CN113491793A

CN113491793A - Line structure compound and preparation method and application thereof

Info

Publication number: CN113491793A
Application number: CN202010190511.XA
Authority: CN
Inventors: 刘宣勇; 欧阳丽萍; 陈宝会; 马小涵
Original assignee: Shanghai Institute of Ceramics of CAS
Current assignee: Shanghai Institute of Ceramics of CAS
Priority date: 2020-03-18
Filing date: 2020-03-18
Publication date: 2021-10-12

Abstract

The invention relates to a line structure compound and a preparation method and application thereof, wherein the line structure compound is a compound with a line structure formed by solid-phase reaction of chitosan and puerarin; preferably, the diameter of the wire structure composite is 60-200 nm.

Description

Line structure compound and preparation method and application thereof

Technical Field

The invention relates to a line structure compound and a preparation method and application thereof, in particular to a compound material which is obtained by taking chitosan and puerarin as raw materials through a solid-phase synthesis method and has the functions of antibiosis and osteogenesis promotion, belonging to the field of biological materials.

Background

Chitosan is also called chitosan, and is obtained by deacetylation of chitin widely existing in nature, and belongs to macromolecule straight-chain polysaccharide. Chitosan has a certain antibacterial property, but high concentrations of chitosan are toxic to cells. Puerarin is a Chinese medicinal component extracted from radix Puerariae, and has effects of eliminating endotoxin (LPS) and regulating immune response. However, puerarin has no antibacterial effect and is toxic to cells when used at high concentrations. LPS plays an important role in the formation of bacterial membranes and contributes to the proliferation of bacteria. LPS can promote macrophage differentiation to type 1, which is not favorable for tissue healing. In addition, LPS induces persistent high fever, causing a variety of complications.

Disclosure of Invention

In order to solve the problem of complications caused by LPS release after antibiosis in the prior art, the invention aims to provide a line structure compound (or antibacterial osteogenesis promoting compound) and a preparation method and application thereof, so as to fill the blank in the prior art.

In one aspect, the present invention provides a linear structure complex, which is a complex having a linear structure formed by a solid-phase reaction of chitosan and puerarin.

In the disclosure, the chitosan and puerarin are compounded to prepare the thread structure compound, so that not only bacteria can be killed, but also endotoxin can be eliminated, and the composite material with immunoregulation, antibiosis and osteogenesis can be obtained.

Preferably, the wire structure diameter of the wire structure composite is 60 to 200 nm.

Preferably, the mass ratio of chitosan to puerarin in the linear structure compound is (0.25-5): 1, preferably (0.5-2): 1.

in another aspect, the present invention provides a method for preparing the above wire structure composite, comprising:

(1) adding chitosan and puerarin into a reaction vessel, and adding an acid solution as a solvent to obtain a mixture;

(2) and carrying out solid phase reaction on the obtained mixture under the physical mixing action, adding deionized water or acid solution for dissolving, and finally carrying out freeze drying to obtain the wire structure compound.

In the invention, chitosan and puerarin are added into a reaction vessel, and an acid solution is used as a solvent to obtain a mixture. The mixture is subjected to physical external force generated by physical mixing action to increase the local temperature in the mixture, so that the chitosan and the puerarin undergo solid phase reaction, the solid phase reaction is essentially that the puerarin and the chitosan undergo chemical reaction at local high temperature provided by the external force, and the possible chemical reaction is that amino of the chitosan and hydroxyl of the puerarin undergo condensation reaction or undergo nucleophilic substitution reaction with carbonyl. Meanwhile, in the presence of an acid solution, a reaction product of puerarin and chitosan is separated out and crystallized to form a linear structure compound. And then adding deionized water or acid solution for dissolving to obtain a mixed solution. And freeze-drying the obtained mixed solution to obtain the wire structure compound.

Preferably, the deacetylation degree of the chitosan is 85-100%; the puerarin has a purity of 90-100%.

Preferably, the solid-phase reaction is realized by external action, and the external physical mixing action comprises grinding, mechanical ball milling, stirring or mixing centrifugation;

the grinding time is 2-20 minutes;

the rotating speed of the mechanical ball milling is 60-600 revolutions per minute, and the time is 2-5 hours;

the stirring speed is 100-1000 r/min, and the stirring time is 2-5 hours;

the rotating speed of the mixing centrifugation is 50 g-12000 g of gravity acceleration, and the time is 2 minutes-20 minutes.

Preferably, the acid solution is a solution containing an organic acid and/or an inorganic acid; preferably, the organic acid is at least one of acetic acid, tartaric acid, oxalic acid, malic acid, citric acid, ascorbic acid, benzoic acid, salicylic acid and caffeic acid, and the inorganic acid is at least one of hydrofluoric acid, hydrochloric acid, nitric acid, sulfuric acid and phosphoric acid; more preferably, the concentration of the acid solution is 0.1-10M.

Preferably, the time of the solid phase reaction is 2 to 20 minutes. Wherein the diameter of the wireform complex increases as the time for complex synthesis increases.

Preferably, the drying mode is natural air drying, drying or freeze drying; the drying temperature is 40-80 ℃, and the drying time is 2-12 hours; the temperature of the freeze drying is-30 to-50 ℃, and the time is 5 to 48 hours.

In a third aspect, a biomedical material, comprising: a biomedical substrate and a coating which is loaded on the surface of the biomedical substrate and consists of the thread structure compound; preferably, the biomedical substrate is selected from medical gauze, medical dressing, implant titanium-based metal, anodized surface-treated titanium-based metal (abbreviated as anodized titanium-based metal), micro-arc oxidized surface-treated titanium-based metal (abbreviated as micro-arc oxidized titanium-based metal), polyetheretherketone PEEK, sulfonated surface-treated PEEK (abbreviated as sulfonated PEEK), or zirconia ceramic.

In a fourth aspect, the invention also provides the use of a composite of the thread structure described above in the preparation of an antimicrobial dressing and bone implant material.

Has the advantages that:

(1) in the invention, the prepared antibacterial bone-promoting compound can be loaded on the surface of a hard tissue implant, and in vitro experiments prove that the antibacterial bone-promoting compound has the following characteristics: good gram-negative bacteria and gram-positive bacteria resisting effects; promoting the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells; regulate macrophage phenotype and factor secretion. In vivo experiments show that the antibacterial osteogenesis promoting compound synthesized by the invention can effectively reduce high thermal reaction caused by endotoxin release and promote the regeneration of infected bones, and provides a bone implant material with antibacterial osteogenesis and postoperative complication reduction;

(2) the preparation process of the antibacterial osteogenesis-promoting compound is stable and controllable, simple to operate, low in cost, easy to implement and convenient to popularize and apply.

Drawings

FIG. 1 is a scanning electron micrograph of a lyophilized linear structure complex synthesized in example 1 using acetic acid as an acidic solvent, wherein (a) chitosan, (b) puerarin, and (c) chitosan complex puerarin lyophilized complex show no distinct structures in the single components of chitosan and puerarin, and the complex shows a distinct linear structure after the two are lyophilized by solid phase reaction, and the diameter of the linear structure is 60-200 nm;

FIG. 2 is a scanning electron micrograph of a lyophilized composite synthesized using HF acid (a), hydrochloric acid (b) and nitric acid (c) as solvents, from which it can be seen that various inorganic acids are used as solvents and that the lyophilized composite has a line structure despite its difference in morphology;

FIG. 3 is a comparison of antibacterial activity of chitosan, puerarin and air-dried compound obtained in example 5, wherein the bacterial colony of Escherichia coli cultured by culturing and re-transplanting the chitosan/puerarin line structure compound to an agar plate shows that the antibacterial activity of chitosan and puerarin is poor, and the line structure compound obtained by solid phase reaction of chitosan and puerarin has excellent Escherichia coli resistance;

FIG. 4 is a photograph showing the proliferation of bacteria in the line structure composite synthesized in example 1, which was loaded with gauze, (a) Escherichia coli, (b) Staphylococcus aureus, and it can be seen that, after being loaded with gauze, chitosan has a certain anti-Staphylococcus aureus ability but promotes the proliferation of Escherichia coli, compared with the blank control group; puerarin has no antibacterial ability; chitosan and puerarin with different mass ratios are compounded to show excellent antibacterial performance;

FIG. 5 is a photograph showing toxicity test of the content of the line-structured complex prepared in example 1 on mesenchymal stem cells, and it can be seen that the complex obtained by directly mixing chitosan and puerarin and performing solid phase reaction has good cell compatibility, and has almost no influence on cell proliferation at a lower content;

FIG. 6 is a scanning electron microscope photograph showing the loading of the bone implant material with the raw material and the linear structure composite, respectively, in example 7, wherein (a) sulfonated PEEK, (b) sulfonated PEEK loaded with chitosan, (C) sulfonated PEEK loaded with puerarin, and (d) sulfonated PEEK loaded with the antibacterial osteogenesis-promoting composite, it can be seen from the figure that the linear structure composite can be loaded into the three-dimensional pore structure of the sulfonated PEEK (PEEK: polyether ether ketone group; SP: sulfonated PEEK group; SPP: sulfonated PEEK loaded with puerarin; SPC: sulfonated PEEK loaded with chitosan; SPP @ C: sulfonated PEEK loaded with the linear structure composite;

fig. 7 shows the effect of the sulfonated PEEK loaded with the wire-structured composite prepared in example 7 against escherichia coli, and it can be seen that the sulfonated PEEK loaded with the wire-structured composite has an excellent bactericidal effect on escherichia coli: the cell membrane of the surface of the sulfonated PEEK loaded by the line structure in the scanning electron microscope picture is cracked; the plate coating result shows that the sulfonated PEEK loaded by the line structure has no obvious colony (PEEK: polyether ether ketone group; SP: sulfonated PEEK group; SPP: puerarin loaded sulfonated PEEK group; SPC: chitosan loaded sulfonated PEEK group; SPP @ C: sulfonated PEEK loaded by the line structure compound);

FIG. 8 shows the effect of sulfonated PEEK against Staphylococcus aureus of the load-line structured composite prepared in example 7, in which it can be seen that the line-structured load sulfonated PEEK has a certain inhibitory effect on Staphylococcus aureus (PEEK: polyether ether ketone group; SP: sulfonated PEEK group; SPP: puerarin load sulfonated PEEK group; SPC: chitosan load sulfonated PEEK group; SPP @ C: line-structured composite load sulfonated PEEK);

FIG. 9 shows the anti-Pseudomonas aeruginosa effect of the sulfonated PEEK loaded with the wire-structured composite prepared in example 7, from which it can be seen that the wire-structured composite has an excellent antibacterial effect against Pseudomonas aeruginosa (PEEK: polyether ether ketone group; SP: sulfonated PEEK group; SPP: puerarin loaded sulfonated PEEK group; SPC: chitosan loaded sulfonated PEEK group; SPP @ C: wire-structured composite loaded sulfonated PEEK);

FIG. 10 shows toxicity of sulfonated PEEK loaded with a linear structure complex prepared in example 7 on bone marrow mesenchymal stem cells (a) and alkaline phosphatase expression (b), from which it can be seen that the trance APEEK material loaded with a linear structure complex can promote proliferation of stem cells and expression of osteogenic differentiation index ALP (PEEK: polyether ether ketone group; SP: sulfonated PEEK group; SPP: puerarin loaded sulfonated PEEK group; SPC: chitosan loaded sulfonated PEEK group; SPP @ C: linear structure complex loaded sulfonated PEEK);

FIG. 11 is a graph of the modulation of macrophage phenotype (a) and cytokine modulation (b) by the thread structure complex prepared in example 7, CCR7 and TNF- α being surface specific proteins and cytokines of M1-type macrophages; CD206 and IL-10 are surface specific protein and cytokine of M2 type macrophage, and it is shown that the macrophage with line structure can promote the differentiation of macrophage to M2 type;

FIG. 12 shows LPS scavenging action of the wire structure complex prepared in example 7, from which it can be seen that LPS can be effectively scavenged by sulfonated PEEK loaded on the wire structure complex (PEEK: polyether ether ketone group; SP: sulfonated PEEK group; SPP: sulfonated PEEK loaded on puerarin group; SPC: sulfonated PEEK loaded on chitosan group; SPP @ C: sulfonated PEEK loaded on the wire structure complex);

FIG. 13 is a temperature measurement of rats implanted with the sulfonated PEEK loaded with the linear structure compound prepared in example 7, which shows that the sulfonated PEEK loaded with the linear structure compound can effectively lower the body temperature of mice in a osteomyelitis model (PEEK: polyetheretherketone group; SP: sulfonated PEEK group; SPP: puerarin loaded sulfonated PEEK group; SPC: chitosan loaded sulfonated PEEK group; SPP @ C: linear structure compound loaded sulfonated PEEK);

FIG. 14 is a graph showing the osteogenesis of sulfonated PEEK loaded with a wire-structured composite prepared in example 7 implanted in rats, in which the top row shows trichromatic fluorescence after implantation; the left side of the middle row is a quantitative structure of a three-color fluorescence pair, and the right side is BV/TV of a trabecula ossis; the bottom row is the micro-CT two-dimensional reconstruction result, and it can be further seen that the sulfonated PEEK loaded by the wire structure compound can promote the in-vivo new bone generation of the osteomyelitis model mouse. (PEEK: polyether ether ketone group; SP: sulfonated PEEK group; SPP: puerarin-loaded sulfonated PEEK group; SPC: chitosan-loaded sulfonated PEEK group; SPP @ C: wire structure complex-loaded sulfonated PEEK);

FIG. 15 is a scanning electron microscope image of the wire structure composite prepared in example 8, wherein (a) grinding is performed for 0min, (b) grinding is performed for 5min, (c) grinding is performed for 20min, (d) 60 ℃ heating is performed for 1h after 5min grinding, and (e) 100 ℃ heating is performed for 1h after 5min grinding, and it can be seen from the figure that the wire structure is more obvious as the grinding time is prolonged, and the number and the wire diameter are increased, and the wire structure is damaged by heating;

fig. 16 is a graph showing the bacterial growth of the wire-structured composite prepared in example 8 after adding the bacterial solution, wherein (a) escherichia coli, (b) staphylococcus aureus, and the abscissa "1-6" corresponds to "control", "5 min", "0 min", "20 min", "5 min-60", "5 min-100" in sequence, as can be seen from fig. 16, after the composite is ground for 5min, the composite has intentional antibacterial activity against both bacteria, and the antibacterial activity is poor after grinding for 0min, and after grinding for 20min, the composite loses antibacterial activity, which may be related to the local high temperature damage of material components and performance during grinding for an excessively long time. After grinding for 5min and heating (60 ℃ and 100 ℃), the material also has excellent antibacterial ability.

Detailed Description

The present invention is further illustrated by the following examples, which are to be understood as merely illustrative and not restrictive.

In the present disclosure, chitosan and puerarin are used as raw materials, and a thread structure compound having antibacterial and bone-promoting effects is obtained by a solid phase synthesis method. Wherein the compound has a linear structure, and a large number of hydrogen bonds are contained in the compound system. In an alternative embodiment, the wire structure in the wire structure composite may have a diameter of 60-200 nm.

In one embodiment of the invention, chitosan and puerarin are used as raw materials, acid solution is used as a solvent, and solid-phase synthesis is realized through external physical mixing, so as to obtain the linear structure compound.

In an alternative embodiment, the degree of deacetylation of chitosan may be 85 to 100%. The purity of the puerarin used can be 90-100%. In the raw materials, the mass ratio of chitosan to puerarin is 0.25-5, preferably 0.5-2.

In alternative embodiments, the solvent used for solid phase synthesis is a diluted mineral acid, a diluted organic acid, or a mixed solution of a diluted mineral acid and an organic acid. The inorganic acid may be at least one of hydrofluoric acid, hydrochloric acid, sulfuric acid, nitric acid, and the like. The organic acid can be at least one of acetic acid, tartaric acid, oxalic acid, malic acid, citric acid, ascorbic acid, benzoic acid, salicylic acid, caffeic acid, etc. The concentration of the acid solution used can be 0.1-10M. Adding acid solution and chitosan at a ratio of (100-300) mu L: (25-100) mg, and the compound has shear thinning performance.

In alternative embodiments, the external physical mixing action may be a solid phase milling process (i.e., milling), mechanical ball milling, agitation, and mixing centrifugation. The reason why the color of the online structure compound changes gradually from light white to milky white after being synthesized is that the compound gradually precipitates and crystallizes in the reaction process to generate a series of color changes. The time for physical mixing can be from 2 minutes to 5 hours. For example, the grinding time may be 2 to 20 minutes. The rotating speed of the mechanical ball milling can be 60-600 revolutions per minute, and the time can be 2-5 hours. The rotating speed of the stirring can be 100-1200 r/min, and the time can be 2-5 hours. The rotating speed of the mixing centrifugation is 50-12000 g of gravitational acceleration, and the time can be 2-20 minutes. Moreover, the antibacterial effect and the bioactivity of the obtained wire structure compound are reduced due to the overlarge acting force or/and the overlong acting time of the external physical mixing action, mainly because the reaction cannot be normally carried out due to the overhigh local temperature generated by the overlarge external force of the obtained wire structure under the action of the external force, and the obtained compound loses the bioactivity.

After the solid phase reaction is finished, deionized water or the acid solution is added to dissolve the obtained high molecular structure compound to obtain a mixed solution.

And drying the mixed solution to obtain the wire structure compound. In alternative embodiments, the drying may be natural air drying, oven drying, or freeze drying. Wherein, the drying temperature can be 40-80 ℃ and the drying time can be 2-12 hours. The temperature of freeze drying can be-30 to-50 ℃, and the time can be 5 to 24 hours.

In addition, the resulting wireform composites can be renamed according to different drying modes. For example, the resulting wire structure composite after natural air drying may be designated as an air dried composite. The resulting wire structure composite after drying may be designated as a dried composite. The obtained line structure compound after freeze drying can be named as freeze-dried compound, freeze-dried sponge, etc.

In the present invention, the resulting thread structure complex can be directly applied to the body in vivo and on the body surface. For example, the thread structure compound (antibacterial bone-promoting compound) is further loaded or coated on the surface of a biomedical substrate such as an implant or gauze to form a biomedical coating, and the biomedical material (e.g., antibacterial dressing, bone implant, etc.) loaded with the thread structure compound is prepared. The surface of the biomedical coating can improve the antibacterial property and the osteogenesis property of the biomaterial. Methods for supporting the wire structure composite on the surface of the substrate include, but are not limited to, dipping, spin coating, spraying, direct coating, and the like.

The prepared antibacterial bone-promoting compound has good antibacterial effect, can be loaded on the surface of a biomedical substrate applied to the body surface or in the body (for example, the surfaces of materials such as medical gauze, medical dressing, implant titanium-based metal, anode oxidation, micro-arc oxidation and other surface-treated titanium-based metal, PEEK, sulfonated PEEK, zirconia ceramics and the like), meets the requirements of antibacterial and bone-promoting in different fields, is compounded by chitosan and traditional Chinese medicines, has good biological safety, and is expected to be clinically applied.

In the invention, the linear structure compound has a nanowire structure and good antibacterial performance, and can promote osteogenic differentiation of bone marrow mesenchymal stem cells and regulate the secretion of macrophage phenotype and related cytokines. For example, the obtained thread structure compound has obvious antibacterial effect on both gram-negative bacteria and gram-positive bacteria. The obtained thread structure compound can remove endotoxin released by bacteria. The obtained thread structure compound can promote macrophage to present M2 type, and promote macrophage to secrete anti-inflammatory factor. The mesenchymal stem cells can be rapidly differentiated to osteoblasts on the surface of the coating consisting of the linear structure compound, and show excellent osteogenic compatibility.

The present invention will be described in detail by way of examples. It is also to be understood that the following examples are illustrative of the present invention and are not to be construed as limiting the scope of the invention, and that certain insubstantial modifications and adaptations of the invention by those skilled in the art may be made in light of the above teachings. The specific process parameters and the like of the following examples are also only one example of suitable ranges, i.e., those skilled in the art can select the appropriate ranges through the description herein, and are not limited to the specific values exemplified below.

Example 1

Dripping 200 μ L0.875M acetic acid solution to wet 50mg chitosan in deionized water solution, adding 52mg puerarin (the mass ratio of chitosan to puerarin is 1: 1.04), grinding for 5min, and adding 10mL 0.175M acetic acid solution to dissolve the obtained antibacterial bone-promoting compound;

the obtained antibacterial osteogenesis promoting compound dissolved in 0.175M acetic acid is freeze-dried for 24 hours by a freeze-dryer (-40 ℃), and the freeze-dried sponge of the antibacterial osteogenesis promoting compound is obtained and marked as C @ P₁。

Example 2

Dripping 200 μ L2.85M HF acid solution wetting 50mg chitosan in deionized water solution, adding 52mg puerarin (the mass ratio of chitosan to puerarin is 1: 1.04), grinding for 5min, and adding 10ml 0.57M HF acid solution to dissolve the obtained antibacterial bone-promoting compound;

the obtained antibacterial osteogenesis promoting compound obtained by dissolving the obtained 0.57M HF acid is freeze-dried for 24 hours by a freeze dryer (-40 ℃), and the freeze-dried sponge of the antibacterial osteogenesis promoting compound is obtained and marked as C @ P₂。

Example 3

a) 200 mul of 1.632M hydrochloric acid is dripped into deionized water solution of 50mg of chitosan, and then 52mg of puerarin (the mass ratio of chitosan to puerarin is 1: 1.04), grinding for 5 minutes, then adding 10ml of 0.324M HCl acid to dissolve the obtained antibacterial bone-promoting complex;

b) the obtained antibacterial osteogenesis promoting compound obtained by dissolving the obtained 0.324M HCl acid is freeze-dried for 24 hours by a freeze dryer (-40 ℃), and the freeze-dried sponge of the antibacterial osteogenesis promoting compound is obtained and is marked as C @ P₃。

Example 4

Dripping 200 μ L of 1.127M nitric acid to wet 5mg/ml chitosan solution in deionized water, adding 5.2mg/ml puerarin (the mass ratio of chitosan to puerarin is 1: 1.04), grinding for 5min, and adding 0.225M acetic acid to dissolve the obtained antibacterial bone-promoting compound;

the obtained antibacterial osteogenesis promoting compound dissolved by the obtained 0.225M nitric acid is frozen and dried for 24 hours by a freezing dryer (-40 ℃), and the freeze-dried sponge of the antibacterial osteogenesis promoting compound is obtained and marked as C @ P₄。

Example 5

Dripping 200 μ L0.875M acetic acid to wet 50mg chitosan, adding 52mg puerarin (the mass ratio of chitosan to puerarin is 1: 1.04), grinding for 5min, and adding 10ml 0.175M acetic acid to dissolve the obtained antibacterial bone-promoting compound;

dripping the antibacterial osteogenesis promoting compound obtained by dissolving the obtained 0.175M acetic acid into a 24-hole plate, placing in a fume hood, and air-drying for 8 hours to obtain the wire structure compound marked as C @ P₅。

Example 6

a) 200 mul of 0.875M acetic acid is dripped to wet 50mg of chitosan, and then 5.2mg/ml of puerarin is added (the mass ratio of chitosan to puerarin is 1: 1.04), grinding for 5 minutes, then adding 0.175M acetic acid to dissolve the obtained antibacterial bone-promoting complex;

b) applying the obtained antibacterial bone promoting compound dissolved in 0.175M acetic acid on the surface of medical gauze, and drying to obtain antibacterial dressing marked as C @ P₆。

Example 7

Polishing the medical PEEK material, sequentially and respectively cleaning the medical PEEK material for three times by using ethanol, acetone and deionized water, sulfonating the medical PEEK material by using concentrated sulfuric acid, performing hydrothermal treatment at 120 ℃ for 4 hours, taking out the medical PEEK material, and naturally drying the medical PEEK material;

dripping 200 μ L of 0.875M acetic acid to wet 5mg/ml chitosan, adding 5.2mg/ml puerarin (the mass ratio of chitosan to puerarin is 1: 1.04), grinding for 5min, and adding 10ml of 0.175M acetic acid to dissolve the obtained antibacterial bone-promoting compound;

soaking sulfonated PEEK (labeled as PEEK) in 500 μ L of antibacterial osteogenesis promoting compound dissolved in 0.175M acetic acid in a 24-well plate, and reacting at 37.5 deg.C for 24 hr by using a shaker;

and taking out the sample, washing the sample twice by PBS, taking out the sample, and naturally airing the sample to obtain the sulfonated PEEK material loaded with the line structure compound. The sulfonated PEEK material loaded with chitosan and the sulfonated PEEK material loaded with puerarin are prepared by the method, and are marked as SPP: puerarin loaded sulfonated PEEK group; SPC: a sulfonated PEEK group loaded with chitosan; SPP @ C: sulfonated PEEK loaded with a wire-structured composite.

Example 8

a) 200 mul of 0.875M acetic acid is dripped into deionized water solution of 50mg of chitosan, and then 52mg of puerarin is added (the mass ratio of chitosan to puerarin is 1: 1.04), ground for 0min (i.e. no grinding), 5min and 20min, respectively, and then 10mL of 0.175M acetic acid was added to dissolve the obtained antibacterial-precipitating bone complex;

b) the obtained 0.175M acetic acid dissolved antibacterial bone-promoting compound was heated in an oven at 60 deg.C and 100 deg.C for 1 hr to obtain compound labels of 5min-60 and 5 min-100.

Claims

1. The line structure compound is characterized in that the line structure compound is a compound with a line structure formed by the solid-phase reaction of chitosan and puerarin; preferably, the diameter of the wire structure composite is 60-200 nm.

2. The linear structure compound of claim 1, wherein the mass ratio of chitosan to puerarin in the linear structure compound is (0.25-5): 1, preferably (0.5-2): 1.

3. a method of preparing a wire structure composite as claimed in claim 1 or 2, comprising:

4. The preparation method according to claim 3, wherein the degree of deacetylation of chitosan is 85-100%; the puerarin has a purity of 90-100%.

5. The method of claim 3 or 4, wherein the physical mixing is performed by a method comprising grinding, mechanical ball milling, stirring, or mixing centrifugation;

the grinding time is 2-20 minutes;

the stirring speed is 100-1000 r/min, and the stirring time is 2-5 hours;

6. The production method according to any one of claims 3 to 5, wherein the acid solution is a solution containing an organic acid and/or an inorganic acid; preferably, the organic acid is at least one of acetic acid, tartaric acid, oxalic acid, malic acid, citric acid, ascorbic acid, benzoic acid, salicylic acid and caffeic acid, and the inorganic acid is at least one of hydrofluoric acid, hydrochloric acid, nitric acid, sulfuric acid and phosphoric acid; more preferably, the concentration of the acid solution is 0.1-10M; most preferred is a 0.875M acetic acid solution.

7. The method according to any one of claims 3 to 6, wherein the time for the solid phase reaction is 0.05 to 24 hours.

8. The method according to any one of claims 3 to 7, wherein the drying is performed by natural air drying, oven drying, or freeze drying; the drying temperature is 40-80 ℃, and the drying time is 2-12 hours; the temperature of the freeze drying is-30 to-50 ℃, and the time is 5 to 48 hours.

9. A biomedical material, comprising: a biomedical substrate, and a coating layer composed of the wire structure composite of claim 1 or 2, which is supported on the surface of the biomedical substrate; preferably, the biomedical substrate is selected from the group consisting of medical gauze, medical dressings, implant titanium-based metal, anodized surface treated titanium-based metal, or micro-arc oxidized surface treated titanium-based metal, polyetheretherketone PEEK, sulfonated surface treated PEEK, or zirconia ceramic.

10. Use of a wire structural composite as claimed in claim 1 or 2 for the preparation of an antimicrobial dressing and bone implant material.