CN111729132A - Polyether-ether-ketone bone repair material with antibacterial property and preparation method thereof - Google Patents

Abstract

The invention discloses a polyether-ether-ketone bone repair material with antibacterial property and a preparation method thereof₃C₂Compounding and delaying Ti₃C₂The combination of electrons and holes in the energy band increases the transferred electrons to form singlet oxygen, thereby solving the problem of Ti₃C₂The antibacterial effect is limited, thereby preventing the SPEEK implant from being infected with bacteria and inhibiting the generation of drug-resistant bacteria. In addition, the invention also provides polyetherThe ether ketone is placed in concentrated sulfuric acid for sulfonation to obtain sulfonated polyether ether ketone with rough and porous surface and sulfonic acid groups, and Ti is added₃C₂The contact area of the-CoNWs composite material and the SPEEK is increased, and the Ti content is improved₃C₂-the bonding force between the connws composite and SPEEK; sulfonic acid group and Ti of SPEEK surface₃C₂The surface hydroxyl groups react to form sulfonate esters to react SPEEK with Ti₃C₂Chemical bonds are also formed between the-CoNWs composite materials, so that Ti is further strengthened₃C₂The binding force between the CoNWs composite material and the SPEEK, avoiding Ti₃C₂The problem of the CoNWs composite material falling off from the SPEEK during use.

Description

Polyether-ether-ketone bone repair material with antibacterial property and preparation method thereof

Technical Field

The invention relates to the field of biological materials, in particular to a polyether-ether-ketone bone repair material with antibacterial property and a preparation method thereof.

Background

Because bone defects caused by various tumors, wounds, congenital malformations and other diseases are difficult to heal, how to improve bone defect healing is still a troublesome problem facing researchers and clinicians today.

The special engineering plastic Polyetheretherketone (PEEK) becomes a hard tissue repair material which has the most development prospect and can replace the traditional metal implant due to the excellent thermal stability, mechanical property, ray permeability and biocompatibility. However, PEEK implantation surgery inevitably involves bacterial infection during the surgical procedure and post-operative rehabilitation, and in order to avoid bacterial infection, antibiotics are usually used to prevent bacterial infection and reduce the incidence of inflammation. Antibiotics such as penicillin, streptomycin, trimethoprim, tetracycline and the like greatly reduce the morbidity and mortality of pathogenic bacterial infection diseases, and are widely applied clinically. However, the long-term abuse of antibiotics can make many pathogenic bacteria easily generate drug resistance to various antibiotics, which can lead to the long-term treatment of pathogen infection diseases, reduce the treatment effect and lead to the deterioration and even death of patients.

In recent years, optical technology has rapidly developed, and photosensitive materials can cooperate with photodynamic therapyMethods and photothermal therapies have found widespread use in the biomedical field. In particular of two-dimensional structure, of a semiconductor Ti₃C₂The material can be coated on the surface of PEEK, and is excited by laser to generate heat and singlet oxygen¹O₂) Can lead bacteria to rupture and die, has great potential in the treatment of bacterial infection, and can replace antibiotic treatment so as to solve the drug resistance problem of pathogenic bacteria. Ti₃C₂The antimicrobial properties and the ability of the coating to harm bacteria are largely dependent on the level of heat generated and the amount of singlet oxygen that it produces in a short time. However, Ti₃C₂The generation of singlet oxygen from the coating, although damaging the integrity of the bacterial membrane, is due to Ti₃C₂The coating layer itself is electron-transferred to generate singlet oxygen too little, so that the antibacterial effect is limited. Therefore, how to increase Ti₃C₂The singlet oxygen generated by self electron transfer solves the problem of Ti₃C₂The antibacterial effect is limited, so that the PEEK implant is prevented from being infected with bacteria, the generation of drug-resistant bacteria is inhibited, the normal bone repair is ensured, and the PEEK bone defect repair material becomes a main research direction of PEEK bone defect repair materials.

Disclosure of Invention

In order to overcome the defects that the polyether-ether-ketone bone defect repairing material has no antibacterial property and is easy to generate drug-resistant bacteria depending on antibiotic bacteriostasis, and the like, the invention aims to provide the polyether-ether-ketone bone repairing material with antibacterial property.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

the polyether-ether-ketone bone repair material with antibacterial performance comprises sulfonated polyether-ether-ketone with rough and porous surface and sulfonic acid groups, and Ti coated on the surface of the sulfonated polyether-ketone₃C₂-a CoNWs composite.

Preferably, the Ti₃C₂-the preparation of a CoNWs composite material comprises the following steps:

step S1: firstly, Ti₃C₂And CoNWs are respectively dispersed in deionized water to obtain Ti₃C₂Suspension and CoNWs suspension, then Ti₃C₂The suspension and the CoNWs suspension are mixed in equal volumes to obtain a mixed suspension, wherein Ti is₃C₂The molar concentration ratio of the suspension to the CoNWs suspension is 1 (0.5-1);

step S2: performing ultrasonic treatment on the mixed solution obtained in the step S1 for 20-30min to obtain Ti₃C₂-a CoNWs composite.

Preferably, the Ti₃C₂Ti being in the form of a monolayer₃C₂。

Preferably, the preparation method of the sulfonated polyether ether ketone comprises the following steps:

firstly, the polyether-ether-ketone base material is placed in concentrated sulfuric acid and is subjected to ultrasonic treatment for 5-15min, and then the polyether-ether-ketone after ultrasonic treatment is washed by acetone and absolute ethyl alcohol to obtain the sulfonated polyether-ether-ketone.

Preferably, the single layer of Ti₃C₂The preparation method comprises the following steps:

firstly, adding LiF into hydrochloric acid solution with the molar concentration of 9-10mol/L, uniformly stirring, and then adding Ti into the solution added with LiF/HCl₃AlC₂Obtaining a mixture, sequentially stirring and reacting the mixture in an ice bath for 30min at 35 ℃ for 24-48h, and then washing the stirred and reacted mixture with deionized water to obtain Ti₃C₂Tx suspension, Ti to be obtained₃C₂The Tx suspension was centrifuged and washed to pH 5-6 to obtain Ti in a multi-layered form₃C₂Tx; obtaining a multilayer Ti₃C₂Re-dispersing Tx in deionized water to obtain Ti₃C₂Tx dispersion, placing the dispersion in ice bath for ultrasonic treatment for 1h, then centrifuging the dispersion after ultrasonic treatment and removing supernatant to obtain monolayer Ti₃C₂Nano-flake dispersed Ti in water₃C₂Suspension of Ti₃C₂Drying the suspension to obtain a single layer of Ti₃C₂。

Preferably, the LiF is mixed with Ti₃AlC₂The mass ratio of (1): (0.5-1).

Preferably, the method for preparing the CoNWs is as follows:

firstly, CoCl is added₂·6H₂Adding O and EDTA-2Na into deionized water according to the molar ratio of 1:1, uniformly stirring to obtain a raw material solution, adjusting the pH of the raw material solution to 13-14 to obtain a reaction solution, then placing the reaction solution in a magnetic field cage at 80 ℃ for reaction, and adding 600-650 mu L of N-doped sodium chloride into the reaction solution₂H_4·H₂O is stirred uniformly, and then 240-300 mu L of H is added into the reaction solution without stirring₂PtCl·6H₂And O, collecting the generated CoNWs, washing the generated CoNWs with deionized water and absolute ethyl alcohol in sequence, and finally performing vacuum freeze drying on the obtained product to obtain the CoNWs.

Preferably, the pH of the feedstock is adjusted with NaOH.

The invention also aims to provide a preparation method of the polyether-ether-ketone bone repair material with antibacterial property, which comprises the following steps:

step C1 first Ti₃C₂Dipping the CoNWs composite material on the surface of sulfonated polyether-ether-ketone, and drying the dipped sulfonated polyether-ether-ketone at the temperature of 50-60 ℃;

step C2, repeating the step C1 for 3-5 times, and finally obtaining the polyether-ether-ketone bone repair material Ti with antibacterial property₃C₂-CoNWs/SPEEK。

Compared with the prior art, the invention has the following beneficial effects:

(1) the invention selects the single-layer Ti with larger specific surface area₃C₂Ti formed by complexing with CoNWs₃C₂-CoNWs composite material, wherein the CoNWs metal material in the composite material is a one-dimensional geometrical structure with excellent electron mobility and is Ti₃C₂The photoelectron transfer provides a relatively straight and fast transmission channel, thereby enhancing the charge transfer and separation capability of electron-hole, and delaying Ti₃C₂The combination of electrons and holes in the energy band increases the electron transfer to increase the formation of singlet oxygen, and solves the problem of Ti₃C₂The antibacterial effect is limited, thereby preventing the SPEEK implant from being infected with bacteria, inhibiting the generation of drug-resistant bacteria and ensuring the bone repairAnd (5) repeating the normal operation. And, Ti₃C₂The photothermal produced by the-CoNWs composite material is also unaffected.

(2) The invention relates to sulfonated polyether ether ketone (SPEEK) with rough and porous surface and sulfonic acid groups, which is obtained by sulfonating polyether ether ketone in concentrated sulfuric acid. When suspended Ti₃C₂When the-CoNWs composite material is dipped on the SPEEK, Ti is added due to the rough and porous characteristics of the surface of the SPEEK₃C₂The contact area of the-CoNWs composite material and the SPEEK is increased, and the Ti content is improved₃C₂The binding force between the CoNWs composite material and the SPEEK, avoiding Ti₃C₂The CoNWs composite is exfoliated from SPEEK. In addition, since the sulfonic acid group on SPEEK can react with Ti₃C₂-Ti in CoNWs composite₃C₂Reacting the hydroxyl groups on the surface to form sulfonate ester, reacting SPEEK with Ti₃C₂Chemical bonds exist among the-CoNWs composite materials, so that Ti is further increased₃C₂The binding force between the CoNWs composite material and the SPEEK, avoiding Ti₃C₂The problem of the CoNWs composite material falling off from the SPEEK during use.

(3) Ti prepared by the invention₃C₂The CoNWs composite material is formed by retarding Ti₃C₂The combination of electrons and holes in the energy band increases Ti₃C₂The photoelectron transfer promotes the electron transfer, thereby increasing the formation of singlet oxygen and enabling Ti₃C₂the-CoNWs/SPEEK has antibacterial property, and solves the problem of drug resistance of germs caused by using antibiotics. In addition, all the materials in the invention have excellent biocompatibility, lower cytotoxicity and durable antibacterial performance, and can be used for coating on the surface of a medical implant, bone repair implant materials, in-vitro use of antibacterial auxiliary materials and the like.

(4) Ti prepared by the invention₃C₂The CoNWs composite material can generate singlet oxygen and can also generate photothermal action under the irradiation of near infrared light; the light and heat generated by the composite material can increase the permeability of the bacterial membrane, and the singlet oxygen utilizes the permeability of the bacterial membrane under the synergistic action of the light and heat and the photodynamicThe change of permeability penetrates through a bacterial membrane into bacteria to cause outbreak of bacteriostatic oxidation and severe penetration of bacterial protein, thereby killing the bacteria, and the Ti of the invention is realized₃C₂The antibacterial action of the CoNWs composite.

Drawings

FIG. 1 is a SEM image of a material of the present invention, wherein a is a material Ti₃C₂SEM image of/SPEEK, panel b is material Ti₃C₂SEM picture of CoNWs/SPEEK;

FIG. 2 is a graph showing the photothermal properties of the material of the present invention, wherein a is SPEEK and Ti₃C₂/SPEEK、Ti₃C₂-photoconversion thermogram of three materials CoNWs/SPEEK, panel b shows SPEEK and Ti₃C₂/SPEEK、Ti₃C₂-photothermal stability plots of three materials, CoNWs/SPEEK;

FIG. 3 is a singlet oxygen evolution diagram of the material of the present invention, wherein a is a diagram for detecting SPEEK and Ti by an electron spin resonance apparatus₃C₂/SPEEK、Ti₃C₂A singlet oxygen yield diagram of three materials of CoNWs/SPEEK, and a diagram b shows that 1, 3-diphenyl iso-furan is adopted as singlet oxygen (¹O₂) The labeling substance (2) detects SPEEK and Ti₃C₂/SPEEK、Ti₃C₂-singlet oxygen generation profile of three materials CoNWs/SPEEK;

FIG. 4 is a graph showing the bacteriostatic effect of the material of the present invention, wherein a is SPEEK and Ti₃C₂/SPEEK、Ti₃C₂The bacteriostatic effect of the-CoNWs/SPEEK three materials on staphylococcus aureus respectively, and the graph b shows SPEEK and Ti₃C₂/SPEEK、Ti₃C₂-CoNWs/SPEEK three materials respectively show the bacteriostatic effect on escherichia coli;

FIG. 5 shows SPEEK and Ti of the present invention₃C₂/SPEEK、Ti₃C₂Bacterial maps of Staphylococcus aureus, Escherichia coli on three materials CoNWs/SPEEK.

Wherein 1 in FIGS. 2 and 3 is material SPEEK, and 2 is material Ti₃C₂The material is Ti₃C₂-CoNWs/SPEEK。

Detailed Description

The technical solution of the present invention will be further clearly and completely described with reference to the following examples, wherein the raw materials used in the examples of the present invention are all commercially available.

Example 1

The preparation method of the polyether-ether-ketone bone repair material with the antibacterial property specifically comprises the following steps:

step 1: ti₃C₂Preparation of

Firstly, 1g of LiF (lithium fluoride) is added into 20mL of hydrochloric acid solution with the molar concentration of 9mol/L and stirred uniformly, and then 1g of Ti is added into the LiF/HCl solution₃AlC₂Obtaining a mixture, stirring and reacting the mixture in ice bath for 30min under the condition of 35 ℃ for 24h, and then washing the stirred and reacted mixture with deionized water to obtain Ti₃C₂Tx suspension, Ti to be obtained₃C₂The Tx suspension was centrifuged and washed to a pH of 6 to obtain Ti in a multi-layered form₃C₂Tx. Ti to be obtained₃C₂Re-dispersing Tx in deionized water to obtain Ti₃C₂Tx dispersion, then placing the dispersion in ice bath for ultrasonic treatment for 1h, then centrifuging the dispersion after ultrasonic treatment and removing supernatant fluid to obtain monolayer Ti₃C₂Nano-flake dispersed Ti in water₃C₂Suspension, finally Ti₃C₂Vacuum freeze drying the suspension to obtain single-layer Ti₃C₂。

Step 2: preparation of cobalt nanowires

Adding CoCl₂·6H₂Adding O and EDTA-2Na into 120mL of deionized water according to the molar ratio of 1:1, uniformly stirring to obtain a raw material solution, adding NaOH into the raw material solution to adjust the pH value of the raw material solution to 14 to obtain a reaction solution, placing the reaction solution in a water bath kettle covered by a magnetic field at 80 ℃ for reaction, and adding 600 mu L of N into the reaction solution₂H_4·H₂O is stirred uniformly and then added into the reaction solution without stirringAdd 240. mu.L of H₂PtCl·6H₂And O, collecting the generated CoNWs, washing the CoNWs by using deionized water and absolute ethyl alcohol in sequence, and finally performing vacuum freeze drying on the CoNWs to obtain the CoNWs.

And step 3: ti₃C₂Preparation of-CoNWs composite Material

Firstly, Ti obtained in the step 1₃C₂And step 2, respectively dispersing the CoNWs in deionized water to obtain Ti₃C₂Suspension and CoNWs suspension, then Ti₃C₂The suspension and the CoNWs suspension are mixed in equal volume to obtain mixed suspension, and then the mixed suspension is subjected to ultrasonic reaction for 20-30min to obtain suspended Ti₃C₂-a CoNWs composite; wherein, Ti₃C₂The molar ratio of the suspension to the CoNWs suspension was 1: 0.5.

And 4, step 4: preparation of sulfonated polyether ether ketone

And (3) putting the polyether-ether-ketone (PEEK) substrate into concentrated sulfuric acid, performing ultrasonic treatment for 10min, taking out the ultrasonically treated PEEK, washing with acetone and absolute ethyl alcohol, and washing to obtain the sulfonated polyether-ether-ketone (SPEEK) with rough and porous surface and sulfonic acid groups.

And 5: polyether-ether-ketone bone repair material (Ti)₃C₂Preparation of-CoNWs/SPEEK)

Firstly, the suspended Ti obtained in the step 3₃C₂-soaking and dripping the CoNWs composite material on the surface of the SPEEK obtained in the step 4, and then drying the soaked SPEEK at the temperature of 60 ℃; repeating the operations of dipping, dripping and drying for 5 times to finally obtain the polyether-ether-ketone bone repair material Ti with antibacterial property₃C₂-CoNWs/SPEEK。

Example 2

A polyetheretherketone bone repair material having antibacterial properties was prepared according to the preparation method described in example 1, except that:

in the step 1, the concentration of hydrochloric acid is 10mol/L, the mixture is stirred at the temperature of 35 ℃ for reaction time of 48h, and Ti₃C₂Tx suspension was centrifuged andwashing to pH 5-6.

In the step 2, N is added into the reaction liquid₂H₄·H₂O、H₂PtCl·6H₂The amounts of O were 650. mu.L and 300. mu.L, respectively.

In said step 3, Ti₃C₂The molar ratio of the suspension to the CoNWs suspension was 1: 0.75, and the ultrasonic time of the mixed solution is 30 min.

In the step 4, the ultrasonic time of the polyether-ether-ketone substrate (PEEK) in concentrated sulfuric acid is 15 min.

In the step 5, the drying temperature of the soaked and dripped SPEEK is 55 ℃, and the repeated times of the soaking, dripping and drying operation cycle is 4.

Example 3

A polyetheretherketone bone repair material having antibacterial properties was prepared according to the preparation method described in example 1, except that:

in the step 1, the concentration of hydrochloric acid is 9.5mol/L, the mixture is stirred at the temperature of 35 ℃ for reaction for 36h, and Ti₃C₂The Tx suspension was centrifuged and washed to a pH of 5.5.

In the step 2, the pH value of the raw material liquid is adjusted to 13 by NaOH, and N is added into the reaction liquid₂H₄·H₂O、H₂PtCl·6H₂The amounts of O were 620. mu.L and 240. mu.L, respectively.

In said step 3, Ti₃C₂The molar ratio of the suspension to the CoNWs suspension was 1: 1.

in the step 4, the ultrasonic time of the polyether-ether-ketone substrate (PEEK) in concentrated sulfuric acid is 15 min.

In the step 5, the drying temperature of the soaked and dripped SPEEK is 50 ℃, and the repeated times of the soaking, dripping and drying operation cycle is 3.

Comparative example 1

A preparation method of an antibacterial polyether-ether-ketone bone repair material specifically comprises the following steps:

step 1: ti₃C₂Preparation of

First 1g of LiF (lithium fluoride) was added to 20mL of a molar concentrationAdding the solution into hydrochloric acid solution with the degree of 9mol/L, uniformly stirring, and then adding 1g of Ti into the LiF/HCl solution₃AlC₂Obtaining a mixture, stirring and reacting the mixture in ice bath for 30min under the condition of 35 ℃ for 24h, and then washing the reacted mixture with deionized water to obtain Ti₃C₂Tx suspension, Ti to be obtained₃C₂The Tx suspension is centrifuged and washed to a pH of 5-6 to obtain Ti in a multi-layered form₃C₂Tx. Ti to be obtained₃C₂Re-dispersing Tx in deionized water to obtain Ti₃C₂Tx dispersion, then placing the dispersion in ice bath for ultrasonic treatment for 1h, then centrifuging the dispersion after ultrasonic treatment and removing supernatant fluid to obtain monolayer Ti₃C₂Nano-flake dispersed Ti in water₃C₂Suspension, finally Ti₃C₂Vacuum freeze drying the suspension to obtain single-layer Ti₃C₂。

Step 2: preparation of sulfonated polyether ether ketone

And (3) putting the polyether-ether-ketone substrate (PEEK) into concentrated sulfuric acid and carrying out ultrasonic treatment for 10min, taking out the ultrasonically treated PEEK, washing the PEEK by using acetone and absolute ethyl alcohol, and washing to obtain the sulfonated polyether-ether-ketone (SPEEK) with a rough and porous surface and sulfonic acid groups.

And step 3: polyether-ether-ketone bone repair material (Ti)₃C₂Preparation of/SPEEK)

Firstly, the single-layer Ti obtained in the step 1₃C₂Dispersing the nano-thin sheets in deionized water to obtain Ti₃C₂Suspending the solution, then adding Ti₃C₂Dipping the suspension on the surface of the SPEEK obtained in the step 2, and then drying the dipped SPEEK at the temperature of 60 ℃; repeating the cycle of dipping, dripping and drying for 5 times to finally obtain the polyether-ether-ketone bone repair material Ti with antibacterial property₃C₂/SPEEK。

And (3) spectrum characterization and analysis:

1. SEM image analysis:

ti prepared in example 1₃C₂-CoNWs/SPEEK and Ti prepared in comparative example 1₃C₂[ SPEEK ] was analyzed by electron microscopy (SEM), as shown in FIG. 1, wherein a is Ti₃C₂SEM image of/SPEEK, panel b is material Ti₃C₂SEM image of CoNWs/SPEEK. As can be seen from FIG. 1, since Ti₃C₂Compounding with CoNW to obtain composite material Ti₃C₂-CoNW dipped on the surface of SPEEK does not resemble Ti₃C₂Dipped and dropped on the surface of SPEEK to be as soft as that of composite material Ti₃C₂The CoNW is able to flood the surface of the SPEEK.

2. And (3) photo-thermal performance analysis:

first, the 48 wells on the well plate were divided equally into 3 groups to form 3 experimental groups. Then, SPEEK and Ti prepared in example 1 are added₃C₂-CoNWs/SPEEK and Ti prepared in comparative example 1₃C₂The SPEEK is put in each corresponding experimental group, wherein the experimental group where the SPEEK is taken as a control group. Then, 500. mu.L of PBS buffer was added to each well, followed by a near-infrared laser (808nm, 1.5W/cm)²) Each well was irradiated until temperature fluctuation was small, and temperature change was captured by an LIFR infrared detector every 15 seconds, and the result is shown in FIG. 2 a.

First, the 48 wells on the well plate were divided equally into 3 groups to form 3 experimental groups. Then, SPEEK and Ti prepared in example 1 are added₃C₂-CoNWs/SPEEK and Ti prepared in comparative example 1₃C₂The SPEEK is put in each corresponding experimental group, wherein the experimental group where the SPEEK is taken as a control group. Then, 500. mu.L of PBS buffer was added to each well, followed by a near-infrared laser (808nm, 1.5W/cm)²) After irradiating each well for 5 minutes, the infrared laser was turned off and the sample was cooled for 5 minutes, and the temperature change was captured by the LIFR infrared detector every 30 seconds, and the laser was turned on/off repeatedly for 5 cycles to observe the photo-thermal stability of the sample, and the results are shown in FIG. 2 b.

As shown in FIG. 2, the material Ti₃C₂The photothermal property of-CoNWs/SPEEK is improved, and the photothermal cycle stability is good, thereby being beneficial to stable photothermal antibiosis.

3. Singlet oxygen release analysis:

first, the 48 wells on the well plate were divided equally into 3 groups to form 3 experimental groups. Then, SPEEK and Ti prepared in example 1 are added₃C₂-CoNWs/SPEEK and Ti prepared in comparative example 1₃C₂The SPEEK is put in each corresponding experimental group, wherein the experimental group where the SPEEK is taken as a control group. Using 2,2,6, 6-tetramethylpiperidin-1-oxy as singlet oxygen (¹O₂) The labeling substance of (1.5) was applied to each experimental group using a near-infrared laser (808nm, 1.5W/cm)²) The irradiation was carried out for 20 minutes and the generation of singlet oxygen was recorded every 5 minutes using an electron spin resonance apparatus, and the results are shown in FIG. 3 a.

First, the 48 wells on the well plate were divided equally into 3 groups to form 3 experimental groups. Then, SPEEK and Ti prepared in example 1 are added₃C₂-CoNWs/SPEEK and Ti prepared in comparative example 1₃C₂The SPEEK is put in each corresponding experimental group, wherein the experimental group where the SPEEK is taken as a control group. Using 1, 3-diphenyliso-furan as singlet oxygen: (¹O₂) Then each experimental group was placed in the dark and irradiated with a near infrared laser (808nm, 1.5W/cm)²) After 20 minutes of irradiation, 100. mu.L of 1, 3-diphenyliso-benzofuran was taken every 5 minutes and the absorbance thereof was measured in a microplate reader apparatus, and the measurement results are shown in FIG. 3 b.

As shown in FIG. 3, the material Ti₃C₂-CoNWs/SPEEK compared to the materials SPEEK and Ti₃C₂The SPEEK is more beneficial to the generation of singlet oxygen under the condition of laser excitation, thereby achieving better bacteriostatic effect.

4. And (3) bacteriostatic analysis:

(1) bacteriostatic analysis of staphylococcus aureus

First, the 48 wells on the well plate were divided equally into 3 groups to form 3 experimental groups. Then, SPEEK and Ti prepared in example 1 are added₃C₂-CoNWs/SPEEK and Ti prepared in comparative example 1₃C₂The SPEEK is put in each corresponding experimental group, wherein the experimental group where the SPEEK is taken as a control group. To each well, 100. mu.L of LB liquid medium and 100. mu.L of 10⁶CFU/mL Staphylococcus aureus, then perHalf of the experimental groups were incubated in the dark for 20 minutes, and the remaining half were irradiated with a near-infrared laser (808nm, 1.5W/cm)²) Culturing for 20 minutes under the conditions of (1); then, the cultured broth was taken out and uniformly applied to LB solid medium, and after culturing at 37 ℃ for 24 hours, the bactericidal effect of each material was observed, and the experimental results are shown in FIG. 4 a.

(2) Bacteriostatic analysis of E.coli

The method for analyzing the escherichia coli bacteriostasis is the same as that for staphylococcus aureus, except that the staphylococcus aureus added into the holes is replaced by the escherichia coli, the sterilization effect of various materials is observed, and the experimental result is shown in fig. 4 b.

As can be seen from FIG. 4, since the material Ti₃C₂Under the condition of laser excitation, CoNWs/SPEEK can generate photothermal and singlet oxygen, and influence the growth of bacteria, thereby inhibiting the growth of the bacteria; and Ti₃C₂The growth inhibition effect of SPEEK on bacteria is limited, and SPEEK has no bacteriostatic effect. Thus material Ti₃C₂The CoNWs/SPEEK has good bacteriostatic effect under the condition of laser excitation.

5. And (3) analyzing the appearance of the bacteria:

(1) morphological analysis of staphylococcus aureus

First, the 48 wells on the well plate were divided equally into 3 groups to form 3 experimental groups. Then, SPEEK and Ti prepared in example 1 are added₃C₂-CoNWs/SPEEK and Ti prepared in comparative example 1₃C₂The SPEEK is put in each corresponding experimental group, wherein the experimental group where the SPEEK is taken as a control group. To each well 100. mu.L of liquid medium and 100. mu.L of 10⁶CFU/mL Staphylococcus aureus, then half of each experimental group was incubated in the dark for 20 minutes, and the remaining half was exposed to a near infrared laser (808nm, 1.5W/cm)²) Was incubated for 20 minutes under the conditions of (1). The cultured Staphylococcus aureus was removed and placed in 500. mu.L of 2.5% glutaraldehyde, stored at 4 ℃ for 12 hours, and then dehydrated with gradient ethanol (30%, 50%, 70%, 90% and 100% v/v) for 10 minutes each, and finally the Staphylococcus aureus was self-treatedAfter air drying, the bacterial morphology was observed by scanning electron microscopy and the results are shown in FIG. 5.

(2) Morphological analysis of Escherichia coli

The bacteriostatic analysis on escherichia coli is the same as the shape analysis method on staphylococcus aureus, except that the staphylococcus aureus added into the holes is replaced by the escherichia coli, finally, the escherichia coli sample is naturally air-dried, and then the shape of the bacteria is observed by a scanning electron microscope, and the result is shown in fig. 5.

As can be seen from FIG. 5, the material Ti₃C₂The CoNWs/SPEEK can effectively destroy the integrity of bacteria through photo-thermal and singlet oxygen, thereby killing the bacteria.

6. Evaluation analysis of cytotoxicity:

first, the 48 wells on the well plate were divided equally into 3 groups to form 3 experimental groups. Then, SPEEK and Ti prepared in example 1 are added₃C₂-CoNWs/SPEEK and Ti prepared in comparative example 1₃C₂the/SPEEK is placed in each corresponding experimental group. Culturing MC with DMEM containing 10% fetal bovine serum culture medium₃T₃Mouse embryonic osteoblasts; after adherent growth of the cells, fresh medium was replaced and when the cells reached a degree of aggregation of 80%, the cells 104/well density was seeded in 48 wells and cultured for 24 h. In addition, a control group and an experimental group are also required to be set, wherein the control group is only added with the culture medium and the cells which are the same as those in the experimental group, and the blank group is only added with the culture medium which is the same as that in the experimental group.

Cytotoxicity was detected using the CCK-8 kit. The absorbance of each set was measured by a microplate reader at a wavelength of 450 nm.

Relative cell survival (%) — (absorbance value of experimental group-absorbance value of blank)/(absorbance value of control group-absorbance value of blank) × 100%.

The experimental results are as follows: material Ti₃C₂The survival rate of cells corresponding to CoNWs/SPEEK was about 80%, although lower than that of other groups.

In summary, the present invention solves the technical deficiencies of the prior art. The invention uses cobalt nano-wire and Ti₃C₂Compounding and delaying Ti₃C₂The combination of electrons and holes in the energy band increases Ti₃C₂The photoelectron transfer promotes the electron transfer to form singlet oxygen, thereby solving the problem of Ti₃C₂The antibacterial effect is limited, thereby preventing the SPEEK implant from being infected with bacteria and inhibiting the generation of drug-resistant bacteria. In addition, the invention also prepares the sulfonated polyether-ether-ketone with rough and porous surface and sulfonic groups by sulfonating the polyether-ether-ketone in concentrated sulfuric acid, and Ti is added₃C₂The contact area of the-CoNWs composite material and the SPEEK is increased, and the Ti content is improved₃C₂-the bonding force between the connws composite and SPEEK; furthermore, sulfonic acid groups on the SPEEK surface are bonded to Ti₃C₂The surface hydroxyl groups react to form sulfonate esters to react SPEEK with Ti₃C₂Chemical bonds are also formed between the-CoNWs composite materials, so that Ti is further strengthened₃C₂The binding force between the CoNWs composite material and the SPEEK, avoiding Ti₃C₂The problem of the CoNWs composite material falling off from the SPEEK during use.

The foregoing is only a partial embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims (9)

1. The polyether-ether-ketone bone repair material with antibacterial property is characterized by comprising sulfonated polyether-ether-ketone with rough and porous surface and sulfonic acid groups and Ti coated on the surface of the sulfonated polyether-ketone₃C₂-a CoNWs composite.

2. The PEEK bone repair material having antibacterial properties of claim 1, wherein the Ti is Ti₃C₂-the preparation of a CoNWs composite material comprises the following steps:

step S1: firstly, Ti₃C₂And CoNWs are respectively dispersed in deionized water to obtain Ti₃C₂Suspension and CoNWs suspension, then Ti₃C₂The suspension and the CoNWs suspension are mixed in equal volumes to obtain a mixed suspension, wherein Ti is₃C₂The molar concentration ratio of the suspension to the CoNWs suspension is 1 (0.5-1);

step S2: performing ultrasonic treatment on the mixed solution obtained in the step S1 for 20-30min to obtain Ti₃C₂-a CoNWs composite.

3. The PEEK bone repair material having antibacterial properties as claimed in claim 2, wherein the Ti is selected from the group consisting of Ti, Ti₃C₂Ti being in the form of a monolayer₃C₂。

4. The bone repair material of polyetheretherketone having antibacterial properties according to any of claims 1 to 3, wherein the sulfonated polyetheretherketone is prepared by the following method:

firstly, the polyether-ether-ketone base material is placed in concentrated sulfuric acid and is subjected to ultrasonic treatment for 5-15min, and then the polyether-ether-ketone after ultrasonic treatment is washed by acetone and absolute ethyl alcohol to obtain the sulfonated polyether-ether-ketone.

5. The PEEK bone repair material having antibacterial properties of claim 3, wherein the single layer of Ti is Ti₃C₂The preparation method comprises the following steps:

firstly, adding LiF into hydrochloric acid solution with the molar concentration of 9-10mol/L, uniformly stirring, and then adding Ti into the solution added with LiF/HCl₃AlC₂Obtaining a mixture, sequentially stirring and reacting the mixture in an ice bath for 30min at 35 ℃ for 24-48h, and then washing the stirred and reacted mixture with deionized water to obtain Ti₃C₂Tx suspension, Ti to be obtained₃C₂The Tx suspension was centrifuged and washed to pH 5-6 to obtain Ti in a multi-layered form₃C₂Tx; obtaining a multilayer Ti₃C₂Re-dispersing Tx in deionized water to obtain Ti₃C₂Tx dispersion, placing said dispersion in iceWhen the ultrasonic treatment is carried out for 1h in the bath, the dispersion liquid after the ultrasonic treatment is centrifuged, and the supernatant is removed to obtain single-layer Ti₃C₂Nano-flake dispersed Ti in water₃C₂Suspension of Ti₃C₂Drying the suspension to obtain a single layer of Ti₃C₂。

6. The PEEK bone repair material of claim 5, wherein the LiF and Ti are selected from the group consisting of₃AlC₂The mass ratio of (1): (0.5-1).

7. The polyetheretherketone bone repair material having antibacterial properties according to any of claims 1 to 3, wherein the CoNWs is prepared by the following method:

firstly, CoCl is added₂·6H₂Adding O and EDTA-2Na into deionized water according to the molar ratio of 1:1, uniformly stirring to obtain a raw material solution, adjusting the pH of the raw material solution to 13-14 to obtain a reaction solution, then placing the reaction solution in a magnetic field cage at 80 ℃ for reaction, and adding 600-650 mu L of N-doped sodium chloride into the reaction solution₂H_4·H₂O is stirred uniformly, and then 240-300 mu L of H is added into the reaction solution without stirring₂PtCl·6H₂And O, collecting the generated CoNWs, washing the generated CoNWs with deionized water and absolute ethyl alcohol in sequence, and finally performing vacuum freeze drying on the obtained product to obtain the CoNWs.

8. The bone repair material according to claim 7, wherein the pH of the raw material solution is adjusted with NaOH.

9. The preparation method of the polyether-ether-ketone bone repair material with the antibacterial property is characterized in that the preparation of the polyether-ether-ketone bone repair material with the antibacterial property according to the claims 3-8 specifically comprises the following steps:

step C1 first Ti₃C₂-CoNWs composite material dipped in sulfonated polyetherDrying the dipped sulfonated polyether-ether-ketone at 50-60 ℃ on the surface of the ether-ketone;

step C2, repeating the step C1 for 3-5 times, and finally obtaining the polyether-ether-ketone bone repair material Ti with antibacterial property₃C₂-CoNWs/SPEEK。

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