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Heat-treated human platelet pellet lysate modulates microglia activation, favors wound healing and promotes neuronal differentiation in vitro

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Pages 226-237 | Received 07 Dec 2019, Accepted 13 Feb 2020, Published online: 27 Feb 2020
 

Abstract

The neurorestorative efficacy of human platelet lysates in neurodegenerative disorders is still under investigation. Platelets prepared from standard and pathogen reduced platelet concentrates were pelletized, washed, concentrated, and subjected to freeze-thawing. The lysate was heated to 56°C for 30 min and characterized. Toxicity was evaluated using SH-SY5Y neuroblastoma, BV-2 microglial, and EA-hy926 endothelial cells. Inflammatory activity was tested by examining tumor necrosis factor (TNF) and cyclooxygenase (COX)-2 expressions by BV-2 microglia with or without stimulation by lipopolysaccharides (LPS). The capacity to stimulate wound healing was evaluated by a scratch assay, and the capacity to differentiate SH-SY5Y into neurons was also examined. Platelet lysates contained a range of neurotrophins. They were not toxic to SH-SY5Y, EA-hy926, or BV-2 cells, did not induce the expression of TNF or COX-2 inflammatory markers by BV-2 microglia, and decreased inflammation after LPS stimulation. They stimulated the wound closure in the scratch assay and induced SH-SY5Y differentiation as revealed by the increased length of neurites as well as β3-tubulin and neurofilament staining. These data confirm the therapeutic potential of platelet lysates in the treatment of disorders of the central nervous system and support further evaluation as novel neurorestorative biotherapy in preclinical models.

Authors’ Contributions

ON, DB, & TB conceived the experimental design, discussed the results, and wrote the manuscript; FK performed the collection of platelet concentrates and provided the platelet material; ON performed the characterization of the platelet lysates and did the cell experiments; LBa and Y-W W prepared the platelet lysates; DD commented the data; LBu discussed the results and provided recommendations; all authors read and approved the manuscript.

Acknowledgements

O. Nebie was supported by a PhD fellowship from TMU and a CABRI grant from the Université de Lille, France. We thank the Taipei Blood Center and the Uppsala University Hospital blood bank for the supply of the platelet concentrates.

Disclosure statement

Authors do not have any financial or other disclosures

Additional information

Funding

This study was supported in part by grant 108-2314-B-038-009 from the Ministry of Science and Technology (MOST) of Taiwan and Taipei Medical University (TMU) Higher Education Sprout Project MoE (DP2-108-21121-01-N-09-01 to T. Burnouf’s laboratory). The cooperation between T. Burnouf’s laboratory of TMU and Inserm UMR-S1172 (Dr. David Blum) was supported by a bilateral Orchid research project (MOST and French Association of Taiwan-Campus France; 109-2927-I-038-504). The funding bodies played no role in the design of the study, collection, analysis, interpretation of the data, or in writing the manuscript.

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