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CN-121971402-A - Gelled cell, preparation method and application thereof

CN121971402ACN 121971402 ACN121971402 ACN 121971402ACN-121971402-A

Abstract

The invention discloses a gelled cell, a preparation method and application thereof. Relates to the fields of synthetic biology and biological medicine. The gelled cell is a cell structure body without cell activity, and comprises a carrier cell frame, an intracellular hydrogel and an antibacterial substance, wherein the carrier cell frame comprises a cell membrane, the cell membrane retains the membrane protein content, the type and the lipid sequence of an original carrier cell, the intracellular hydrogel is formed in situ in the carrier cell frame through supermolecule host-guest interaction or cross-linking reaction and is fused with the inner surface of the cell membrane, the antibacterial substance is loaded in the intracellular hydrogel, pores are distributed on the cell membrane, and the intracellular hydrogel is communicated with the external environment through the pores and exposes the loaded antibacterial substance. The gelation cell can realize targeted delivery, contact sterilization and endotoxin adsorption simultaneously, and the problems of secondary inflammation caused by endotoxin release in the traditional antibiotic treatment, membrane protein loss or disordered arrangement in the cell membrane coating technology and the like are avoided.

Inventors

  • GAO CHENG
  • Hou Hengchuo
  • LI YIZHEN
  • HUA QIAN
  • YANG YING

Assignees

  • 深圳大学

Dates

Publication Date
20260505
Application Date
20260128

Claims (10)

  1. 1. A gelled cell is characterized by comprising a cell structure having no cell activity, and comprising: A carrier cell frame comprising a cell membrane, wherein the cell membrane retains the membrane protein content, species and lipid order of the original carrier cell; Intracellular hydrogels formed in situ within the carrier cell frame by supramolecular host-guest interactions or cross-linking reactions and fused with the inner surface of the cell membrane; An antibacterial substance supported in the intracellular hydrogel; wherein, the cell membrane is distributed with pores, and the intracellular hydrogel is communicated with the external environment through the pores and exposes the loaded antibacterial substances.
  2. 2. A gelled cell according to claim 1, wherein the carrier cell frame is derived from a primary carrier cell, preferably the primary carrier cell is an immune cell, more preferably the immune cell comprises at least one of a monocyte, a macrophage, a dendritic cell, a neutrophil and a mast cell.
  3. 3. A gelled cell according to claim 2, wherein the immune cell is a macrophage.
  4. 4. A gelled cell according to claim 1, wherein the antibacterial substance is a cationic monomer, which is one of the cross-linking monomers of the intracellular hydrogel.
  5. 5. A gelled cell according to claim 4, wherein the cationic monomer is at least one selected from the group consisting of 2-aminoethylmethacrylate hydrochloride and (3-acrylamidopropyl) trimethylammonium chloride.
  6. 6. A method for producing a gelled cell according to any one of claims 1 to 5, comprising the steps of: S1, mixing a gel precursor solution comprising a hydrogel monomer, an antibacterial substance and a photoinitiator with original carrier cells to obtain a mixed system; s2, performing at least one freeze thawing operation on the mixed system to enable the gel precursor to permeate into the original carrier cells and form pores on cell membranes; S3, triggering polymerization reaction in ice bath, so that gel precursors permeated into cells of the original carrier are crosslinked in situ to form intracellular hydrogel fused with the inner surface of a cell membrane, and thus, the gelled cells without cell activity are obtained.
  7. 7. The method of claim 6, wherein the freezing temperature of the freeze-thaw operation is no higher than-20 ℃.
  8. 8. The method of claim 6, wherein the concentration of the hydrogel monomer in the gel precursor solution is from 10 to 30 wt%.
  9. 9. The method of claim 6, wherein the freezing time of the freeze-thawing operation is 15min or longer, preferably 15-60 min.
  10. 10. A pharmaceutical composition comprising the gelled cell of any one of claims 1 to 5, and a pharmaceutically acceptable carrier or excipient.

Description

Gelled cell, preparation method and application thereof Technical Field The invention relates to the technical fields of synthetic biology and biological medicine, in particular to a gelled cell, a preparation method and application thereof. Background Bacterial infections are one of the major threats facing human health, and are widely and profoundly compromised. The pathogenic mechanisms of bacteria are complex and diverse, ranging from the initiation of acute, virulent infectious diseases to chronic infections leading to prolonged disunion, from local tissue damage to the initiation of systemic sepsis or even multiple organ failure. It can not only directly destroy the structure and function of host cells by secreting exotoxins, but also can release a large amount of exotoxins after bacterial death and lysis by self structural components such as lipopolysaccharide (i.e. endotoxin) in the cell wall, and continuously and strongly activate the natural immune system of the host. The activation causes excessive activation of immune cells such as macrophages, releases massive inflammatory mediators such as tumor necrosis factor-alpha, interleukin-1 beta, interleukin-6 and the like, and forms a 'cytokine storm', thereby causing systemic inflammatory reaction syndrome and causing secondary damage to tissues and organs. Traditional antibiotic therapy plays a key role in killing bacteria, however, the traditional antibiotic therapy cannot eliminate endotoxin released after bacterial death, and even a large amount of endotoxin is released possibly due to the fact that a large amount of bacteria are killed in a short time, so that the illness state is aggravated. Clinically, although various therapeutic strategies aimed at neutralizing or eliminating endotoxin are tried, the existing methods have obvious limitations on curative effects due to complex molecular structure and various action mechanisms of endotoxin and wide interaction with a host immune system, and are difficult to fundamentally block inflammatory cascade reactions. On the other hand, for the excessive inflammatory response itself, the field of modern biological medicine has developed targeting agents such as monoclonal antibodies directed against specific inflammatory cytokines. The preparations can precisely block the signal paths of key inflammatory factors such as interleukin-6 or tumor necrosis factor-alpha, and the like, and show good effects in the treatment of partial inflammatory diseases. However, in complex systemic inflammatory environments caused by severe infections (such as severe pneumonia), tissue damage is often the result of networking, synergistic effects of a variety of inflammatory mediators. After a single pathway is inhibited, other inflammatory factors may continue to drive the inflammatory process through a bypass or feedback mechanism, resulting in a greatly compromised therapeutic effect. Therefore, developing a therapeutic agent that can adsorb or neutralize a wide variety of different inflammatory mediators with a broad spectrum and high efficiency, thereby more comprehensively regulating the inflammatory network from upstream, has become a very challenging and urgent need in the anti-inflammatory therapeutic field. Under the background, the cell-derived preparation provides a new idea for anti-inflammatory treatment, namely, natural cell membranes are coated on the surfaces of the nano materials by using a cell membrane coating technology, so that the natural cell membranes inherit biological functions (such as selective permeation, cell identification and the like) of the cell membranes, and drug targeted delivery is realized. For example, there are researchers using Macrophage (MA) enriched cell membranes of cytokine binding receptors to coat reactive oxygen species responsive nanoparticles that mimic MA adsorption to neutralize inflammatory cytokines. However, this technical path faces fundamental technical bottlenecks in the actual preparation and application. The preparation process usually involves multiple complex steps of separation, purification, vesicular formation of cell membranes, fusion with or coating of nanoparticles, etc. In this series of in vitro procedures, the cell membrane structure is inevitably subject to physical and chemical disturbances, leading to loss, denaturation or loss of activity of the membrane proteins. More particularly, the recombination process of the cell membrane on the surface of the nanoparticle is difficult to control accurately, and the original transmembrane topology structure, spatial orientation and lateral distribution of the membrane protein in the membrane plane are extremely easy to be disordered. The function of a protein is highly dependent on its precise three-dimensional conformation and the correct anchoring and alignment on the membrane, and this spatial structure misalignment will directly lead to a significant decrease in its binding affinity and specificity to