CN-121971702-A - Artificial cell and preparation method thereof, bionic tissue and preparation method thereof

CN121971702ACN 121971702 ACN121971702 ACN 121971702ACN-121971702-A

Abstract

The invention discloses an artificial cell and a preparation method thereof, and a bionic tissue and a preparation method thereof, by a gas-liquid shearing micro-fluidic technology, the aqueous dispersion of negative electric nanometer material is sprayed into water phase receiving bath containing cationic polyelectrolyte and surfactant to realize one-step construction of artificial cell membrane and cytoskeleton. The method utilizes the electrostatic interaction of negative electric nanometer materials and cationic polyelectrolyte to construct artificial cells with reinforced internal skeletons in a one-step method in water phase, and has the characteristics of convenient preparation and excellent stability. By introducing different functional materials, the multifunctional integration of the artificial cells can be realized. Meanwhile, the adhesion inducer is added to further construct a bionic tissue, and the bionic tissue integrates excellent mechanical properties of artificial cells and can bear a certain degree of disturbance without disintegration.

Inventors

SHI SHAOWEI
FENG WEIXIAO
LI PEIFAN

Assignees

北京化工大学

Dates

Publication Date: 20260505
Application Date: 20241218

Claims (10)

1. The preparation method of the artificial cell is characterized by comprising the following steps: S11) preparing extrusion phase solution A1 by utilizing negative electric nano materials and ultrapure water; S12) preparing a receiving bath solution C1 by utilizing a cationic polyelectrolyte, a surfactant and ultrapure water; S13) pumping the extrusion phase solution A1 prepared in the step S11) into the receiving bath solution C1 by using a syringe pump and a coaxial needle to prepare artificial cells, wherein the artificial cells are specifically: Pumping the extrusion phase solution A1 into an inner layer needle of a coaxial needle by using a syringe pump, when the extrusion phase solution A1 forms liquid drops D1 at the outlet of the inner layer needle, breaking the liquid drops D1 into uniform micro liquid drops by using the shearing action of gas blown out from an outer layer needle, and enabling the micro liquid drops to fall into a receiving bath solution C1 to form artificial Cell microcapsules Cel-1, wherein the artificial Cell microcapsules Cel-1 are artificial cells, the gas blown out from the outer layer needle is air or inert gas, and the flow rate of the gas blown out from the outer layer needle is controlled by a rotor flowmeter.
2. The method for preparing artificial cells according to claim 1, wherein the negatively charged nanomaterial is a surface-modified negatively charged water-dispersible nanomaterial comprising cellulose nanofibers, and the weight of the negatively charged nanomaterial in the extrusion phase solution A1 is 0.05 to 5% of the weight of the extrusion phase solution A1.
3. The method of preparing artificial cells according to claim 2, wherein the water-dispersible nanomaterial further comprises one or more of graphene oxide, MXene, fe3O4 nanoparticles, gold nanoparticles, and silica nanoparticles.
4. The method for preparing artificial cells according to claim 1, wherein the cationic polyelectrolyte is one or more of polydimethyl diallyl ammonium chloride, chitosan, polyethyleneimine, polylysine, cationic polyacrylamide and polyquaternium type acrylate/methacrylate, and the weight of the cationic polyelectrolyte in the receiving bath solution C1 is 0.05-5% of the weight of the receiving bath solution C1.
5. The method according to claim 1, wherein the surfactant is an ionic surfactant or/and a nonionic surfactant in the receiving bath solution C1, and the weight of the surfactant is 0.01-1% of the weight of the receiving bath solution C1.
6. The method for preparing artificial cells according to claim 1, wherein the injection rate of the extrusion phase solution A1 is 10-30 mL/h, the flow rate of the gas blown out from the outer needle is 0.5-1.0L/min, the outer needle gauge is 12G-18G, and the inner needle gauge is 17G-30G.
7. An artificial cell, wherein the artificial cell is prepared by the artificial cell preparation method according to claim 1.
8. The bionic tissue preparation method is characterized by comprising the following steps of: S21) preparing an extrusion phase solution B using the extrusion phase solution A2 and the adhesion inducer, wherein the extrusion phase solution A2 is the extrusion phase solution A1 of claim 1; S22) pumping the extrusion phase solution B prepared in the step S21) into the receiving bath solution C2 by using a syringe pump and a coaxial needle to prepare bionic tissues, wherein the bionic tissues are specifically operated as follows: Pumping the extrusion phase solution B into an inner layer needle of a coaxial needle by using a syringe pump, when the extrusion phase solution B forms liquid drops D2 at the outlet of the inner layer needle, breaking the liquid drops D2 into uniform micro-liquid drops by using the shearing action of gas blown out from the outer layer needle, wherein the micro-liquid drops fall into a receiving bath solution C2 to form artificial Cell microcapsules Cel-2, and the artificial Cell microcapsules Cel-2 spontaneously complete adhesion in the receiving bath solution C2 under the action of an adhesion inducer to form bionic tissues, wherein the gas blown out from the outer layer needle is air or inert gas, the flow rate of the gas blown out from the outer layer needle is controlled by a rotameter, and the receiving bath solution is the receiving bath solution C1 in claim 1.
9. The method for preparing the bionic tissue according to claim 8, wherein the extrusion phase solution B is prepared by mixing the extrusion phase solution A2 and the adhesion inducer according to a volume ratio of 4:1, wherein the adhesion inducer is a strong electrolyte solution with a concentration of 0.1-1.0M or a hydrophobic protein pure water solution with a concentration of 0.5-5 mg/mL.
10. The bionic tissue is characterized in that the bionic tissue is prepared by the bionic tissue preparation method according to claim 8.

Description

Artificial cell and preparation method thereof, bionic tissue and preparation method thereof Technical Field The invention belongs to the field of artificial cell and bionic tissue preparation, and relates to a method for constructing artificial cells and bionic tissues in situ in a water phase by utilizing a gas-liquid shearing microfluidic technology. Background The artificial cell, as a micro biological simulation system for simulating the structure and the function of natural cells, not only provides a simplified model for exploring complex physiological processes in cells, but also shows great application potential in the aspects of life origin research, cell engineering, biosensors, drug delivery and the like, and becomes a research front in the field of life science. The existing artificial cell construction is mainly based on two methods, namely a top-down method and a bottom-up method. The top-down method is to remove or replace some components in cells by physical or chemical means, and to obtain a minimum unit capable of performing basic vital activities. This approach allows to preserve maximally the biological functions of the real cells and has good biocompatibility, but the high precision and complex operations involved therein, the high costs and the potential ethical problems limit the wide application of the method. Thus, the current research is mainly focused on the bottom-up method, i.e., the structure with cellular characteristics is gradually constructed using the most basic chemicals, which provides extremely high design flexibility for the construction of artificial cells. Construction of various artificial cell assemblies such as liposomes, polymers, colloids, and aggregates has been achieved based on the bottom-up approach, but its lower mechanical strength and stability limit further development and application of artificial cells. To solve this problem, researchers have acquired inspiration from the cytoskeleton, a complex network structure inside the cell. The cytoskeleton is not only responsible for maintaining cell morphology, but also provides the necessary mechanical strength for the cells. By constructing the cytoskeleton within the artificial cells, researchers have successfully improved the long-term stability of the artificial cells. However, existing artificial cytoskeletal mimics, such as DNA, proteins, metal-polyphenol frameworks, synthetic polymers, etc., often have problems of poor mechanical stability, complicated construction methods, high cost, etc., which make them difficult to meet the demands of mass production. Therefore, the development of a simple and efficient method for preparing artificial cells with excellent mechanical properties has become a problem to be solved. Disclosure of Invention Therefore, the technical problem to be solved by the invention is to provide an artificial cell, a preparation method thereof, a bionic tissue and a preparation method thereof, wherein an aqueous dispersion of negative electric nano materials is sprayed into an aqueous phase receiving bath containing cationic polyelectrolyte and a surfactant by a gas-liquid shearing microfluidic technology, one-step construction of an artificial cell membrane and a cytoskeleton is realized, the prepared artificial cell has excellent mechanical stability and functional modification, and the artificial cell is subjected to interface bonding by adding an adhesion inducer, so that the bionic tissue with a specific structure and excellent mechanical strength is constructed by a one-step method. In order to solve the technical problems, the invention provides the following technical scheme: The preparation method of the artificial cell comprises the following steps: S11) preparing extrusion phase solution A1 by utilizing negative electric nano materials and ultrapure water; S12) preparing a receiving bath solution C1 by utilizing a cationic polyelectrolyte, a surfactant and ultrapure water; S13) pumping the extrusion phase solution A1 prepared in the step S11) into the receiving bath solution C1 by using a syringe pump and a coaxial needle to prepare artificial cells, wherein the artificial cells are specifically: Pumping the extrusion phase solution A1 into an inner layer needle of a coaxial needle by using a syringe pump, when the extrusion phase solution A1 forms liquid drops D1 at the outlet of the inner layer needle, breaking the liquid drops D1 into uniform micro liquid drops by using the shearing action of gas blown out from an outer layer needle, and enabling the micro liquid drops to fall into a receiving bath solution C1 to form artificial Cell microcapsules Cell-1, wherein the artificial Cell microcapsules Cell-1 are artificial cells, the gas blown out from the outer layer needle is air or inert gas, and the flow rate of the gas blown out from the outer layer needle is controlled by a rotameter. According to the preparation method of the artificial cell, the negative-charge nano m