Synthesis method and application of urushiol modified mesoporous silica microspheres containing catechol groups
Technical Field
The invention relates to a urushiol modified mesoporous silica microsphere containing catechol groups, and belongs to the technical field of functional polymer materials.
Background
In war and civil trauma, uncontrolled massive hemorrhage is the main cause of casualties, and relevant data suggest: nearly 50% of deaths in the military are due to uncontrolled blood loss, and 15-25% of civilian injuries are deaths due to lack of timely hemostasis. Therefore, the development of hemostatic materials is very important to reduce casualties. The mesoporous silica has a large specific surface area and good biocompatibility, and is widely studied as a hemostatic material. Meanwhile, the surface of the blood coagulation factor XII is provided with a large amount of negative charges of alcohol hydroxyl groups, and the blood coagulation factor XII is activated after the blood coagulation factor XII is contacted with blood, so that the blood coagulation waterfall reaction is accelerated, and the hemostasis performance is improved. However, mesoporous silica has a characteristic of poor adhesion to tissues, and is easily permeated into blood and oozed out.
In recent years, the surface chemistry inspired by the super-strong adhesion property of mussel byssus protein is widely concerned by researchers, and becomes a cross-research hotspot in the fields of bionics, chemistry, materials science, biology, biomedicine and the like. At the same time, researchers have demonstrated that catechol (catechol) is a key group for this class of materials with strong adhesion properties. Urushiol is a derivative having a catechol structure, which is extracted from raw lacquer and is contained in the raw lacquer in an amount of about 80%. The raw lacquer has better biocompatibility, can dredge the channels, stop bleeding, relieve cough and the like, and is recorded as a medicament for use in China for a long time. In order to solve the problems of poor adhesiveness and blood seepage of mesoporous silica, urushiol with a catechol structure is selected for surface modification. The current method of directly modifying silica with urushiol (patent publication No. CN 101891205A) is to graft urushiol onto the silica surface by reacting phenolic hydroxyl groups in the catechol structure with alcoholic hydroxyl groups on the silica surface, which also causes catechol groups having tissue adhesion properties to react and lose the adhesion properties. The catechol group with the adhesion characteristic is reserved through the free radical reaction of unsaturated double bonds of a urushiol side chain, so that the tissue is adhered in the hemostasis process; the urushiol long side chain is crosslinked and intertwined to form a film, has a certain hydrophobic property, and can prevent the permeation of blood and further accelerate the hemostasis.
Disclosure of Invention
The invention aims to provide a synthetic method of urushiol modified mesoporous silica microspheres which are simple to operate, easy to control conditions, capable of being industrially produced, uniform in particle size and tissue adhesion and application thereof, aiming at overcoming the defects of the prior art.
The invention is realized by the following technical scheme:
a. synthesizing mesoporous silica microspheres:
adding Cetyl Trimethyl Ammonium Bromide (CTAB) and sodium hydroxide into a water/Ethylene Glycol (EG) solution, stirring for 1 h at 80 ℃, adding Tetraethoxysilane (TEOS), continuously stirring for 6 h, obtaining a precursor of the mesoporous silica microsphere by a sol-gel method, and calcining at high temperature to obtain the mesoporous silica microsphere; wherein the sodium hydroxide and CTAB are calculated by mass parts, TEOS, EG and water are calculated by volume parts, and the ratio of sodium hydroxide, CTAB, EG, water and TEOS is (0.01-2): (0.10-2): (5-30): (70-85): 0.1-5).
b. Urushiol-modified mesoporous silica microspheres:
dispersing the mesoporous silica microspheres prepared in the step a) in ethanol, adding gamma-Methacryloxypropyltrimethoxysilane (MPS) at 70 ℃, stirring for 24 hours, washing with ethanol, dispersing in the ethanol, adding Azobisisobutyronitrile (AIBN) and urushiol at 70 ℃, and reacting for 24 hours to obtain the urushiol modified mesoporous silica microspheres containing catechol groups; wherein the mesoporous silica microspheres, the AIBN and the urushiol are calculated by mass parts, and the proportion of the mesoporous silica microspheres, the AIBN, the urushiol and the ethanol is (0.1-10): (0.001-1): (0.1-30): (10-100) by volume parts.
The sodium hydroxide and CTAB are calculated by parts by mass, when the parts by mass are grams or kilograms, the parts by volume of TEOS, EG and water in the parts by volume are mL or L, and the ratio of the sodium hydroxide, CTAB, EG, water and TEOS is (0.01-2), (0.10-2), (5-30), (70-85) and (0.1-5).
When the mass parts of the mesoporous silica microspheres, the AIBN and the urushiol are counted by mass parts of grams or kilograms, the volume parts of the ethanol in the volume parts are correspondingly mL or L, and the proportion of the mesoporous silica microspheres, the AIBN, the urushiol and the ethanol is (0.1-10): (0.001-1): 0.1-30): 10-100).
Urushiol-modified mesoporous silica microspheres containing catechol groups synthesized by the method and application thereof in hemostatic materials.
Compared with the prior art, the invention has the following beneficial effects:
1. can conveniently synthesize the urushiol modified mesoporous silica microspheres containing catechol groups, has simple operation, easily controlled conditions, industrialized production and uniform particle size.
2. The urushiol modified mesoporous silica microspheres synthesized by the method of the invention keep catechol groups in urushiol, and can form a good adhesion effect with tissues.
3. The urushiol-modified mesoporous silica microsphere containing catechol groups prepared by the method has the specific surface area as high as 448.91 m2The/g can be self-assembled into an amphiphilic Janus film, can rapidly generate a hemostatic effect and is used as a hemostatic, and the in-vivo liver hemostasis time is reduced to 22 s.
Drawings
Other features, objects and advantages of the invention will become more apparent upon reading of the detailed description of non-limiting embodiments with reference to the following drawings:
fig. 1 is a transmission electron microscope image of the morphology of the urushiol-modified mesoporous silica microspheres containing catechol groups prepared in example 1 of the present invention.
FIG. 2 shows how urushiol-modified mesoporous silica microspheres containing catechol groups according to example 1 of the present invention are solidified13C-NMR chart.
FIG. 3 is a graph showing the in vivo liver hemostasis time of urushiol-modified mesoporous silica microspheres containing catechol groups prepared in example 1 of the present invention in example 6.
Detailed Description
Example 1
Step one
0.1 g of sodium hydroxide is dissolved in 240 mL of ethylene glycol/water (the volume ratio of the ethylene glycol to the water is 1: 5), 0.5 g of Cetyl Trimethyl Ammonium Bromide (CTAB) and 3 mL of Tetraethoxysilane (TEOS) are added, the mixture is stirred for 6 hours at the temperature of 80 ℃, and the precursor of the mesoporous silica microsphere is obtained by centrifugation, water washing and freeze drying. Then calcining the mixture at the high temperature of 600 ℃ for 6 hours to obtain the mesoporous silica microspheres.
Step two
And (3) dispersing 0.5 g of the mesoporous silica microspheres in ethanol, adding 3 mL of gamma-Methacryloxypropyltrimethoxysilane (MPS), reacting at 70 ℃ for 24 hours, centrifuging, and washing with ethanol for several times. And dispersing the porous silica microspheres in ethanol, adding 0.1 g of Azobisisobutyronitrile (AIBN) and 1 g of urushiol, reacting for 24h at 70 ℃, centrifuging, washing with ethanol/water for several times, and freeze-drying to obtain the urushiol modified mesoporous silica microspheres containing catechol groups.
FIG. 1 shows a transmission electron microscope that the urushiol-modified mesoporous silica microsphere containing catechol groups is of a core-shell structure, the shell thickness is about 17 nm, the inside of the urushiol-modified mesoporous silica microsphere is provided with rich pores, and the outside of the urushiol-modified mesoporous silica microsphere is provided with a denser urushiol organic layer; FIG. 2 13The peak of 140-120 ppm in C-NMR corresponding to the benzene ring and side chain double bond of urushiol shows that the synthesized urushiol modified mesoporous silica microsphere is obtained.
Example 2
Step one
Dissolving 0.15 g of sodium hydroxide in 360 mL of ethylene glycol/water (the volume ratio of ethylene glycol to water is 1: 5), adding 1.5 g of CTAB and 2 mL of TEOS, stirring for 6 h at 80 ℃, centrifuging, washing with water and freeze-drying to obtain a precursor of the mesoporous silica microsphere. Then calcining the mixture at the high temperature of 600 ℃ for 6 hours to obtain the mesoporous silica microspheres.
Step two
And (3) dispersing 1 g of the mesoporous silica microspheres in ethanol, adding 2 mL of MPS, reacting at 70 ℃ for 24h, centrifuging, and washing with ethanol for several times. And dispersing the porous silica microspheres in ethanol, adding 0.1 g of AIBN and 2 g of urushiol, reacting for 24h at 70 ℃, centrifuging, washing with ethanol/water for several times, and freeze-drying to obtain the urushiol modified mesoporous silica microspheres containing catechol groups.
Example 3
Step one
Dissolving 0.3 g of sodium hydroxide in 500 mL of ethylene glycol/water (the volume ratio of ethylene glycol to water is 1: 5), adding 2.5 g of CTAB and 3 mL of TEOS, stirring for 6 h at 80 ℃, centrifuging, washing with water and freeze-drying to obtain a precursor of the mesoporous silica microsphere. Then calcining the mixture at the high temperature of 600 ℃ for 6 hours to obtain the mesoporous silica microspheres.
Step two
And (3) dispersing 1 g of the mesoporous silica microspheres in ethanol, adding 5 mL of MPS, reacting at 70 ℃ for 24h, centrifuging, and washing with ethanol for several times. And dispersing the porous silica microspheres in ethanol, adding 0.2 g of AIBN and 3 g of urushiol, reacting for 24h at 70 ℃, centrifuging, washing with ethanol/water for several times, and freeze-drying to obtain the urushiol modified mesoporous silica microspheres containing catechol groups.
Example 4
Step one
0.1 g of sodium hydroxide is dissolved in 240 mL of ethylene glycol/water (the volume ratio of ethylene glycol to water is 1: 5) solution, 1.5 g of CTAB and 2 mL of TEOS are added, the mixture is stirred for 6 hours at the temperature of 70 ℃, and then the precursor of the mesoporous silica microsphere is obtained by centrifugation, water washing and freeze drying. Then calcining the mixture at the high temperature of 600 ℃ for 6 hours to obtain the mesoporous silica microspheres.
Step two
3 g of the mesoporous silica microspheres are dispersed in ethanol, 3 mL of MPS is added, the reaction is carried out for 24h at 70 ℃, and the mixture is centrifuged and washed by ethanol for several times. And dispersing the porous silica microspheres in ethanol, adding 0.1 g of AIBN and 3 g of urushiol, reacting for 24h at 70 ℃, centrifuging, washing with ethanol/water for several times, and freeze-drying to obtain the urushiol modified mesoporous silica microspheres containing catechol groups.
Example 5
Step one
Dissolving 0.3 g of sodium hydroxide in 300 mL of ethylene glycol/water (the volume ratio of ethylene glycol to water is 1: 5), adding 2.5 g of CTAB and 2 mL of TEOS, stirring for 6 h at 80 ℃, centrifuging, washing with water and freeze-drying to obtain a precursor of the mesoporous silica microsphere. Then calcining the mixture at the high temperature of 600 ℃ for 6 hours to obtain the mesoporous silica microspheres.
Step two
And (3) dispersing 2 g of the mesoporous silica microspheres in ethanol, adding 1 mL of MPS, reacting at 70 ℃ for 24h, centrifuging, and washing with ethanol for several times. And dispersing the porous silica microspheres in ethanol, adding 0.1 g of AIBN and 5 g of urushiol, reacting for 24h at 70 ℃, centrifuging, washing with ethanol/water for several times, and freeze-drying to obtain the urushiol modified mesoporous silica microspheres containing catechol groups.
Example 6
10 mg of the mesoporous silica microspheres prepared in the embodiment 1 of the present invention and urushiol-modified mesoporous silica microspheres are respectively sprinkled on a rat liver cut wound (length 1 cm, depth 0.5 cm), timing is started until blood does not seep, timing is stopped, the time is the liver hemostasis time, and each sample is repeated for 5 times to obtain the average value. The liver hemostasis time is shown in figure 3.
The data in figure 3 show that the liver hemostasis time of the urushiol modified mesoporous silica microsphere containing the catechol group only needs 22 s, and compared with the hemostasis time (54 s) of the mesoporous silica microsphere, the hemostasis time is shortened by 59%. The urushiol modified mesoporous silicon dioxide microspheres containing catechol groups can become a safe and efficient hemostatic material by comprehensively considering the biocompatibility and the human degradability.
The foregoing description of specific embodiments of the present invention has been presented. It is to be understood that the present invention is not limited to the specific embodiments described above, and that various changes and modifications may be made by one skilled in the art within the scope of the appended claims without departing from the spirit of the invention.