5/6 Nephrectomy Using Sharp Bipolectomy Via Midline Laparotomy in Rats

Summary

This manuscript introduces a standardized protocol for a 5/6 nephrectomy by sharp bipolectomy via midline laparotomy in a rat model, aiming to induce a state of renal insufficiency via renal parenchyma reduction with a great degree of methodical precision and low risk of technical error.

Abstract

Chronic kidney disease (CKD) affects over 10% of the global population, amounting to more than 800 million individuals worldwide. The advances in the treatment of CKD have had a significant impact on patient outcomes. While in the past, CKD was often considered a death sentence, with many patients succumbing to the complications of end-stage renal disease, it is now increasingly being managed as a chronic condition with the availability of dialysis and kidney transplantation, as well as new pharmaceutical developments such as SGLT2 inhibitors or nonsteroidal mineralocorticoid receptor antagonists.

Yet, there continues to be a growing demand for further exploration into the pathophysiological processes and potential therapeutic interventions. Reliable biological models play a crucial role in facilitating this research. Given the multifaceted nature of renal disease, which encompasses not only cell biology but also vascular microanatomy and endocrine signaling, an appropriate model must possess a level of biological complexity that only an animal model can offer, rendering rodents an obvious choice.

This manuscript, therefore, provides an intricate, systematic protocol for surgically reducing renal parenchyma through midline laparotomy and combined total and partial nephrectomy in rats for survival as well as non-survival applications. It emphasizes the critical role of precise surgical techniques in ensuring consistent and reliable outcomes. Prime examples of potential applications for this model include biomolecular and pharmaceutical studies as well as the development of innovative intraoperative imaging modalities, such as hyperspectral imaging, to objectively visualize and differentiate renal malperfusion.

Introduction

Chronic kidney disease (CKD) is a progressive condition that affects a significant portion of the global population. It is characterized by the gradual loss of endocrine and filtrative kidney function over time, leading to the accumulation of waste products and fluid in the body and an imbalance in the endocrine system. Recent data suggest that 9.1% to 13.4% of the worldwide population (between 700 million and one billion people) has CKD¹. The prevalence of CKD increases with age, affecting around 34% of people aged 65 years or older in the United States, compared to 12% in those aged 45-64 years and 6% in those aged 18-44 years

Protocol

All animal procedures outlined in this document were carried out within accredited facilities and have been granted approval by the institutional animal care and use committee (IACUC) of the Baden-Württemberg Regional Council in Karlsruhe, Germany (35-9185.81/G-62/23). Experimental animals were handled in accordance with institutional protocols and in compliance with German legislation governing animal welfare, as well as adhering to the guidelines set forth by the European Community Council (2010/63/EU) and the ARRIVE guidelines. Male Sprague Dawley rats with an initial weight of 400 g were utilized following a 1-week acclimatization period.

Results

This protocol was conducted in 10 male rats (mean weight 398 ± 35 g) in a non-survival setting and the procedure was performed by a third-year surgical resident. The success rate defined by survival over 20 min after abdominal wall closure was 100%. The mean duration of the preparation from skin incision until skin closure was 18 min 34 s ± 7 min 31 s.

Unfortunately, due to the non-survival nature of this manuscript, there are no data on the postoperative renal function. Future animal studies .......

Discussion

CKD is defined by kidney damage or reduced kidney function for at least 3 months, regardless of the cause^28,29. Kidney damage encompasses pathologic anomalies in the native or transplanted kidney, identified via imaging, biopsy, or deduced from clinical markers like increased albuminuria (albumin-to-creatinine ratio > 30 mg/g or 3.4 mg/mmol) or urinary sediment alterations. Reduced kidney function implies a reduced glomerular filtration rate, which is usually.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
atraumatic preparation forceps	Aesculap	FB395R	DE BAKEY ATRAUMATA atraumatic forceps, straight
blunt overholt clamp	Aesculap	BJ012R	BABY-MIXTER preparation and ligature clamp, bent, 180 mm
cannula	BD (Beckton, Dickinson)	301300	BD Microlance 3 cannula 20 G
fixation rods	legefirm	‎500343896	tuning forks used as y-shaped metal fixation rods
heating pad	Royal Gardineer	IP67	Royal Gardineer Heating Pad Size S, 20 Watt
plastic perfusor tube	M. Schilling GmbH	S702NC150	connecting tube COEX 150 cm
polyfilament suture	Covidien	CL-769	Covidien Polysorb Braided Absorbable Suture 2-0 75 cm
preparation scissors	Aesculap	BC177R	JAMESON preparation scissors, bent, fine model, blunt/blunt, 150 mm (6")
sealing hemostat patch	Baxter	1506257	Hemopatch Sealing Hemopatch Baxter 45 x 90 mm
silicone vessel loop tie	SERAG WIESSNER	SL26	silicone vessel loop tie 2.5 mm red
Spraque Dawley rat	Janvier Labs	RN-SD-M	Spraque Dawley rat
steel plate	Maschinenbau Feld GmbH	C010206	Galvanized sheet plate, 40 x 50 cm, thickness 4.0 mm
Yasargil clip	Aesculap	FE795K	YASARGIL Aneurysm Clip System Phynox Temporary (Standard) Clip
Yasargil clip applicator	Aesculap	FE558K	YASARGIL Aneurysm Clip Applicator Phynox (Standard)