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Mesenchymal stem cell membrane-coated bionic nanoparticles with overexpressed PD-L1 molecules on surface as well as preparation and application of mesenchymal stem cell membrane-coated bionic nanoparticles

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 treatment methods, etc., to achieve enhanced immunosuppressive effect, high biocompatibility, Improve the effectiveness of targeting

Active Publication Date: 2020-06-12
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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  • Mesenchymal stem cell membrane-coated bionic nanoparticles with overexpressed PD-L1 molecules on surface as well as preparation and application of mesenchymal stem cell membrane-coated bionic nanoparticles
  • Mesenchymal stem cell membrane-coated bionic nanoparticles with overexpressed PD-L1 molecules on surface as well as preparation and application of mesenchymal stem cell membrane-coated bionic nanoparticles
  • Mesenchymal stem cell membrane-coated bionic nanoparticles with overexpressed PD-L1 molecules on surface as well as preparation and application of mesenchymal stem cell membrane-coated bionic nanoparticles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1: Induction of PD-L1 molecule overexpression 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, change to the fresh medium containing 100ng / mL interferon gamma, and continue to cultivate 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 obtained by trypsinization.

[0076] The PD-L1 expression level of the obtained cells was analyzed by flow cytometry, and the results showed that the PD-L1 expression level of the mesenchymal stem cells treated with gamma interferon was significantly increased ( figure 1 ).

Embodiment 2

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

[0078] (1) The mesenchymal stem cells treated with interferon-gamma are harvested, resuspended in a cell disruption solution, and subjected to ultrasonic treatment to obtain a cell disruption mixture.

[0079] (2) Centrifuge the mixture in step (1) at 4000 rpm for 10 minutes, collect the supernatant, and continue to centrifuge at 14,800 rpm for 20 minutes to obtain cell membrane pellets for use.

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

Embodiment 3

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

[0082] (1) The harvested PD-L1 molecule-overexpressed mesenchymal stem cell membranes were resuspended in PBS, sonicated until the system had no obvious particles, and ice cubes were added to keep the temperature at a low temperature during the sonication.

[0083] (2) repeatedly extruding the membrane suspension obtained in step (1) with a homogenizer, first using a 400nm filter membrane, and then changing to a 200nm filter membrane to continue extrusion after the resistance is significantly reduced to obtain uniform nanoplasma membrane vesicles.

[0084] (3) According to the ratio of the total membrane protein / PLGA mass ratio of 2:1, the nanoplasma membrane vesicles obtained in step (2) and the prepared PLGA nanonuclei are uniformly mixed.

[0085] (4) repeatedly extruding the mixture obtained in step (3) with a homogenizer, first us...

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Abstract

The invention relates to mesenchymal stem cell membrane-coated bionic nanoparticles with overexpressed PD-L1 molecules on the surface as well as preparation and application of the mesenchymal stem cell membrane-coated bionic nanoparticles. The mesenchymal stem cell membrane-coated bionic nanoparticle with the overexpressed PD-L1 molecules on the surface comprises a nano-core and a mesenchymal stemcell membrane wrapping the nano-core, the nano core comprises a polymer with biocompatibility, and PD-L1 molecules are overexpressed on the membrane surface of the mesenchymal stem cell membrane. Theinvention also discloses an application of the mesenchymal stem cell membrane-coated bionic nano-particle with over-expressed PD-L1 molecules on the surface in preparation of an inflammation treatment preparation. The bionic nanoparticle has a remarkable immunosuppression effect, is high in biocompatibility and simple and mature in preparation process, can be used for preparing an inflammation treatment preparation, can be effectively enriched in an inflammation part, and shows an excellent curative effect in inflammation treatment by inhibiting excessively activated immune response and cytokine storm of the inflammation part.

Description

technical field [0001] The invention relates to the field of biomedical materials, in particular to a biomimetic nanoparticle coated with a membrane of mesenchymal stem cells overexpressing PD-L1 molecules on the surface, and preparation and application thereof. Background technique [0002] Inflammation is a defense-based pathological response of the body 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 high inflammatory response is actually an over-activated immune response, which induces a cytokine storm, which in turn leads to severe inflammatory damage. The pathogenesis of cytokine storm is complex, but it progresses rapidly and has a high mortality ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K47/34A61K47/36A61K9/16A61P29/00A61P31/04A61P31/10A61P31/12A61P33/02A61P33/10A61P37/06A61P25/00A61P9/00A61P1/16A61P1/00A61P11/00A61P13/12C12N5/0775
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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