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Switch type liposome nano fluorescent probe as well as preparation method and application thereof

A nano-fluorescent probe and liposome technology, which is applied in the field of nanotechnology and fluorescence diagnosis research, can solve the problem of distance between dye molecules with limited switching performance, achieve good in vivo imaging effect, convenient use, and improve blood stability Effect

Active Publication Date: 2021-01-05
XIDIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the existing ACQ-liposome probes are all realized by loading high-concentration hydrophilic dyes in the hydrophilic phase, or encapsulating high-concentration hydrophobic dyes in the hydrophobic phase, and their switching performance is limited by the entrapment. rate and distance between dye molecules

Method used

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  • Switch type liposome nano fluorescent probe as well as preparation method and application thereof
  • Switch type liposome nano fluorescent probe as well as preparation method and application thereof
  • Switch type liposome nano fluorescent probe as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The preparation method of DiR-Lipo described in this embodiment is as follows:

[0041] Distearoylphosphatidylcholine (DSPC), cholesterol, distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-PEG2000) in a molar ratio of 55:40:5 (total lipid molar concentration 5mM) Dissolve in chloroform, then remove the chloroform by rotary evaporation to form a lipid film;

[0042] Add 350 mM ammonium sulfate aqueous solution into the lipid film, and rotate in a 60° C. water bath to hydrate the lipid film. After the lipid film to be formed is hydrated to become an emulsion liposome suspension, extrude 10 times on a 100nm polycarbonate film with an extruder;

[0043] The extruded liposome solution was dialyzed 3 times with 100 mM acetate buffer to form a liposome preparation with ammonium sulfate gradient (inside: 350 mM ammonium sulfate; outside: 100 mM acetate buffer)

[0044]Mix the liposome solution obtained in the previous step with the DMSO solution of DiR (DiR account...

Embodiment 2D

[0049] Morphology comparison of embodiment 2DiR-Lipo and DiR-Lipo-TH

[0050] The DiR-Lipo prepared in Example 1 is indeed encapsulated in the aqueous core of liposomes, and the morphology of DiR-Lipo and DiR-Lipo-TH is characterized by cryo-transmission electron microscopy (Cryo-TEM). The results are shown in figure 2 . Such as figure 2 As shown in A, the DiR-Lipo core exhibits a bilayer conformation with an electron-dense structure, indicating that the ammonium ion forms a complex with DiR and is retained in the aqueous core of the liposome. However, the detection result of DiR-Lipo-TH (2B) did not show a bilayer structure, indicating that DiR entered the hydrophobic phase.

Embodiment 3D

[0051] Example 3 Fluorescence comparison of DiR-Lipo and DiR-Lipo-TH before and after membrane rupture

[0052] The DiR-Lipo prepared in Example 1 is indeed encapsulated in the aqueous core of liposomes, and has in vitro fluorescence activatable properties. This example will use 0.05% Triton X-100 to treat DiR-Lipo and DiR-Lipo -TH perform membrane rupture respectively. First dilute DiR-Lipo and DiR-Lipo-TH with PBS to measure the fluorescence intensity of liposomes in the intact state, then add 0.05% Triton X-100 to disrupt the membrane of liposomes, and measure the dye DiR in monomeric state Fluorescence intensity, compare the fluorescence intensity before and after membrane rupture, the results are shown in image 3 . Depend on image 3 A shows that the fluorescence intensity of DiR-Lipo after membrane rupture (f) is much greater than the fluorescence intensity before membrane rupture (e), and the fluorescence enhancement factor is 70 times, indicating that the fluoresce...

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Abstract

The invention discloses a switch type liposome nano fluorescent probe. The probe comprises a near-infrared fluorescent dye and a liposome phospholipid membrane wrapping the exterior of the near-infrared fluorescent dye, wherein the liposome phospholipid membrane is composed of choline derivatives, cholesterol and distearoyl phosphoethanolamine-polyethylene glycol 2000. According to the switch typelipidosome nano fluorescent probe, the hydrophobic dye is wrapped in a lipidosome inner core; and compared with a traditional method of loading the hydrophobic dye on a lipid bilayer, the probe has the advantages that the load rate of the dye is higher, the stability is better, and the advantage that fluorescence can be activated is combined. The probe is simple in preparation process, low in cost, convenient to use and good in in-vivo imaging effect.

Description

technical field [0001] The invention belongs to the field of nanotechnology and fluorescent diagnosis research, and in particular relates to a switch-type liposome nanometer fluorescent probe and its preparation method and application. Background technique [0002] The near-infrared fluorescence imaging technology for tumors has excellent resolution and sensitivity, and the penetration depth of near-infrared fluorescence imaging in biological tissues is large, and the imaging signal is less affected by the tissue. It is a kind developed in recent years. new imaging methods. Fluorescent contrast agents (such as indocyanine green (ICG), methylene blue (MB), etc.) themselves have poor tumor selectivity, resulting in a very low tumor signal-to-background ratio (TBR). TBR is considered to be an important criterion for evaluating and selecting fluorescent contrast agents, so improving the TBR of fluorescent probes is very important for tumor fluorescence imaging. Encapsulating t...

Claims

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Application Information

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IPC IPC(8): A61K49/00
CPCA61K49/0032A61K49/0084
Inventor 夏玉琼许春仲张象涵王忠良张瑞丽吴延坤李采儒
Owner XIDIAN UNIV
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