Journal Article

Neuroprotective effects of intranasal extracellular vesicles from human platelet concentrates supernatants in traumatic brain injury and Parkinson’s disease models

Nebie, Ouada, Burnouf, Thierry, Delila, Liling, Le, Nhi Thao Ngoc, Chou, Ming-Li, Devos, David, Timmerman, Kelly, Buée, Luc, Lee, Deng-Yao, Wu, Yu-Wen, Blum, David, Chou, Szu-Yi

2024

Overview

Background The burgeoning field of regenerative medicine has significantly advanced with recent findings on biotherapies using human platelet lysates (HPLs), derived from clinical-grade platelet concentrates (PCs), for treating brain disorders. These developments have opened new translational research avenues to explore the neuroprotective effects of platelet-extracellular vesicles (PEVs). Their potential in managing neurodegenerative conditions like traumatic brain injury (TBI) and Parkinson’s disease (PD) warrants further exploration. We aimed here to characterize the composition of a PEV preparation isolated from platelet concentrate (PC) supernatant, and determine its neuroprotective potential and neurorestorative effects in cellular and animal models of TBI and PD. Methods We isolated PEVs from the supernatant of clinical-grade PC collected from healthy blood donors utilizing high-speed centrifugation. PEVs were characterized by biophysical, biochemical, microscopic, and LC–MS/MS proteomics methods to unveil biological functions. Their functionality was assessed in vitro using SH-SY5Y neuronal cells, LUHMES dopaminergic neurons, and BV-2 microglial cells, and in vivo by intranasal administration in a controlled cortical impact (CCI)-TBI model using 8-weeks-old male C57/BL6 mice, and in a PD model induced by MPTP in 5-month-old male C57/BL6 mice. Results PEVs varied in size from 50 to 350 nm, predominantly around 200 nm, with concentrations ranging between 10 10 and 10 11 /mL. They expressed specific platelet membrane markers, exhibited a lipid bilayer by cryo-electron microscopy and, importantly, showed low expression of pro-coagulant phosphatidylserine. LC–MS/MS indicated a rich composition of trophic factors, including neurotrophins, anti-inflammatory agents, neurotransmitters, and antioxidants, unveiling their multifaceted biological functions. PEVs aided in the restoration of neuronal functions in SH-SY5Y cells and demonstrated remarkable neuroprotective capabilities against erastin-induced ferroptosis in dopaminergic neurons. In microglial cells, they promoted anti-inflammatory responses, particularly under inflammatory conditions. In vivo , intranasally delivered PEVs showed strong anti-inflammatory effects in a TBI mouse model and conserved tyrosine hydroxylase expression of dopaminergic neurons of the substantia nigra in a PD model, leading to improved motor function. Conclusions The potential of PEV-based therapies in neuroprotection opens new therapeutic avenues for neurodegenerative disorders. The study advocates for clinical trials to establish the efficacy of PEV-based biotherapies in neuroregenerative medicine. Graphical Abstract

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Administration, Intranasal / Animal models / Animals / Anti-inflammatory agents / Antibodies / Antioxidants / Biomedical and Life Sciences / Biomedicine / Blood / Blood & organ donations / Blood donors / Blood platelets / Blood Platelets - metabolism / Brain / Brain Injuries, Traumatic - metabolism / Brain research / Brain-derived neurotrophic factor / Central Nervous System / Centrifugation / Clinical trials / Coagulants / Cytokines / Disease / Disease Models, Animal / Diseases / Dopamine receptors / Electron microscopy / Exosomes / Extracellular vesicles / Extracellular Vesicles - metabolism / Extracellular Vesicles in Cancer and Regenerative Medicine / Ferroptosis / France / Head injuries / Homeopathy / Humans / In vitro methods and tests / In vivo methods and tests / Injuries / Intranasal administration / Life Sciences / Ligands / Lipid bilayers / Lipids / Lysates / Male / Males / Massachusetts / Materia medica and therapeutics / Medical research / Medicine, Experimental / Membrane lipids / Mice / Mice, Inbred C57BL / Microglia / Microscopy / Movement disorders / Neurodegenerative diseases / Neurodevelopmental disorders / Neurological disorders / Neurons / Neurophysiology / Neuroprotection / Neuroprotective Agents - administration & dosage / Neuroprotective Agents - pharmacology / Neurotrophic factors / Parkinson Disease - therapy / Parkinson's disease / Phosphatidylserine / Platelets / Proteins / Proteomics / Reagents / Regenerative medicine / Scientific equipment and supplies industry / Substantia nigra / Taiwan / Therapeutics / Traumatic brain injury / Trophic factors / Tyrosine / United States / Vascular endothelial growth factor / Vesicles