CN114796582A - Preparation method of bacteriostatic gel containing graphene oxide - Google Patents
Preparation method of bacteriostatic gel containing graphene oxide Download PDFInfo
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- CN114796582A CN114796582A CN202210380379.8A CN202210380379A CN114796582A CN 114796582 A CN114796582 A CN 114796582A CN 202210380379 A CN202210380379 A CN 202210380379A CN 114796582 A CN114796582 A CN 114796582A
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 57
- 229910021389 graphene Inorganic materials 0.000 title claims abstract description 56
- 238000002360 preparation method Methods 0.000 title claims abstract description 28
- 230000003385 bacteriostatic effect Effects 0.000 title claims description 28
- 229920001661 Chitosan Polymers 0.000 claims abstract description 28
- 230000000844 anti-bacterial effect Effects 0.000 claims abstract description 28
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 25
- 235000010413 sodium alginate Nutrition 0.000 claims abstract description 25
- 239000000661 sodium alginate Substances 0.000 claims abstract description 25
- 229940005550 sodium alginate Drugs 0.000 claims abstract description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 20
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 20
- 239000008367 deionised water Substances 0.000 claims description 20
- 229910021641 deionized water Inorganic materials 0.000 claims description 20
- 239000011812 mixed powder Substances 0.000 claims description 18
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 16
- 239000000843 powder Substances 0.000 claims description 16
- 238000005303 weighing Methods 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 12
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- 238000002156 mixing Methods 0.000 claims description 11
- 238000001132 ultrasonic dispersion Methods 0.000 claims description 11
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 10
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 10
- 239000001103 potassium chloride Substances 0.000 claims description 10
- 235000011164 potassium chloride Nutrition 0.000 claims description 10
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- 238000003756 stirring Methods 0.000 claims description 10
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- 238000012986 modification Methods 0.000 claims description 5
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- 239000006228 supernatant Substances 0.000 claims description 3
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- 238000006243 chemical reaction Methods 0.000 claims description 2
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- 230000001105 regulatory effect Effects 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 6
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- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 abstract description 2
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- AEMOLEFTQBMNLQ-AZLKCVHYSA-N (2r,3s,4s,5s,6r)-3,4,5,6-tetrahydroxyoxane-2-carboxylic acid Chemical compound O[C@@H]1O[C@@H](C(O)=O)[C@@H](O)[C@H](O)[C@@H]1O AEMOLEFTQBMNLQ-AZLKCVHYSA-N 0.000 description 1
- AEMOLEFTQBMNLQ-SYJWYVCOSA-N (2s,3s,4s,5s,6r)-3,4,5,6-tetrahydroxyoxane-2-carboxylic acid Chemical compound O[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@@H]1O AEMOLEFTQBMNLQ-SYJWYVCOSA-N 0.000 description 1
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- 231100000956 nontoxicity Toxicity 0.000 description 1
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- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/28—Polysaccharides or their derivatives
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
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Abstract
The invention discloses a preparation method of a graphene oxide-containing antibacterial gel, which comprises the steps of preparing a graphene oxide suspension and a buffer solution, and reacting with sodium alginate and chitosan to obtain the gel. According to the invention, the biochemical graphene/chitosan single-network structure is prepared by functionalizing natural high-molecular chitosan and neutralizing the electronegativity of graphene, the mechanical strength and porosity optimization is realized by controlling a microstructure, the functional defects of alginate gel are solved, and the chitosan which is a low-cost and wide-source natural high-molecular material is adopted in the preparation process, so that the toxicity and environmental protection problems are solved, the graphene dispersion is promoted in the gel, a double-network reinforced structure is formed with sodium alginate, and the mechanical property and the anti-cracking property of the gel are improved. Due to the antibacterial function of the graphene, the gel has long-term antibacterial capacity, and the effect of controlling bacterial infection for a long time is achieved. The gel provided by the invention has high mechanical properties, inhibits the growth of microorganisms, and has the advantages of simple preparation method and no biotoxicity.
Description
Technical Field
The invention belongs to the field of medical materials, relates to a preparation method of gel, and particularly relates to a preparation method of antibacterial gel containing graphene oxide, which is simple to prepare, free of biotoxicity and environment-friendly in raw material.
Background
Tissue engineering achieves the therapeutic goal by combining biological materials with specific biologically active cells to construct tissues/organs with functional activity. Among the many effective biomaterials applied to tissue engineering, sodium alginate is a polysaccharide substance extracted from seaweed plants, derived from marine algae, is a block linear polymer consisting of 2 monomers of β -D-mannuronic acid (M) and α -L-guluronic acid (G), can be rapidly dissolved in water, and has many excellent properties, such as: no toxicity, hydrophilicity, biocompatibility, biodegradability, low price and the like, so the biological membrane is widely applied to the field of biological medicine. The fiber material with the characteristics of the raw material can be prepared by taking sodium alginate as the raw material and utilizing spinning technology and equipment, and the fiber material is researched and applied to the technical fields of wound dressings and the like.
Sodium alginate and chitosan are natural polysaccharides which are widely researched at present, and have the potential of being developed into a drug carrier by virtue of excellent bioadhesion, biocompatibility and no toxic or side effect. In addition, the two polysaccharides respectively have carboxylate ions and amino groups, show pH sensitivity, and have medicinal prospects which are mainly focused on the aspects of pH-sensitive and sustained-release drug carriers.
However, the existing sodium alginate chitosan gel has the defects of low strength, easy breakage and the like, and the moisture absorption, retention and permeability performances of the gel are required to be further improved.
Disclosure of Invention
In order to solve the defects of poor mechanical strength, no environmental protection and poor antibacterial activity of the existing sodium alginate chitosan gel, the invention provides a preparation method of the antibacterial gel containing graphene oxide.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of a bacteriostatic gel containing graphene oxide comprises the following steps:
1) adding graphene oxide into deionized water for ultrasonic dispersion to obtain a solution A;
2) dissolving sodium chloride, potassium chloride, disodium hydrogen phosphate and dipotassium hydrogen phosphate in deionized water, and regulating the pH value of the solution by using hydrochloric acid to obtain a solution B;
3) weighing sodium alginate powder and chitosan powder, and fully and uniformly mixing to obtain mixed powder a;
4) adding the mixed powder a of the step 3) into the solution B of the step 2), then adding the solution A of the step 1), and heating and stirring the whole system to obtain a solution C.
In the invention, the graphene has a very large specific surface area and a large number of surface functional groups, such as hydroxyl, carboxyl, epoxide and carbon, has very strong adsorption capacity on protein, can enhance the adhesion of a material interface and cells, and can be subjected to various functional modifications. According to the invention, the biochemical graphene/chitosan single-network structure is prepared by functionalizing natural high-molecular chitosan and neutralizing the electronegativity of graphene, the mechanical strength and porosity optimization is realized by controlling a microstructure, the functional defects of alginate gel are solved, and the chitosan which is a low-cost and wide-source natural high-molecular material is adopted in the preparation process, so that the toxicity and environmental protection problems are solved, the graphene dispersion is promoted in the gel, a double-network reinforced structure is formed with sodium alginate, and the mechanical property and the anti-cracking property of the gel are improved.
Meanwhile, due to the antibacterial function of the graphene, the gel has long-term bacteriostatic ability, and the effect of controlling germ infection for a long time is achieved.
The invention prepares the gel biological composite material which has high mechanical property, inhibits the growth of microorganisms, promotes the attachment, proliferation and differentiation of cells, has simple preparation method and no biological toxicity.
As a preferable scheme of the invention, in the step 1), the number of times of ultrasonic dispersion is 2-5, the time of each ultrasonic dispersion is 15-25min, the gap time is 25-35min, and the ultrasonic power is 250-.
In a preferable embodiment of the invention, in step 1), the concentration of graphene oxide in the solution a is 8-12 mg/mL.
In the technical scheme, the content of the graphene oxide is too much, so that the graphene oxide is unevenly dispersed in a system, and the cost is increased; and too little addition can result in low degree of combination of the graphene oxide, the chitosan and the sodium alginate, so that the mechanical property and the antibacterial property required by the invention cannot be achieved.
As a preferable scheme of the present invention, in the step 2), the mass ratio of sodium chloride, potassium chloride, disodium hydrogen phosphate, dipotassium hydrogen phosphate and deionized water is 8: 0.2: 1.44: 0.24: 1000.
as a preferable mode of the present invention, in the step 2), the pH value is 7 to 8.
As a preferable scheme of the invention, in the step 3), the mass ratio of sodium alginate to chitosan is 1: 0.1-1: 10.
in a preferable embodiment of the invention, in the step 4), the mixed powder a is 2 to 20 parts by weight, the solution B is 94 parts by weight, and the solution A is 0 to 20 parts by weight.
As a preferable scheme of the invention, in the step 4), the reaction temperature is 40-60 ℃, and the stirring time is 2-12 h.
As a preferable embodiment of the present invention, the graphene oxide used in step 1) is surface-modified.
As a preferred embodiment of the present invention, a surface modification method of graphene oxide comprises: heating the solution A to 65-70 ℃, adding 2, 2' - (1, 2-ethanediylbis oxo) bisethanethiol, wherein the addition amount is 1% -2% of the mass of graphene oxide; and continuing ultrasonic treatment for 2-3min, centrifuging, removing the supernatant, adding the lower-layer substance into deionized water, and ultrasonically mixing to obtain a modified solution A.
In the technical scheme, the graphene oxide is subjected to surface modification firstly, so that the dispersibility of the graphene oxide in a system is further improved, and secondly, after the graphene oxide is dispersed in the system, the subsequently prepared gel is convenient for drug loading or targeted delivery.
Compared with the prior art, the invention has the following beneficial effects:
1) the chitosan in the raw materials is low in cost and wide in source, solves the problems of toxicity and environmental protection, and accords with the concept of environmental protection;
2) according to the invention, the antibacterial function of the graphene oxide is utilized, so that the hydrogel has long-term antibacterial capability, and the effect of controlling bacterial infection for a long time is achieved;
3) according to the invention, abundant hydroxyl, carboxyl, epoxy group and other groups on the surface of graphene oxide are utilized, and a good interface effect is formed with a hydrophilic matrix through hydrogen bonds and chemical bonds, so that the interface compatibility is improved, and the swelling performance of the gel is improved; in addition, the mechanical strength of the hydrogel is also improved due to the mechanical properties of graphene oxide;
4) the bacteriostatic gel prepared by the invention can be used for: 1) the medical beauty field has great potential value in developing wound dressing and artificial skin; 2) the field of biological tissue engineering, which can be used for developing artificial implanted cartilage tissues; 3) the drug-carrying and targeted transportation field can provide reference value for drug-carrying materials; 4) the medicine packaging field can be further developed as an oral medicine capsule material.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The embodiment provides a preparation method of a bacteriostatic gel containing graphene oxide, which comprises the following steps:
(1) preparation of antibacterial gel
a) Weighing a proper amount of graphene oxide, and ultrasonically dispersing in deionized water, wherein the ultrasonic power is 285W, the ultrasonic dispersion time is 20 minutes each time and 3 times in total, and the intermittent time is 30 minutes, so that a uniform solution A of 10mg/mL is finally obtained;
b) dissolving 8g of sodium chloride, 0.2g of potassium chloride, 1.44g of disodium hydrogen phosphate and 0.24g of dipotassium hydrogen phosphate in 1000g of deionized water, and adjusting the pH value of the solution to 7.4 by using hydrochloric acid to prepare a solution B;
c) according to the weight percentage of sodium alginate: chitosan 1: 1, weighing sodium alginate powder and chitosan powder, and fully and uniformly mixing to obtain mixed powder a;
d) adding 0.6g of the mixed powder a into 9.4g of the solution B, then adding 0.01g of the solution A, and heating and stirring the whole system at 50 ℃ for 8 hours to obtain a solution C;
(2) gel bacteriostatic performance test
(a) Transferring the solution C obtained in (1) to a container, and then placing the container in an autoclave for sterilization at 80 ℃ for 40 min.
(b) According to the formula C: liquid medium ═ 1: 1, removing the solution C and the liquid culture medium and adding the solution C and the liquid culture medium into the conical flask A. At the same time, an equal total amount of liquid medium was removed and added to Erlenmeyer flask B as a blank comparison.
(c) Erlenmeyer flask A, B was then placed in a cell growth curve (OD value) on-line detection bioreactor (Shanghai Zhicheng) for calibration.
(d) After the calibration, the strain was added in an amount of 0.2 OD600 to the flask, incubated at 37 ℃ for 12 hours, and the OD value was measured in real time.
(e) The method is used for testing the bacteriostasis rate of the antibacterial gel on escherichia coli and staphylococcus aureus.
Example 2
The embodiment provides a preparation method of a bacteriostatic gel containing graphene oxide, which comprises the following steps:
(1) preparation of antibacterial gel
a) Weighing a proper amount of graphene oxide, and ultrasonically dispersing in deionized water, wherein the ultrasonic power is 285W, the ultrasonic dispersion time is 20 minutes each time and 3 times in total, and the intermittent time is 30 minutes, so that a uniform solution A of 10mg/mL is finally obtained;
b) dissolving 8g of sodium chloride, 0.2g of potassium chloride, 1.44g of disodium hydrogen phosphate and 0.24g of dipotassium hydrogen phosphate in 1000g of deionized water, and adjusting the pH value of the solution to 7.4 by using hydrochloric acid to prepare a solution B;
c) according to the weight percentage of sodium alginate: chitosan ═ 1: 1, weighing sodium alginate powder and chitosan powder, and fully and uniformly mixing to obtain mixed powder a;
d) adding 0.6g of the mixed powder a into 9.4g of the solution B, then adding 0.05g of the solution A, and heating and stirring the whole system at 50 ℃ for 8 hours to obtain a solution C;
(2) gel bacteriostatic performance test
(a) Transferring the solution C obtained in (1) to a container, and then placing the container in an autoclave for sterilization at 80 ℃ for 40 min.
(b) According to the formula C: liquid medium ═ 1: 1, removing the solution C and the liquid culture medium and adding the solution C and the liquid culture medium into the conical flask A. At the same time, an equal total amount of liquid medium was removed and added to Erlenmeyer flask B as a blank comparison.
(c) Erlenmeyer flask A, B was then placed in a cell growth curve (OD value) on-line detection bioreactor (Shanghai Zhicheng) for calibration.
(d) After the calibration, the strain was added in an amount of 0.2 OD600 to the flask, incubated at 37 ℃ for 12 hours, and the OD value was measured in real time.
(e) The method is used for testing the bacteriostasis rate of the antibacterial gel on escherichia coli and staphylococcus aureus.
Example 3
The embodiment provides a preparation method of a bacteriostatic gel containing graphene oxide, which comprises the following steps:
(1) preparation of antibacterial gel
a) Weighing a proper amount of graphene oxide, and ultrasonically dispersing in deionized water, wherein the ultrasonic power is 285W, the ultrasonic dispersion time is 20 minutes each time and 3 times in total, and the intermittent time is 30 minutes, so that a uniform solution A of 10mg/mL is finally obtained;
b) dissolving 8g of sodium chloride, 0.2g of potassium chloride, 1.44g of disodium hydrogen phosphate and 0.24g of dipotassium hydrogen phosphate in 1000g of deionized water, and adjusting the pH value of the solution to 7.4 by using hydrochloric acid to prepare a solution B;
c) according to the weight percentage of sodium alginate: chitosan ═ 1: 1, weighing sodium alginate powder and chitosan powder, and fully and uniformly mixing to obtain mixed powder a;
d) adding 0.6g of the mixed powder a into 9.4g of the solution B, then adding 0.1g of the solution A, and heating and stirring the whole system at 50 ℃ for 8 hours to obtain a solution C;
(2) gel bacteriostatic property test
(a) Transferring the solution C obtained in (1) to a container, and then placing the container in an autoclave for sterilization at 80 ℃ for 40 min.
(b) According to the formula C: liquid medium ═ 1: 1, adding the solution C and the liquid culture medium into the conical flask A. At the same time, an equal total amount of liquid medium was removed and added to Erlenmeyer flask B as a blank comparison.
(c) The Erlenmeyer flask A, B was then placed in a cell growth curve (OD) on-line detection bioreactor (Shanghai Zhicheng) for calibration.
(d) After the calibration, the strain was added in an amount of 0.2 OD600 to the flask, incubated at 37 ℃ for 12 hours, and the OD value was measured in real time.
(e) The method is used for testing the bacteriostasis rate of the antibacterial gel on escherichia coli and staphylococcus aureus.
Example 4
The embodiment provides a preparation method of a bacteriostatic gel containing graphene oxide, which comprises the following steps:
(1) preparation of antibacterial gel
a) Weighing a proper amount of graphene oxide, and ultrasonically dispersing in deionized water, wherein the ultrasonic power is 285W, the ultrasonic dispersion time is 20 minutes each time and 3 times in total, and the intermittent time is 30 minutes; then heating to 65-70 ℃, adding 2, 2' - (1, 2-ethanediylbis oxo) bisethanethiol, wherein the adding amount is 1-2% of the mass of the graphene oxide; continuing to perform ultrasonic treatment for 2-3min, centrifuging, removing supernatant, adding lower layer substance into deionized water, and performing ultrasonic mixing to obtain uniform 10mg/mL modified solution A
b) Dissolving 8g of sodium chloride, 0.2g of potassium chloride, 1.44g of disodium hydrogen phosphate and 0.24g of dipotassium hydrogen phosphate in 1000g of deionized water, and adjusting the pH value of the solution to 7.4 by using hydrochloric acid to prepare a solution B;
c) according to the weight percentage of sodium alginate: chitosan ═ 1: 1, weighing sodium alginate powder and chitosan powder, and fully and uniformly mixing to obtain mixed powder a;
d) adding 0.6g of the mixed powder a into 9.4g of the solution B, then adding 0.1g of the solution A, and heating and stirring the whole system at 50 ℃ for 8 hours to obtain a solution C;
(2) gel bacteriostatic performance test
(a) Transferring the solution C obtained in (1) to a container, and then placing the container in an autoclave for sterilization at 80 ℃ for 40 min.
(b) According to the formula C: liquid medium ═ 1: 1, removing the solution C and the liquid culture medium and adding the solution C and the liquid culture medium into the conical flask A. At the same time, an equal total amount of liquid medium was removed and added to Erlenmeyer flask B as a blank comparison.
(c) Erlenmeyer flask A, B was then placed in a cell growth curve (OD value) on-line detection bioreactor (Shanghai Zhicheng) for calibration.
(d) After the calibration, the strain was added in an amount of 0.2 OD600 to the flask, incubated at 37 ℃ for 12 hours, and the OD value was measured in real time.
(e) The method is used for testing the bacteriostasis rate of the antibacterial gel on escherichia coli and staphylococcus aureus.
Comparative example 1
(1) Preparation of antibacterial gel
a) Dissolving 8g of sodium chloride, 0.2g of potassium chloride, 1.44g of disodium hydrogen phosphate and 0.24g of dipotassium hydrogen phosphate in 1000g of deionized water, and adjusting the pH value of the solution to 7.4 by using hydrochloric acid to prepare a solution B;
b) according to the weight percentage of sodium alginate: chitosan ═ 1: 1, weighing sodium alginate powder and chitosan powder, and fully and uniformly mixing to obtain mixed powder a;
c) adding 0.6g of mixed powder a into 9.4g of solution B, heating the whole system at 50 ℃, and stirring for 8 hours to obtain solution C;
(2) gel bacteriostatic performance test
(a) Transferring the solution C obtained in (1) to a container, and then placing the container in an autoclave for sterilization at 80 ℃ for 40 min.
(b) According to the proportion of solution C: liquid medium ═ 1: 1, removing the solution C and the liquid culture medium and adding the solution C and the liquid culture medium into the conical flask A. At the same time, an equal total amount of liquid medium was removed and added to Erlenmeyer flask B as a blank comparison.
(c) Erlenmeyer flask A, B was then placed in a cell growth curve (OD value) on-line detection bioreactor (Shanghai Zhicheng) for calibration.
(d) After the calibration, the strain was added in an amount of 0.2 OD600 to the flask, incubated at 37 ℃ for 12 hours, and the OD value was measured in real time.
(e) The method is used for testing the bacteriostasis rate of the antibacterial gel on escherichia coli and staphylococcus aureus.
Comparative example 2
(1) Preparation of antibacterial gel
a) Weighing a proper amount of graphene oxide, and ultrasonically dispersing in deionized water, wherein the ultrasonic power is 285W, the ultrasonic dispersion time is 20 minutes each time and 3 times in total, and the intermittent time is 30 minutes, so that a uniform 20mg/mL solution A is finally obtained;
b) dissolving 8g of sodium chloride, 0.2g of potassium chloride, 1.44g of disodium hydrogen phosphate and 0.24g of dipotassium hydrogen phosphate in 1000g of deionized water, and adjusting the pH value of the solution to 7.4 by using hydrochloric acid to prepare a solution B;
c) according to the weight percentage of sodium alginate: chitosan ═ 1: 1, weighing sodium alginate powder and chitosan powder, and fully and uniformly mixing to obtain mixed powder a;
d) adding 0.6g of the mixed powder a into 9.4g of the solution B, then adding 0.1g of the solution A, and heating and stirring the whole system at 50 ℃ for 8 hours to obtain a solution C;
(2) gel bacteriostatic performance test
(a) Transferring the solution C obtained in (1) to a container, and then placing the container in an autoclave for sterilization at 80 ℃ for 40 min.
(b) According to the formula C: liquid medium ═ 1: 1, removing the solution C and the liquid culture medium and adding the solution C and the liquid culture medium into the conical flask A. At the same time, an equal total amount of liquid medium was removed and added to Erlenmeyer flask B as a blank comparison.
(c) Erlenmeyer flask A, B was then placed in a cell growth curve (OD value) on-line detection bioreactor (Shanghai Zhicheng) for calibration.
(d) After the calibration, the strain was added in an amount of 0.2 OD600 to the flask, incubated at 37 ℃ for 12 hours, and the OD value was measured in real time.
(e) The method is used for testing the bacteriostasis rate of the antibacterial gel on escherichia coli and staphylococcus aureus.
To test the long-lasting bacteriostatic activity of the antibacterial gel, the bacterial species having an OD600 of 0.2 was added again to the gels of examples 1-3 and comparative example 1 after 36 hours and 72 hours, respectively, and the results are shown in table 1.
TABLE 1 results of bacteriostatic properties of examples 1-3 and comparative example 1
| Group of | Rate of inhibition of bacteria | Bacteriostasis rate after 36 hours | Bacteriostasis rate after 72 hours |
| Example 1 | 89％～95％ | 88％～94％ | 85％～92％ |
| Example 2 | 97％～100％ | 95％～99％ | 93％～97％ |
| Example 3 | 100％ | 99％ | 95％～97％ |
| Comparative example 1 | 65％～75％ | 50％～55％ | 30％～45 |
As can be seen from table 1, the bacteriostatic activity of the gels prepared in examples 1 to 3 is better than that of the gel prepared in comparative example 1, while the bacteriostatic activity of the gels prepared in examples 2 and 3 is better than that of example 1, and it can be seen that the bacteriostatic activity of the gels is increased with the increase of the content of graphene oxide;
with the increase of time, the antibacterial performance of the gel prepared in the examples 1-3 can be seen to be kept for a long time, while the antibacterial performance of the gel prepared in the comparative example 1 is also reduced with the increase of time, and the invention can be seen in that the gel biological composite material which has high mechanical property, inhibits the growth of microorganisms, promotes the attachment, proliferation and differentiation of cells, has a simple preparation method and no biological toxicity is prepared, so that the hydrogel has long-term antibacterial capability, and can achieve the effect of controlling the bacterial infection for a long time.
The raw materials and equipment used in the invention are common raw materials and equipment in the field if not specified; the methods used in the present invention are conventional in the art unless otherwise specified.
While the invention has been described with respect to a preferred embodiment, it will be understood by those skilled in the art that the foregoing and other changes, omissions and deviations in the form and detail thereof may be made without departing from the scope of this invention. Those skilled in the art can make various changes, modifications and equivalent arrangements, which are equivalent to the embodiments of the present invention, without departing from the spirit and scope of the present invention, and which may be made by utilizing the techniques disclosed above; meanwhile, any changes, modifications and variations of the above-described embodiments, which are equivalent to those of the technical spirit of the present invention, are within the scope of the technical solution of the present invention.
Claims (10)
1. A preparation method of a bacteriostatic gel containing graphene oxide is characterized by comprising the following steps:
1) adding graphene oxide into deionized water for ultrasonic dispersion to obtain a solution A;
2) dissolving sodium chloride, potassium chloride, disodium hydrogen phosphate and dipotassium hydrogen phosphate in deionized water, and regulating the pH value of the solution by using hydrochloric acid to obtain a solution B;
3) weighing sodium alginate powder and chitosan powder, and fully and uniformly mixing to obtain mixed powder a;
4) adding the mixed powder a of the step 3) into the solution B of the step 2), then adding the solution A of the step 1), and heating and stirring the whole system to obtain a solution C.
2. The method for preparing bacteriostatic gel containing graphene oxide according to claim 1, wherein in step 1), the number of ultrasonic dispersion is 2-5, the time of each ultrasonic dispersion is 15-25min, the gap time is 25-35min, and the ultrasonic power is 250-.
3. The method for preparing the bacteriostatic gel containing graphene oxide according to claim 1, wherein in the step 1), the concentration of the graphene oxide in the solution A is 8-12 mg/mL.
4. The preparation method of the bacteriostatic gel containing graphene oxide according to claim 1, wherein in the step 2), the mass ratio of the sodium chloride, the potassium chloride, the disodium hydrogen phosphate, the dipotassium hydrogen phosphate and the deionized water is 8: 0.2: 1.44: 0.24: 1000.
5. the method for preparing the bacteriostatic gel containing the graphene oxide according to claim 1, wherein in the step 2), the pH value is 7-8.
6. The preparation method of the graphene oxide-containing antibacterial gel according to claim 1, wherein in the step 3), the mass ratio of sodium alginate to chitosan is 1: 0.1-1: 10.
7. the method for preparing an antibacterial gel containing graphene oxide according to claim 1, wherein in the step 4), the mixed powder a is 2-20 parts, the solution B is 94 parts, and the solution A is 0-20 parts by weight.
8. The preparation method of the bacteriostatic gel containing the graphene oxide according to claim 1, wherein in the step 4), the reaction temperature is 40-60 ℃ and the stirring time is 2-12 h.
9. The preparation method of the graphene oxide-containing bacteriostatic gel according to any one of claims 1 to 8, characterized in that the graphene oxide used in the step 1) is subjected to surface modification.
10. The preparation method of the bacteriostatic gel containing graphene oxide according to claim 9, wherein the surface modification method of graphene oxide comprises the following steps: heating the solution A to 65-70 ℃, adding 2, 2' - (1, 2-ethanediylbis oxo) bisethanethiol, wherein the addition amount is 1% -2% of the mass of graphene oxide; and continuing ultrasonic treatment for 2-3min, centrifuging, removing the supernatant, adding the lower-layer substance into deionized water, and ultrasonically mixing to obtain a modified solution A.
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