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Membrane-coated biomimetic nanoparticles of mesenchymal stem cells with overexpressed PD-L1 molecules on the surface and their preparation and application

A PD-L1, bionic nanotechnology, applied in the field of biomedical materials, can solve the problems of poor immunosuppressive effect of nanoparticles, limited sources, complicated processing methods, etc., and achieve simple and fast extraction methods, high biological safety, and avoid side effects

Active Publication Date: 2022-03-01
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

All of the above technologies use substances derived from biological cells as carriers, their sources are limited, and the processing methods are complicated
US2012 / 039411, WO2013 / 052167 and CN103857387A disclose a membrane-encapsulated nanoparticle and a method of use, in which the surface of the inner core of the non-cellular substance is coated with a cell membrane derived from a cell or a membrane derived from a virus, but these membrane-encapsulated nanoparticles Nanoparticles are less effective for immunosuppression
[0006] Therefore, in order to avoid controversy and reduce side effects, it would be a safer, more convenient and effective strategy to develop a preparation based on the cell membrane of MSCs with high expression of PD-L1, which has a simple preparation process and low cost, and is used in the treatment of inflammation. No reports of such preparations

Method used

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  • Membrane-coated biomimetic nanoparticles of mesenchymal stem cells with overexpressed PD-L1 molecules on the surface and their preparation and application
  • Membrane-coated biomimetic nanoparticles of mesenchymal stem cells with overexpressed PD-L1 molecules on the surface and their preparation and application
  • Membrane-coated biomimetic nanoparticles of mesenchymal stem cells with overexpressed PD-L1 molecules on the surface and their preparation and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1: Induction of overexpression of PD-L1 molecules on the surface of mesenchymal stem cells

[0073] (1) The purified and expanded mesenchymal stem cells are normally cultured in an incubator to make them adhere to the wall.

[0074](2) Remove the cell culture supernatant, replace it with fresh medium containing 100ng / mL gamma interferon, and continue culturing in the incubator for 24 hours.

[0075] (3) After 24 hours, the cell culture supernatant was removed, the cells were washed three times with PBS, and the cells were digested with trypsin.

[0076] Using flow cytometry to analyze the PD-L1 expression level of the obtained cells, the results showed that the PD-L1 expression level of interferon-gamma-treated mesenchymal stem cells increased significantly ( figure 1 ).

Embodiment 2

[0077] Example 2: Extraction and characterization of PD-L1 overexpressed mesenchymal stem cell membranes

[0078] (1) Harvest interferon-gamma-treated mesenchymal stem cells, resuspend in cell disruption solution, and sonicate to obtain a cell disruption mixture.

[0079] (2) Centrifuge the mixture in step (1) at 4000rpm for 10 minutes, collect the supernatant, and continue to centrifuge at 14800rpm for 20 minutes to obtain a cell membrane pellet for future use.

[0080] (3) Extract the membrane-precipitated protein in step (2) using a membrane protein extraction reagent, and perform Western blot analysis on it. The PD-L1 signal on the interferon-treated mesenchymal stem cell membrane was significantly stronger, indicating the overexpression of PD-L1 ( figure 2 ).

Embodiment 3

[0081] Example 3: Preparation and characterization of PD-L1 overexpressed mesenchymal stem cell membrane-coated biomimetic nanoparticles

[0082] (1) Resuspend the harvested PD-L1 overexpressed mesenchymal stem cell membranes in PBS, sonicate until the system has no obvious particles, and add ice cubes during the sonication process to keep the temperature low.

[0083] (2) The membrane suspension obtained in step (1) is repeatedly extruded with a homogenizer, first using a 400nm filter membrane, and after the resistance becomes significantly smaller, the membrane suspension is replaced with a 200nm filter membrane to continue extrusion to obtain uniform nanoplasmic membrane vesicles.

[0084] (3) According to the ratio of the total amount of membrane protein / PLGA mass ratio of 2:1, the nanoplasmic membrane vesicles obtained in step (2) are mixed evenly with the prepared PLGA nano-core.

[0085] (4) Extrude the mixture obtained in step (3) repeatedly with a homogenizer. First, ...

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Abstract

The invention relates to a biomimetic nanoparticle coated with a mesenchymal stem cell membrane with PD-L1 molecules overexpressed on the surface, and its preparation and application. The mesenchymal stem cell membrane-coated biomimetic nanoparticle with PD-L1 molecule overexpression on the surface of the present invention includes a nano-core and a mesenchymal stem cell membrane wrapped outside the nano-core; the nano-core includes a biocompatible polymer, The membrane surface of mesenchymal stem cell membranes overexpresses PD‑L1 molecules. The present invention also discloses the application of the membrane-coated biomimetic nanoparticles of mesenchymal stem cells overexpressed on the surface of PD-L1 molecules in the preparation of inflammation treatment preparations. The biomimetic nanoparticle of the present invention has significant immunosuppressive effect, good biocompatibility, simple and mature preparation process, and can be used to prepare inflammation treatment preparations, which can be effectively enriched in the inflammatory site, by inhibiting the excessively activated immune response of the inflammatory site and cytokine storm, showing excellent curative effect in the treatment of inflammation.

Description

technical field [0001] The present invention relates to the field of biomedical materials, in particular to a biomimetic nanoparticle coated with a mesenchymal stem cell membrane with overexpressed PD-L1 molecules on the surface, and its preparation and application. Background technique [0002] Inflammation is a defense-based pathological response to various physical, chemical, biological and other harmful stimuli. It is a complex response involving a variety of cells and factors, including various immune cells in the immune system. and cytokines. A moderate inflammatory response helps the body recover, while an excessive inflammatory response can lead to tissue damage and even death. The essence of this state of hyperinflammatory response is actually an overactivated immune response, which induces a cytokine storm, which in turn leads to severe inflammatory damage. The pathogenesis of cytokine storm is complex, but the progression is rapid and the mortality rate is extre...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/28A61K47/34A61K47/36A61K9/16A61P29/00A61P31/04A61P31/10A61P31/12A61P33/02A61P33/10A61P37/06C12N5/0775
CPCA61K35/28A61K9/1676A61K9/1682A61P29/00A61P31/04A61P31/10A61P31/12A61P33/02A61P33/10A61P37/06A61P25/00A61P9/00A61P1/16A61P1/00A61P11/00A61P13/12C12N5/0662C12N2509/10
Inventor 汪超沈淑芳
Owner SUZHOU UNIV
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