CN115414386B - Preparation method of bioactive glass nanocomposite with catalytic antibacterial property - Google Patents

Preparation method of bioactive glass nanocomposite with catalytic antibacterial property Download PDF

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CN115414386B
CN115414386B CN202211122821.3A CN202211122821A CN115414386B CN 115414386 B CN115414386 B CN 115414386B CN 202211122821 A CN202211122821 A CN 202211122821A CN 115414386 B CN115414386 B CN 115414386B
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bioactive glass
auncs
nanocomposite
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bgn
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CN115414386A (en
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薛语萌
徐子琪
尚利
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Northwestern Polytechnical University
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Abstract

The invention belongs to the technical field of nano biological material preparation, and discloses a preparation method of a bioactive glass nano composite material with catalytic antibacterial property, which comprises the following preparation processes: preparing bioactive glass nano particles by adopting a microemulsion method; and (3) carrying out positive electrification on the gold nanoclusters, and then reacting with bioactive glass nanoparticles under stirring. The invention utilizes the positive electrochemical gold nanocluster to modify the bioactive glass nanomaterial for the first time, and endows the bioactive glass nanomaterial with high-efficiency enzyme-like catalytic property and antibacterial property. The preparation method of the bioactive glass nanocomposite is simple and easy to implement, the reaction condition is mild, no complex instrument or equipment is needed, and the prepared bioactive glass nanocomposite not only has excellent biocompatibility, catalase-like catalysis and antibacterial activity, but also has the functions of fluorescent imaging, cell behavior regulation and the like, and has great application potential in the biomedical fields of tissue regeneration and the like.

Description

Preparation method of bioactive glass nanocomposite with catalytic antibacterial property
Technical Field
The invention belongs to the technical field of nano biological material preparation, and relates to a preparation method of a bioactive glass nano composite material with catalytic antibacterial property.
Background
The bioactive glass is made of SiO 2 、CaO、P 2 O 5 Typical silicate materials of equal composition, which are found to play an important role in various phases of wound healing, are currently available in a variety of commercial products based on bioactive glass for wound healing. However, the existing novel dressing based on bioactive glass has the disadvantages of insignificant antibacterial performance and single function, and is difficult to meet the application requirements of complex biological system environments. Therefore, there is a need for further functionalization and modification of bioactive glass to enhance its efficacy in complex wound treatments.
In order to improve the performance of bioactive glass in the treatment process of infectious wounds, the antibacterial performance of bioactive glass is generally realized in two ways: (1) In the process of preparing bioactive glass, metal ions with antibacterial capability are directly doped into a glass network, and the aim of killing bacteria is achieved along with continuous dissolution of the ions in the later period. Common dopant ions include Ag + 、Cu 2 + 、Zn 2+ And the like, these metal ions can kill bacteria by various mechanisms such as disruption of bacterial membrane structure, inhibition of protein synthesis, damage to DNA and RNA, and generation of Reactive Oxygen Species (ROS). (2) The bioactive glass is filled with medicines such as antibiotics to improve the antibacterial activity. The bioactive glass nano material with mesoporous structure and the Si-OH with rich surface can realize gradual release of the medicine in physiological environment through physical adsorption, hydrogen bond action and Van der Waals force and medicine action, thereby playing a role in sterilization.
Although ion doped bioactive glass has certain antibacterial activity, the ion release process is uncontrollable and potentially toxic, which greatly limits its further application and clinical transformation in infectious wound treatment. The bioactive glass is an ideal choice for loading the antibacterial drug, and has obvious burst release when releasing the antibiotic, so that the long-acting load and slow release of the antibiotic cannot be realized, and the safe and long-acting antibacterial effect cannot be realized. Moreover, as the antibiotics are widely used for a long time, the bacteria are easy to generate drug resistance, so that the therapeutic effect of the medicine is obviously reduced. Therefore, a novel bioactive glass nanomaterial with high biosafety and excellent antibacterial performance has not been developed yet.
Disclosure of Invention
Aiming at the defects of an antibacterial strategy of a bioactive glass nano material in the prior art, the invention aims to provide a preparation method of the bioactive glass nano composite material with catalytic antibacterial performance, which endows the gold nanoclusters (BSA-AuNCs) with excellent enzyme-like catalytic antibacterial activity by carrying out amination modification on the gold nanoclusters, and simultaneously realizes high-efficiency functionalization on BGN through rich positive charges on the surface of the cBSA-auNCs, so that the BGN can show remarkably enhanced antibacterial activity at a lower material concentration; the preparation method has mild reaction conditions, does not need complex instruments and equipment, and is easy to operate. The bioactive glass nanocomposite synthesized by the invention also has excellent fluorescence imaging, fibroblast proliferation promoting and other functions, and has wider application scene than the traditional bioactive glass material.
In order to achieve the above purpose, the technical scheme adopted in the application is as follows:
a preparation method of a bioactive glass nanocomposite with catalytic antibacterial performance comprises the following steps:
1) Preparing bioactive glass nano particles by adopting a microemulsion method;
2) And (3) reacting the bioactive glass nano particles with the positive electrochemical gold nanoclusters in a reaction solvent to obtain the bioactive glass nano composite material with catalytic antibacterial performance.
Further, the bioactive glass nanoparticle comprises silicon element and calcium element in a molar ratio of 4:1-4:2.
Further, the positive-electrical gold nanoclusters are prepared by the following process: and (3) carrying out amination modification reaction on the gold nanocluster, ethylenediamine hydrochloride and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide to obtain the positive-electrification gold nanocluster.
Further, the usage ratio of the gold nanoclusters to ethylenediamine hydrochloride is 10mmol:1-5mol.
Further, the ratio of the gold nanoclusters to the 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide was 10mmol:4-20mg.
Further, the amination modification reaction is carried out for 0.5 to 6 hours.
Further, the mass ratio of the bioactive glass nano-particles to the positive-powered gold nanoclusters is 1:1-3:1.
Further, the reaction solvent in the step 2) is ultrapure water, and the reaction time is 12-48 hours.
Compared with the prior art, the invention has the beneficial effects that:
the invention utilizes the positive electric gold nanocluster to remove the modified bioactive glass nanoparticle, so that the bioactive glass nanocomposite BGN-AuNCs has excellent catalase-like antibacterial activity, and can obviously reduce the concentration threshold of the antibacterial material of the bioactive glass. In trace amounts of H 2 O 2 In the presence of the nano composite material, the nano composite material of the bioactive glass with 75 mug/mL has excellent antibacterial performance, and the traditional antibacterial bioactive glass which relies on ion doping and antibiotic loading can generally show ideal antibacterial effect at the concentration level of milligrams per milliliter. The preparation method of the bioactive glass nanocomposite is simple and easy to implement, the reaction condition is mild, no complex instrument or equipment is needed, and the prepared bioactive glass nanocomposite not only has excellent biocompatibility, catalase-like catalysis and antibacterial activity, but also has the functions of fluorescent imaging, cell behavior regulation and the like, and has great application potential in the biomedical fields of tissue regeneration and the like.
Furthermore, the gold nanoclusters (BSA-AuNCs) protected by bovine serum albumin are utilized for the first time to modify bioactive glass, the surface of the bioactive glass is provided with rich positive charges through amination modification on the bovine serum albumin, and the electropositive modification technology can effectively enhance the binding force of the BSA-AuNCs and BGN with negative charges on the surface on one hand, and can also enhance the catalytic capability of the BSA-AuNCs to catalase under physiological environment and promote the killing effect of the BSA-AuNCs to bacteria on the other hand, so that the bioactive glass can generate excellent sterilizing effect (100% killing of escherichia coli and staphylococcus aureus) under extremely low concentration (2.5 mug/mL).
Drawings
FIG. 1 is a fluorescence spectrum of a bioactive glass nanocomposite of the present invention, wherein samples BA1, BA2, BA3 and BA4 are BGN-AuNCs composite materials prepared in the mass ratio of BGN to cBSA-AuNCs of 3:1,2:1,1.5:1 and 1:1 in sequence, and cBSA-AuNCs adopted in the preparation of samples BA1-BA4 are positively-charged cBSA-AUNCs products with ethylenediamine hydrochloride concentration of 5 mol/L;
FIG. 2 is a Zeta potential diagram of BA1, BA2, BA3 and BA4 of the bioactive glass nanocomposites of the present invention;
FIG. 3 is a transmission electron microscope image and element distribution of the bioactive glass nanocomposite BA4 of the invention, wherein the image (a) is a transmission electron microscope image of the sample BA4, the image (b) is an energy spectrum scanning area of the sample BA4, and the images (c), (d), (e), (f) and (g) are distribution diagrams of Si, O, ca, au and S elements respectively;
FIG. 4 is a catalase-like catalytic effect study of the bioactive glass nanocomposite BA4 of the invention;
FIG. 5 is a graph showing the antibacterial effect of the bioactive glass nanocomposite BA4 of the present invention, wherein (a) is the survival rate of E.coli, and (b) is the survival rate of Staphylococcus aureus.
Detailed Description
In order to make the technical means, the creation characteristics, the achievement of the purpose and the effect of the present invention easy to understand, the technical solution of the present invention will be clearly and completely described below with reference to the specific embodiments and the accompanying drawings of the present invention.
The invention utilizes the positive electric gold nanoclusters to remove modified bioactive glass to prepare the bioactive glass nanocomposite with catalytic antibacterial performance. Gold nanoclusters protected by bovine serum albumin with good biocompatibility are selected and subjected to amination modification. Bovine serum albumin gold clusters have excellent biocompatibility, but have poor antibacterial effect and poor catalase-like activity under neutral and weak alkaline conditions. The enzyme-like catalytic antibacterial activity of the bovine serum albumin gold cluster can be improved through amination modification, and on the other hand, the binding force of the bovine serum albumin gold cluster and the bioactive nano particles can be obviously enhanced through electropositive modification due to electronegativity of the bioactive glass nano material in a physiological environment, so that the efficient functionalization of the bioactive glass is realized, and the bioactive glass nano composite material with catalytic antibacterial performance is prepared.
The invention relates to a preparation method of a bioactive glass nanocomposite with catalytic antibacterial performance, which comprises the following steps:
1) Preparation of bioactive glass: the preparation method comprises the steps of taking bromocetyl pyridine as a surfactant and a template agent, fully hydrolyzing bromocetyl pyridine and urea in water, adding cyclohexane and n-butanol, vigorously stirring to obtain a uniform emulsion, gradually adding tetraethyl orthosilicate and calcium nitrate tetrahydrate, reacting to obtain milky bioactive glass sol-gel, washing, drying and calcining to obtain bioactive glass nano particles (BGN).
Wherein the mass ratio of urea to bromocetyl pyridine is 0.6:1; the volume ratio of cyclohexane to water is 1:1; the volume ratio of cyclohexane to n-butanol is 30:1, and the mol ratio of tetraethyl orthosilicate to calcium nitrate tetrahydrate is 4:1-4:2; the reaction temperature is 65-75 ℃ and the reaction time is 8-16 hours.
2) Preparation of electropositive gold nanoclusters: preparing gold nanoclusters (BSA-AuNCs) by taking bovine serum albumin as a template agent and a reducing agent, performing ultrafiltration purification, performing amination modification reaction on the gold nanoclusters and ethylenediamine hydrochloride and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) for 0.5-6 hours, performing ultrafiltration purification on a reaction final solution to obtain the positive-electricity gold nanoclusters (cBSA-auNCs), and storing the positive-electricity gold nanoclusters in an environment of 4 ℃.
The concentration of BSA-AuNCs was 10mmol/L and the volume was 5mL.
The concentration of the ethylenediamine hydrochloride is 1-5mol/L, the volume is 5mL, and the ethylenediamine hydrochloride is slowly added into BSA-AuNCs under the condition of intense stirring.
The EDC is used in an amount of 20-100mg.
3) Preparation of bioactive glass nanocomposite: the prepared BGN was reacted with cBSA-auNCs for 12-48 hours under vigorous stirring, centrifuged at 8000rpm for 15 minutes and the precipitate was washed 3 times to remove the non-effectively bonded cBSA-auNCs, thereby obtaining bioactive glass nanocomposite (BGN-AuNCs) having catalytic antibacterial properties.
Wherein 15mg BGN should be dispersed in 500. Mu.L deionized water solution and then sonicated for 5 minutes, cBSA-auNCs were added.
The mass ratio of BGN to cBSA-auNCs is 1:1-3:1, preferably 3:1,2:1,1.5:1 and 1:1.
Example 1
1) Preparation of bioactive glass: 0.6g of urea and 1g of cetylpyridinium bromide were taken and dissolved well in 30mL of deionized water for 20 minutes, then 30mL of cyclohexane and 1.1mL of n-butanol were slowly added and vigorously stirred for 30 minutes to form a uniform emulsion. 2.7mL of TEO (tetraethyl orthosilicate) was then slowly added dropwise, the temperature was raised to 70℃after 30 minutes of hydrolysis, and the reaction was continued with vigorous stirring for 8 hours. Finally, 0.7mg of calcium nitrate tetrahydrate was added, and the reaction was continued for 4 hours. The reaction final solution was washed 3 times with acetone, ethanol and water, respectively. The precipitate obtained was freeze-dried and then calcined at 600℃for 5 hours (heating rate 1 ℃/min). The prepared sample BGN is stored at room temperature for standby.
2) Preparation of electropositive gold nanoclusters: BSA-AuNCs are prepared by taking bovine serum albumin as a template agent and a reducing agent, and the specific preparation steps are as follows: xie J, zheng Y, ying J Y.protein-directed Synthesis of Highly Fluorescent Gold Nanoclusters [ J ]. Journal of the American Chemical Society,2009,131 (3): 888-889. Ethylenediamine hydrochloride solution (5 mol/L,5 mL) was added to the purified BSA-AuNCs (10 mmol/L,5 mL) with stirring at 1000rpm, and after stirring for 2 minutes, 100mg EDC was added and the reaction was continued with stirring at 1000rpm for 2 hours. The reaction final solution was subjected to 5 ultrafiltration centrifugation operations using a 30kDa ultrafiltration tube to purify the amino-modified cBSA-auNCs. The prepared sample cBSA-auNCs was stored at 4℃for further use.
3) Preparation of bioactive glass nanocomposite: the mass ratio of BGN to cBSA-auNCs is 3:1. 15mg of BGN was taken and slowly added to 3.3mL of cBSA-auNCs (1.5 mg/mL) after sufficiently sonicating with a small amount of deionized water, the reaction was vigorously stirred at room temperature for 48 hours, the reaction final solution was centrifuged at 8000rpm for 15 minutes and the precipitate was washed 3 times to remove the non-efficiently bound cBSA-auNCs.
Example 2
1) Preparation of BGN: 0.6g of urea and 1g of cetylpyridinium bromide were taken and dissolved well in 30mL of deionized water for 20 minutes, then 30mL of cyclohexane and 1.1mL of n-butanol were slowly added and vigorously stirred for 30 minutes to form a uniform emulsion. 2.7mL TEOS was then slowly added dropwise, hydrolyzed for 30 minutes, then warmed to 70℃and reacted for 8 hours with continued vigorous stirring. Finally, 0.7mg of calcium nitrate tetrahydrate was added, and the reaction was continued for 4 hours. The reaction final solution was washed 3 times with acetone, ethanol and water, respectively. The precipitate obtained was freeze-dried and then calcined at 600℃for 5 hours (heating rate 1 ℃/min). The prepared sample BGN is stored at room temperature for standby.
2) Preparation of cBSA-auNCs: BSA-AuNCs are prepared by taking bovine serum albumin as a template agent and a reducing agent, and the specific preparation steps are as follows: xie J, zheng Y, ying J Y.protein-directed Synthesis of Highly Fluorescent Gold Nanoclusters [ J ]. Journal of the American Chemical Society,2009,131 (3): 888-889. Ethylenediamine hydrochloride solution (5 mol/L,5 mL) was added to the purified BSA-AuNCs (10 mmol/L,5 mL) with stirring at 1000rpm, and after stirring for 2 minutes, 100mg EDC was added and the reaction was continued with stirring at 1000rpm for 2 hours. The reaction final solution was subjected to 5 ultrafiltration centrifugation operations using a 30kDa ultrafiltration tube to purify the amino-modified cBSA-auNCs. The prepared sample cBSA-auNCs was stored at 4℃for further use.
3) Preparation of bioactive glass nanocomposite: the mass ratio of BGN to cBSA-auNCs is 2:1. 15mg of BGN was taken and sufficiently dispersed in a small amount of deionized water by sonication and slowly added to 5mL of cBSA-auNCs (1.5 mg/mL), reacted with vigorous stirring at room temperature for 48 hours, and the reaction final solution was centrifuged at 8000rpm for 15 minutes and the precipitate was washed 3 times to remove cBSA-auNCs which were not effectively bonded.
Example 3
1) Preparation of BGN: 0.6g of urea and 1g of cetylpyridinium bromide were taken and dissolved well in 30mL of deionized water for 20 minutes, then 30mL of cyclohexane and 1.1mL of n-butanol were slowly added and vigorously stirred for 30 minutes to form a uniform emulsion. 2.7mL TEOS was then slowly added dropwise, hydrolyzed for 30 minutes, then warmed to 70℃and reacted for 8 hours with continued vigorous stirring. Finally, 0.7mg of calcium nitrate tetrahydrate was added, and the reaction was continued for 4 hours. The reaction final solution was washed 3 times with acetone, ethanol and water, respectively. The precipitate obtained was freeze-dried and then calcined at 600℃for 5 hours (heating rate 1 ℃/min). The prepared sample BGN is stored at room temperature for standby.
2) Preparation of cBSA-auNCs: BSA-AuNCs are prepared by taking bovine serum albumin as a template agent and a reducing agent, and the specific preparation steps are as follows: xie J, zheng Y, ying J Y.protein-directed Synthesis of Highly Fluorescent Gold Nanoclusters [ J ]. Journal of the American Chemical Society,2009,131 (3): 888-889. Ethylenediamine hydrochloride solution (5 mol/L,5 mL) was added to the purified BSA-AuNCs (10 m mol/L,5 mL) with stirring at 1000rpm, and after stirring for 2 minutes, 100mg EDC was added and the reaction was continued with stirring at 1000rpm for 2 hours. The reaction final solution was subjected to 5 ultrafiltration centrifugation operations using a 30kDa ultrafiltration tube to purify the amino-modified cBSA-auNCs. The prepared sample cBSA-auNCs was stored at 4℃for further use.
3) Preparation of bioactive glass nanocomposite: the mass ratio of BGN to cBSA-auNCs is 1.5:1. 15mg of BGN was taken and sufficiently dispersed in a small amount of deionized water by sonication and slowly added to 6.7mL of cBSA-auNCs (1.5 mg/mL), the reaction was vigorously stirred at room temperature for 48 hours, the reaction final solution was centrifuged at 8000rpm for 15 minutes and the precipitate was washed 3 times to remove the non-efficiently bound cBSA-auNCs.
Example 4
1) Preparation of BGN: 0.6g of urea and 1g of cetylpyridinium bromide were taken and dissolved well in 30mL of deionized water for 20 minutes, then 30mL of cyclohexane and 1.1mL of n-butanol were slowly added and vigorously stirred for 30 minutes to form a uniform emulsion. 2.7mL TEOS was then slowly added dropwise, hydrolyzed for 30 minutes, then warmed to 70℃and reacted for 8 hours with continued vigorous stirring. Finally, 0.7mg of calcium nitrate tetrahydrate was added, and the reaction was continued for 4 hours. The reaction final solution was washed 3 times with acetone, ethanol and water, respectively. The precipitate obtained was freeze-dried and then calcined at 600℃for 5 hours (heating rate 1 ℃/min). The prepared sample BGN is stored at room temperature for standby.
2) Preparation of cBSA-auNCs: BSA-AuNCs are prepared by taking bovine serum albumin as a template agent and a reducing agent, and the specific preparation steps are as follows: xie J, zheng Y, ying J Y.protein-directed Synthesis of Highly Fluorescent Gold Nanoclusters [ J ]. Journal of the American Chemical Society,2009,131 (3): 888-889. Ethylenediamine hydrochloride solution (5 mol/L,5 mL) was added to the purified BSA-AuNCs (10 mmol/L,5 mL) with stirring at 1000rpm, and after stirring for 2 minutes, 100mg EDC was added and the reaction was continued with stirring at 1000rpm for 2 hours. The reaction final solution was subjected to 5 ultrafiltration centrifugation operations using a 30kDa ultrafiltration tube to purify the amino-modified cBSA-auNCs. The prepared sample cBSA-auNCs was stored at 4℃for further use.
3) Preparation of bioactive glass nanocomposite: the mass ratio of BGN to cBSA-auNCs is 1:1. 15mg of BGN was taken and sufficiently dispersed in a small amount of deionized water by sonication and slowly added to 10mL of cBSA-auNCs (1.5 mg/mL), reacted for 48 hours with vigorous stirring at room temperature, and the reaction final solution was centrifuged at 8000rpm for 15 minutes and the precipitate was washed 3 times to remove cBSA-auNCs which were not effectively bonded.
Example 5
1) Preparation of BGN: 0.6g of urea and 1g of cetylpyridinium bromide were taken and dissolved well in 30mL of deionized water for 20 minutes, then 30mL of cyclohexane and 1.1mL of n-butanol were slowly added and vigorously stirred for 30 minutes to form a uniform emulsion. 2.7mL TEOS was then slowly added dropwise, hydrolyzed for 30 minutes, then warmed to 65deg.C, and reacted with vigorous stirring for 16 hours. Finally, 0.7mg of calcium nitrate tetrahydrate was added, and the reaction was continued for 4 hours. The reaction final solution was washed 3 times with acetone, ethanol and water, respectively. The precipitate obtained was freeze-dried and then calcined at 600℃for 5 hours (heating rate 1 ℃/min). The prepared sample BGN is stored at room temperature for standby.
2) Preparation of cBSA-auNCs: BSA-AuNCs are prepared by taking bovine serum albumin as a template agent and a reducing agent, and the specific preparation steps are as follows: xie J, zheng Y, ying J Y.protein-directed Synthesis of Highly Fluorescent Gold Nanoclusters [ J ]. Journal of the American Chemical Society,2009,131 (3): 888-889. Ethylenediamine hydrochloride solution (1 mol/L,5 mL) was added to the purified BSA-AuNCs (10 mmol/L,5 mL) with stirring at 1000rpm, and after stirring for 2 minutes, 20mg EDC was added and the reaction was continued with stirring at 1000rpm for 6 hours. The reaction final solution was subjected to 5 ultrafiltration centrifugation operations using a 30kDa ultrafiltration tube to purify the amino-modified cBSA-auNCs. The prepared sample cBSA-auNCs was stored at 4℃for further use.
3) Preparation of bioactive glass nanocomposite: the mass ratio of BGN to cBSA-auNCs is 1:1. 15mg of BGN was taken and sufficiently dispersed in a small amount of deionized water by sonication and slowly added to 10mL of cBSA-auNCs (1.5 mg/mL), reacted for 48 hours with vigorous stirring at room temperature, and the reaction final solution was centrifuged at 8000rpm for 15 minutes and the precipitate was washed 3 times to remove cBSA-auNCs which were not effectively bonded.
Example 6
1) Preparation of BGN: 0.6g of urea and 1g of cetylpyridinium bromide were taken and dissolved well in 30mL of deionized water for 20 minutes, then 30mL of cyclohexane and 1.1mL of n-butanol were slowly added and vigorously stirred for 30 minutes to form a uniform emulsion. 2.7mL TEOS was then slowly added dropwise, hydrolyzed for 30 minutes, then warmed to 75deg.C and reacted with vigorous stirring for 8 hours. Finally, 0.7mg of calcium nitrate tetrahydrate was added, and the reaction was continued for 4 hours. The reaction final solution was washed 3 times with acetone, ethanol and water, respectively. The precipitate obtained was freeze-dried and then calcined at 600℃for 5 hours (heating rate 1 ℃/min). The prepared sample BGN is stored at room temperature for standby.
2) Preparation of cBSA-auNCs: BSA-AuNCs are prepared by taking bovine serum albumin as a template agent and a reducing agent, and the specific preparation steps are as follows: xie J, zheng Y, ying J Y.protein-directed Synthesis of Highly Fluorescent Gold Nanoclusters [ J ]. Journal of the American Chemical Society,2009,131 (3): 888-889. Ethylenediamine hydrochloride solution (2.5 mol/L,5 mL) was added to the purified BSA-AuNCs (10 mmol/L,5 mL) with stirring at 1000rpm, and after stirring for 2 minutes, 50mg EDC was added and the reaction was continued with stirring at 1000rpm for 4 hours. The reaction final solution was subjected to 5 ultrafiltration centrifugation operations using a 30kDa ultrafiltration tube to purify the amino-modified cBSA-auNCs. The prepared sample cBSA-auNCs was stored at 4℃for further use.
3) Preparation of bioactive glass nanocomposite: the mass ratio of BGN to cBSA-auNCs is 1:1. 15mg of BGN was taken and sufficiently dispersed in a small amount of deionized water by sonication and slowly added to 10mL of cBSA-auNCs (1.5 mg/mL), reacted for 48 hours with vigorous stirring at room temperature, and the reaction final solution was centrifuged at 8000rpm for 15 minutes and the precipitate was washed 3 times to remove cBSA-auNCs which were not effectively bonded.
Example 7
1) Preparation of BGN: 0.6g of urea and 1g of cetylpyridinium bromide were taken and dissolved well in 30mL of deionized water for 20 minutes, then 30mL of cyclohexane and 1.1mL of n-butanol were slowly added and vigorously stirred for 30 minutes to form a uniform emulsion. 2.7mL TEOS was then slowly added dropwise, hydrolyzed for 30 minutes, then warmed to 70℃and reacted for 13 hours with continued vigorous stirring. Finally, 0.7mg of calcium nitrate tetrahydrate was added, and the reaction was continued for 4 hours. The reaction final solution was washed 3 times with acetone, ethanol and water, respectively. The precipitate obtained was freeze-dried and then calcined at 600℃for 5 hours (heating rate 1 ℃/min). The prepared sample BGN is stored at room temperature for standby.
2) Preparation of cBSA-auNCs: BSA-AuNCs are prepared by taking bovine serum albumin as a template agent and a reducing agent, and the specific preparation steps are as follows: xie J, zheng Y, ying J Y.protein-directed Synthesis of Highly Fluorescent Gold Nanoclusters [ J ]. Journal of the American Chemical Society,2009,131 (3): 888-889. Ethylenediamine hydrochloride solution (3 mol/L,5 mL) was added to the purified BSA-AuNCs (10 mmol/L,5 mL) with stirring at 1000rpm, and after stirring for 2 minutes, 60mg EDC was added and the reaction was continued with stirring at 1000rpm for 5 hours. The reaction final solution was subjected to 5 ultrafiltration centrifugation operations using a 30kDa ultrafiltration tube to purify the amino-modified cBSA-auNCs. The prepared sample cBSA-auNCs was stored at 4℃for further use.
3) Preparation of bioactive glass nanocomposite: the mass ratio of BGN to cBSA-auNCs is 1:1. 15mg of BGN was taken and sufficiently dispersed in a small amount of deionized water by sonication and slowly added to 10mL of cBSA-auNCs (1.5 mg/mL), the reaction was vigorously stirred at room temperature for 24 hours, the reaction final solution was centrifuged at 8000rpm for 15 minutes and the precipitate was washed 3 times to remove cBSA-auNCs which were not effectively bonded.
Example 8
1) Preparation of BGN: 0.6g of urea and 1g of cetylpyridinium bromide were taken and dissolved well in 30mL of deionized water for 20 minutes, then 30mL of cyclohexane and 1.1mL of n-butanol were slowly added and vigorously stirred for 30 minutes to form a uniform emulsion. 2.7mL TEOS was then slowly added dropwise, hydrolyzed for 30 minutes, then warmed to 70℃and reacted for 8 hours with continued vigorous stirring. Finally, 0.7mg of calcium nitrate tetrahydrate was added, and the reaction was continued for 4 hours. The reaction final solution was washed 3 times with acetone, ethanol and water, respectively. The precipitate obtained was freeze-dried and then calcined at 600℃for 5 hours (heating rate 1 ℃/min). The prepared sample BGN is stored at room temperature for standby.
2) Preparation of cBSA-auNCs: BSA-AuNCs are prepared by taking bovine serum albumin as a template agent and a reducing agent, and the specific preparation steps are as follows: xie J, zheng Y, ying J Y.protein-directed Synthesis of Highly Fluorescent Gold Nanoclusters [ J ]. Journal of the American Chemical Society,2009,131 (3): 888-889. Ethylenediamine hydrochloride solution (4 mol/L,5 mL) was added to the purified BSA-AuNCs (10 mmol/L,5 mL) with stirring at 1000rpm, and after stirring for 2 minutes, 80mg EDC was added and the reaction was continued with stirring at 1000rpm for 2 hours. The reaction final solution was subjected to 5 ultrafiltration centrifugation operations using a 30kDa ultrafiltration tube to purify the amino-modified cBSA-auNCs. The prepared sample cBSA-auNCs was stored at 4℃for further use.
3) Preparation of bioactive glass nanocomposite: the mass ratio of BGN to cBSA-auNCs is 1:1. 15mg of BGN was taken and sufficiently dispersed in a small amount of deionized water by sonication and slowly added to 10mL of cBSA-auNCs (1.5 mg/mL), the reaction was vigorously stirred at room temperature for 24 hours, the reaction final solution was centrifuged at 8000rpm for 15 minutes and the precipitate was washed 3 times to remove cBSA-auNCs which were not effectively bonded.
The bioactive glass nanocomposite obtained in the preparation process is light yellow powder after freeze drying, and has obvious red fluorescence under an ultraviolet lamp.
The effective cBSA-AUNCs loaded in the bioactive glass nanocomposite prepared by the invention has obvious dependence on the feeding ratio. FIG. 1 is a fluorescence spectrum of a bioactive glass nanocomposite, wherein samples BA1, BA2, BA3 and BA4 were BGN-AuNCs composites prepared with a BGN to cBSA-auNCs mass ratio of 3:1 (example 1), 2:1 (example 2), 1.5:1 (example 3) and 1:1 (example 4) in that order. From FIG. 1, it can be seen that when the mass ratio of BGN to cBSA-auNCs is gradually reduced within the range of 3:1-1:1, i.e., the input amount of cBSA-auNCs in the reaction system is gradually increased, the effective load amount of BGN to cBSA-auNCs is also continuously increased, which is manifested in that the fluorescence intensity of BGN-auNCs is continuously enhanced.
FIG. 2 is a Zeta potential diagram of a bioactive glass nanocomposite, BGN itself shows electronegativity, the Zeta potential is-12.9+ -0.1 mV, as the amount of cBSA-auNCs fed in a reaction system increases, the amount of cBSA-auNCs loaded by BGN also increases, so that the potential shows electropositivity and gradually increases, and when the mass ratio of BGN to cBSA-auNCs is 1:1, the potential of the BGN-auNCs bioactive glass nanocomposite reaches 30.8+ -0.3 mV. Therefore, when the reaction mass ratio of BGN to cBSA-auNCs is 1:1, the recombination efficiency of the two is optimal, and sample BA4 has excellent fluorescence properties and the surface has rich positive charges.
Fig. 3 (a) is a transmission electron microscope image of a bioactive glass nanocomposite BA4, wherein fig. 3 (a) is a transmission electron microscope image of a sample BA4, fig. 3 (b) is an energy spectrum scanning area of the sample BA4, and fig. 3 (c), (d), (e), (f), and (g) are distribution diagrams of Si, O, ca, au and S elements, respectively. It can be seen that when the reaction mass ratio of BGN to cBSA-AuNCs is 1:1, the morphology and structure of BGN are not destroyed after the BGN and the cBSA-AuNCs are combined efficiently, and the BGN still has good monodispersity, and the distribution of Au and S elements can judge that the cBSA-AuNCs are uniformly distributed in the BGN.
FIG. 4 is a graph of enzyme-like catalytic effect of bioactive glass nanocomposite BA 4. The substrate 3,3', 5' -tetramethyl benzidine (TMB) was used to detect the catalase-like properties of the bioactive glass nanocomposite BGN-AuNCs. Taking 20 mu L of BGN-AuNCs (BA 4) with certain concentration and uniformly dispersing by ultrasonic, adding 150 mu L of Phosphate Buffer Solution (PBS) with pH=6.0, then sequentially adding 10 mu L of TMB (final concentration of 800 mu mol/L) and 20 mu L H 2 O 2 (final concentration 50 mmol/L), after 15 minutes incubation at 37℃BGN-AuNCS was removed by centrifugation and absorbance changes of each set of supernatants were recorded by UV-visible absorption spectroscopy. At H 2 O 2 BGN-AuNCs catalyze H in the presence of 2 O 2 Decomposing to generate hydroxyl radicalThereby oxidizing TMB to oxidized TMB, with distinct characteristic absorption peaks at 370nm and 650 nm. Thus, as shown in fig. 4, BGN does not have obvious catalytic performance, but the bioactive glass nanocomposite BGN-AuNCs of the present invention has obvious characteristic absorption peaks at 370nm and 650nm, which indicates that BGN has excellent catalase-like catalytic activity after being compounded with cBSa-AuNCs.
Fig. 5 is a graph of antibacterial effect of bioactive glass nanocomposite BA 4. By 10 -4 mol/L H 2 O 2 Together with different concentrations of BGN-AuNCs 10 6 CFU/mL escherichia coli or staphylococcus aureus were incubated on a shaker for 6 hours (37 ℃,200 rpm), 100 μl of the diluted co-incubation solution was smeared on an agar plate, and incubated at 37 ℃ for 16 hours for colony counting. From the results of FIG. 5, it was found that BGN-AuNCs were able to effectively kill 95.9% of E.coli (see (a) in FIG. 5) and 56.4% of Staphylococcus aureus (see (b) in FIG. 5) at a low concentration of 75. Mu.g/mL. Further, since BGN-AuNCs have excellent peroxidase-like activity, it is possible to produce a low concentration of H 2 O 2 Under the condition, good synergistic antibacterial performance is exerted by generating active oxygen. At 10 -4 M H 2 O 2 And after 75 mug/mL BGN-AuNCs, the effective killing rate of the material against escherichia coli is 100% (see (a) in figure 5), the effective killing rate against staphylococcus aureus is 81.6% (see (b) in figure 5), and the bioactive glass nanocomposite material provided by the invention has excellent antibacterial effect and can kill bacteria through various ways such as positive surface charges, active oxygen generation and the like.
The invention carries out amination modification on gold nanoclusters (BSA-AuNCs) synthesized in one step, then reacts with BGN prepared by a microemulsion method according to a certain proportion, finally removes the cBSA-auNCs which are not effectively bonded through centrifugation, and the obtained bioactive glass nano-composite (BGN-AuNCs) is stored at room temperature after freeze-drying. The technology for carrying out positive electricity on BSA-AuNCs can promote the efficient combination of the BSA-AuNCs and bioactive glass nano particles on one hand, and is the key for improving the catalytic antibacterial performance of BGN-AuNCs on the other hand.
The preparation method of the bioactive glass nanocomposite is simple, the reaction condition is mild, and complex instruments and equipment are not needed. Meanwhile, the bioactive glass nano composite material prepared by the method has good biocompatibility, excellent enzyme-like catalytic and antibacterial activities, can greatly reduce the action dose of the bioactive glass nano material in the treatment of infectious diseases, and is widely applied to the biomedical fields such as tissue repair and reconstruction.
The invention is different from the previous synthetic strategies (ion doping and antibiotic loading) of the antibacterial bioactive glass, so that adverse factors such as high potential toxicity of ions, uncontrollable release of antibiotics and the like can be effectively eliminated, and the application limit of the antibacterial bioactive glass in the field of regenerative medicine is broken through. The invention utilizes mesoporous radial bioactive glass nano particles (BGN) to efficiently load aminated bovine serum albumin protected gold clusters (cBSA-auNCs), and the prepared nano composite not only has good biocompatibility, but also has excellent enzyme-like catalytic antibacterial activity.
The above is only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited by this, and any modification made on the basis of the technical scheme according to the technical idea of the present invention falls within the protection scope of the claims of the present invention.

Claims (7)

1. The preparation method of the bioactive glass nanocomposite with catalytic antibacterial performance is characterized by comprising the following steps:
1) Preparing bioactive glass nano particles by adopting a microemulsion method;
2) Reacting the bioactive glass nano-particles with the positive electrochemical gold nanoclusters in a reaction solvent to obtain a bioactive glass nano-composite material with catalytic antibacterial performance;
wherein, the positive electrification gold nanocluster is prepared by the following processes: preparing gold nanoclusters by taking bovine serum albumin as a template agent and a reducing agent; and (3) carrying out amination modification reaction on the gold nanocluster, ethylenediamine hydrochloride and 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide to obtain the positive-electrification gold nanocluster.
2. The method for preparing a bioactive glass nanocomposite with catalytic antibacterial properties according to claim 1, wherein the bioactive glass nanoparticles comprise silicon element and calcium element in a molar ratio of 4:1-4:2.
3. The method for preparing the bioactive glass nanocomposite with catalytic antibacterial performance according to claim 1, wherein the dosage ratio of gold nanoclusters to ethylenediamine hydrochloride is 10mmol:1-5mol.
4. The method for preparing the bioactive glass nanocomposite with catalytic antibacterial performance according to claim 1, wherein the ratio of the gold nanoclusters to the 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide is 10mmol:4-20mg.
5. The method for preparing a bioactive glass nanocomposite with catalytic antibacterial properties according to claim 1, wherein the amination modification reaction is carried out for 0.5-6 hours.
6. The method for preparing the bioactive glass nanocomposite with catalytic antibacterial performance according to claim 1, wherein the mass ratio of bioactive glass nanoparticles to the gold nanoclusters subjected to positive electrification is 1:1-3:1.
7. The method for preparing a bioactive glass nanocomposite with catalytic antibacterial properties according to claim 1, wherein the reaction solvent in step 2) is ultrapure water, and the reaction time is 12-48 hours.
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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102078617A (en) * 2010-12-28 2011-06-01 中国科学院上海硅酸盐研究所 Method for synthesizing mesoporous silica-based nanocomposite embedded with gold nanorods
CN105452185A (en) * 2013-06-17 2016-03-30 康宁股份有限公司 Antimicrobial glass articles with improved strength and methods of making and using same
RO132343A2 (en) * 2016-07-20 2018-01-30 Universitatea "Babeş-Bolyai" Din Cluj-Napoca Porous bioactive glass doped with gold nanoparticles
WO2021011398A1 (en) * 2019-07-12 2021-01-21 The Board Of Trustees Of The Leland Stanford Junior University Functionalized nanoparticles and their use in treating bacterial infections
CN112472665A (en) * 2020-12-04 2021-03-12 上海交通大学医学院附属第九人民医院 Self-oxygen-generating photodynamic light therapy enhanced transdermal drug delivery nano gel and preparation method thereof
CN114191548A (en) * 2021-11-19 2022-03-18 三峡大学 Tumor microenvironment responsive nano-assembly and application thereof in tumor combined treatment

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070259101A1 (en) * 2006-05-02 2007-11-08 Kleiner Lothar W Microporous coating on medical devices

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102078617A (en) * 2010-12-28 2011-06-01 中国科学院上海硅酸盐研究所 Method for synthesizing mesoporous silica-based nanocomposite embedded with gold nanorods
CN105452185A (en) * 2013-06-17 2016-03-30 康宁股份有限公司 Antimicrobial glass articles with improved strength and methods of making and using same
RO132343A2 (en) * 2016-07-20 2018-01-30 Universitatea "Babeş-Bolyai" Din Cluj-Napoca Porous bioactive glass doped with gold nanoparticles
WO2021011398A1 (en) * 2019-07-12 2021-01-21 The Board Of Trustees Of The Leland Stanford Junior University Functionalized nanoparticles and their use in treating bacterial infections
CN112472665A (en) * 2020-12-04 2021-03-12 上海交通大学医学院附属第九人民医院 Self-oxygen-generating photodynamic light therapy enhanced transdermal drug delivery nano gel and preparation method thereof
CN114191548A (en) * 2021-11-19 2022-03-18 三峡大学 Tumor microenvironment responsive nano-assembly and application thereof in tumor combined treatment

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
Antibacterial activity of nanostructured Ti−45S5 bioglass−Ag composite against Streptococcus mutans and Staphylococcus aureus;K. JURCZYK et al.;《Trans. Nonferrous Met. Soc. China》;第26卷;第118-125页 *

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