CN114225089B - Preparation method of electroactive antibacterial hemostatic dressing - Google Patents
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Abstract
A preparation method of an electroactive antibacterial hemostatic dressing, and relates to a preparation method of a hemostatic dressing. Aiming at solving the problem of unstable structure when the existing chitosan and MXene are compounded. The preparation method comprises the following steps: the chitosan fiber dressing base material is placed in a vacuum drying oven to be dried, mxene water dispersion liquid is prepared, MXene is modified by Poly Dopamine (PDA) to obtain a PDA modified MXene nanosheet solution, and finally the PDA modified MXene nanosheet solution is mixed with the chitosan fiber dressing to be subjected to suction filtration, so that the chitosan/MXene electroactive antibacterial hemostatic dressing is obtained. According to the invention, MXene is modified by oxidizing and polymerizing polydopamine on the surface of MXene, so that the tissue wet adhesion capability of the obtained dressing is improved, and the obtained dressing has good water absorption performance, excellent biocompatibility, hemostatic performance, conductivity and mechanical strength. The invention is suitable for preparing the hemostatic dressing.
Description
Technical Field
The invention relates to a preparation method of a hemostatic dressing.
Background
Uncontrolled bleeding and wound infections are the leading causes of death in the field of wound care. Improperly bandaged wounds can prolong healing time and pose a high risk of infection, which can seriously affect the quality of life of patients and even be a continuously growing cause of death in ill patients. In addition, the treatment of infections places a heavy burden on the medical system and even the entire society. For example, north america costs up to $ 100 billion per year for complex wound treatment, and by 2020, costs for wound care worldwide are expected to reach $ 220 billion. Despite advances in the development of advanced hemostatic materials over the past few decades, two challenges remain to be addressed, excessive blood loss during clot formation, strong clot adhesion to hemostatic dressings, pain, secondary bleeding, and possible infection upon dressing removal. Therefore, there is an urgent need to develop biomaterials for treating bacterially infected wounds. Although antibiotic dressings have been developed for the treatment of bacterial infected wounds, the side effects caused by antibiotics and the cytotoxicity of antibiotics themselves are not negligible. In addition, due to the existence of microbial biofilms, antibiotics have weak infiltration capacity on bacterial infection wound surfaces, and the treatment effect is influenced.
An ideal wound dressing needs to have: 1. is suitable for and actively adapts to various bleeding wounds; 2. can quickly reduce bleeding and meet the requirement of hemostasis for a long time; 3. easy preparation, disinfection and carrying; 4. can be preserved under various atmospheric and environmental conditions; 5. non-professional personnel should be easy to use if desired; 6. has reasonable biocompatibility to avoid causing any short-term or long-term adverse effects in vivo. To date, almost no wound dressing can completely meet the requirements of ideal wound dressings in clinic. Therefore, there is an urgent need to develop a multifunctional wound dressing for inhibiting wound infection and promoting wound healing.
Chitosan is a biodegradable polysaccharide with positive charge, has good biocompatibility, adhesion and impressive therapeutic functions, comprises hemostatic, antibacterial, antitumor and anti-inflammatory activities, and has good application effect in the fields of tissue engineering and drug delivery. The hemostatic properties of chitosan are believed to be due to its electrostatic interaction with the negatively charged cell membrane of erythrocytes, which leads to agglutination of the erythrocytes and formation of a tampon at the site of injury. Currently, chitosan-based wound dressings such as hydrogels, films, scaffolds and sponges are reported for different shaping methods, and some chitosan-based wound dressing products such as Syvek-Patch, chitopackC, tegasorb, hemCon bandage and KytoCel are already on the market.
Conductive polymers have been shown to modulate cellular activityIncluding promoting cell adhesion, proliferation, migration, and electrically stimulating cell differentiation. Since the skin is very sensitive to electrical stimulation, biomaterials with excellent electrical conductivity can construct a cellular communication network by increasing electrical transmission, thereby improving the quality of wound healing and skin regeneration. Therefore, designing a novel functional wound dressing with conductivity is more helpful for promoting wound healing. MXene, as a two-dimensional material consisting of transition metal carbides, nitrides or carbonitrides, is widely used in the biomedical field like other two-dimensional materials. MXene has many attractive properties such as hydrophilicity derived from its surface functional groups (e.g. hydroxyl, oxygen or fluorine), excellent conductivity, mechanical flexibility, easy functionalization, and strong absorption in the Near Infrared (NIR) region. In addition, MXene exhibits good biocompatibility and biodegradability due to its main elements of C, N and the inertness of transition metals (e.g., ti, nb, ta) to living organisms. In addition, MXene (Ti) 3 C 2 T x ) The antibacterial activity to gram-negative bacteria and gram-positive bacteria is higher than that of Graphene Oxide (GO) materials. Currently, research is mainly focused on biosensors, bio-imaging probes, therapeutic, diagnostic, and antibacterial materials, etc. MXene lacks adhesion capability, and the adhesion capability of the material is important to enhance tissue adhesion during surgery.
The existing chitosan and MXene compounding method adopts a blending method, such as CN111617309A, CN112546286A, and the blending method has the problems that MXene is easy to fall off and powder, and the prepared composite material is unstable in structure.
Disclosure of Invention
The invention provides a preparation method of an electroactive antibacterial hemostatic dressing, which aims to solve the problem of unstable structure of the existing chitosan and MXene during compounding.
The preparation method of the electroactive antibacterial hemostatic dressing comprises the following steps:
the method comprises the following steps: placing the chitosan fiber dressing base material in a vacuum drying oven for drying;
step two: adding 0.4-3g LiF into 20mL of HCl solution with the concentration of 5-13mol/L, and magnetically stirring for 30min to obtain uniform and stable LiF/HCl solution; then adding 1g of MAX powder into LiF/HCl solution, stirring for 18-24h at 35-40 ℃ to obtain suspension, and then repeatedly centrifuging and washing the suspension by deionized water until the pH value of the supernatant is 6 to obtain aqueous MXene precipitate; ultrasonically treating the aqueous MXene precipitate for 0.5-2h, and centrifuging at 3500rpm for 0.5-1h to obtain an Mxene aqueous dispersion liquid; MXene in the Mxene water dispersion liquid is a monolayer structure nanosheet, is dark green, and shows that the yield of the obtained MXene is high;
step three: modification of MXene with Polydopamine (PDA):
taking 10-30mg of MXene aqueous dispersion, washing with a Tris buffer solution for 3 times, dispersing in 40mL of Tris buffer solution to obtain an MXene solution, adding 80mg of dopamine into the MXene solution, and reacting at room temperature for 1h to obtain a PDA modified MXene nanosheet solution;
step four: placing the chitosan fiber dressing on a filter membrane in a suction cup, pouring a certain volume of PDA modified MXene nanosheet solution into the suction cup, carrying out vacuum suction filtration to obtain a chitosan/MXene composite dressing, repeatedly washing with deionized water to remove unreacted PDA modified MXene nanosheets, and finally carrying out vacuum freeze drying to obtain the chitosan/MXene electroactive antibacterial hemostatic dressing;
the mass ratio of the chitosan fiber dressing to the PDA modified MXene nanosheet in the PDA modified MXene nanosheet solution is (1.27-4.07): 1.
the principle and the beneficial effects of the invention are as follows:
1. the polydopamine molecular structure adopted by the invention is similar to that of mussel adhesive protein, and can generate phenolic hydroxyl chemical reaction on dopamine molecules in the similar adhesive protein, so that polydopamine has mussel induction characteristics, MXene is modified by oxidation and polymerization of polydopamine on the surface of the MXene, the modified Mxene can improve the adhesive force with chitosan fibers, the MXene is prevented from falling off from the surface or gaps of the chitosan fibers, the tissue wet adhesion capability of the obtained dressing is improved, the problem that the traditional dressing is not completely attached to a wound surface is solved, the dressing can be cut according to the shape and size of the wound surface, and the requirements of various types of wound surfaces are met.
2. The MXene and the chitosan fiber dressing have synergistic effect, so that the antibacterial performance of the composite dressing is better improved, the dressing has certain electric activity, and the differentiation, proliferation and migration of wound surface cells can be promoted, so that the healing of the wound surface is accelerated; the composite dressing also has good water absorption performance, excellent biocompatibility and hemostatic performance, and has potential application prospect in the field of wound repair. MXene is taken as a typical two-dimensional nano material, the M-X valence bond binding energy in the structure is strong, so that the MXene is endowed with excellent mechanical property, the bending rigidity of the MXene is high, and the MXene can be used as a reinforcing material in a composite material, so that the mechanical strength of the composite dressing prepared by the method is high.
3. The method has the advantages of simple preparation, short preparation process, mild conditions, low raw material cost, easy product conversion and contribution to industrial large-scale production, and no toxic or harmful substance emission in the preparation process belongs to a green pollution-free processing process.
Drawings
FIG. 1 shows MXene dispersion prepared in example 1, wherein a is MXene dispersion before dilution and b is MXene dispersion after dilution;
FIG. 2 is a chitosan fiber dressing;
FIG. 3 is the chitosan/MXene electroactive antimicrobial hemostatic dressing prepared in example 1;
FIG. 4 is a scanning electron microscope image of chitosan fibers;
FIG. 5 is a scanning electron microscope image of the chitosan/MXene electro-active anti-bacterial hemostatic dressing prepared in example 1;
fig. 6 is a diagram showing the hemostatic effect of the chitosan/MXene electroactive antibacterial hemostatic dressing prepared in example 1.
Detailed Description
The technical scheme of the invention is not limited to the specific embodiments listed below, and any reasonable combination of the specific embodiments is included.
The first embodiment is as follows: the preparation method of the electroactive antibacterial hemostatic dressing of the embodiment is carried out according to the following steps:
the method comprises the following steps: placing the chitosan fiber dressing base material in a vacuum drying oven for drying;
step two: adding 0.4-3g LiF into 20mL of HCl solution with the concentration of 5-13mol/L, and magnetically stirring for 30min to obtain uniform and stable LiF/HCl solution; then adding 1g of MAX powder into LiF/HCl solution, stirring for 18-24h at 35-40 ℃ to obtain suspension, and then repeatedly centrifuging and washing the suspension by deionized water until the pH value of the supernatant is 6 to obtain aqueous MXene precipitate; ultrasonically treating the aqueous MXene precipitate for 0.5-2h, and centrifuging at 3500rpm for 0.5-1h to obtain an Mxene aqueous dispersion; MXene in the Mxene water dispersion liquid is a monolayer structure nanosheet, is dark green, and shows that the yield of the obtained MXene is high;
step three: modification of MXene with Polydopamine (PDA):
taking 10-30mg of MXene aqueous dispersion, washing with a Tris buffer solution for 3 times, dispersing in 40mL of Tris buffer solution to obtain an MXene solution, adding 80mg of dopamine into the MXene solution, and reacting at room temperature for 1h to obtain a PDA modified MXene nanosheet solution;
step four: placing the chitosan fiber dressing on a filter membrane in a suction cup, pouring a certain volume of PDA modified MXene nanosheet solution into the suction cup, carrying out vacuum suction filtration to obtain a chitosan/MXene composite dressing, repeatedly washing with deionized water to remove unreacted PDA modified MXene nanosheets, and finally carrying out vacuum freeze drying to obtain the chitosan/MXene electroactive antibacterial hemostatic dressing;
the mass ratio of the chitosan fiber dressing to the PDA modified MXene nanosheet in the PDA modified MXene nanosheet solution is (1.27-4.07): 1.
1. the poly-dopamine molecular structure adopted by the embodiment is similar to the structure of mussel adhesive protein, and phenolic hydroxyl chemical reaction on dopamine molecules in the similar adhesive protein can occur, so that poly-dopamine has mussel induction characteristics, MXene is modified by oxidation and polymerization of poly-dopamine on the surface of the MXene, the modified Mxene can improve the adhesive force between the modified Mxene and chitosan fibers, MXene is prevented from falling off from the surface or gaps of the chitosan fibers, the tissue wet adhesion capability of the obtained dressing is improved, the problem that the traditional dressing is not completely attached to a wound surface is solved, the dressing can be cut according to the shape and size of the wound surface, and the requirements of various types of wound surfaces are met.
2. The MXene and the chitosan fiber dressing have synergistic effect, so that the antibacterial performance of the composite dressing of the embodiment is better improved, the dressing of the embodiment has certain electric activity, and the differentiation, proliferation and migration of wound cells can be promoted, so that the healing of the wound is accelerated; the composite dressing of the embodiment also has good water absorption performance, excellent biocompatibility and hemostatic performance, and has potential application prospect in the field of wound repair. MXene is taken as a typical two-dimensional nano material, the M-X valence bond binding energy in the structure of the MXene is strong, so that the MXene is endowed with excellent mechanical property, the bending rigidity of the MXene is high, the MXene can be used as a reinforcing material in a composite material, and the mechanical strength of the composite dressing prepared by the embodiment is high.
3. The method of the embodiment has the advantages of simple preparation, short preparation process, mild conditions, low raw material cost, easy product conversion and contribution to industrial large-scale production, and no toxic or harmful substance emission in the preparation process belongs to a green pollution-free processing process.
The second embodiment is as follows: the first difference between the present embodiment and the specific embodiment is: step one, the drying temperature is 30-60 ℃, and the chitosan fiber dressing is dried until the water content of the chitosan fiber dressing is lower than 0.01%.
The third concrete implementation mode: the present embodiment differs from the first or second embodiment in that: step one, the size of the chitosan fiber dressing is 10cm multiplied by 10cm.
The fourth concrete implementation mode: the difference between this embodiment and one of the first to third embodiments is: step one, the chitosan fiber dressing is chitosan fiber spunlace non-woven fabric with the gram weight of 50g/m 2 。
The fifth concrete implementation mode: the difference between this embodiment and one of the first to fourth embodiments is: step two, the molecular formula of the MAX powder is M n+1 AX n (Ti 3 AlC 2 ) In the molecular formula, n =1, 2 or 3,M is Ti, V, cr, zr,Nb, mo, hf or Ta, A is Al, si, P, S, ga, ge, as, cd, in, sn, tl or Pb, and X is C or N.
The sixth specific implementation mode: the difference between this embodiment and one of the first to fifth embodiments is: and the concentration of the Mxene aqueous dispersion in the second step is 2-10mg/mL.
The seventh embodiment: the difference between this embodiment and one of the first to sixth embodiments is: and step three, the concentration of the Tris buffer solution is 10mmol/L, and the pH value is 8.5.
The specific implementation mode is eight: the present embodiment differs from one of the first to seventh embodiments in that: fourthly, the mass ratio of the chitosan fiber dressing to the PDA modified MXene nanosheets in the PDA modified MXene nanosheet solution is 1.27:1.
the specific implementation method nine: the present embodiment differs from the first to eighth embodiments in that: step four, the mass ratio of the chitosan fiber dressing to the PDA modified MXene nanosheets in the PDA modified MXene nanosheet solution is 4.07:1.
the detailed implementation mode is ten: the present embodiment differs from one of the first to ninth embodiments in that: fourthly, the mass ratio of the chitosan fiber dressing to the PDA modified MXene nanosheets in the PDA modified MXene nanosheet solution is 3:1.
example 1:
the preparation method of the electroactive antibacterial hemostatic dressing comprises the following steps:
the method comprises the following steps: placing the chitosan fiber dressing base material in a vacuum drying oven for drying;
the drying temperature is 60 ℃, and the chitosan fiber dressing is dried until the water content of the chitosan fiber dressing is lower than 0.01%;
the size of the chitosan fiber dressing is 10cm multiplied by 10cm;
the chitosan fiber dressing is chitosan fiber spunlace non-woven fabric with the gram weight of 50g/m 2 ；
Step two: 1g LiF is added into 20mL of 9mol/L HCl solution, and the uniform and stable LiF/HCl solution is obtained after magnetic stirring for 30 min. Then 1g of Ti 3 AlC 2 The powder was added to LiF/HCl solutionStirring the solution for 24 hours at 35 ℃ to obtain a suspension, and then repeatedly centrifuging and washing the suspension by using deionized water until the pH value of a supernatant is 6 to obtain an aqueous MXene precipitate; ultrasonically treating the aqueous MXene precipitate for 1h, and then centrifuging at 3500rpm for 1h to obtain an Mxene aqueous dispersion;
the concentration of the Mxene water dispersion liquid is 6mg/mL;
step three: modifying MXene by adopting PDA:
taking 20mg of MXene aqueous dispersion, washing with a Tris buffer solution for 3 times, dispersing in 40mL of Tris buffer solution to obtain an MXene solution, adding 80mg of PDA into the MXene solution, and reacting at room temperature for 1h to obtain a PDA modified MXene nanosheet solution; the concentration of the Tris buffer solution is 10mmol/L, and the pH value is 8.5;
step four: placing the chitosan fiber dressing on a filter membrane in a suction cup, pouring 15ml of PDA modified MXene nanosheet solution into the suction cup, carrying out vacuum suction filtration until no solution is on the surface of the chitosan fiber dressing, repeatedly washing with deionized water to remove unreacted PDA modified MXene nanosheets, and finally carrying out vacuum freeze drying to obtain the chitosan/MXene electroactive antibacterial hemostatic dressing; the mass ratio of the chitosan fiber dressing to the PDA modified MXene nanosheet in the PDA modified MXene nanosheet solution is 1.45:1;
fig. 1 shows an MXene dispersion prepared in step one of example 1, a before dilution and b after dilution, by the following dilution methods: the MXene dispersion liquid is added with deionized water and then is subjected to ultrasonic dispersion for dilution, the Tyndall phenomenon generated by the colloid of the diluted MXene dispersion liquid can show that MXene in the MXene aqueous dispersion liquid is stripped and dispersed in a nanoscale form, a large number of oxygen-containing functional groups exist on the surface of the prepared MXene, the surface of an MXene lamella is hydrophilic and can be negatively charged in water, and MXene can be suspended in water by virtue of electrostatic repulsion between adjacent layers to form a colloidal solution. FIG. 2 is a chitosan fiber dressing; FIG. 3 is the chitosan/MXene electroactive antimicrobial hemostatic dressing prepared in example 1; fig. 2 and 3 can see that MXene is uniformly compounded with chitosan fiber dressing. FIG. 4 is a scanning electron microscope image of chitosan fibers; FIG. 5 is a scanning electron microscope image of the chitosan/MXene electro-active anti-bacterial hemostatic dressing prepared in example 1; by comparing fig. 4 and fig. 5, it can be clearly seen that MXene is supported in the chitosan fiber dressing.
FIG. 6 is a graph showing the hemostatic effect of the chitosan/MXene electroactive antimicrobial hemostatic dressing prepared in example 1; in the figure, a is the chitosan/MXene electroactive antibacterial hemostatic dressing prepared in example 1, and b is the chitosan fiber dressing; as can be seen from fig. 6, the chitosan/MXene electroactive antibacterial hemostatic dressing prepared in example 1 has a lower bleeding amount than the chitosan fiber dressing, which indicates that the hemostatic effect is better than that of the chitosan fiber dressing.
The chitosan/MXene electroactive antibacterial hemostatic dressing prepared by the embodiment has a bacteriostasis rate of over 90% on staphylococcus aureus and escherichia coli.
Example 2:
the difference of this example from example 1 is that 20ml of PDA modified MXene nanosheet solution was poured into a suction cup and vacuum filtered in step four. The other steps and process parameters were the same as in example 1.
The chitosan/MXene electroactive antibacterial hemostatic dressing prepared by the embodiment has a bacteriostasis rate of over 90% on staphylococcus aureus and escherichia coli.
Example 3:
this example differs from example 1 in that 15mg of MXene aqueous dispersion was taken in step three and washed 3 times with Tris buffer, and then dispersed in 40mL of Tris buffer to obtain MXene solution. The other steps and process parameters were the same as in example 1.
The chitosan/MXene electroactive antibacterial hemostatic dressing prepared by the embodiment has a bacteriostasis rate of over 90% on staphylococcus aureus and escherichia coli.
Table 1 shows the strength, conductivity, and biocompatibility of the dressings prepared in examples 1 to 3 and the chitosan fiber dressing, and it can be seen from table 1 that the strength, conductivity, and biocompatibility of the dressings prepared in examples 1 to 3 are significantly improved as compared to the chitosan fiber dressing.
TABLE 1
| Tensile Strength (MPa) | Conductivity (Scm) -1 ) | Cell Activity after 3d (%) | |
| Chitosan fiber dressing | 223.38±23.58 | 0 | 97.52±3.78 |
| Example 1 | 444.92±37.55 | 0.028±0.006 | 103.4±7.38 |
| Example 2 | 531.54±56.39 | 0.063±0.009 | 101.3±8.21 |
| Example 3 | 355.75±21.79 | 0.023±0.005 | 102.1±6.33 |
Claims (7)
1. A preparation method of an electroactive antibacterial hemostatic dressing is characterized by comprising the following steps: the preparation method of the electroactive antibacterial hemostatic dressing comprises the following steps:
the method comprises the following steps: placing the chitosan fiber dressing base material in a vacuum drying oven for drying;
step two: adding 0.4-3g LiF into 5-13mol/L HCl solution of 20mL, and magnetically stirring for 30min to obtain uniform and stable LiF/HCl solution; then adding 1g of MAX powder into LiF/HCl solution, stirring for 18-24h at 35-40 ℃ to obtain suspension, and then repeatedly centrifuging and washing the suspension by deionized water until the pH value of the supernatant is 6 to obtain aqueous MXene precipitate; ultrasonically treating the aqueous MXene precipitate for 0.5-2h, and then centrifuging at 3500rpm for 0.5-1h to obtain an Mxene aqueous dispersion liquid;
step three: MXene was modified with polydopamine:
taking 10-30mg of MXene aqueous dispersion, washing with a Tris buffer solution for 3 times, dispersing in 40mL of Tris buffer solution to obtain an MXene solution, adding 80mg of dopamine into the MXene solution, and reacting at room temperature for 1h to obtain a PDA modified MXene nanosheet solution;
step four: placing the chitosan fiber dressing on a filter membrane in a suction cup, pouring a certain volume of PDA modified MXene nanosheet solution into the suction cup, carrying out vacuum suction filtration to obtain a chitosan/MXene composite dressing, repeatedly washing with deionized water to remove unreacted PDA modified MXene nanosheets, and finally carrying out vacuum freeze drying to obtain the chitosan/MXene electroactive antibacterial hemostatic dressing;
the mass ratio of the chitosan fiber dressing to the PDA modified MXene nanosheet in the PDA modified MXene nanosheet solution is 3:1.
2. the method of making an electroactive antimicrobial hemostatic dressing of claim 1, wherein: step one, the drying temperature is 30-60 ℃, and the chitosan fiber dressing is dried until the water content of the chitosan fiber dressing is lower than 0.01%.
3. The method of making an electroactive antimicrobial hemostatic dressing of claim 1, wherein: step one, the size of the chitosan fiber dressing is 10cm multiplied by 10cm.
4. The method of making an electroactive antimicrobial hemostatic dressing of claim 1, wherein: step one, the chitosan fiber dressing is chitosan fiber spunlace non-woven fabric with the gram weight of 50g/m 2 。
5. The method of making an electroactive antimicrobial hemostatic dressing of claim 1, wherein: step two, the molecular formula of the MAX powder is M n+1 AX n （Ti 3 AlC 2 ) In the molecular formula, N =1, 2 or 3,M is Ti, V, cr, zr, nb, mo, hf or Ta, A is Al, si, P, S, ga, ge, as, cd, in, sn, tl or Pb, and X is C or N.
6. The method of making an electroactive antimicrobial hemostatic dressing of claim 1, wherein: and the concentration of the Mxene aqueous dispersion in the second step is 2-10mg/mL.
7. The method of making an electroactive antimicrobial hemostatic dressing of claim 1, wherein: and step three, the concentration of the Tris buffer solution is 10mmol/L, and the pH value is 8.5.
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