Monash Health

9 ARTICLES PUBLISHED IN JoVE

Biology

Anterior Cervical Discectomy and Fusion in the Ovine Model

Tony Goldschlager^1,2, Jeffrey V. Rosenfeld², Ian R. Young¹, Graham Jenkin¹

¹Monash Immunology and Stem Cell Laboratories (MISCL), Monash University, ²Department of Surgery, Monash University

This video demonstrates the technique of anterior cervical discectomy and fusion in the ovine model.

Biology

Undecalcified Bone Preparation for Histology, Histomorphometry and Fluorochrome Analysis

Tony Goldschlager¹, Amany Abdelkader¹, Jeffrey Kerr², Ian Boundy², Graham Jenkin¹

¹Monash Immunology and Stem Cell Laboratories, Monash University, ²Anatomy and Developmental Biology, Monash University

Undecalcified bone histology provides important information for a variety of clinical and research applications. It is technically challenging, particularly with large size specimens. This video illustrates the process of producing good quality sections and demonstrates the technical difficulties and methods with which to overcome them.

Medicine

Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury

Kai-Zheong Lim^*1, Christopher D. Daly^*1,2,3, Peter Ghosh⁴, Graham Jenkin³, David Oehme⁵, Justin Cooper-White^6,7, Taryn Naidoo⁶, Tony Goldschlager^1,8

¹Department of Surgery, Monash University, ²Department of Neurosurgery, Monash University, ³The Ritchie Centre, Hudson Institute of Medical Research, ⁴Proteobioactives, Pty Ltd, ⁵Department of Neurosurgery, St Vincent's Hospital, ⁶Australian Institute for Bioengineering and Nanotechnology, University of Queensland, ⁷School of Chemical Engineering, University of Queensland, ⁸Department of Neurosurgery, Monash Health

Intervertebral disc degeneration is a significant contributor to back pain and a leading cause of disability worldwide. Numerous animal models of intervertebral disc degeneration exist. We demonstrate an ovine model of intervertebral disc degeneration, utilizing a drill bit, which achieves a consistent disc injury and reproducible level of disc degeneration.

Immunology and Infection

Visualizing Macrophage Extracellular Traps Using Confocal Microscopy

Roleen Sharma^1,2, Kim M. O'Sullivan², Stephen R. Holdsworth², Philip G. Bardin^1,2, Paul T. King^1,2

¹Monash Lung and Sleep, Monash Medical Centre, ²Monash University

Macrophage extracellular traps are a newly described entity. This article will concentrate on confocal microscopy methods and how they are visualized in vitro and in vivo from lung samples.

Medicine

Optical Clearing and Imaging of Immunolabeled Kidney Tissue

Turgay Saritas¹, Victor G. Puelles^1,2,3, Xiao-Tong Su⁴, David H. Ellison^4,5,6, Rafael Kramann^1,7

¹Division of Nephrology and Clinical Immunology, University Hospital RWTH Aachen, ²III. Department of Medicine, University Medical Center, Hamburg-Eppendorf, ³Department of Nephrology, Monash Health, ⁴Division of Nephrology and Hypertension, Oregon Health and Science University, ⁵Renal Section, Veterans Affairs Portland Health Care System, ⁶Fondation LeDucq Transatlantic Networks of Excellence, ⁷Department of Internal Medicine, Nephrology and Transplantation, Erasmus Medical Center

The combination of antibody labeling, optical clearing, and advanced light microscopy allows three-dimensional analysis of complete structures or organs. Described here is a simple method to combine immunolabeling of thick kidney slices, optical clearing with ethyl cinnamate, and confocal imaging that enables visualization and quantification of three-dimensional kidney structures.

Behavior

Radiotracer Administration for High Temporal Resolution Positron Emission Tomography of the Human Brain: Application to FDG-fPET

Sharna D. Jamadar^1,2,3, Phillip G.D. Ward^1,2,3, Alexandra Carey^1,4, Richard McIntyre^1,4, Linden Parkes^1,3, Disha Sasan¹, John Fallon¹, Edwina Orchard^1,2,3, Shenpeng Li^1,5, Zhaolin Chen^1,5, Gary F Egan^1,2,3

¹Monash Biomedical Imaging, Monash University, ²Australian Research Council Centre of Excellence for Integrative Brain Function, ³Turner Institute for Brain and Mental Health, Monash University, ⁴Department of Medical Imaging, Monash Health, ⁵Department of Electrical and Computer Systems Engineering, Monash University

This manuscript describes two radiotracer administration protocols for FDG-PET (constant infusion and bolus plus infusion) and compares them to bolus administration. Temporal resolutions of 16 s are achievable using these protocols.

Immunology and Infection

Supervised Machine Learning for Semi-Quantification of Extracellular DNA in Glomerulonephritis

Kim Maree O'Sullivan¹, Sarah Creed², Poh-Yi Gan¹, Stephen R. Holdsworth^1,3

¹Department of Medicine, Monash University, ²Monash Micro Imaging, Monash University, ³Immunology Department, Monash Health

Extracellular DNA (ecDNA) released during cell death is proinflammatory and contributes to inflammation. Measurement of ecDNA at the site of injury can determine the efficacy of therapeutic treatment in the target organ. This protocol describes the use of a machine learning tool to automate measurement of ecDNA in kidney tissue.

Medicine

Regenerative Peripheral Nerve Interface: Surgical Protocol for a Randomized Controlled Trial in Postamputation Pain

Emily Pettersen^1,2,3,4, Paolo Sassu⁵, Francesca Alice Pedrini^1,5, Hannes Granberg^1,2, Carina Reinholdt^2,6, Juan Manuel Breyer⁷, Aidan Roche⁸, Andrew Hart^9,10, Adil Ladak¹¹, Hollie A. Power¹¹, Michael Leung¹², Michael Lo¹², Ian Valerio¹³, Kyle R. Eberlin¹³, Jason Ko¹⁴, Gregory A. Dumanian¹⁴, Theodore A. Kung¹⁵, Paul Cederna¹⁵, Max Ortiz-Catalan^1,4,16,17

¹Center for Bionics and Pain Research, ²Center for Advanced Reconstruction of Extremities, Sahlgrenska University Hospital, ³Department of Electrical Engineering, Chalmers University of Technology, ⁴Bionics Institute, ⁵IV Clinica Ortoplastica, IRCCS Istituto Ortopedico Rizzoli, ⁶Department of Hand Surgery, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, Sahlgrenska University Hospital, ⁷Department of Orthopedic Surgery, Hand Unit, Worker Hospital, ⁸College of Medicine and Veterinary Medicine, The Queen's Medical Research Institute, The University of Edinburgh, ⁹Canniesburn Plastic Surgery Unit, Glasgow Royal Infirmary, ¹⁰College of Medicine, Veterinary & Life Sciences, The University of Glasgow, ¹¹Division of Plastic Surgery, Department of Surgery, Faculty of Medicine and Dentistry, University of Alberta, ¹²Plastics and Reconstructive Surgery, Dandenong Hospital, Monash Health, ¹³Division of Plastic and Reconstructive Surgery, Massachusetts General Hospital & Harvard Medical School, ¹⁴Division of Plastic Surgery, Department of Surgery, Northwestern Feinberg School of Medicine, ¹⁵Section of Plastic Surgery, Department of Surgery, Michigan Medicine, ¹⁶Medical Bionics Department, University of Melbourne, ¹⁷Prometei Pain Rehabilitation Center

Here, we describe the surgical procedure to perform Regenerative Peripheral Nerve Interface (RPNI) surgery for treating postamputation neuropathic pain in the context of an international, randomized controlled trial (RCT) (ClinicalTrials.gov, NCT05009394). The RCT compares RPNI with two other surgical techniques, namely, Targeted Muscle Reinnervation (TMR) and neuroma excision combined with intra-muscular transposition.

Medicine

Targeted Muscle Reinnervation: Surgical Protocol for a Randomized Controlled Trial in Postamputation Pain

Emily Pettersen^1,2,3,4, Paolo Sassu⁵, Francesca Alice Pedrini^1,5, Hannes Granberg^1,2, Carina Reinholdt^2,6, Juan Manuel Breyer⁷, Aidan Roche⁸, Andrew Hart^9,10, Adil Ladak¹¹, Hollie A. Power¹¹, Michael Leung¹², Michael Lo¹², Ian Valerio¹³, Kyle R. Eberlin¹³, Theodore A. Kung¹⁴, Paul Cederna¹⁴, Jason M. Souza^15,16, Oskar Aszmann¹⁷, Jason Ko¹⁸, Gregory A. Dumanian¹⁸, Max Ortiz-Catalan^1,4,19,20

¹Center for Bionics and Pain Research, ²Center for Advanced Reconstruction of Extremities, Sahlgrenska University Hospital, ³Department of Electrical Engineering, Chalmers University of Technology, ⁴Bionics Institute, ⁵IV Clinica Ortoplastica, IRCCS Istituto Ortopedico Rizzoli, ⁶Department of Hand Surgery, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, Sahlgrenska University Hospital, ⁷Department of Orthopedic, Hand Unit, Worker Hospital, ⁸College of Medicine and Veterinary Medicine, The Queen's Medical Research Institute, The University of Edinburgh, ⁹Canniesburn Plastic Surgery Unit, Glasgow Royal Infirmary, ¹⁰College of Medicine, Veterinary & Life Sciences, The University of Glasgow, ¹¹Division of Plastic Surgery, Department of Surgery, Faculty of Medicine and Dentistry, University of Alberta, ¹²Plastics and Reconstructive Surgery, Dandenong Hospital, Monash Health, ¹³Division of Plastic and Reconstructive Surgery, Massachusetts General Hospital &, Harvard Medical School, ¹⁴Section of Plastic Surgery, Department of Surgery, Michigan Medicine, ¹⁵Department of Surgery, Uniformed Services University of the Health Sciences and Walter Reed National Military Medical Center, ¹⁶Department of Plastic and Reconstructive Surgery, Ohio State University, ¹⁷Clinical Laboratory for Bionic Extremity Reconstruction, Department of Plastic, Reconstructive and Aesthetic Surgery, Medical University Vienna, ¹⁸Division of Plastic Surgery, Department of Surgery, Northwestern Feinberg School of Medicine, ¹⁹Medical Bionics Department, University of Melbourne, ²⁰Prometei Pain Rehabilitation Center

The protocol outlines the surgical procedure for the treatment of postamputation pain using Targeted Muscle Reinnervation (TMR). TMR will be compared with two other surgical techniques, specifically Regenerative Peripheral Nerve Interface (RPNI) and neuroma excision, followed by immediate burying within muscle under the context of an international, randomized controlled trial.