En murin model af arteriel restenose: Tekniske aspekter af Femoral Wire Skade
Summary
Musen femorale arterie wire skade model af restenose er teknisk udfordrende. I denne protokol viser vi de vigtigste tekniske detaljer afgørende for en vellykket udførelse af wire skade at fremkalde ensartet neointima for studier af restenose.
Abstract
Kardiovaskulær sygdom forårsaget af aterosklerose er den førende dødsårsag i den udviklede verden. Indsnævring af karlumenet grund af aterosklerotisk plaque udvikling eller bristning af etablerede plaques, afbryder normal blodgennemstrømning fører til forskellige følgesygdomme såsom myokardieinfarkt og slagtilfælde. I klinikken endovaskulære procedurer, såsom ballonudvidelse er almindeligt udført for at genåbne lumen. Men disse behandlinger uundgåeligt beskadige karvæggen samt det vaskulære endotel, udløser en overdreven healing reaktion og udvikling af en neointimal plak, der strækker sig ind i hulrummet forårsager fartøj restenose (re-forsnævring). Restenose fortsat en væsentlig årsag til svigt af endovaskulære behandlinger for åreforkalkning. Således prækliniske dyremodeller for restenose er af vital betydning for at undersøge patofysiologiske mekanismer samt translationel tilgange til vaskulære indgreb. Blandt flere murine experimental modeller er lårarterien wire skade bredt accepteret som den mest egnede til studier af post-angioplastik restenose, fordi det ligner angioplastiproceduren der skader både endotel og karvæggen. Men mange forskere har svært ved at udnytte denne model på grund af sin høje grad af tekniske vanskeligheder. Dette skyldes primært en metaltråd skal indsættes i den femorale arterie, der er ca. tre gange tyndere end wiren, for at generere tilstrækkelig skade til at inducere fremtrædende neointima. Her beskriver vi de vigtigste kirurgiske detaljer til effektivt at overvinde de store tekniske vanskeligheder ved denne model. Ved at følge de præsenterede procedurer udfører musen lårarterien wire skade bliver lettere. Når bekendt, kan hele proceduren være afsluttet inden for 20 min.
Introduction
In the era of expanded application of endovascular treatments for various cardiovascular diseases, restenosis after angioplasty is one of the major problems for patients undergoing such treatments. Damage to the vascular endothelium at the time of angioplasty, in concert with the atherosclerotic background, induces excessive smooth muscle cell proliferation in the medial layer, resulting in neointimal hyperplasia.1,2 A viable animal model that recapitulates post-angioplasty neointimal hyperplasia is, thus, important not only for the investigation of disease mechanisms but also for the development of effective therapeutics to treat this pathology.
Mice represent an excellent model animal to recapitulate neointimal hyperplasia for the following reasons: the genetic backgrounds of experimental mice are well established; a wide variety of genetically-modified strains are available;2 obtaining littermates of the same background is easy; and the cost of the animals is relatively low. Arterial ligation model and wire injury model are the two most common mouse models of mechanically-induced neointimal hyperplasia. The arterial ligation model is easy to create, but physiologically dissimilar to the actual angioplasty procedure. The wire injury model closely mimics actual angioplasty procedures but is technically difficult due to the small size of mouse arteries.2,3 Sata et al. first described a wire injury method for mouse femoral arteries based on the anatomical structure of the vasculature and the use of proper-sized flexible wire. Utilizing this technique, they succeeded in reproducibly inducing neointimal hyperplasia in various strains of mice.4
Although femoral wire injury is a well-established model, some of the technical aspects of the technique are highly challenging compared to other models such as ligation.5 The purpose of this paper is to describe our mouse wire injury model procedures in detail, which is a modified version of Sata’s original method. We have made two main modifications: 1) Looping only the arteries, and 2) No lidocaine use.
Protocol
Representative Results
Discussion
Proceduren Tråden skade er gældende for alle musestammer, så længe deres anatomiske strukturer er ens. 4 I dette papir, anvendte vi C57BL / 6-mus med en alder på 12-16 uger. Som vi har lært fra vores indledende undersøgelser, lårarterierne af mus yngre end 10 uger er ofte så små, at wire indsættelse er ganske udfordrende. På den anden side, wire indsættelse i mus ældre end 16 uger er teknisk lettere, men har tendens til at resultere i mere variabel neointimal hyperplasi. Hunmus tendens til at p?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
This work was supported by a Wisconsin Partnership Program New Investigator Award (ID 2832), a National Heart, Lung, Blood Institute R01 Grant (HL-068673) and a T32 training Grant (HL-110853). We thank Dr. Melina Kibbe’s group at Northwestern University for providing helpful information.
Materials
| Name of Reagent/ Equipment | Company | Catalog Number | Comments/Description |
| Fixed Core Wire Guide | Cook | G02426 | Diameter 0.015 inch, Straight |
| Dilation Forceps | Roboz | RS-4927 | Curved, blunt tips |
| Dumont Tweesers #5 | World Precision Instruments | 14095 | Straight, sharp tips |
| Dumont Vessel Cannulation Forceps | World Precision Instruments | 503373 | |
| McPherson-Vannas Scissors | World Precision Instruments | 501234 | |
| Mosquito Forceps | World Precision Instruments | 501291 | |
| Ethilon Nylon Suture 9-0 | Ethicon | 7717G | 9-0, Black Nylon Monofilament |
| Micro AROSuture, Sterile 11-0, 70 Microns, MET Point | AROSurgical | VT4A00N07 | 11-0, Black Nylon Monofilament |
| 3M Precise Multi-Shot DS Disposable Skin Stapler | 3M | DS-25 |
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