The Experimental Exploration of the Microbiome in the Intervertebral Disc
Abstract: Low back pain (LBP) is a burden to both individuals and societies worldwide. Lumbar disc degeneration (LDD), disc herniations and Modic changes have been identified as possible pain sources, but these structural changes can be found in asymptomatic people as well. Vertebral bone marrow lesions visualised as Modic changes (MC) on magnetic resonance imaging have been associated with disc degeneration. For MC type I two different aetiologies have been identified. An autoimmune response of the bone marrow to the intervertebral disc (IVD) or infection of the IVD. The presence of bacteria in the IVDs and their role in disc degeneration is an area of controversy. Even though there is increasing evidence for bacterial presence within the herniated disc, there are limited studies which determine whether bacteria are present in the intact IVD in vivo or whether they represent contamination.
Immunohistochemistry (IHC) is a standard technique used for the localization and semi quantification for protein detection. The standard to perform semiquantitative analysis of the IHC results in manually counting 200 cells within a region of interest. However, manual subjective scoring cannot ensure reproducible and objective analysis essential for the quantification and correlation of proteins in biological tissue. The ability to acquire high resolution digital scans of entire microscopic slides with whole slide scanning and the use of new bioimage analysis tools such as QuPath enable the performance of semi-automatic digital image analysis. QuPath is an open-source software for bioimage analysis and is often used for digital pathology. With its user-friendly interface, built-in algorithms for tissue and cell detection, the ability for interactive machine learning and the possibility for groovy scripting it is a powerful tool for whole slide image analysis. The aim of this project was to create a script for semi-automatic cell counting in QuPath that produces more reproducible immunopositivity rates, shows a higher accuracy, a reduced time to evaluate the samples and is a non-black box approach.
Biosketch: Andrea Nüesch is a final year PhD student at the University of Sheffield, UK. Her research focuses on the intervertebral disc (IVD), with a specific emphasis on the experimental exploration of the microbiome in the IVD. Her work seeks to investigate the presence of bacteria within the IVD, the influence of bacteria on IVD cells and the route of bacteria into the immune privileged tissue.
Andrea completed her master’s Thesis at the AO foundation Davos, focusing on the effect of mesenchymal stromal cells secretome on human disc cells. She graduated in 2020 in Health Sciences and Technology, specialised in Medical Technology from the federal Institute of Technology (ETH) Zurich.
