A novel organotypic cortical slice culture model for traumatic brain injury: molecular changes induced by injury and mesenchymal stromal cell secretome treatment

Frontiers in Cellular Neuroscience

Neuroscience

Elisa R. Zanier¹  Francesca Tribuzio¹  Francesca Pischiutta¹  Federico Moro¹  Noemi Di Marzo²  Helena Cavaleiro³  Enrico Caruso⁴  António J. Salgado⁵  Firas Kobeissy⁶  Kevin K. Wang⁶ 

[1] Department of Acute Brain and Cardiovascular Injury, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy;Department of Acute Brain and Cardiovascular Injury, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy;Centro Ricerca Tettamanti, Clinica Pediatrica, Università di Milano-Bicocca, Fondazione MBBM, Monza, Italy;Department of Acute Brain and Cardiovascular Injury, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy;Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, Braga, Portugal;ICVS/3B’s–PT Government Associate Laboratory, Braga/Guimarães, Portugal;Department of Acute Brain and Cardiovascular Injury, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy;Neuroscience Intensive Care Unit, Department of Anesthesia and Critical Care, Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, Milan, Italy;Life and Health Sciences Research Institute (ICVS), School of Medicine, University of Minho, Braga, Portugal;ICVS/3B’s–PT Government Associate Laboratory, Braga/Guimarães, Portugal;Program for Neurotrauma, Neuroproteomics and Biomarkers Research, Departments of Emergency Medicine, Psychiatry, Neuroscience and Chemistry, University of Florida, Gainesville, FL, United States;Department of Neurobiology, Center for Neurotrauma, Multiomics and Biomarkers (CNMB), Neuroscience Institute, Morehouse School of Medicine, Atlanta, GA, United States;

关键词: traumatic brain injury;    in vitro;    organotypic cortical brain slices;    mesenchymal stromal cells;    secretome;   

DOI  :  10.3389/fncel.2023.1217987

 received in 2023-05-06, accepted in 2023-06-23,  发布年份 2023

来源: Frontiers

【 摘 要 】

Traumatic brain injury (TBI) is a major worldwide neurological disorder with no neuroprotective treatment available. Three-dimensional (3D) in vitro models of brain contusion serving as a screening platform for drug testing are lacking. Here we developed a new in vitro model of brain contusion on organotypic cortical brain slices and tested its responsiveness to mesenchymal stromal cell (MSC) derived secretome. A focal TBI was induced on organotypic slices by an electromagnetic impactor. Compared to control condition, a temporal increase in cell death was observed after TBI by propidium iodide incorporation and lactate dehydrogenase release assays up to 48 h post-injury. TBI induced gross neuronal loss in the lesion core, with disruption of neuronal arborizations measured by microtubule-associated protein-2 (MAP-2) immunostaining and associated with MAP-2 gene down-regulation. Neuronal damage was confirmed by increased levels of neurofilament light chain (NfL), microtubule associated protein (Tau) and ubiquitin C-terminal hydrolase L1 (UCH-L1) released into the culture medium 48 h after TBI. We detected glial activation with microglia cells acquiring an amoeboid shape with less ramified morphology in the contusion core. MSC-secretome treatment, delivered 1 h post-injury, reduced cell death in the contusion core, decreased NfL release in the culture media, promoted neuronal reorganization and improved microglia survival/activation. Our 3D in vitro model of brain contusion recapitulates key features of TBI pathology. We showed protective effects of MSC-secretome, suggesting the model stands as a tractable medium/high throughput, ethically viable, and pathomimetic biological asset for testing new cell-based therapies.

【 授权许可】

Unknown   
Copyright © 2023 Pischiutta, Cavaleiro, Caruso, Tribuzio, Di Marzo, Moro, Kobeissy, Wang, Salgado and Zanier.