I am working on organ bath using aortic ring and most of the researchers do cleaning on the aortic ring before fixing it to the organ bath. However, in my point of view, after hypercholesterolemia induction on the rat, cleaning may cause plaque removal thus, sounds impractical if I want to use hypercholesterolemic aortic ring for tension study...
I used to perform some experiments with aortic ring and I dont used to clean it. If you clean, maybe you can remove the endothelium or even cause some damage on the vessel.
Yes absolutely, you need to clean the aorta. If you are a novice, then try to remove the as much fat as you can. Increased fat deposition on the aorta could result in the compromised contraction and relaxation. So it is best to remove fat without disrupting the endothelium. The trick is to remove the fat when the aorta is in mice/rat itself.
I did not study specifically hypercholesterolemic rats before, but I have some experience with obese zucker rats. In these experiments we cleaned the vessels the same way we use to do for other vessels.
Dear Doctor NOOR FAZILA,
Hypercholesterolemic, obese or other experimental animals, it is better the protocol to be as follows:
(i) aorta or other blood vessel with TOTALLY PRESERVED WALL including all 4 (four) coats/tunica (intima, media, adventitia, and adiposa),
(ii) the same vessels with REMOVED TUNICA ADIPOSA (that is, periadvetitial adipose tissue).
Attached are 3 of our related papers...
2015 best wishes!!!!
George Chaldakov, cordiallly yours
Removing luminal fat deposited on the endothelial layer does not make sense, for two reasons 1) you are likely to denude the endothelium and alter the Endothelium dependent responses, and 2) teleologically removing this cholesterol buildup would not reflect the in vivo condition of the animal.
Usually, the aorta is used in rogan bath to measure the effect of vasoconstrictor and relaxant agents (endothelium- and endothelium-independent responses). In my experience of almost 30 years, the aorta needs to be cleaned out from connective and fat very gently so one does not damage the media or endothelium; and in order to assess the viability of the preparation, KCL (50-100 mM) is added to organ bath to generate maxiumum myogenic effect. Goodluck
If you want to study the effect of hypercholesterolemia on some physiopathological factor , do not clean the endothelium of the aortic ring , because you will modify your experiment and lose one of the most important results of that model
hello,
I worked with mice feed with high fat diet and we cleaned the fat from the Aorta. And of course you should be very carefull to protect the endothelium, it is essential for the vascular reactivity. And usually, we test endothelium integrity with Acetylcholine.
Regards
I always use clean aorta but if the control group and hypercholesterolemic are under the same conditions ... no problem
I do not think you should clean aorta. As others have suggested if you clean you will not be providing in vivo condition and you are not answereing the question. However, I think if you get variability in your results you need to use enough animal to solve this matter.
During my MSc in human physiology we used to clean the aorta carefully to remove the blood clot with a clean glass rod, definitely without affecting the endothelium. However, when we worked when we worked on hyperchelesteromic rabbit or rat model we never clean the aorta because that may disturb the fatty streak/foamcells and results may vary.
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