intrarenal drug delivery in rats: a method to deliver therapeutic drugs in rat kidney via intra-arterial injection

Intrarenal Drug Delivery in Rats: A Method to Deliver Therapeutic Drugs in Rat Kidney via Intra-arterial Injection

To begin surgery, ensure that the depth of sedation is adequate by monitoring physical reflexes, such as a withdrawal from a toe pinch. Use a No. 10 surgical scalpel blade to perform a laparotomy through a small midline incision. With cotton swabs, pull the intestine and colon to the right side of the abdomen and cover them with sterile gauze soaked in 0.9% normal saline to keep the organs moist. Gently retract upward the spleen, liver, stomach, and pancreas to expose the aorta and the left kidney artery.
Next, under a surgical microscope, use blunt dissecting curved forceps with a repeated open-close motion along the length of the vessels and sterile cotton swabs to carefully separate the abdominal aorta above and below the left kidney and the left renal artery from the veins, the fat, and the surrounding connective tissue. The use of cotton swabs helps avoid damage to nerves and lymphatic or blood vessels, in particular, to the thin-walled veins.
Then, place a 4-0 silk suture underneath the aorta, and using microvascular clips, clamp the aorta above and below the renal artery bifurcation. Now, using a 24-gauge intravenous catheter, puncture the aorta at the level of the left kidney artery bifurcation and carefully advance the catheter into the renal artery. Because of the small size of the renal artery, the catheter might puncture through the vessel. It is important to work under a surgical microscope and to insert the needle as parallel to the artery as possible.
Then, connect the syringe filled with the drug solution or saline to the catheter and perfuse the kidney. Immediately after perfusion, use a microvascular clip to clamp the left renal vein and the left ureter before removing the catheter. Then, place a piece of absorbable hemostat gelatin sponge with a small drop of tissue adhesive over the punctured area of the aorta, and with a cotton swab, gently apply pressure. At the same time, release the clamp from the aorta below the left renal artery bifurcation.
After 5 minutes, release the clamp from the renal vein and ureter. Carefully release the clamp from the aorta above the left renal artery bifurcation and allow the kidney to reperfuse. The total renal ischemia should last no longer than 7 minutes. After ensuring that no active bleeding occurs and closely observing the area for 10 more minutes, use 4-0 absorbable sutures and a continuous pattern to prevent infection. Enclose the abdominal incision in two layers.
Apply topical antibiotic ointment over the incision area to prevent infections. Then, transfer the rat into the observation cage on a warm pad until complete recovery.

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Universidad de Cartagena UDEC

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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Surgical Procedure
<ol>
	<li>Anesthetize the mouse and ensure that the depth of sedation is adequate by monitoring physical reflexes, such as withdrawal from toe pinch, palpebral reflex, jaw tone, and respiration rate/pattern.</li>
	<li>Perform a laparotomy through a small midline incision (2&#8211;2.5 cm in length) using a surgical scalpel blade No. 10.</li>
	<li>Pull the intestine and colon to the right side of the abdomen by using cotton swabs and cover them with sterile gauze soaked into 0.9% normal saline to maintain the organs moist.</li>
	<li>Gently retract upward the spleen, liver, stomach, and pancreas to expose the aorta and the left kidney artery.</li>
	<li>With the help of a surgical microscope, carefully separate the abdominal aorta above and below the left kidney and the left renal artery from the veins, the fat and the surrounding connective tissue with blunt dissecting curved forceps and sterile cotton swabs.
	<ol>
		<li>Use the forceps with a repeated open-close motion (blunt dissection) along the length of the vessels to remove the connective tissue and the cotton swabs with a lateral rolling motion to remove the fat. 
		NOTE: The dissection of the peri-aortic region is a very delicate step as nerves and lymphatic vessels might be damaged. Make sure to keep the arteries moist with saline during the dissection procedure.</li>
	</ol>
	</li>
	<li>Place a 4-0 silk suture underneath the aorta.</li>
	<li>Using microvascular clips, clamp the aorta above (just below the superior mesenteric artery) and below the renal artery bifurcation.</li>
	<li>Puncture the aorta at the level of the left kidney artery bifurcation with a 24G intravenous catheter and advance the catheter into the renal artery. 
	NOTE: This is a critical step as puncture through the renal artery may occur.</li>
	<li>Connect a syringe filled with the drug solution or saline (up to 500 &#956;L) to the catheter and perfuse the kidney.</li>
	<li>Immediately after perfusion, clamp the left renal vein and the left ureter with a microvascular clip and remove the catheter. Then, place a piece of absorbable hemostat gelatin sponge, with a small drop of tissue adhesive, over the punctured area of the aorta and gently apply pressure with a cotton swab.</li>
	<li>At the same time, release the clamp from the abdominal aorta, below the left renal artery bifurcation. After 5 min, release the clamp from the renal vein and ureter.</li>
	<li>Carefully release the clamp from the aorta, above the left renal artery bifurcation, and allow for kidney reperfusion. The total renal ischemia should last no longer than 7 min.</li>
	<li>Ensure that no active bleeding occurs and closely observe the area for 10 more min.</li>
	<li>Close the abdominal incision in two layers (muscle and skin), using 4-0 absorbable sutures and a continuous pattern to prevent infection.&#160;In addition to the continuous pattern suture technique, another option would be to use a simple, interrupted technique, especially for body wall closure to prevent dehiscence.</li>
	<li>Apply topical antibiotic ointment over the incision area to prevent infections.</li>
	<li>Transfer the rat into a bedding-free observation cage on a warm pad until complete recovery with a temperature range set at 35&#8211;37 &#176;C. Loose bedding should be covered (e.g.&#160;with a drape or paper towel) or removed from the cage until animals are fully recovered to prevent suffocation or aspiration of bedding.</li>
	<li>After surgery, observe the animals continuously until breathing spontaneously, then hourly for a few hours. Re-dose the analgesic Buprenorphine SR 72 hr later if signs of discomfort are observed, such as lethargy, hunched and scruffy, grimace, not resuming normal activities.</li>
</ol>

<table><tbody><tr><td>Surgical Microscope</td><td>Leica</td><td>M125</td><td></td></tr><tr><td>Isoflurane 100 ml</td><td>Cardinal Healthcare</td><td>PI23238</td><td>Anesthetic</td></tr><tr><td>Buprenorphine HCL SR LAB 1 mg/ml, 5 ml</td><td>ZooPharm Pharmacy</td><td></td><td>Buprenorphine narcotic analgesic formulated in a polymer that slows absorption extending duration of action (72 hr duration of activity). Liquid is viscous, warming to RT aids in drawing into syringe.&amp;nbsp;Recommended dosage: 1-1.2 mg/kg SC. DO NOT DILUTE.</td></tr><tr><td>Lactated Ringer&#039;s Injection, USP, 250 ml VIAFLEX Plastic Container</td><td>Baxter Healthcare Corp.</td><td>NDC0338-0117-02</td><td>For body fluids replacement</td></tr><tr><td>Sol Povidone-Iodine&amp;nbsp; Swabstick, 3&#039;&amp;nbsp;</td><td>Cardinal Heatlhcare</td><td>23405-010B</td><td></td></tr><tr><td>Sterile cotton tipped applicators</td><td>Kendall</td><td>8884541300</td><td></td></tr><tr><td>4-0 silk suture (without needle)&amp;nbsp;</td><td>Cardinal Heatlhcare</td><td>A183H</td><td></td></tr><tr><td>Vessel Clip, Straight, 0.75 mm x 4 mm Jaw</td><td>World Precision Instruments&amp;nbsp;</td><td>501779-G</td><td></td></tr><tr><td>I.V. Catheter, Straight Hub, Radiopaque, 24 g x 3/4&quot;, FEP Polymer</td><td>Jelco</td><td>4053</td><td></td></tr><tr><td>Phosphate Buffered Saline</td><td>Life Technologies</td><td>10010023</td><td></td></tr><tr><td>SURGIFOAM Absorbable Gelatin Sponge</td><td>Cardinal Healthcare</td><td>179082</td><td></td></tr><tr><td>4-0 VICRYL PLUS (ANTIBACTERIAL) VIOLET 27&quot; RB-1 TAPER</td><td>Ethicon</td><td>VCP304H</td><td>For muscle layer suturing</td></tr><tr><td>4-0 VICRYL PLUS (ANTIBACTERIAL) UNDYED 18&quot; PC-3 CUTTING</td><td>Ethicon</td><td>VCP845G</td><td>For skin layer suturing</td></tr><tr><td>Triple antibiotic ointment</td><td>Actavis</td><td>NDC0472-0179-56</td><td>For topical use on the site of the incision</td></tr></tbody></table>

Source: Franchi, et al. <a href="https://jove.unicartagenaproxy.elogim.com/video/54418" target="_blank">Intravascular Delivery of Biologics to the Rat Kidney.</a> J. Vis. Exp.&#160; (2016)
In this video, we demonstrate intra-renal drug administration in rats via the renal artery. This method is beneficial in improving drug targeting, drug potency, and minimizing systemic toxicity.

Source: Franchi, et al. Intravascular Delivery of Biologics to the Rat Kidney. J. Vis. Exp.&#160; (2016)
In this video, we demonstrate intra-renal drug ...

Intra-arterial injection facilitates localized delivery to target organs like the kidney while minimizing drug accumulation in other organs. To begin, prep an anesthetized rat in the supine position. Make a small midline incision to expose the peritoneal cavity.
Exteriorize the underlying organs to locate the abdominal aorta. Around the kidney, the aorta branches into the renal artery, between the renal vein and the ureter. Now, remove the connective tissue surrounding the aorta and renal artery.
Clamp the posterior and anterior segments of the aorta to briefly obstruct the blood supply during injection. Gently insert a catheter into the aorta and advance it parallelly into the renal artery. Additionally, connect a syringe filled with the desired drug suspension to the catheter.
Following syringe attachment, inject the suspension into the renal artery. Immediately thereafter, clamp the renal vein and ureter to allow drug distribution within the kidney. A gradual color change in the kidney confirms efficient delivery of the drug suspension.
Next, remove the catheter and apply tissue adhesive to seal the punctured site. Subsequently, remove the clamps from the surrounding vasculature. Return the organs into the body cavity and suture the incision. Allow the rat to recover and regularly monitor for further analysis.&#160;

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Intrarenal Drug Delivery in Rats: A Method to Deliver Therapeutic Drugs in Rat Kidney via Intra-arterial Injection

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Intrarenal Drug Delivery in Rats: A Method to Deliver Therapeutic Drugs in Rat Kidney via Intra-arterial Injection