Abstract: SA-OR38
Light Sheet Fluorescence Microscopy: A Novel Tool for Three-Dimensional Optical Reconstructions of Mouse Arteriovenous Fistulae in Control and Uremic Animals
Session Information
- Dialysis Vascular Access: Research Advances
October 26, 2024 | Location: Room 8, Convention Center
Abstract Time: 04:40 PM - 04:50 PM
Category: Dialysis
- 803 Dialysis: Vascular Access
Authors
- Uriyanghai, Unimunkh, The University of North Carolina at Chapel Hill Kidney Center, Chapel Hill, North Carolina, United States
- Bahnson, Edward Moreira, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States
- Wai, Christine, The University of North Carolina at Chapel Hill Kidney Center, Chapel Hill, North Carolina, United States
- Li, Lianxia, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States
- Livingston, Eric W., The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States
- Sudarsanam, Vinay A., The University of North Carolina at Chapel Hill Kidney Center, Chapel Hill, North Carolina, United States
- Haddad, Samuel, The University of North Carolina at Chapel Hill Kidney Center, Chapel Hill, North Carolina, United States
- Su, Huanjuan, The University of North Carolina at Chapel Hill Kidney Center, Chapel Hill, North Carolina, United States
- Arteaga, Eyla C., The University of North Carolina at Chapel Hill Kidney Center, Chapel Hill, North Carolina, United States
- Griffith, Boyce E., The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States
- Roy-Chaudhury, Prabir, The University of North Carolina at Chapel Hill Kidney Center, Chapel Hill, North Carolina, United States
- Xi, Gang, The University of North Carolina at Chapel Hill Kidney Center, Chapel Hill, North Carolina, United States
Background
The arteriovenous fistula (AVF) is currently the gold standard for dialysis vascular access. Unfortunately over 50% of AVFs fail to mature, most often as a result of a venous segment stenosis. We and others have hypothesized that this is likely due to two interactive upstream pathogenetic pathways (hemodynamic stress and uremic vascular biology) which then results in two interacting downstream pathological pathways (neointimal hyperplasia and inadequate outward remodeling) that result in venous segment stenosis.To better elucidate the mechanisms involved in these processes, we have developed a novel approach termed Light Sheet Fluorescence Microscopy (LSFM), which can generate 3D optical reconstructions of AVF vessel architecture, with regard to volume, geometry, and the presence or absence of neointimal hyperplasia. The goal of this study was to describe the use of LSFM in normal and uremic animals.
Methods
Carotid-jugular AVFs were created in 16-week-old wild-type and uremic mice on a C57BL/6 background. Uremia was induced by a 5/6 nephrectomy on day 14 and Day 7 using a two-step surgical approach. Animals were sacrificed at 21d and the AVF was harvested for LSFM and conventional histomorphometry.
Results
Uremic mice had a decrease in the average (6 sections at 100 micron intervals across the venous segment) percentage luminal patency as compared to control animals on histomorphometric analysis (12±1.4% vs 36.1±11.5%, p<0.05) at 21 days post-surgery. Consistent with this data, the LSFM analysis documented that lesion volume within the venous segment was increased in the uremic mouse model (0.79 cubic mm versus 0.25 cubic mm) and the length of lesion was also doubled in the uremic mouse model (1mm vs 2mm) (see Fig 1).
Conclusion
Our results suggest that LSFM could be a particularly useful tool for obtaining a holistic assessment of both 3D anatomy and stenosis across the entire venous segment as opposed to looking at discreet sections every 100-200 microns. While the latter is the current technical standard, it is unfortunately prone to miss short regions of maximal stenosis.
Funding
- NIDDK Support