CN-121987830-A - Vesicle permeation enhancement method based on membrane fluidity regulation and control and characterization system thereof

Abstract

The invention discloses a vesicle permeation enhancement method based on membrane fluidity regulation and a characterization system thereof, and relates to the field of biomaterial modification and tumor delivery; wherein, according to the vesicle membrane composition difference, the membrane fluidity is detected by adopting a fluorescence bleaching recovery method and a fluorescence anisotropy method respectively. Further, the osmotic behavior change of the membrane fluidity enhanced vesicles in tumor tissue was verified by in vivo experiments in mice. The method takes the fluidity of the membrane as a unified physical variable, establishes a technical framework which can be compared and verified in vesicle systems with different sources, and provides a general method for regulating and evaluating the behavior in the vesicle.

Inventors

• QIAO HAI
• BI PENGYU
• YANG SONG

Assignees

• 重庆医科大学

Dates

Publication Date

20260508

Application Date

20260210

Claims (4)

1. 1. The vesicle permeation enhancement method based on membrane fluidity regulation is characterized by comprising the following steps: s1, performing membrane fluidity regulation treatment on vesicles by using n-butanol to obtain modified vesicles with membrane fluidity changed; S2, characterizing the membrane fluidity of vesicles before and after modification, wherein the characterization comprises fluorescence bleaching recovery detection and fluorescence anisotropy detection; s3, using the vesicle with the changed membrane fluidity for in vivo experiments to evaluate the permeation behavior of the vesicle in tumor tissues.
2. 2. The method of claim 1, wherein FRAP detection is performed using fluorescent-labeled lipopolysaccharide FITC-LPS as a membrane marker molecule when the vesicle membrane is free or substantially free of lipopolysaccharide.
3. 3. The method of claim 1, wherein the membrane fluidity is characterized by adopting TMA-DPH fluorescence anisotropy for vesicles naturally containing lipopolysaccharide, and the membrane fluidity is characterized by adopting FRAP for introducing FITC-LPS as a membrane marker molecule for vesicles not containing lipopolysaccharide, so that the unified evaluation of the change of the membrane fluidity of vesicles of different sources is realized.
4. 4. The method of claim 1, wherein the effect of changes in membrane fluidity on the behavior of vesicles in a mouse is assessed by comparing the distribution or penetration of vesicles in tumor tissue in the mouse before and after regulation of membrane fluidity.

Description

Vesicle permeation enhancement method based on membrane fluidity regulation and control and characterization system thereof Technical Field The invention relates to the field of biological material modification and tumor delivery, in particular to a vesicle permeation enhancement method based on membrane fluidity regulation and control and a characterization system thereof. Background Vesicles, which are a typical lipid bilayer structure, have important effects on vesicle behavior due to the physical properties of their membranes during in vivo delivery and tissue interactions. Wherein membrane fluidity is directly related to lateral diffusion of membrane lipid molecules, membrane structural rearrangement, and interaction of vesicles with tissue interfaces. However, the prior art focuses on the size, charge or surface modification of vesicles, and lacks systematic methods of regulation and validation of this physical parameter of membrane fluidity. In addition, vesicles of different sources differ in membrane composition and structural complexity, resulting in difficulty in comprehensively reflecting membrane fluidity changes by a single detection means. Therefore, there is a need to propose a solution that allows to characterize the membrane fluidity variations in different vesicle systems and to further correlate the in vivo permeation behaviour. Disclosure of Invention The invention aims to provide a vesicle permeation enhancement method taking membrane fluidity as a unified physical variable, and the influence of membrane fluidity change on the permeation behavior of tumor tissues in a vesicle is evaluated through membrane fluidity regulation and differentiation characterization means. In order to achieve the aim of the invention, the invention adopts the following technical scheme: the vesicle permeation enhancement method based on membrane fluidity regulation is provided, and comprises the following specific steps: s1, performing membrane fluidity regulation treatment on vesicles by using n-butanol to obtain modified vesicles with membrane fluidity changed; S2, characterizing the membrane fluidity of vesicles before and after modification, wherein the characterization comprises fluorescence bleaching recovery detection and fluorescence anisotropy detection; s3, using the vesicle with the changed membrane fluidity for in vivo experiments to evaluate the permeation behavior of the vesicle in tumor tissues. Further, when the vesicle membrane does not contain or does not substantially contain lipopolysaccharide, FRAP detection is performed using fluorescence-labeled lipopolysaccharide FITC-LPS as a membrane-labeling molecule. Furthermore, TMA-DPH fluorescence anisotropy is adopted for characterizing the fluidity of the membrane for the vesicles containing lipopolysaccharide, FITC-LPS is introduced for the vesicles without lipopolysaccharide as a membrane marking molecule, and FRAP is adopted for characterizing the fluidity of the membrane, so that the uniform evaluation of the fluidity change of the vesicles from different sources is realized. Further, the influence of membrane fluidity changes on the in vivo behavior of vesicles is evaluated by comparing the distribution or penetration of vesicles in tumor tissues in mice before and after membrane fluidity regulation. The beneficial effects of the invention are as follows: 1. the method comprises the steps of carrying out membrane fluidity regulation treatment on vesicles, and characterizing membrane fluidity changes of the vesicles before and after modification, wherein the membrane fluidity is detected by adopting a fluorescence bleaching recovery method and a fluorescence anisotropy method according to vesicle membrane composition differences. Further, the osmotic behavior change of the membrane fluidity enhanced vesicles in tumor tissue was verified by in vivo experiments in mice. The method takes the fluidity of the membrane as a unified physical variable, establishes a technical framework which can be compared and verified in vesicle systems with different sources, and provides a general method for regulating and evaluating the behavior in the vesicle. 2. The reliability of vesicle membrane physical property evaluation is improved through a differential but equivalent membrane fluidity characterization means; 3. The membrane fluidity is enhanced under the condition of not damaging the integral structure of the vesicle; 4. the correlation between membrane fluidity changes and vesicle tumor tissue penetration behavior was verified in vivo experiments. Drawings FIG. 1 is a graph showing membrane fluidity control and characterization of artificial vesicles in example 1; FIG. 2 is a graph showing membrane fluidity control and characterization of the natural vesicles of example 2; FIG. 3 shows the fluorescence results of the in vivo tumor tissue penetration experiment of the vesicle of example 3. Detailed Description The following description of the embo