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A kind of chitosan carries siRNA coating vascular stent and preparation method thereof

A vascular stent, chitosan technology, applied in the field of vascular stent, can solve the problems of poor water solubility, poor targeting, low transfection efficiency, etc., to prevent thrombosis and regeneration, induce cell apoptosis, and treat cell proliferation diseases. Effect

Inactive Publication Date: 2017-11-03
李健 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The invention provides a chitosan-carrying siRNA-coated vascular stent and a preparation method thereof, which solves the problems of low transfection efficiency and poor water solubility of the vascular stent in the prior art, which are easily formed in the stent, restenosis and its coating carrier and poor targeting

Method used

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  • A kind of chitosan carries siRNA coating vascular stent and preparation method thereof
  • A kind of chitosan carries siRNA coating vascular stent and preparation method thereof
  • A kind of chitosan carries siRNA coating vascular stent and preparation method thereof

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preparation example Construction

[0036]A kind of preparation method that chitosan carries siRNA coating vascular stent, comprises the following steps:

[0037] (1) Construction of TF-siRNA target sequence: TF-mRNA 167-186bp sequence, the sequence is 5-GCG CTT CAG GCACTA CAA AT-3, labeled with fluorescein to obtain TF-siRNA tissue factor;

[0038] (2) Preparation of hydroxybutyl chitosan-carried tissue factor siRNA nanoparticles:

[0039] (a) Take the layered sample of hydroxybutyl chitosan, tear it into thin slices with clean tweezers, put it in a flat glass petri dish, put it under the ultraviolet lamp and irradiate the front and back sides for 50-80 minutes, then dissolve it in acetic acid solution and filter Sterilize to get hydroxybutyl chitosan acetic acid solution;

[0040] (b) Take sodium tripolyphosphate (TPP), add water to dissolve, stir evenly, filter and sterilize to obtain TPP aqueous solution;

[0041] (c) Take the TF-siRNA tissue factor obtained in step (1) and add it to DEPC (diethylpyrocarbo...

Embodiment 1

[0051] A preparation method of hydroxybutyl chitosan carrying tissue factor siRNA coating vascular stent, comprising the following steps:

[0052] (1) Construction of the TF-siRNA target sequence: First, search for the full-length 2104bp mRNA sequence of coagulation factor III (TF) in NCBI (M16553.1, GI: 339503); the TF-siRNA target sequence screened in this experiment was selected for The 167-186bp sequence of TF-mRNA (5-GCG CTT CAG GCA CTA CAA AT-3) was handed over to the company to design its silencing sequence and labeled with FAM fluorescence to obtain TF-siRNA tissue factor.

[0053] (2) Preparation of hydroxybutyl chitosan-carried tissue factor siRNA nanoparticles:

[0054] (a) Accurately weigh 3 mg of dry hydroxybutyl chitosan sheet-like sample, tear it into thin slices with clean tweezers, lay it flat on a glass petri dish, place it under a UV lamp for 60 minutes on the front and back sides, and then dissolve it in 3 mL Acetic acid solution (concentration of acetic a...

Embodiment 2

[0062] A kind of preparation method that chitosan carries siRNA coating vascular stent, comprises the following steps:

[0063] (1) Construction of the TF-siRNA target sequence: First, search for the full-length 2104bp mRNA sequence of coagulation factor III (TF) in NCBI (M16553.1, GI: 339503); the TF-siRNA target sequence screened in this experiment was selected for The 167-186bp sequence of TF-mRNA (5-GCG CTT CAG GCA CTA CAA AT-3) was submitted to the company to design its silencing sequence and labeled with fluorescein to obtain TF-siRNA tissue factor.

[0064] (2) Preparation of hydroxybutyl chitosan-carried tissue factor siRNA nanoparticles:

[0065] (a) Accurately weigh 1.5 mg of dry hydroxybutyl chitosan sheet-like sample, tear it into thin slices with clean tweezers, lay it flat on a glass petri dish, place it under a UV lamp for 50 minutes on the front and back sides, and then dissolve it in 3mL acetic acid solution (the concentration of acetic acid is 0.2M, the pH v...

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Abstract

The invention discloses a chitosan-carried siRNA-coated vascular stent and a preparation method thereof, belonging to the technical field of vascular stents. The invention includes a stent body, the outer surface of the stent body is provided with a nano-coating, and the nano-coating is a nano-coating of hydroxybutyl chitosan carrying tissue factor siRNA; the invention also provides a preparation method of the above-mentioned vascular stent, comprising constructing Tissue factor siRNA, preparation of hydroxybutyl chitosan nanoparticles carrying tissue factor siRNA, surface alkalization of bare metal vascular stents and preparation of nano-coating of hydroxybutyl chitosan carrying tissue factor siRNA. The present invention uses nanotechnology to make hydroxybutyl chitosan into a nanoparticle complex, and attaches the hydroxybutyl chitosan nanocomposite carrying tissue factor siRNA to the vascular stent, and releases tissue factor to silence the RNA gene. Inhibit excessive proliferation of smooth muscle cells, prevent stent thrombosis and restenosis.

Description

technical field [0001] The invention belongs to the technical field of vascular stents, in particular to a hydroxybutyl chitosan-carrying tissue factor siRNA-coated vascular stent and a preparation method thereof. Background technique [0002] After coronary artery stenting, it stimulates the excessive proliferation and migration of vascular smooth muscle cells (VSMC), stimulates the expression of local tissue factor in the body to increase significantly, makes the body in a state of high thrombus load, and easily forms in-stent thrombosis and in-stent restenosis. For hyperproliferation of vascular smooth muscle cells (VSMCs), local direct drug delivery strategies, such as drug-eluting stents (DES), can be used to reduce the incidence of restenosis. At the same time, vascular endothelial cells are inhibited to varying degrees. During this period, smooth muscle cells (SMC) are prone to excessive proliferation, migration, and release of tissue factor, causing platelet aggregat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61L31/10A61L31/16
Inventor 李健李丹安毅万康
Owner 李健
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