CN115137842A - Silicon dioxide microsphere capable of adsorbing nano vesicles and used for treating ulcerative colitis and preparation method and application thereof - Google Patents

Silicon dioxide microsphere capable of adsorbing nano vesicles and used for treating ulcerative colitis and preparation method and application thereof Download PDF

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CN115137842A
CN115137842A CN202210573390.6A CN202210573390A CN115137842A CN 115137842 A CN115137842 A CN 115137842A CN 202210573390 A CN202210573390 A CN 202210573390A CN 115137842 A CN115137842 A CN 115137842A
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孙坚
刘旭伟
臧涛
李伟雄
蒋刚彪
刘雅红
廖晓萍
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South China Agricultural University
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Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a silica microsphere capable of adsorbing nano vesicles and used for treating ulcerative colitis, and a preparation method and application thereof. The electronegativity of the surface of the silicon dioxide microsphere is changed by adding phenyl triethoxysilane and aminopropyl triethoxysilane step by step in a sol-gel method and introducing a required functional group through a silane coupling agent with an organic functional group, and the silicon dioxide microsphere prepared by the method has uniform particle size and good dispersibility. According to the invention, the silicon dioxide loaded with the aromatic aldehyde compound is used as a carrier, the nano vesicles are adsorbed and protected, the vesicles are more efficiently utilized to relieve inflammation, and the preparation method can be applied to treating ulcerative colitis.

Description

Silicon dioxide microsphere capable of adsorbing nano vesicles and used for treating ulcerative colitis and preparation method and application thereof
Technical Field
The invention relates to the technical field of biological medicines, in particular to a silicon dioxide microsphere capable of adsorbing nano vesicles and used for treating ulcerative colitis, and a preparation method and application thereof.
Background
Ulcerative Colitis (UC) is a nonspecific inflammatory bowel disease that develops in the colonic mucosa and submucosa with severe complications, and tends to transform colon cancer if left untreated for a long time, seriously jeopardizing human public health safety. Meanwhile, colitis gradually becomes a common digestive tract disease in large-scale breeding animals such as pigs, cows and the like. The pathogenesis of ulcerative enteritis is complicated, but according to the existing research results, the immune response is abnormal due to a plurality of factors such as genetic susceptibility factors of a host, the change of the environment, intestinal microbial disorder and the like. The aim of relieving symptoms is achieved clinically by using corticosteroids, salicylates, immunosuppressants and microbial agents. The study found that by taking probiotics and a microbial preparation derived therefrom, and to a process for the preparation thereof, can effectively regulate intestinal flora, repair mucous membrane and slow down persistent inflammation. However, the long-term administration of the live bacteria can easily cause secondary infection of patients with low immunity and even deficiency.
In response to the problem, researchers at home and abroad find that Outer Membrane Vesicles (OMVs) produced by Nissle 1917 also have the effect of relieving inflammation. The EcN-OMVs, as metagens of EcN, have the ability to perform intercellular signaling, and can activate cytokines by triggering the intestinal immune barrier system, induce the expression of inflammatory factors and do not cause the risk of probiotic drug resistance. The OMVs have the problems of small particle size, small contact area with intestinal epithelial cells, easy damage by the intestinal environment and the like. How to efficiently utilize OMVs has also become a research hotspot. The main way is also to protect it by means of nanocarriers.
Silicon is an essential trace element in the body of mammals and is mainly present in the epidermis and tissues of mammals. The present research shows that silicon is mainly involved in the calcification process of bone, can promote growth, maintain elasticity of arterial wall and protect inner wall membrane, promote normal growth of bone, cartilage and connective tissue, participate in the metabolism of polysaccharide, and is the main component of some glucose amino polysaccharide carboxylic acids. Based on the advantages of silicon element, the application of biological materials using silicon as the main synthetic raw material in biomedicine is increasingly common.
The silicon dioxide nano material is widely applied to the aspect of microbial preparations as an oral medicament carrier for resisting the damage of gastric acid and intestinal juice. Werner in the beginning of the fifties of the twentieth century
Figure BDA0003661160060000021
The monodisperse silicon dioxide microspheres are successfully prepared by a sol-gel method for the first time by using ethanol as a solvent, tetraethoxysilane as a silicon source and ammonia water as a catalyst. Monodisperse silica has become one of the most studied monodisperse systems. However, the existing silica microspheres still need to be improved in drug loading, mainly because the surface of the silica microspheres synthesized by the traditional method is mainly negatively charged, which is not beneficial to loading most of negatively charged drugs; on the other hand, in the prior art, the adsorption of the aromatic aldehyde compound is irreversible, which is not beneficial to the release of the aromatic aldehyde compound.
Aromatic aldehydes are compounds in which two single bonds on the carbonyl group, one to the aromatic radical and one to the hydrogen, are present. Common aromatic aldehydes are benzaldehyde (also known as benzoin aldehyde), cumin aldehyde, trans-cinnamaldehyde, and vanillin. The aromatic aldehyde compounds have wide sources and are mainly extracted from plants. In the present researches, the aromatic aldehyde can be used as a plant feed additive, has multiple functions of regulating intestinal flora, improving the immunity and the production performance of animals, improving the oxidation resistance and the meat quality and the like, and even some researches show that the aromatic aldehyde can resist virus infection, help digestion and relieve pain.
Disclosure of Invention
The invention aims to solve the problems and provides a silicon dioxide microsphere capable of adsorbing nano vesicles and used for treating ulcerative colitis, and a preparation method and application thereof.
The invention firstly provides a silicon dioxide microsphere capable of adsorbing nano vesicles for treating ulcerative colitis.
The invention also provides a preparation method of the silicon dioxide microsphere capable of adsorbing the nano vesicles for treating ulcerative colitis.
In order to realize the purpose, the invention is carried out by the following technical scheme:
a preparation method of silicon dioxide microspheres capable of adsorbing nano vesicles for treating ulcerative colitis comprises the following steps:
s1, mixing potassium chloride, ethanol, water, ethyl orthosilicate and ammonia water to form a solution I, mixing the ethyl orthosilicate and the ethanol to form a solution II, stirring the solution I to turn into milk white, and adding the solution II into the solution I;
s2, preparing ethyl orthosilicate and aminopropyltriethoxysilane into a solution III, preparing phenyltriethoxysilane and toluene into a solution IV, supplementing ammonia water and potassium chloride into the solution I after the solution II is added in the step S1, respectively adding the solution III and the solution IV to fully react, and then washing to obtain silicon dioxide microspheres;
and S3, dispersing the silicon dioxide microspheres and the aromatic aldehyde obtained in the step S2 by using a solvent, performing ultrasonic treatment, stirring overnight, then performing centrifugal washing, dispersing the washed microspheres by using a tris (hydroxymethyl) aminomethane hydrochloride solution, adding hydrochloric acid to adjust the pH value of the solution to be acidic, continuously reacting, removing excessive hydrochloric acid, and storing at low temperature.
The method is mainly characterized in that phenyl triethoxysilane and aminopropyl triethoxysilane are added step by step in the traditional sol-gel method. The electronegativity of the surface of the silica microsphere is changed by introducing a needed functional group through a silane coupling agent with an organic functional group. The silicon dioxide microspheres prepared by the method have uniform particle size and good dispersibility.
According to the preferable technical scheme of the invention, the solution I in the step S1 comprises the following substances in parts by mass: 0.02-0.03% of potassium chloride, 10-12% of water, 1.5-2.0% of ethyl orthosilicate, 4.0-4.2% of ammonia water and the balance of ethanol, wherein the mass fraction of the ethyl orthosilicate in the solution II is 12-14%.
Preferably, the solution II is added to the solution I in the step S1 at an injection rate of 0.3-0.4mL/min.
Preferably, the reaction mass ratio of the ethyl orthosilicate and the aminopropyltriethoxysilane in the step S2 is 5:1-5:2, the mass ratio of the tetraethoxysilane to the phenyltriethoxysilane is 5:1-5:2.
preferably, the amount of the supplemented ammonia and the potassium chloride in the step S2 is 20% of the mass of the ammonia and the potassium chloride in the solution I in the step S1, the dropping speed of the adding solution III is 0.4-0.6mL/min, and the dropping speed of the adding solution IV is 0.08-0.12mL/min.
Preferably, the aromatic aldehyde in step S3 comprises cinnamaldehyde, benzaldehyde, cuminaldehyde, vanillin.
Preferably, the centrifugal washing in step S3 is centrifugally washed three times with anhydrous ethanol and a tris (hydroxymethyl) aminomethane hydrochloride solution, respectively.
The invention also provides application of the silicon dioxide microspheres adsorbing the nano vesicles in preparation of a preparation for treating ulcerative colitis. Preferably, the nanovesicles are EcN-OMVs.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention provides a preparation method of silicon dioxide microspheres capable of adsorbing nano vesicles and used for treating ulcerative colitis, which mainly comprises the step-by-step addition of phenyltriethoxysilane and aminopropyltriethoxysilane in a traditional sol-gel method. The electronegativity of the surface of the silica microspheres is changed by introducing a desired functional group through a silane coupling agent with an organic functional group. The silicon dioxide microspheres prepared by the method have uniform particle size and good dispersibility.
(2) The invention utilizes the functionalized silicon dioxide microspheres to adsorb the nano vesicles for protection, and can efficiently utilize the vesicles to relieve inflammationIn use, SAP-NH was found by treatment in a mouse colitis model 2 The CHO-OMVs can enhance the mucous membrane barrier, maintain the stability of the intestinal flora Akkermansia (Akkermansia) and Lactobacillus (Lactobacillus) and avoid the intestinal flora from intensifying the intestinal inflammatory reaction to a certain extent by increasing the expression of ZO-1.
Drawings
FIG. 1 is a schematic view of a preparation process according to the technical scheme of the present invention.
Fig. 2 is a morphological diagram of the supported cinnamaldehyde silica microspheres prepared in example 1 of the present invention under different magnifications.
FIG. 3 is a scanning electron microscopy spectroscopy analysis chart of cinnamaldehyde-loaded silica prepared in example 1 of the present invention, wherein a is an EDS layered picture of cinnamaldehyde-loaded silica; b is Si element of silica carrying cinnamaldehyde; c is element C of silica carrying cinnamaldehyde; d is the N element of the silica carrying cinnamaldehyde; e is the O element of the silica carrying cinnamaldehyde; the f picture is the P element of the silica carrying cinnamaldehyde.
FIG. 4 is a graph of the change in body weight (a) and survival rate (b) of mice.
FIG. 5 shows the colon length (a) and the statistical graph (b) of mice, where p <0.05, p-woven fabric 0.01, and P-woven fabric 0.0001.
FIG. 6 is a statistical graph showing the expression levels of the cytokines TNF-. Alpha., IL-1. Beta. And ZO-1 proteins, wherein P <0.01, P < -0.001, and P < -0.0001, respectively.
FIG. 7 is a photograph of HE stained tissue sections of mouse colon, wherein a represents CON group, b represents DSS group, c represents SAPO group, d represents OMVs group, e represents SAP group, observed at 15 times.
Figure 8 is a graph of the effect on intestinal flora levels in mice with acute colitis, a: abundance analysis of microorganisms at the genus level (Family); b: (ii) an interclass multiple comparison of the flora level alpha diversity Shannon index; c: an NMDS analysis chart of the flora structure among the sample groups; d: comparing beta diversity among sample groups; * And represents p <0.05, p and the straw bundle of 0.01.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood 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.
The test methods used in the examples of the present invention are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.
Example 1
The silicon dioxide microsphere capable of adsorbing the nano vesicles and used for treating ulcerative colitis is prepared by the following steps:
weighing 0.0248g of potassium chloride, 75.39g of absolute ethyl alcohol, 9.45g of deionized water, 1.645g of Tetraethoxysilane (TEOS) and 4mL of ammonia water, respectively adding the materials into a 250mL three-neck flask, magnetically stirring the materials at 240rpm at room temperature for 20min to form a solution I, and uniformly mixing 2.06g of TEOS and 13.12g of absolute ethyl alcohol to form a solution II; when the solution I in the three-neck flask begins to turn into milk white, slowly and uniformly injecting the solution II into the three-neck flask by using a micro-injection pump at the speed of 0.3 mL/min; simultaneously weighing 2.06g TEOS, 0.412g aminopropyl triethoxysilane (APTES) to prepare a solution III, and 0.412g Phenyltriethoxysilane (PTOES) and 5g toluene to prepare a solution IV; after the solution II is completely dripped, 0.8mL of ammonia water and 0.0062g of potassium chloride are supplemented into a three-neck flask, and the solution III and the solution IV are respectively dripped at the speed of 0.4mL/min and the speed of 0.08 mL/min; after the dropwise addition is finished, the magnetic stirring is continued for 30min to make the reaction continue to be fully performed. After the reaction is finished, washing the silica microspheres twice by using absolute ethyl alcohol and deionized water respectively to remove the residual solvent.
Weighing 5g of the precipitate SAP microspheres and 5mL of cinnamaldehyde, adding into a 50mL beaker, dispersing with 20mL of absolute ethanol, performing ultrasonic treatment in an ultrasonic instrument for 20min, and magnetically stirring overnight at room temperature; and (2) respectively centrifuging and washing the microspheres for three times by using absolute ethyl alcohol and a Tris-HCl solution, dispersing the washed microspheres by using 20mL of a Tris-HCl solution with the pH of 6.0, adding 5mL of HCl (0.5 mol/l) in the process of magnetic stirring to enable the final pH of the solution to be =5.0 in a meta-acid environment, continuously stirring for 30min, washing off excessive HCl by using a Tris-HCl solution with the pH of =6.0 after the reaction is finished, and storing at 4 ℃ for later use.
The morphology of the obtained silica microspheres loaded with cinnamaldehyde is shown in figure 2, and the prepared silica microspheres have uniform particle size and good dispersibility.
Example 2
The cinnamaldehyde-supporting silica microspheres (SAP-NH) prepared in example 1 were mixed 2 -CHO) adsorbing nanovesicles to prepare SAP-NH 2 The — CHO-OMVS method is as follows:
30mg of cinnamaldehyde-loaded silica (SAP-NH) was weighed out 2 -CHO), and 20mL Outer Membrane Vesicles (OMVs) were added to a 50mL Erlenmeyer flask and magnetically stirred at 200rpm at 4 ℃ for 18h. Centrifuging at 4 deg.C and 4000rpm every other day to obtain precipitate, re-suspending the precipitate with 5mL PBS buffer solution, and centrifuging at 4 deg.C and 4000rpm to obtain microsphere (SAP-NH) for adsorbing outer membrane vesicle 2 -CHO-OMV S )。
Example 3
Treatment of functionalized silica supports after adsorption of EcN-OMVs in mouse colitis models
1 Experimental conditions:
mice were bred adaptively for one week in an experimental environment to avoid stress reactions during transportation, and were randomly divided into 5 groups of 5 mice each, namely, a CON group (normal control group), a DSS group (DSS model group), and a SAPO group (SAP-NH group) 2 -CHO-OMV S Group), OMVs group (to OMVs group), SAP group (to SAP-NH) 2 -CHO group). The SAPO, OMVs and SAP groups were pre-dosed via gavage. SAP-NH with a load of about 40 mug OMVs per mouse was administered to the SAP groups 2 -CHO-OMV S Suspension, OMVs group mice each drench OMVs solution of about 40. Mu.g mass, SAP group mice each drench SAP-NH of 25mg mass 2 -CHO microsphere suspension, CON group, DSS group were also given 200 μ L PBS for 10 consecutive days by gavage; 11d, the other four groups except the normal control group are given the aqueous solution with the Dextran Sodium Sulfate (DSS) content of 3% by a drinking water mode, and the water is freely drunk for 7d; 18d recovery of Normal Drinking Water with SAPOThe group, OMVs group and SAP group were administered by gavage for 5 days continuously, and the normal control group and model control group were also pre-administered with an equal volume of PBS by gavage.
2 analysis of results
2.1 growth conditions in mice
The test method comprises the following steps: during the 7D molding period, dextran sulfate (DSS) was dosed freely from day 1 to day 7, labeled D1, D2, D3, D4, D5, D6, D7, respectively. The mice of each group were weighed daily, and the growth, weight change and mental status of the mice of different groups were observed during the period. The statistical results are shown in FIG. 4.
As shown in FIG. 4-a, mice in the CON group remained stable during the DSS treatment period from 0-7d, and the SAP group tended to gain weight from 0-3 d. From 4d, except for CON group, the mice in DSS, SAP, OMVs and SAPO groups began to lose weight, the weight of DSS group decreased fastest and decreased to less than 80% of the initial weight in 7d, and the weight of the mice in SAP, OMVs and SAPO groups decreased slowly. The weight of mice in the DSS treatment groups 4-7d,OMVs was reduced to the minimum, and the weight of mice in the groups 7d,SAPO and OMVs was reduced to the initial 90%; in contrast, mice in the SAP group lost body weight to around the initial 85%. The SAP group, OMVs group, SAPO group were significantly different from the DSS group.
After 7d DSS treatment, DSS-containing drinking water of DSS group, SAPO group, OMVs group and SAP-group was changed to sterile drinking water, observed for 3d, and the survival rate of mice was recorded. As shown in figure 4-b, mice in the DSS-treated 7d, DSS group began to die, and mice in the SAPO and SAP groups began to die at 8 d. The survival rate of mice in the 10d, DSS group was only 40%, and the survival rate of mice in the SAPO group and the SAP group was 70%. Among them, mice in the OMVs group had 90% mortality compared to the DSS group. Although the survival rate of mice in the SAPO group and the SAP group is 70%, the mice still have the effects of relieving inflammation and prolonging survival time compared with the DSS group.
2.2 Scent recording of Colon Length
The test method comprises the following steps: after the experiment, the mice were sacrificed by cervical dislocation, dissected, and the length of the colon was measured with a ruler and recorded, and then the colon was washed with a syringe containing 1mL of PBS solution to remove feces and other contents in the intestinal tract. The recorded results are shown in FIG. 5.
The change of the colon length of the mouse can visually reflect an important index of the pathological change degree of the colitis of the mouse. At 10d, all mice were euthanized and the colon length was measured to be shortest, on average below 4cm; the colon length shortening of mice in SAPO group, OMVs group and SAP group is relieved and is about 5 cm. The colon length of mice in the DSS group and the CON group is obviously shortened and has a significant difference (p < 0.05), and the colon length of the DSS group is also significantly different from that of the SAPO group and that of the OMVs group (p <0.01 and p-woven cover 0.05).
2.3 extraction and quality control analysis of inflammatory factor mRNA in colonic tissue
The test method comprises the following steps: the post-mortem mice were dissected and the colons were removed, washed with a syringe containing 1mL of PBS solution, and the intestinal feces and other contents were removed, and 0.5cm tissue fractions were collected and total tissue mRNA was extracted by Trizol.
As shown in fig. 6-a, the expression level of TNF- α mRNA in colon tissue of mouse was lower in CON group, and the expression level of TNF- α mRNA was significantly increased in colon tissue of DSS group (p < 0.05) and was down-regulated in colon tissue of SAPO group, OMVs group and SAP group, compared to CON group, wherein the expression level of TNF- α mRNA was significantly different between SAPO group and DSS group (p < 0.05); IL-beta is a member of the interleukin IL-1 family, belongs to a proinflammatory cytokine and has a proinflammatory function. As shown in fig. 6-b, the IL- β expression of DSS group was significantly different from that of CON group, and the expression levels of SAPO group, OMVs group, and SAP group were also decreased from DSS group, which was significantly different from that of DSS group; the zonulin ZO-1 is used as a main component of the intestinal barrier, and the expression quantity of the zonulin ZO-1 directly influences the permeability of the intestinal mucosa. As shown in FIG. 6-c, the colon tissue of DSS group had the least amount of ZO-1mRNA expression and was significantly different (p < 0.05) from that of CON group, while the other three groups had increased amounts of ZO-1mRNA expression, wherein the OMVs group had significant difference (p < 0.05) from that of DSS group.
2.4 Colon tissue sections H & E staining
The test method comprises the following steps: dissecting the dead mouse, taking out the colon, washing the colon with an injector containing 1mL PBS solution, washing away the feces and other contents in the intestinal tract, taking a tissue part with a length of about 0.5cm, and adding into a prepared fixing solution to rapidly denature and solidify the proteins of the tissue and cells, so as to maintain the original morphological structure of the cells. Paraffin embedding was performed, and then pathological lesions of the colon of mice were observed by HE staining of pathological sections and scored.
From fig. 7-a colon H & E pathological section, it can be seen that the colon mucosal epithelial cells of CON group are well-arranged and intact, and no deformation or even necrosis occurs, and abundant goblet cells and clear crypt structure can be observed, but there is little inflammatory cell infiltration in the mucosa layer, and edema occurs in submucosa layer, probably due to long-term perfusion with PBS solution. Compared with the CON group, the DSS group has serious structural damage to colon tissues, disappearance of crypts, deletion of goblet cells, infiltration of inflammatory cells deep into mucosa and mucosal muscularis, extensive necrotic deletion of mucosa tissues with extensive ulcer formation, and necrotic exudates in cavities (fig. 7-e); partial ulceration of the epithelium in the SAPO group, infiltration of part of the mucosal layer by inflammatory cells, and reduction of crypts (FIG. 7-c); the OMVs group also had significant crypt presence, but there was necrosis of the local ulcerated tissue and a reduction in goblet cells (FIG. 7-b); the SAP group had only a small amount of mucosal tissue and extensive ulceration appeared, with the disease cells infiltrating the mucosal layer in large amounts, but with some relief effect relative to the DSS control group (fig. 7-d).
2.5 fecal sample Collection and 16S rDNA high throughput sequencing
The test method comprises the following steps: at the end of the experiment, each group of mice was transferred to 75% alcohol-wiped squirrel cages, left for 1h, and with a disposable sterile inoculating loop, mouse feces were collected, loaded into 2mL centrifuge tubes sterilized at high temperature and high pressure, numbered and quick-frozen with liquid nitrogen, and then transferred to a-80 ℃ low-temperature refrigerator for storage for use.
As shown in fig. 8-a, the composition of the intestinal flora level of mice with acute colitis was analyzed, and the flora abundance was ranked as Akkermansia (Akkermansia), lactobacillus (Lactobacillus), streptococcus (Streptococcus), bacteroides (Bacteroides), allobacteroides (allobactum), and backing bacilli (odonobacter). The relative abundance of Lactobacillus (Lactobacillus), allobacteroides (Allobaculum) was reduced by 27%, 10.8% for the DSS-treated group relative to the CON group, respectively, while Akkermansia (Akkermansia) was increased by up to 61.6%. OMVs are mainly increases of Akkermansia (Akkermansia) and Streptococcus (Streptococcus) by 36.6% and 18.2% relative to CON. The increase in SAPO group was only 10.8% in Akkermansia (Akkermansia).
In 16S rRNA sequencing, the Alpha diversity index is a sample diversity analysis, including two factors of abundance and uniformity of species composition in a sample, and the diversity of a sample is usually evaluated by using indexes such as ObservedOTU, shannon and Faith' S Phylogenetic, and the higher the index is, the more complex the diversity of the sample is. Comparing Shannon indexes of different groups, the Shannon indexes of the floras are reduced after DSS treatment compared with those of CON groups. With the DSS group dropping most significantly (p < 0.01); the diversity of SAP group, SAPO group and OMVs group is similar, and the diversity is not obviously different compared with DSS group. Taken together, the SAP group, SAPO group, and OMVs group DSS group all reduced the diversity (diversity) of the mouse intestinal flora after DSS treatment, with the Shannon index of the DSS group decreasing most significantly.
The non-metric multidimensional scaling (NMDS) statistic is a beta diversity analysis similar to PCoA microbial colony studies, and NMDS reflects only the distance between samples, and does not reflect true numerical differences. Sample differences between groups and within groups can be seen by NMDS analysis of the samples. As shown in fig. 8-c, the samples in the DSS group were concentrated and showed a more similar flora structure compared to the CON group, while the relative abundance of the samples in the OMVs group was significantly different from that in the SAPO group. From fig. 8-d, the β diversity ratio of the SAP group samples to the CON group samples was significantly different (p < 0.05), while the β diversity difference among the SAP group samples, DSS group samples, and OMVs group samples was significantly different (p < 0.01). The samples differed significantly in beta diversity (p < 0.05) compared to the SAP group and DSS group.
In conclusion, the mouse colitis model was used for SAP-NH 2 Comparison of the preventive effects of-CHO-OMVs and OMVs themselves. In certain aspects SAP-NH 2 CHO-OMVs have a comparable effect to OMVs, SAP-NH 2 CHO-OMVs and OMVs have the effects of relieving inflammation and relieving changes of flora abundance in an acute colitis model. SAP-NH 2 CHO-OMVs can enhance the mucosal barrier, maintain the stability of the intestinal flora Akkermansia (Akkermansia) and Lactobacillus (Lactobacillus) and avoid the intestinal flora from intensifying the intestinal inflammatory reaction by increasing ZO-1 expression to a certain extent.
It should be understood that the above-mentioned embodiments of the present invention are only examples for clearly illustrating the technical solutions of the present invention, and are not intended to limit the specific embodiments of the present invention. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention claims should be included in the protection scope of the present invention claims.

Claims (10)

1. A preparation method of a silicon dioxide microsphere capable of adsorbing nano vesicles and used for treating ulcerative colitis is characterized by comprising the following steps:
s1, mixing potassium chloride, ethanol, water, ethyl orthosilicate and ammonia water to form a solution I, mixing the ethyl orthosilicate and the ethanol to form a solution II, stirring the solution I to turn into milk white, and adding the solution II into the solution I;
s2, preparing ethyl orthosilicate and aminopropyltriethoxysilane into a solution III, preparing ethyl orthosilicate, phenyltriethoxysilane and toluene into a solution IV, supplementing ammonia water and potassium chloride into the solution I after the solution II is added in the step S1, then respectively adding the solution III and the solution IV to react sufficiently, and washing to obtain silicon dioxide microspheres;
and S3, dispersing the silicon dioxide microspheres and the aromatic aldehyde obtained in the step S2 by using a solvent, performing ultrasonic treatment, stirring overnight, then performing centrifugal washing, dispersing by using a tris (hydroxymethyl) aminomethane hydrochloride solution, adding hydrochloric acid to adjust the pH value of the solution to be acidic, continuing to react, removing excessive hydrochloric acid, and then storing.
2. The method for preparing nano vesicle-adsorbable silica microspheres according to claim 1, wherein in step S1, the solution i contains the following substances in parts by mass: 0.02-0.03% of potassium chloride, 10-12% of water, 1.5-2.0% of ethyl orthosilicate, 4.0-4.2% of ammonia water and the balance of ethanol, wherein the mass fraction of the ethyl orthosilicate in the solution II is 12-14%.
3. The method for preparing silica microspheres capable of adsorbing nanovesicles for treating ulcerative colitis according to claim 1, wherein the injection rate of solution II into solution I in step S1 is 0.3-0.4mL/min.
4. The method for preparing the silicon dioxide microspheres capable of adsorbing nanovesicles for treating ulcerative colitis according to claim 1, wherein the mass ratio of ethyl orthosilicate to aminopropyltriethoxysilane in step S2 is 5:1-5:2, the mass ratio of the ethyl orthosilicate to the phenyltriethoxysilane is 5:1-5:2.
5. the method for preparing nano vesicle-absorbable silica microspheres for the treatment of ulcerative colitis according to claim 1, wherein the amount of ammonia and potassium chloride supplemented in step S2 is 20% of the amount of ammonia and potassium chloride in solution i in step S1, the dropping speed of solution iii is 0.4-0.6mL/min, and the dropping speed of solution iv is 0.08-0.12mL/min.
6. The method for preparing nano vesicle-adsorptive silica microspheres according to claim 1, wherein the aromatic aldehydes in step S3 comprise cinnamaldehyde, benzaldehyde, cumin aldehyde and vanillin.
7. The method for preparing silica microspheres capable of adsorbing nanovesicles for treating ulcerative colitis according to claim 1, wherein the centrifugal washing in step S3 is performed with three times of centrifugal washing with absolute ethanol and tris (hydroxymethyl) aminomethane hydrochloride solution, respectively.
8. Silica microspheres capable of adsorbing nanovesicles and used for treating ulcerative colitis, prepared by the preparation method of any one of claims 1-7.
9. Use of the silica microspheres capable of adsorbing nanovesicles according to claim 8 in the preparation of a formulation for treating ulcerative colitis.
10. The use according to claim 9, wherein the nanovesicles are EcN-OMVs.
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CN108892144A (en) * 2018-06-22 2018-11-27 南京亘闪生物科技有限公司 A kind of preparation method of the porous partial size monodisperse silica sphere particle shape material of functionalization
CN111116311A (en) * 2019-12-17 2020-05-08 南京科技职业学院 Method for synthesizing cinnamyl alcohol by selective hydrogenation of cinnamyl aldehyde

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CN103030752A (en) * 2011-09-29 2013-04-10 比亚迪股份有限公司 Polymer microsphere, preparation method and application of polymer microsphere, composition, light diffusion plate or light diffusion film, lamp and backlight module
CN104010722A (en) * 2011-12-01 2014-08-27 Les创新材料公司 Silica microcapsules, process of making the same and uses thereof
CN103657726A (en) * 2013-12-23 2014-03-26 湖南大学 Preparation method and application of silica-microsphere-immobilized nanometer metal compound catalyst
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