Use of electrogenic microbial vesicles in the preparation of a medicament or biological agent for modulating the functioning of an organism

Abstract

The application discloses applications of electricity-generating microbial vesicles, in particular, applications in preparation of medicines or biological preparations for regulating functions of organisms. The electricity-active microbial vesicles provided by the application have a natural biocompatibility and degradability of a lipid bilayer shell, can establish close electrical communication with host cells or tissues, and can be safely metabolized by the body after completing the electrical regulation function, fundamentally avoiding safety hazards caused by low electrical communication efficiency of synthetic materials and long-term retention.

Description

Technical Field

[0001] This application relates to the field of new materials and their application technology, specifically to the application of electrogenic microbial vesicles in the preparation of drugs or biological agents for regulating the functions of organisms. Background Technology

[0002] Microbial vesicles, as natural nanoscale lipid bilayer structures, have shown great potential in the biomedical field due to their unique physicochemical properties and biological functions. These spherical particles carry a variety of bioactive molecules derived from microorganisms, such as lipopolysaccharides, outer membrane proteins, periplasmic proteins, and nucleic acids, and can serve as natural intercellular communication carriers, precisely regulating host functions at the molecular and cellular levels. However, single-component vesicles often cannot cope with the complexity of the disease microenvironment and lack the ability to integrate multiple functions according to treatment needs, resulting in low treatment efficiency.

[0003] In nature, there exists a special group of microorganisms known as electroactive microorganisms, including species such as *Shewanella* and *Geobacterium*, which possess unique extracellular electron transport capabilities. These microorganisms transfer electrons generated intracellularly to extracellular solid-state acceptors through multiple electron transport pathways, including cytochrome proteins, conductive nanowires, and redox-active small molecules. Vesicles secreted by these electroactive microorganisms inevitably encapsulate electron transport elements such as cytochrome proteins and conductive appendages from the parent bacterial membrane during their formation, endowing them with unique electrochemical activity. However, current technology lacks understanding of this subgroup of microbial vesicles with redox activity and conductivity, failing to distinguish them from ordinary microbial vesicles and failing to specifically develop and utilize them.

[0004] Numerous studies have confirmed that endogenous electrical signals play a crucial guiding role in tissue development and regeneration. Damage to tissues such as skin, bone, nerves, and myocardium generates changes in transepithelial potential differences, forming an endogenous electric field that guides immune cell chemotaxis, stem cell migration and differentiation, and orderly tissue reconstruction. However, current strategies for simulating or enhancing this electrical microenvironment using biomaterials to promote tissue regeneration primarily rely on non-biosynthetic systems such as conductive polymers, carbon-based nanomaterials, or piezoelectric materials. These exogenous conductive materials generally suffer from inherent drawbacks such as non-biodegradability, chronic inflammatory responses, and low electrical coupling efficiency with host tissues. Currently, there is a lack of naturally derived, highly biocompatible bioactive materials capable of establishing effective electrical communication with host tissues to respond to or regulate the endogenous electrical signaling network of tissue regeneration.

[0005] Therefore, the targeted extraction of electroactive microbial vesicles from electroactive microorganisms, the systematic characterization of their electron transfer and electroregulation functions, and the development of them into natural bioelectronic materials that can respond to and regulate the tissue electromicroenvironment are the technological gaps and key bottlenecks that urgently need to be overcome in the current research and application of microbial vesicles. Summary of the Invention

[0006] Based on this, this application provides at least the use of electrogenic microbial vesicles in the preparation of drugs or biological agents for regulating the functions of organisms.

[0007] In a first aspect of this application, the use of electrogenic microbial vesicles in the preparation of drugs or biological agents for regulating the function of organisms is provided.

[0008] In a second aspect of this application, a method for regulating cell function in vitro is provided, the method comprising:

[0009] Mix electrogenic microbial vesicles as defined above with cells to allow them to interact;

[0010] Assess changes in cell function.

[0011] The electrogenic microbial vesicles of this application for regulating the functions of organisms have at least the following advantages compared to existing technologies:

[0012] 1. This application is the first to extract and isolate bacterial exovesicles with stable electrochemical activity from electroactive microorganisms, and systematically characterize their electron transfer capabilities. It innovatively expands microbial vesicles from simple "biochemical carriers" to "bioelectronic functional units", providing a new natural source material for bioelectronic medicine.

[0013] 2. The electroactive microbial vesicles provided in this application have a lipid bilayer shell with natural biocompatibility and degradability, enabling them to establish close electrical communication with host cells or tissues. After completing their electroregulation function, they can be safely metabolized by the body, fundamentally avoiding the safety hazards caused by the low electrical communication efficiency and long-term retention of synthetic materials.

[0014] 3. Existing microbial vesicle technologies can only regulate the biochemical signaling pathways of target cells by loading molecules such as proteins and nucleic acids. Their functional dimensions are limited, making it difficult to meet the needs of tissue engineering applications highly sensitive to electrical signals, such as nerve regeneration and myocardial repair. This application utilizes the naturally occurring "dual-mode signaling" capability of electroactive microbial vesicles—they can not only deliver biochemical signaling molecules but also act as nanoscale "charge shuttles," directly providing electrons to target cells or the tissue microenvironment and regulating transmembrane potential, achieving spatiotemporal synergistic regulation of electrical and biochemical signals. This multimodal synergistic effect demonstrates therapeutic potential in promoting targeted tissue regeneration and functional reconstruction that is unparalleled by traditional vesicle technologies.

[0015] 4. The electroactive microbial vesicles provided in this application can simulate and enhance the endogenous electrical signal network after tissue damage, guide the directional migration and differentiation of stem cells, promote functional neural synaptic connections and myocardial reconstruction, and elevate the scope of tissue repair application of microbial vesicles from "structural repair" to a new level of "functional electrointegrative repair". Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments and examples of this application, and to more completely understand this application and its beneficial effects, the drawings used in the description of the embodiments or examples will be briefly introduced below. Obviously, the drawings described below are merely some embodiments of this application. Those skilled in the art can obtain other drawings based on these drawings without creative effort. It should also be noted that the drawings are all drawn in a simplified form and are only used to conveniently and clearly assist in illustrating this application.

[0017] Figure 1 This is a schematic diagram illustrating the vesicle generation of electrogenic microorganisms under different antibiotic concentrations in one embodiment of this application.

[0018] Figure 2 This is a scanning electron microscope image of an electrogenic microbial vesicle in one embodiment of this application.

[0019] Figure 3 This diagram shows the test results of the reactive oxygen species scavenging capacity of electrogenic microbial vesicles in one embodiment of this application.

[0020] Figure 4 Electrochemical impedance spectroscopy of electrogenic microorganisms and their vesicles.

[0021] Figure 5 Cyclic voltammetry curves of electrogenic microorganisms and their vesicles.

[0022] Figure 6 This diagram illustrates the evaluation of the antifibrotic ability of electrogenic microbial vesicles in one embodiment of this application.

[0023] Figure 7 This diagram illustrates the EGF expression effect of engineered bacterial cells in the presence of electrogenic microbial vesicles according to an embodiment of this application.

[0024] Figure 8 This diagram illustrates a rat cerebral infarction treatment evaluation in the presence of electrogenic microbial vesicles according to an embodiment of this application. Detailed Implementation

[0025] To facilitate understanding of this application, a more complete description will be provided below with reference to the accompanying drawings. Preferred embodiments of this application are shown in the drawings. However, this application can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of this application.

[0026] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0027] In this application, unless otherwise specified, "one or more" means any one of the listed items or any combination of the listed items. Similarly, "one or more" and other instances that otherwise indicate "one or more" shall be understood in the same way unless otherwise specified.

[0028] The terms “combinations thereof,” “any combination thereof,” and “any combination thereof” as used in this application include all suitable combinations of any two or more of the listed items.

[0029] In this application, the word "suitable" in "suitable combination", "suitable method", "any suitable method" etc., shall be defined as being able to implement the technical solution of this application, solve the technical problem of this application, and achieve the expected technical effect of this application.

[0030] In this application, terms such as "further," "even more," "particularly," "for example," "like," "example," and "exemplary" are used for descriptive purposes to indicate that different technical solutions preceding and following each other are related in terms of their coverage, but should not be construed as limiting the preceding technical solution or restricting the scope of protection of this application. In this application, unless otherwise specified, A (e.g., B) indicates that B is a non-limiting example of A, and it can be understood that A is not limited to B.

[0031] In this application, "optionally," "optionally," and "optional" mean that something is optional, that is, it refers to either "with" or "without" a parallel solution. If multiple "options" appear in a technical solution, unless otherwise specified and there are no contradictions or mutual constraints, each "option" is independent. Unless otherwise specified, the descriptions such as "optionally include" and "optionally contain" in this application, taking "optionally include" as an example, mean "may include or not include."

[0032] The terms “containing,” “comprising,” and “including” as used in this application are synonyms and are inclusive or open-ended, not excluding additional, uncited members or features. Members or features include, for example, materials or components, structures, elements, instruments, etc.; non-limiting examples of members or features include actions, conditions under which actions occur, timing, states, etc.

[0033] In this application, the technical features or solutions described in open-ended language include both closed-ended technical features or solutions consisting of the listed contents and open-ended technical features or solutions that include the listed contents.

[0034] In this application, the exemplary descriptions such as "in some implementations (or embodiments)" and "in one implementation (or embodiment)" may cover, but are not limited to, the following meanings: these solutions can be combined with other solutions in a suitable manner to form new technical solutions.

[0035] In this application, the terms "first aspect," "second aspect," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first aspect," "second aspect," etc., serve only as a non-exhaustive enumeration and should be understood not to constitute a closed limitation on quantity.

[0036] In this application, when numerical intervals (i.e., numerical ranges) are involved, unless otherwise specified, the distribution of selectable numerical values ​​within the numerical interval is considered continuous, and includes the two endpoints of the numerical interval (i.e., the minimum and maximum values), as well as every numerical value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that numerical interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints, which is equivalent to directly listing every integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be merged. In other words, unless otherwise specified, the numerical ranges disclosed herein should be understood to include any and all subranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, percentage, ratio, etc. The term "numerical interval" can be broadly included to include numerical interval types such as percentage intervals, ratio intervals, and proportion intervals.

[0037] In this application, where the method flow involves multiple steps, unless otherwise explicitly stated herein, there is no strict order restriction on the execution of these steps; they can be executed in any order other than those described. Moreover, any step may include multiple sub-steps or multiple stages, which are not necessarily completed at the same time, but can be executed at different times, and their execution order is not necessarily sequential, but can be performed alternately or simultaneously with other steps or parts of the sub-steps or stages of other steps.

[0038] One aspect of this application provides the use of electrogenic microbial vesicles in the preparation of drugs or biological agents for regulating the function of organisms.

[0039] In some embodiments, the regulation of organism function includes regulating cellular function and / or regulating animal function. Exemplarily, this may involve regulating cellular function, regulating animal function, or simultaneously regulating both cellular and animal functions.

[0040] In some embodiments, the regulation of cellular function includes regulating one or more of the following: cell proliferation, differentiation, protein expression, reactive oxygen species levels, and electrophysiological activities.

[0041] In some implementations, regulating cell differentiation includes inhibiting fibroblast differentiation into myofibroblasts.

[0042] In some embodiments, regulating cellular protein expression includes enhancing the ability to express functional proteins; said functional proteins may optionally include human epidermal growth factor.

[0043] For example, the electrogenic microbial vesicles have been found to have excellent reactive oxygen species scavenging capabilities. While not wishing to be limited by any theory, this capability is considered to indicate a potential for inflammation reduction.

[0044] In some implementations, the regulation of animal function includes one or more of promoting tissue regeneration, repairing organ function, and treating diseases.

[0045] The tissue regeneration mentioned includes, but is not limited to, nerve tissue regeneration, myocardial tissue regeneration, and bone tissue regeneration.

[0046] In some embodiments, the disease includes, but is not limited to, ischemic stroke, myocardial injury, and neurodegenerative diseases.

[0047] In some embodiments, the electrogenic microbial vesicles are electrochemically active and capable of mediating extracellular electron transfer.

[0048] In some embodiments, the electrogenic microbial vesicles possess excellent redox capabilities and electron transport capabilities.

[0049] In some embodiments, the electrogenic microbial vesicles have low electrical impedance, for example, 12.5 Ω to 13 Ω.

[0050] For electrogenic microbial vesicles, low impedance means that they have good electron transport capabilities on their surface or inside, which is conducive to promoting the formation of electroactive biofilms or intercellular electron exchange.

[0051] In some embodiments, the electrogenic microbial vesicles are obtained by extracting electrogenic microorganisms after interventional culture.

[0052] For example, the intervention culture includes stress culture of electrogenic microorganisms using at least one of antibiotics, chelating agents, L-cysteine, elevated temperature, varying pH, varying dissolved oxygen, carbon source starvation, and amino acid starvation.

[0053] In some embodiments, the antibiotic is ampicillin.

[0054] In some embodiments, the extraction method of the electrogenic microbial vesicles includes one or more of the following: tangential flow filtration combined with size exclusion chromatography, differential centrifugation combined with density gradient centrifugation, immunomagnetic bead method, and tangential flow filtration combined with aqueous two-phase extraction.

[0055] In some embodiments, the method for preparing the electrogenic microbial vesicles includes the following steps:

[0056] S100. Activation and amplification culture of strains: Single colonies of electrogenic microorganisms were inoculated into liquid culture medium and cultured under aerobic conditions with shaking until the bacterial concentration reached the middle and late stages of the logarithmic growth phase to obtain seed culture;

[0057] S200. Stress-induced culture: The seed culture is transferred to fresh liquid culture medium at a volume ratio of 1:(50~200), and a certain concentration of β-lactam antibiotics (such as ampicillin) is added to the culture medium as a stress inducer. The culture is shaken at a suitable temperature until the bacterial concentration reaches the middle and late logarithmic growth phase again to obtain the fermentation culture medium.

[0058] S300. Supernatant collection: Centrifuge the fermentation broth at medium to low speed (e.g., 8000 rpm) to remove the cell precipitate and collect the supernatant;

[0059] S400. Sterile filtration: The supernatant is filtered through a microporous membrane (e.g., a 0.45 μm needle filter membrane) to remove residual bacteria and large particulate impurities (including but not limited to residual bacterial fragments, denatured protein aggregates, and insoluble matter in the culture medium) to obtain a sterile filtrate;

[0060] S500. Primary Concentration: The sterile filtrate is placed in an ultrafiltration tube with a molecular weight cutoff of 50 KD~150 KDa and centrifuged at low temperature to obtain a concentrated solution.

[0061] S600. Collection of vesicles by ultracentrifugation: The concentrate is subjected to ultracentrifugation (e.g., 150,000 g) under a hypergravity field to cause the nanoscale vesicles to settle. The supernatant is discarded, and the precipitate is resuspended in buffer solution to obtain an electrogenic microbial vesicle suspension.

[0062] Electrogenic microorganisms are mainly concentrated in the phyla Proteobacteria and Firmicutes, such as Bacillus, Pseudomonas, Rubus, Shewanella, Lactococcus, and Escherichia coli. Electrogenic microorganisms produce electrons during the metabolism of organic matter and transmit them through an electron transport chain. The electrogenic microorganisms on the anode can be considered a mixture of cells with different functions; they serve as the power source for the cells and can be used for power generation and proliferation.

[0063] In some embodiments, the electrogenic microorganisms include, but are not limited to, microorganisms of the genera *Shewanella* and *Geobacter*.

[0064] In some embodiments, the electrogenic microorganism is Shewanella oneidensis MR-1.

[0065] Another aspect of this application provides a method for regulating cell function in vitro, the method comprising:

[0066] S100'. Mix electrogenic microbial vesicles as defined above with cells to allow them to interact;

[0067] S200'. Assess changes in cell function.

[0068] For example, in step S100', an electrogenic microbial vesicle is used to interact with the cell using a co-incubation method.

[0069] In some embodiments, the cells include one or more of mesenchymal stem cells, fibroblasts, cardiomyocytes, and bacterial cells.

[0070] In some implementations, changes in cellular function are assessed from at least one of the cellular, molecular, and bioinformatics analyses.

[0071] Another aspect of this application provides a method for regulating animal function, the method comprising:

[0072] S1100. Using methods such as tail vein injection, in situ injection, and implantation material combination, the electrogenic microbial vesicles described above are interacted with animals;

[0073] S1200. Assess functional changes in animals.

[0074] In some implementations, step S1200 includes: assessing the functional changes in the animal from aspects such as macroscopic observation, medical imaging, histopathology, and immunofluorescence.

[0075] The following are some examples.

[0076] The embodiments of this application will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application. For experimental methods in the following embodiments where conditions are not specified, reference should be made to the guidelines given in this application, or to experimental manuals or conventional conditions in the art, or to the conditions recommended by the manufacturer, or to experimental methods known in the art.

[0077] The practicality of the electrogenic microbial vesicles for regulating the functions of organisms in this application will be described below with reference to Example 1.

[0078] In the following examples, Shewanella MR-1 was selected as the electrogenic microorganism, the culture intervention method was the antibiotic stress method, and the vesicle extraction method was a combination of differential centrifugation and density gradient centrifugation.

[0079] The cells used mainly consist of fibroblasts and bacterial cells capable of expressing functional proteins.

[0080] The cell function regulation involved mainly includes fibroblast differentiation and the expression of functional proteins (human epidermal growth factor) in bacterial cells.

[0081] Animal function regulation mainly includes the treatment of ischemic stroke.

[0082] Example 1

[0083] Pick a single colony from an LB agar plate and inoculate it into 100 mL of LB liquid medium. Incubate overnight at 30 °C and 200 rpm with aerobic culture until OD reaches zero. 600 =0.8-1, approximately 12-16 h. The bacteria were subcultured with the culture medium at a 1:100 volume ratio, and simultaneously subjected to stress culture with 8 μg / ml ampicillin. The culture was carried out at 30°C on a shaker at 200 rpm until OD reached [value missing]. 600=1-1.6. Centrifuge at 8000 rpm for 15 min and collect the supernatant. Filter the supernatant through a 0.45 μm needle filter membrane and concentrate the filtrate using a 100 kDa, 15 mL ultrafiltration tube, centrifuging at 3500 rpm and 4℃ for 20 min. Centrifuge the concentrate at 150000 g and 4℃ for 2 h, discarding a portion of the supernatant and resuspending the precipitate in 1×PBS. Quantify the protein using a BCA kit. Figure 1 The results showed that the highest yield of microbial vesicles was observed under culture with 8 μg / ml ampicillin stress, which was 1.5 times that of the control group. Transmission electron microscopy revealed that the electrogenic microbial vesicles possessed complete morphological characteristics. Figure 2 ).

[0084] The reactive oxygen species scavenging capacity of electrogenic microbial vesicles was assessed using a UV-Vis absorption spectrometer and a total antioxidant capacity (T-AOC) assay kit (ABTS microplate method) (see [link to relevant documentation]). Figure 3 An aqueous suspension (0.4 mg / mL) of electrogenic microbial vesicles was added to a solution containing ABTS. + Samples were taken from the solution and subjected to UV-Vis absorption spectroscopy measurements. The experimental results showed that electrogenic microbial vesicles have excellent reactive oxygen species scavenging ability, indicating their potential for inflammation elimination.

[0085] The electrical properties of electrogenic microbial vesicles were evaluated using a three-electrode system on an electrochemical workstation. Electrochemical impedance spectroscopy revealed that the electrogenic microbial vesicles possessed low electrical impedance (see [link to electrochemical impedance spectroscopy]). Figure 4 Cyclic voltammetry (CV) revealed that electrogenic microbial vesicles possess excellent redox and electron transport capabilities (see [reference needed]). Figure 5 ).

[0086] Immunofluorescence staining was used to assess the differentiation of fibroblasts into myofibroblasts (collagen secretion and α-SMA (myofibroblast marker)) (see [link]). Figure 6 The results showed that the addition of electrogenic microbial vesicles significantly reduced collagen secretion and decreased the number of myofibroblasts.

[0087] The regulatory role of electrogenic microbial vesicles on microbial metabolism was assessed using ELISA. The results showed that the presence of vesicles significantly increased the ability of engineered microorganisms to express functional proteins (human epidermal growth factor) (see [link to ELISA]). Figure 7 ).

[0088] A cerebral ischemia-reperfusion model was established using middle cerebral artery occlusion. Two hours after reperfusion, the rats were given the first dose of microbial vesicles, administered once daily for seven consecutive days. After the intervention, the rats were re-anesthetized, and the entire brain was removed and placed in PBS at 4 °C. The brain was then rapidly transferred to a -20 °C freezer for 30 min. The olfactory bulb, cerebellum, and brainstem were removed, and coronal sections with a thickness of 2 mm were prepared. The brain sections were incubated in 2% TTC solution at 37 °C for 30 min, with the sections being rotated every 5 min to ensure complete contact with the solution. Subsequently, the brain sections were washed three times with PBS, fixed in 4% paraformaldehyde for 6 h, and then photographed. The experimental results showed that the microbial vesicles exhibited excellent therapeutic effects on cerebral infarction; the results are shown below. Figure 8 .

[0089] In summary, this application provides an electrogenic microbial vesicle for regulating the functions of organisms. By utilizing the electron shuttle capability, immune activation ability, and close electrical communication established with host cells or tissues of the electrogenic microbial vesicle, it is possible to achieve functional regulation of various cells, as well as tissue regeneration, functional regulation, and disease treatment functions in organisms. The electrogenic microbial vesicle proposed in this application holds promise for providing a new application direction for bioelectronic medicine.

[0090] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0091] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. Application of electrogenic microbial vesicles in the preparation of drugs or biological agents for regulating the function of organisms.

2. Use according to claim 1, wherein The regulation of organism functions includes the regulation of cellular functions and / or the regulation of animal functions.

3. Use according to claim 2, wherein the compound is ###0002### The regulation of cellular functions includes one or more of the following: regulation of cell proliferation, differentiation, protein expression, reactive oxygen species levels, and electrophysiological activities. Optionally, regulating cell differentiation includes inhibiting fibroblast differentiation into myofibroblasts; Optionally, regulating cellular protein expression includes enhancing the ability to express functional proteins; said functional proteins may optionally include human epidermal growth factor.

4. The use according to claim 2, wherein The regulation of animal functions includes one or more of the following: promoting tissue regeneration, repairing organ function, and treating diseases.

5. The use according to claim 4, wherein the compound is ###0002### The tissue regeneration includes one or more of nerve tissue regeneration, myocardial tissue regeneration, and bone tissue regeneration; and / or, The diseases mentioned include one or more of ischemic stroke, myocardial injury, and neurodegenerative diseases.

6. The use according to claim 1, wherein The electrogenic microbial vesicles possess both redox and electron transport capabilities; and / or, The electrical impedance of the electrogenic microbial vesicles is 12.5 Ω to 13 Ω.

7. The application as described in claim 1, characterized in that, The electrogenic microbial vesicles were obtained by extracting electrogenic microorganisms after interventional culture. Optionally, the intervention culture includes stress culture of electrogenic microorganisms using at least one of the following methods: antibiotics, chelating agents, L-cysteine, elevated temperature, varying pH, varying dissolved oxygen, carbon source starvation, and amino acid starvation. The antibiotic may optionally include or be ampicillin.

8. The application as described in claim 7, characterized in that, The extraction methods for the electrogenic microbial vesicles include one or more of the following: tangential flow filtration combined with size exclusion chromatography, differential centrifugation combined with density gradient centrifugation, immunomagnetic bead method, and tangential flow filtration combined with aqueous two-phase extraction.

9. The application as described in claim 7 or 8, characterized in that, The electrogenic microorganisms include microorganisms of the genus Shewanella or Geobacter; Optionally, the electrogenic microorganism is Shewanella oneidensis MR-1.

10. A method for regulating cell function in vitro, characterized in that, The method includes: The electrogenic microbial vesicles as defined in any one of claims 1, 6 to 9 are mixed with cells to interact; Assess changes in cell function; Optionally, the cells include one or more of mesenchymal stem cells, fibroblasts, cardiomyocytes, and bacterial cells; Optionally, changes in cell function can be assessed from at least one of the cellular, molecular, and bioinformatics analyses.

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