A Murine Model of Hemodialysis Access-Related Hand Dysfunction

Summary

This protocol details the surgical steps of murine common iliac arteriovenous fistula creation. We developed this model to study hemodialysis access-related limb pathophysiology.

Abstract

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement therapies such as hemodialysis continues to increase. Autogenous arteriovenous fistula (AVF) placement remains a primary vascular access option for ESRD patients. Unfortunately, approximately half of the hemodialysis patients experience dialysis access-related hand dysfunction (ARHD), ranging from subtle paresthesia to digital gangrene. Notably, the underlying biologic drivers responsible for ARHD are poorly understood, and no adequate animal model exists to elucidate the mechanisms and/or develop novel therapeutics for the prevention/treatment of ARHD. Herein, we describe a new mouse model in which an AVF is created between the left common iliac artery and vein, thereby facilitating the assessment of limb pathophysiology. The microsurgery includes vessel isolation, longitudinal venotomy, creation of arteriovenous anastomosis, and venous reconstruction. Sham surgeries include all the critical steps except for AVF creation. Iliac AVF placement results in clinically relevant alterations in central hemodynamics, peripheral ischemia, and impairments in hindlimb neuromotor performance. This novel preclinical AVF model provides a useful platform that recapitulates common neuromotor perturbations reported by hemodialysis patients, allowing researchers to investigate the mechanisms of ARHD pathophysiology and test potential therapeutics.

Introduction

The establishment and preservation of functional vascular access remain an important primary goal for end-stage renal disease (ESRD) patients receiving renal replacement therapy via hemodialysis¹. Repeated hemodialysis treatments are necessary to remove waste products, normalize electrolytes, and maintain fluid balance once kidney function becomes inadequate, and thus are necessary for long-term survival². Therefore, vascular access represents a "lifeline" for patients with ESRD, and autogenous arteriovenous fistula (AVF) placement remains a preferred dialysis access option among this cohort³

Protocol

All procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Florida and Malcom Randall Veterans Affairs Medical Center.

NOTE: Young adult (8-10 weeks old) male C57BL/6J mice were purchased from The Jackson Laboratory and housed in a light (12 h light: 12 h dark cycle), temperature (22 °C ± 1 °C), and humidity (50% ± 10%) controlled animal facility. Five mice were allowed to dwell per cage (W:18 cm x L:29 cm x H:12.5 cm) with nesting materials, food, and water being made available ad libitum. Following 7 days of habitat acclimation with standard chow, the mice were....

Results

Animals exposed to an adenine diet have reduced glomerular filtration rates (control: 441.3 ± 54.2 µL/min vs. CKD: 165.1 ± 118.3 µL/min, p < 0.05) and increased serum blood urea nitrogen levels (control: 20.39 ± 4.2 µL/min vs. CKD: 38.20 ± 10.65 µL/min, p < 0.05) compared to the animals that received casein-based chow, confirming the presence of kidney insufficiency prior to arteriovenous fistula surgery.

Validation of AVF patency
Although intra-ope.......

Discussion

The prevalence of hemodialysis patients with ARHD following AVF creation has continued to increase^30,31. Indeed, unresolved symptomatic complications^4,32,33 such as pain, weakness, paresthesia, and/or reduced range of motion can negatively impact patient wellbeing^4,32,33,

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.15% Adenine diet	ENVIGO	TD.130899	20% casein, 0.15% adenine, 0.9% P
0.2% Adenine diet	ENVIGO	TD.130900	20% casein, 0.2% adenine, 0.9% P
10-0 Nylon suture	AD surgical	XXS-N1005T4
29 G needle syringes	Exel International	14-841-32
31 G needle syringes	Advocate	U-100 insulin syringe
4-0 silk suture	AD surgical	S-S41813
45-degree angled dumont forceps	Fine Science Tools	11253-25
5-0 PGA suture	AD surgical	PSGU-518R13
6-0 silk suture	AD surgical	S-S618R13
Absorbable gelatin sponge	ETHICON	1975
Alcohol preps	Covidien	5110-cs4000	70% isopropyl alcohol
Buprenorphine	NA	NA	0.01 g/mL
C57BL6/J mice	Jaxon Laboratory
Casein diet	ENVIGO	TD.130898	20% casein, 0.9% P
Cotton swabs	CONSTIX	SC-9	Medium single-ended round cotton swab
Cotton swabs	CONSTIX	SC-4	Small double-ended hard, sharp, pointed cotton swab
Curity non-woven sponges (2x2)	Covidien	9022
Curved Vannas spring scissors	Fine Science Tools	15001-08
Doppler ultrasound	VisualSonics	Vevo 2100
Extra fine graefe forceps	Fine Science Tools	11150-10	2 pairs
Eye lubricant	CLCMEDICA	Optixcare eye lube
Heparin (5000 U/mL)	National Drug Codes List	63739-953-25	100 IU/mL
Hot bead sterilizer	Fine Science Tools	18000-50
Low-temperature cautery	Bovie	AA04
Pen trimmer	Wahl	5640-600
Powder-free surgical gloves	Ansell	7824PF
Round handled needle holders	Fine Science Tools	12076-12
Sterile towel drape	Dynarex	DY440-MI
Sterilized 0.9% saline	National Drug Codes List	46066-807-25
Straight dumont forceps	Fine Science Tools	11253-20
Straight needle holder	Fine Science Tools	FST 12001-13
Straight vannas spring scissors	Fine Science Tools	25001-08
TrizChLOR4	National Drug Codes List	17033-279-50