liquid into the bionic calcium phosphate coating liquid to prepare the bionic calcium phosphate coating liquid containing 6mM KSL.
Soaking bone cement in the bionic calcium phosphate coating liquid for 48 hours, precooling for 48 hours at the temperature below-40 ℃ in a freeze dryer, then keeping the temperature, freeze-drying at the vacuum degree below 5Pa, then soaking in the antibacterial peptide bionic calcium phosphate coating liquid for 72 hours, precooling for 48 hours at the temperature below-40 ℃ in the freeze dryer, then keeping the temperature, and freeze-drying at the vacuum degree below 5 Pa.

Claims (10)

  1. The antibacterial peptide bionic calcium phosphate coating is characterized by comprising an amorphous calcium phosphate coating and a crystalline calcium phosphate coating containing antibacterial peptides, wherein the amorphous calcium phosphate coating and the crystalline calcium phosphate coating containing the antibacterial peptides are sequentially coated on the surface layer of a material to be coated from inside to outside.
  2. 2. The biomimetic calcium phosphate coatings according to claim 1, wherein the amorphous calcium phosphate coating is prepared from 650-750 mM NaCl, 15-25 mM NaHCO3，2～7mM Na2HPO4·2H2O，5～10mM MgCl2·2H2O，10～15mM CaCl2·2H2And (4) dipping and coating the solution obtained by the O reaction on the surface layer of the material to be coated.
  3. 3. The biomimetic calcium phosphate coatings according to claim 1, wherein the crystalline calcium phosphate coating containing antibacterial peptide is composed of 1-5 mM Na2HPO4·2H2O，1～5mM HCl，3～7mM CaCl2·2H2Mixing the solution obtained by the O reaction with the antibacterial peptideThe surface layer of the coating material is obtained by dip coating.
  4. The preparation method of the antibacterial peptide bionic calcium phosphate coating is characterized by comprising the following steps:
    step 1, preparing bionic calcium phosphate coating A liquid and bionic calcium phosphate coating B liquid:
    slowly adding NaCl and NaHCO into deionized water in sequence3、Na2HPO4·2H2O、MgCl2·2H2O, then adjusting the pH of the solution for the first time, and slowly adding CaCl2·2H2O, finally, regulating the pH value of the solution again to obtain bionic calcium phosphate coating A liquid;
    slowly adding Na into deionized water in sequence2HPO4·2H2O、HCl、CaCl2·2H2Regulating the pH value of the solution by O to obtain a bionic calcium phosphate coating B solution;
    step 2, preparing the antibacterial peptide bionic calcium phosphate coating liquid:
    adding antibacterial peptide into the bionic calcium phosphate coating B liquid obtained in the step 1 to obtain bionic calcium phosphate coating liquid containing the antibacterial peptide;
    step 3, forming an amorphous calcium phosphate coating:
    soaking the material to be coated in the bionic calcium phosphate coating A solution, taking out, and freeze-drying to form an amorphous calcium phosphate coating on the surface of the material to be coated;
    step 4, forming a crystalline calcium phosphate coating containing antibacterial peptide:
    and (3) soaking the material to be coated with the amorphous calcium phosphate coating obtained in the step (3) in the antibacterial peptide bionic calcium phosphate coating liquid, taking out, and freeze-drying to form the crystalline calcium phosphate coating containing the antibacterial peptide on the amorphous calcium phosphate coating.
  5. 5. The method for preparing biomimetic calcium phosphate coatings according to claim 4, wherein the dosage of each substance in the solution A of the biomimetic calcium phosphate coating is 650-750 mM NaCl and 15-25 mM NaHCO3，2～7mMNa2HPO4·2H2O，5～10mM MgCl2·2H2O，10～15mM CaCl2·2H2O。
  6. 6. The method for preparing biomimetic calcium phosphate coatings according to claim 4, wherein the amount of each substance in the solution B of the biomimetic calcium phosphate coating is 1-5 mM Na2HPO4·2H2O，1～5mM HCl，3～7mM CaCl2·2H2O。
  7. 7. The method for preparing bionic calcium phosphate coatings containing antibacterial peptides according to claim 4, wherein the pH of the primary adjusting solution in the bionic calcium phosphate coating A is adjusted to 2-2.5 by using 36-38% HCl and tris (hydroxymethyl) aminomethane, and the pH of the secondary adjusting solution is adjusted to 6-6.5 by using tris (hydroxymethyl) aminomethane;
    and (3) regulating the pH value of the primary regulating solution in the bionic calcium phosphate coating B solution to 7-7.5 by adopting trihydroxymethyl aminomethane.
  8. 8. The method for preparing antibacterial peptide bionic calcium phosphate coatings according to claim 4, wherein the antibacterial peptide in step 2 is a cationic antibacterial peptide, and the mass-to-volume ratio of the antibacterial peptide to the bionic calcium phosphate coating B liquid is greater than 10-3mg/mL。
  9. 9. The method for preparing biomimetic calcium phosphate coatings with antibacterial peptides according to claim 4, wherein the temperature of the immersion environment in step 3 and step 4 is 36.5-37.5 ℃, the immersion time in step 3 is 24-48 hours, and the immersion time in step 4 is 48-72 hours;
    the specific operation of freeze drying in step 3 and step 4 is as follows: precooling for 2-48 hours at the temperature below-40 ℃ in a freeze dryer, then keeping the temperature, and freeze-drying under the vacuum degree of 5 Pa.
  10. 10. The use of biomimetic calcium phosphate coatings according to claim 1, wherein the biomimetic calcium phosphate coatings of antimicrobial peptides are applied on oral implants or bone substitute materials.
CN201910910453.0A 2019-09-25 2019-09-25 antibacterial peptide bionic calcium phosphate coating and preparation method and application thereof Pending CN110732039A (en)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112604028A (en) * 2020-10-19 2021-04-06 北京联袂义齿技术有限公司 Surface coating for implant and preparation method thereof
CN113304318A (en) * 2021-06-08 2021-08-27 同济大学 Amorphous-crystalline calcium phosphate composite material based on biomimetic mineralization and preparation method and application thereof
WO2022242413A1 (en) * 2021-05-19 2022-11-24 北京荷月顺畅生物科技有限公司 Bionic bone material and preparation method therefor
WO2022242414A1 (en) * 2021-05-19 2022-11-24 北京荷月顺畅生物科技有限公司 Anti-cancer bionic bone material, and pharmaceutical composition comprising bionic bone material and preparation method for pharmaceutical composition

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070213832A1 (en) * 2005-09-28 2007-09-13 Wen Hai B Surface treatments for calcium phosphate-based implants
CN101214394A (en) * 2008-01-10 2008-07-09 中国人民解放军第三军医大学第三附属医院 Bone plate with sustained-release antimicrobial peptide
US20100249900A1 (en) * 2009-03-26 2010-09-30 Biotronik Vi Patent Ag Drug eluting medical implant with porous surface
CN102695751A (en) * 2009-09-28 2012-09-26 组织再生医疗公司 Porous materials coated with calcium phosphate and methods of fabrication thereof
CN107530145A (en) * 2015-02-04 2018-01-02 古萨有限公司 Dental prosthesis surface utilizes the coating of the obvious coating of synthetized hydroxyapatite

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070213832A1 (en) * 2005-09-28 2007-09-13 Wen Hai B Surface treatments for calcium phosphate-based implants
CN101214394A (en) * 2008-01-10 2008-07-09 中国人民解放军第三军医大学第三附属医院 Bone plate with sustained-release antimicrobial peptide
US20100249900A1 (en) * 2009-03-26 2010-09-30 Biotronik Vi Patent Ag Drug eluting medical implant with porous surface
CN102695751A (en) * 2009-09-28 2012-09-26 组织再生医疗公司 Porous materials coated with calcium phosphate and methods of fabrication thereof
CN107530145A (en) * 2015-02-04 2018-01-02 古萨有限公司 Dental prosthesis surface utilizes the coating of the obvious coating of synthetized hydroxyapatite

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112604028A (en) * 2020-10-19 2021-04-06 北京联袂义齿技术有限公司 Surface coating for implant and preparation method thereof
CN112604028B (en) * 2020-10-19 2023-02-17 北京联袂义齿技术有限公司 Surface coating for implant and preparation method thereof
WO2022242413A1 (en) * 2021-05-19 2022-11-24 北京荷月顺畅生物科技有限公司 Bionic bone material and preparation method therefor
WO2022242414A1 (en) * 2021-05-19 2022-11-24 北京荷月顺畅生物科技有限公司 Anti-cancer bionic bone material, and pharmaceutical composition comprising bionic bone material and preparation method for pharmaceutical composition
CN115382016A (en) * 2021-05-19 2022-11-25 北京荷月顺畅生物科技有限公司 Bionic bone material for resisting cancer, medicinal composition containing bionic bone material and preparation method of medicinal composition
CN115382010A (en) * 2021-05-19 2022-11-25 北京荷月顺畅生物科技有限公司 Bionic bone material and preparation method thereof
CN115382016B (en) * 2021-05-19 2023-06-23 北京荷月顺畅生物科技有限公司 Bionic bone material for resisting cancer, medicinal composition containing bionic bone material and preparation method of medicinal composition
CN113304318A (en) * 2021-06-08 2021-08-27 同济大学 Amorphous-crystalline calcium phosphate composite material based on biomimetic mineralization and preparation method and application thereof

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