CN114917259A - Two-dimensional hydrosilylene-microorganism composite material and preparation method and application thereof - Google Patents
Two-dimensional hydrosilylene-microorganism composite material and preparation method and application thereof Download PDFInfo
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Abstract
The inventionProviding a two-dimensional hydrosilylene-microbial composite comprising: microbial carrier and two-dimensional hydrogen-bonded silylene nanosheets loaded on the microbial carrier, wherein the total amount of the two-dimensional hydrogen-bonded silylene nanosheets loaded on each microbial carrier is 10 –6 ‑10 –11 μ g. The invention also provides a preparation method of the two-dimensional hydrosilacene-microorganism composite material. The invention also provides application of the two-dimensional hydrosilylene-microbial composite material as an intestinal delivery carrier and a free radical scavenger in preparation of medicines for treating inflammatory bowel diseases. The two-dimensional hydrosilacene-microorganism composite material provided by the invention has excellent stability under the acidic condition of the stomach, can be effectively coated on the surface of the microorganism for a long time, protects the microorganism from the attack of gastric acid and enzyme, and greatly improves the survival time and survival rate of the microorganism in the stomach.
Description
Technical Field
The invention belongs to the technical field of biological nano materials, and particularly relates to a two-dimensional hydrosilylene-microorganism composite material with intestinal microenvironment response characteristics, a preparation method thereof and related applications in probiotic intestinal delivery and inflammatory bowel disease treatment.
Background
The human intestinal flora is composed of numerous bacteria and other microorganisms, has very complex physiological functions, and is closely related to the occurrence and development of various diseases (Science 2018,362, 776-780). How to regulate the intestinal flora in a safe and effective way to treat diseases is one of the problems which are widely concerned. Probiotic therapy has been a research focus in the field of recent years. Compared with the traditional treatment method, the probiotic therapy has the advantages of high safety, difficulty in generating drug resistance and the like, can assist the reconstruction of the intestinal flora steady state, and can achieve the treatment effect on various diseases. Among them, the treatment of enteritis has also received a great deal of attention (nat. Rev. gastroenterol. hepatol.2019,16, 605-. Probiotic therapy may not only have a beneficial effect on the intestinal epithelium, either directly or indirectly, but may also further propagate in the gut to improve gut flora homeostasis, thereby providing long-term treatment. However, the practical use of probiotics still faces a lot of difficulties, and the most important point is that the probiotics easily lose activity under the acidic condition of the stomach and the impact of enzyme, and are difficult to reach the intestinal tract smoothly for colonization and exert the therapeutic effect.
Therefore, the development of an efficient microbial intestinal delivery strategy is urgently needed, which has an effective protection effect on microbes in the extreme stomach environment, promotes intestinal colonization of probiotics and improves the enteritis treatment effect.
Disclosure of Invention
Aiming at the limitations of the prior art, the invention aims to provide a two-dimensional hydrosilylene-microorganism composite material with intestinal microenvironment response characteristics, a preparation method thereof and relevant application in probiotic intestinal delivery and inflammatory bowel disease treatment, and solves the problems of low survival rate of microorganisms in the stomach, insufficient intestinal colonization efficiency and limited treatment effect.
In a first aspect, the present invention provides a two-dimensional hydrosilylene-microbial composite comprising: microbial carrier and two-dimensional hydrogen-bonded silylene nanosheets loaded on the microbial carrier, wherein the total amount of the two-dimensional hydrogen-bonded silylene nanosheets loaded on each microbial carrier is 10 –6 -10 –11 μg。
Preferably, the microbial carrier is fungus or bacteria, preferably probiotics, including yeast, Trichoderma reesei, Escherichia coli, Bifidobacterium, Lactobacillus, Bacillus subtilis, Staphylococcus aureus and Bacillus proteus, preferably Escherichia coli Nissel 1917 subtype.
Preferably, the two-dimensional hydrogen bonding silicon alkene nano-sheet is of a layered lamellar structure, the transverse dimension is preferably 300-500 nm, and the thickness is preferably 0.5-10 nm.
Preferably, the polymer surface modifier is used for promoting the two-dimensional hydrogen bonding silicon alkene nanometer sheet loading.
Preferably, the polymer surface modifier is a positively charged amphiphilic polymer, preferably vitamin E-polyethylene glycol-polyethyleneimine.
Preferably, in the vitamin E-polyethylene glycol-polyethyleneimine, the molecular weight of polyethylene glycol is 500-10000, preferably 2000; the molecular weight of the polyethyleneimine is 400-10000, preferably 600.
In a second aspect, the present invention also provides a preparation method of the two-dimensional hydrosilylene-microbial composite material, which comprises the following steps:
step (1), providing a two-dimensional hydrogen bonding silicon alkene nano sheet;
mixing the two-dimensional hydrogen bonded silylene nanosheets and the high molecular modifier in a solvent, then blowing the solvent dry by using inert gas, dispersing the product in water, and centrifugally cleaning;
and (3) co-incubating the product obtained in the step (2) and a microbial carrier at a low temperature, centrifuging and washing to obtain the two-dimensional hydrosilylene-microbial composite material.
Preferably, in the step (1), the preparation method of the two-dimensional hydrogen bonding silicon alkene nano material comprises the following steps: mixing CaSi 2 And immersing the powder into concentrated hydrochloric acid solution, stirring and mixing at-30 to-10 ℃, centrifuging, removing supernatant, and washing a centrifugal product to obtain the two-dimensional hydrogen bonded silylene nano material.
Preferably, the CaSi is 2 The particle size of the powder is 5-10 mu m, and the CaSi is 2 The concentration of (a) is 5-20 mg/mL; stirring and mixing at-30 to-10 ℃ to form magnetic stirring, wherein the rotating speed of magnetons of the magnetic stirring is 500 to 1000 revolutions per molecule, the magnetic stirring time is 3 to 10 days, and the magnetic stirring and mixing are carried out under the protection of inert gas, and the inert gas is preferably argon; the rotation speed of the centrifugal treatment is 13000rpm to 20000rpm, and the time is 15 minutes to 20 minutes; the solution adopted by washing is absolute ethyl alcohol or deionized water, and the washing times are 3-4; the method further comprises the step of carrying out ultrasonic crushing on the material after the centrifugal product is washed in absolute ethyl alcohol, wherein the power of the ultrasonic crushing is 500-800W, and the time of the ultrasonic crushing is 8-15 h.
Preferably, in the step (2), the solvent is one or more selected from water, ethanol, acetone and chloroform, preferably absolute ethanol, and the inert gas is nitrogen; the high molecular modifier is vitamin E-polyethylene glycol-polyethyleneimine, and the mass ratio of the vitamin E-polyethylene glycol-polyethyleneimine to the two-dimensional hydrogen bonded silylene nanosheet is preferably (1-20): 1.
preferably, in step (3), the concentration of the microorganism carrier is 10 6 -10 12 CFU/mL, the concentration of the two-dimensional hydrogen bonding silicon alkene nano sheet modified by the macromolecular modifier is 5-200 mug/mL, and the co-incubation temperature is 0-10 ℃, preferably 4 ℃.
In a third aspect, the invention also provides the application of the two-dimensional hydrosilylene-microbial composite material as described above as an intestinal delivery carrier and a free radical scavenger in preparing a medicament for treating inflammatory bowel diseases.
According to the invention, the two-dimensional hydrogen bonded silylene-microorganism composite material is characterized in that two-dimensional hydrogen bonded silylene nanosheets are loaded on a probiotic microorganism carrier, and the probiotic microorganism carrier coated by the two-dimensional hydrogen bonded silylene nanosheets can protect the probiotic microorganism from being supplied by gastric acid and enzyme, so that the survival time and the survival rate of the probiotic microorganism in the stomach are improved. The probiotic microorganism wrapped by the two-dimensional hydrogen bonded silylene nanosheets has good micro-environment response characteristics, the two-dimensional hydrogen silylene-microorganism composite material can be rapidly degraded and release the internally wrapped probiotic microorganism after reaching the intestinal tract, the metabolism and proliferation of the microorganism are not influenced, and the intestinal tract colonization efficiency of the microorganism is greatly improved. In the invention, the two-dimensional hydrogen bonded silylene-microbial composite material also comprises a polymer surface modifier for promoting the loading of the two-dimensional hydrogen bonded silylene nanosheets, so that the loading rate of the two-dimensional hydrogen bonded silylene nanosheets loaded on the probiotic microbial carrier is further improved, and meanwhile, vitamin E-polyethylene glycol-polyethyleneimine is preferably used as the polymer surface modifier, so that the reduction of microbial activity cannot be caused in the modification process.
The invention has the beneficial effects that:
the two-dimensional hydrosilacene-microorganism composite material and the preparation method thereof have the advantages of mild preparation conditions, simple and convenient operation, and no reduction of microorganism activity in the modification process; the two-dimensional hydrosilylene-microorganism composite material provided by the invention has excellent stability under the acidic condition of the stomach, can be effectively coated on the surface of a microorganism for a long time, protects the microorganism from being attacked by gastric acid and enzyme, and greatly improves the survival time and survival rate of the microorganism in the stomach; the two-dimensional hydrosilylene-microorganism composite material provided by the invention has good micro-environment response characteristics, can be rapidly degraded and release internally wrapped microorganisms after reaching the intestinal tract, does not influence the metabolism and proliferation of the microorganisms, and greatly improves the intestinal tract colonization efficiency of the microorganisms.
Drawings
FIG. 1 shows a flow diagram for preparing a two-dimensional hydrosilylene-microbial composite according to one embodiment of the present invention.
Fig. 2 shows two-dimensional hydrosilylene nanosheets (SiH) prepared according to example 1 of the present invention and the morphology of vitamin E-polyethylene glycol-polyethyleneimine (SiH @ TPGS-PEI) modified hydrosilylene nanosheets under a transmission electron microscope.
Fig. 3 shows a scanning electron microscope image of the two-dimensional hydrosilylene-microbial composite prepared according to example 1 of the present invention.
Fig. 4 shows the uv-vis absorption spectrum of the hydrolimonene nanosheets in the artificial gastric juice after modification with vitamin E-polyethylene glycol-polyethyleneimine according to example 2 of the present invention as a function of time.
Fig. 5 shows a graph of the uv-vis absorption spectrum of the hydrogen-silane nanosheet in the artificial intestinal fluid after being modified by vitamin E-polyethylene glycol-polyethyleneimine in example 2 according to the present invention as a function of time.
Fig. 6 shows the survival of bacteria in artificial gastric juice in the two-dimensional hydrosilacene-microorganism composite material according to example 3 of the present invention.
FIG. 7 is a graph showing the evaluation of the survival time of the two-dimensional hydrosilylene-microbial composite in example 4 according to the present invention in artificial gastric juice.
Detailed Description
The present invention is further described below in conjunction with the following embodiments and the accompanying drawings, it being understood that the embodiments and the drawings are illustrative of the invention only and are not limiting.
According to a first aspect of the present invention, there is provided a two-dimensional hydrosilylene-microbial composite comprising: the microbial carrier comprises a microbial carrier and two-dimensional hydrogen bonding silicon alkene nano sheets loaded on the microbial carrier, and optionally further comprises a polymer surface modifier for promoting the loading of the two-dimensional hydrogen bonding silicon alkene nano sheets.
According to the invention, the microbial carrier is fungi or bacteria, in the embodiment of the invention, probiotics, including one or more of yeast, trichoderma reesei, escherichia coli, bifidobacterium, lactobacillus, bacillus subtilis, staphylococcus aureus and proteus, in the embodiment of the invention, escherichia coli belongs to the Nissel 1917 subtype.
The probiotics are fungi or bacteria which are beneficial to human bodies or animal bodies, and the probiotics change the composition of flora at a certain part of a host through colonizing in the human bodies, so that the probiotics are active microorganisms which are beneficial to the host. The intestinal health-care function is realized by regulating the immune function of a host mucous membrane and a system or regulating the flora balance in the intestinal tract, and the nutrition absorption is promoted, so that the single microorganism or the mixed microorganism with definite composition which is favorable for the health is generated.
However, the practical use of probiotics still faces a lot of difficulties, and the most important point is that the probiotics easily lose activity under the acidic condition of the stomach and the impact of enzyme, and are difficult to reach the intestinal tract smoothly for colonization and exert the therapeutic effect. Therefore, the invention also loads the two-dimensional hydrogen bonded silylene nanosheet on the probiotic microorganism carrier on the basis of the original probiotic microorganism carrier. The probiotic microorganism carrier wrapped by the two-dimensional hydrogen bonded silylene nanosheets can protect the probiotic microorganisms from being supplied by gastric acid and enzyme, and improve the survival time and survival rate of the probiotic microorganisms in the stomach. The probiotic microorganism wrapped by the two-dimensional hydrogen bonded silylene nanosheets has good micro-environment response characteristics, the two-dimensional hydrogen silylene-microorganism composite material can be rapidly degraded and release the internally wrapped probiotic microorganism after reaching the intestinal tract, the metabolism and proliferation of the microorganism are not influenced, and the intestinal tract colonization efficiency of the microorganism is greatly improved.
According to the invention, the two-dimensional hydrogen bonding silicon alkene nano sheet is of a layered lamellar structure, the transverse dimension is preferably 300-500 nm, and the thickness is preferably 0.5-10 nm.
In order to promote the loading of the two-dimensional hydrogen bonding silicon alkene nano-sheets, the invention also optionally comprises a macromolecular surface modifier for modifying the surfaces of the two-dimensional hydrogen bonding silicon alkene nano-sheets. The polymer surface modifier is an amphiphilic polymer with positive charges, and vitamin E-polyethylene glycol-polyethyleneimine is adopted in the embodiment of the invention.
The two-dimensional hydrosilylene-microbial composite material also comprises a high molecular surface modifier used for modifying the surface of the two-dimensional hydrogen bonding silylene nanosheet so as to promote the two-dimensional hydrogen bonding silylene nanosheet to be loaded on a probiotic microbial carrier, the loading rate of the two-dimensional hydrogen bonding silylene nanosheet loaded on the probiotic microbial carrier is further improved, and meanwhile, vitamin E-polyethylene glycol-polyethyleneimine is preferably used as the high molecular surface modifier to ensure that the modification process cannot cause the reduction of microbial activity.
In a preferred embodiment, in said vitamin E-PEG-polyethyleneimine the molecular weight of PEG is 500-10000, such as 2000; the molecular weight of the polyethyleneimine is 400-10000, for example 600.
The invention also provides a preparation method of the two-dimensional hydrosilylene-microorganism composite material, which mainly comprises three parts, namely preparation of hydrosilylene nanosheets, modification of the hydrosilylene nanosheets, and assembly of the modified hydrosilylene nanosheets and microorganisms. The preparation method comprises the following steps:
In this step 1, the CaSi 2 The particle size of the powder is 5-10 mu m, and the concentration is 5-20mg/mL, such as 10 mg/mL; the rotating speed of the magnetons of the magnetic stirring is 500-1000 revolutions per molecule,for a period of 3-10 days, e.g., 7 days; the inert gas is preferably argon; the rotation speed of the centrifugal treatment is 13000rpm to 20000rpm, and the time is 15 minutes to 20 minutes; the solution adopted by washing is absolute ethyl alcohol or deionized water, and the washing times are 3-4.
In the preparation process of the two-dimensional hydrosilylene nanosheet, the step of ultrasonically crushing the material after washing the centrifugal product in absolute ethyl alcohol is also included, the power of ultrasonic crushing is 500-800W, and the time of ultrasonic crushing is 8-15 h.
Step 2, modification of two-dimensional hydrogen-silane nanosheets: mixing the two-dimensional hydrogen bonded silylene nanosheet with a polymer modifier in a solvent, then blowing the solvent dry by inert gas, dispersing the product in water, and centrifugally cleaning.
In the step 2, the solvent is one or more selected from water, ethanol, acetone and chloroform, preferably absolute ethanol, and the inert gas is nitrogen; in an embodiment of the present invention, the polymer modifier is vitamin E-polyethylene glycol-polyethyleneimine, and a mass ratio of the vitamin E-polyethylene glycol-polyethyleneimine to the two-dimensional hydrogen-bonded silylene nanosheet is preferably (1-20): 1, e.g., 10: 1.
step 3, assembling the modified two-dimensional hydrosilylene nanosheet and the probiotic microbial carrier: and (3) co-incubating the product obtained in the step (2) and a probiotic microorganism carrier at a low temperature of 4 ℃, centrifuging and washing to obtain the two-dimensional hydrosilylene-microorganism composite material.
In this step 3, the concentration of the microorganism carrier is 10 6 -10 12 CFU/mL, e.g. 10 8 CFU/mL; the concentration of the two-dimensional hydrogen bonding silicon alkene nano sheet modified by the macromolecule modifier is 5-200 mug/mL, such as 30 mug/mL.
The invention also provides application of the two-dimensional hydrosilylene-microbial composite material as an intestinal delivery carrier and a free radical scavenger in preparation of medicines for treating inflammatory bowel diseases. According to the invention, the two-dimensional hydrosilylene-microorganism composite material provided by the invention has excellent stability under the acidic condition of the stomach, can be effectively coated on the surface of the microorganism for a long time, protects the microorganism from being attacked by gastric acid and enzyme, and greatly improves the survival time and survival rate of the microorganism in the stomach; the two-dimensional hydrosilylene-microorganism composite material provided by the invention has good micro-environment response characteristics, can be rapidly degraded and release internally wrapped microorganisms after reaching the intestinal tract, does not influence the metabolism and proliferation of the microorganisms, and greatly improves the intestinal tract colonization efficiency of the microorganisms.
The present invention will be described in detail by way of examples. It is also to be understood that the following examples are illustrative of the present invention and are not to be construed as limiting the scope of the invention, and that certain insubstantial modifications and adaptations of the invention by those skilled in the art in light of the foregoing description are intended to be included within the scope of the invention. The specific process parameters and the like of the following examples are also only one example of suitable ranges, i.e., those skilled in the art can select the appropriate ranges through the description herein, and do not necessarily limit the specific values to the examples below.
Example 1:
preparing a two-dimensional hydrosilylene-microorganism composite material: the preparation steps are mainly divided into three parts, including the preparation of the hydrogen-silane-alkene nano-sheets, the modification of the hydrogen-silane-alkene nano-sheets and the assembly of the modified hydrogen-silane-alkene nano-sheets and the probiotic microorganism. The specific preparation process is shown in fig. 1, and three steps are respectively explained as follows:
preparation of hydrosilylene nanosheets: 1g of calcium silicide was added to 100mL of precooled concentrated HCl and stirred vigorously at-20 ℃ under argon protection for 7 days. After the reaction is finished, centrifuging at 12000rpm for 10min, washing precipitates by acetone and absolute ethyl alcohol in sequence, and then suspending the precipitates in the absolute ethyl alcohol. And (3) performing ultrasonic treatment for 8 hours by using an ice bath probe to obtain the hydrogen-silane nanosheet, wherein the transmission electron microscope result is shown in figure 2.
Modification of hydrogen silicon alkene nano-sheet: 1mg of hydrogen-silane-alkene nano-sheets and 10mg of vitamin E-polyethylene glycol-polyethyleneimine are dispersed in absolute ethyl alcohol together, dried by nitrogen and then re-suspended by ultrapure water, and centrifugally washed for 2 times to obtain the vitamin E-polyethylene glycol-polyethyleneimine modified hydrogen-silane-alkene nano-sheets, and the result of a transmission electron microscope is shown in figure 2.
Assembling the modified hydrogen-silane-alkene nanosheet and the probiotic microorganism: and (3) resuspending the modified hydrogen-silane nano-sheets in ultrapure water, and diluting the ultra-pure water into hydrogen-silane suspension with different concentrations. The probiotic microorganisms were centrifuged at 7000rpm for 5min, washed twice and dispersed in ultrapure water. And mixing the hydrosilicene suspension and the probiotic microorganism suspension, co-incubating for 20-30min at the temperature of 4 ℃, and centrifuging and washing for 3 times to obtain the two-dimensional hydrosilicene-microorganism composite material. The scanning electron microscope results are shown in FIG. 3.
Example 2:
the vitamin E-polyethylene glycol-polyethyleneimine modified hydrogen-graphene nanosheets are dispersed in artificial gastric juice and artificial intestinal juice, and the stability and the responsiveness of the vitamin E-polyethylene glycol-polyethyleneimine modified hydrogen-graphene nanosheets in different physiological environments are judged by monitoring the change of ultraviolet-visible absorption spectra of the vitamin E-polyethylene glycol-polyethyleneimine modified hydrogen-graphene nanosheets. As can be seen from FIGS. 4 and 5, the material has good stability in artificial gastric juice (FIG. 4) and almost no change in the UV-visible absorption spectrum. Whereas in artificial intestinal fluids (fig. 5), the uv-visible absorbance decreased rapidly, indicating that the material can degrade rapidly in artificial intestinal fluids.
Example 3:
and evaluating the bacterial survival rate of the two-dimensional hydrosilacene-microorganism composite material in gastric juice. 1mL of probiotics in the logarithmic growth phase is centrifuged and then resuspended in 100 mu L of ultrapure water, and is mixed with hydrogen-silicon alkene nano-sheets modified by vitamin E-polyethylene glycol-polyethyleneimine with different concentrations in sterile water, and finally the content of hydrogen-silicon alkene is 0, 3, 15 and 30 mu g. And after incubation for half an hour, centrifugally cleaning to obtain the two-dimensional hydrosilylene-probiotic composite material. Dispersing the obtained composite material in artificial gastric juice, incubating for 15min, and counting the survival rate of the composite material by a plate coating method. As shown in fig. 6, the unmodified probiotics have no bacteria to survive after the probiotics react with gastric juice, and the hydrosilane-probiotics composite material has good probiotic protection effect, so that the survival rate of the probiotics in gastric juice is greatly improved.
Example 4:
evaluation of the survival time of the two-dimensional hydrosilaene-microbial composite in gastric juice. 1mL of probiotics in the logarithmic growth phase is centrifuged and then suspended in 100 mu L of ultrapure water, and is mixed with vitamin E-polyethylene glycol-polyethyleneimine modified hydrosilylene nanosheets in sterile water, and finally the content of hydrosilylene is 30 mu g. And after incubation for half an hour, centrifugally cleaning to obtain the two-dimensional hydrosilylene-probiotic composite material. The obtained composite material is dispersed in 1mL artificial gastric juice, after incubation for 15min, 30min and 60min, the survival rate of the composite material is counted by a plate coating method, and the statistical result is shown in figure 7, so that the survival efficiency of probiotics can be greatly improved within 1 hour.
Industrial applicability:
the two-dimensional hydrosilylene-microorganism composite material and the preparation method thereof have the advantages that the preparation condition is mild, the operation is simple and convenient, and the modification process cannot cause the reduction of the activity of microorganisms; the two-dimensional hydrosilylene-microorganism composite material provided by the invention has excellent stability under the acidic condition of the stomach, can be effectively coated on the surface of a microorganism for a long time, protects the microorganism from being attacked by gastric acid and enzyme, and greatly improves the survival time and survival rate of the microorganism in the stomach; the two-dimensional hydrosilylene-microorganism composite material provided by the invention has good micro-environment response characteristics, can be rapidly degraded and release internally wrapped microorganisms after reaching the intestinal tract, does not influence the metabolism and proliferation of the microorganisms, and greatly improves the intestinal tract colonization efficiency of the microorganisms.
Claims (12)
1. A two-dimensional hydrosilylene-microbial composite, comprising: microbial carrier and two-dimensional hydrogen-bonded silylene nanosheets loaded on the microbial carrier, wherein the total amount of the two-dimensional hydrogen-bonded silylene nanosheets loaded on each microbial carrier is 10 –6 -10 –11 μg。
2. The two-dimensional hydrosilylene-microbe composite material of claim 1, wherein the microbe carrier is fungus or bacteria, preferably probiotic bacteria, including one or more of yeast, trichoderma reesei, escherichia coli, bifidobacterium, lactobacillus, bacillus subtilis, staphylococcus aureus and proteus, preferably escherichia coli, Nissel 1917 subtype.
3. The two-dimensional hydrosilylene-microbial composite material as claimed in claim 1 or 2, wherein the two-dimensional hydrogen bonded silylene nanosheet is of a layered lamellar structure, the transverse dimension is preferably 300-500 nm, and the thickness is preferably 0.5-10 nm.
4. The two-dimensional hydrosilylene-microbial composite of any of claims 1-3 further comprising a polymeric surface modifier for promoting the loading of the two-dimensional hydrogen bonded silylene nanoplatelets.
5. The two-dimensional hydrosilylene-microbial composite of claim 4, wherein the polymeric surface modifier is a positively charged amphiphilic polymer, preferably vitamin E-polyethylene glycol-polyethyleneimine.
6. The two-dimensional hydrosilylene-microbe composite material as claimed in claim 5, wherein in the vitamin E-PEG-polyethyleneimine, the molecular weight of PEG is 500-10000, preferably 2000; the molecular weight of polyethyleneimine is 400-10000, preferably 600.
7. A method for preparing a two-dimensional hydrosilylene-microbial composite as claimed in any of claims 1-6, comprising the steps of:
step (1), providing a two-dimensional hydrogen bonding silicon alkene nano sheet;
mixing the two-dimensional hydrogen bonded silylene nanosheets and the high molecular modifier in a solvent, then blowing the solvent dry by using inert gas, dispersing the product in water, and centrifugally cleaning;
and (3) co-incubating the product obtained in the step (2) and a microbial carrier at a low temperature, centrifuging and washing to obtain the two-dimensional hydrosilylene-microbial composite material.
8. The preparation method according to claim 7, wherein in the step (1), the preparation method of the two-dimensional hydrogen-bonded silylene nanomaterial comprises the following steps: mixing CaSi 2 And immersing the powder into concentrated hydrochloric acid solution, stirring and mixing at-30 to-10 ℃, centrifuging, removing supernatant, and washing a centrifugal product to obtain the two-dimensional hydrogen bonded silylene nano material.
9. The method according to claim 8, wherein the CaSi is used as a base material 2 The particle size of the powder is 5-10 mu m, and the CaSi is 2 The concentration of (a) is 5-20 mg/mL; stirring and mixing at-30 to-10 ℃ to form magnetic stirring, wherein the rotating speed of magnetons of the magnetic stirring is 500 to 1000 revolutions per molecule, the magnetic stirring time is 3 to 10 days, and the magnetic stirring and mixing are carried out under the protection of inert gas, wherein the inert gas is preferably argon; the rotation speed of the centrifugal treatment is 13000rpm to 20000rpm, and the time is 15 minutes to 20 minutes; the solution adopted by washing is absolute ethyl alcohol or deionized water, and the washing times are 3-4 times; the method further comprises the step of carrying out ultrasonic crushing on the material after the centrifugal product is washed in absolute ethyl alcohol, wherein the power of the ultrasonic crushing is 500-800W, and the time of the ultrasonic crushing is 8-15 h.
10. The preparation method according to claim 7, wherein in the step (2), the solvent is one or more selected from water, ethanol, acetone and chloroform, preferably absolute ethanol, and the inert gas is nitrogen; the high-molecular modifier is vitamin E-polyethylene glycol-polyethyleneimine, and the mass ratio of the vitamin E-polyethylene glycol-polyethyleneimine to the two-dimensional hydrogen bonded silylene nanosheet is preferably (1-20): 1.
11. the method according to claim 7, wherein the concentration of the microorganism carrier in the step (3) is 10 6 -10 12 CFU/mL, the concentration of the two-dimensional hydrogen bonding silicon alkene nano-sheet modified by the macromolecular modifier is 5-200 mug/mL, and the co-incubation temperatureIs 0 to 10 ℃ and preferably 4 ℃.
12. Use of the two-dimensional hydrosilylene-microbial composite of any of claims 1-6 as an intestinal delivery vehicle and a free radical scavenger for the manufacture of a medicament for the treatment of inflammatory bowel disease.
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