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Preparation method and application of nano material with synovial immunoregulation function

A technology of immune regulation and nanomaterials, applied in the biological field, can solve problems such as pain and loss of function, and achieve the effect of reducing the inflammatory cascade reaction and inhibiting the immune response of the synovium

Pending Publication Date: 2022-02-25
WEST CHINA HOSPITAL SICHUAN UNIV
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Problems solved by technology

[0005] At present, OA is the most common joint disease in the elderly population in the world. OA has become an important cause of the quality of life of the elderly population. Nanomaterials for anti-inflammatory repair

Method used

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  • Preparation method and application of nano material with synovial immunoregulation function
  • Preparation method and application of nano material with synovial immunoregulation function
  • Preparation method and application of nano material with synovial immunoregulation function

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Embodiment 1

[0033] Embodiment 1: The preparation method of the nanomaterial with synovium immunomodulatory function, comprises the following steps:

[0034] S1. Construction of shRNA-LeptinR: Insert the synthetic shRNA-LeptinR fragment into the Plko.1-EGFP-PURO plasmid vector to obtain a recombinant plasmid vector, then transform the recombinant plasmid vector into Escherichia coli, and extract to obtain shRNA-LeptinR plasmid, the shRNA-LeptinR plasmid contains a green fluorescent protein marker fragment, and has ampicillin resistance, and then the shRNA-LeptinR plasmid is transformed into Escherichia coli, and the forward sequence of the primer used is 5' -CACAAAGAAGTTACATCTT-3'; the reverse sequence of the constructed primer is 3'-AAGATGTAACTTCTTTGTG-5'.

[0035] S2. Macrophage membrane preparation:

[0036] A. Take the mouse mononuclear macrophage leukemia cells (M0) in the logarithmic growth phase, and when the density reaches 70%, add 20ng·mL -1 After IL-4 and IL-13 acted for 24 ho...

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Abstract

The invention discloses a preparation method and application of a nano material with a synovial immunoregulation function, and belongs to the technical field of biologication.The key point of the technical scheme is that the preparation method comprises the following steps that S1, constructing shRNA-LeptinR: specifically, inserting a synthesized shRNA-LeptinR fragment into a plasmid vector to obtain a recombinant plasmid vector, then, transforming the recombinant plasmid vector into escherichia coli, and extracting the shRNA-LeptinR plasmid; S2, preparing a macrophage membrane; S3, preparing gene-loaded nanoparticle M2H@RPK: preparing the nanoparticle M2H@RPK by adopting an electrostatic adsorption method; and wrapping the nanoparticle H@RPK with the M2 macrophage membrane extracted in the step S2, and repeatedly passing through a polycarbonate membrane by using a micro extruder to obtain the nanoparticle M2H@RPK. The invention is mainly used for inhibiting synovial immune response and relieving inflammatory cascade reaction in the OA process.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method and application of a nanometer material with synovial immune regulation function. Background technique [0002] Accumulating evidence indicates that osteoarthritis (OA) is an inflammation-related immune disease in which the synovial membrane plays an important role. The synovial membrane of the joint can be divided into two layers, namely the inner synovial layer and the subsynovial layer which are thinner and close to the joint cavity. Clearly demarcated, they are macrophage-like synoviocytes and fibroblast-like synoviocytes. [0003] During synovitis, the number of macrophages increases dramatically. This massive increase is partly due to the escape of the apoptotic pathway by the original macrophages in the synovium, but more from the infiltrating macrophages in the blood circulation. Macrophages are highly variable cells that respond to different microenvi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/54A61K47/59A61K47/61A61K47/62A61K9/51A61K47/46A61K31/7088A61K38/16A61P19/02A61P29/00A61P37/02C12N15/113C12N15/70
CPCA61K9/5176A61K47/62A61K47/549A61K47/59A61K47/61A61K31/7088A61K38/16A61P19/02A61P29/00A61P37/02C12N15/70C12N15/113C12N2310/531A61K2300/00
Inventor 钱志勇周凯杨成莉胡丹蓉
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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