The Effect of Leukocyte- and Platelet-Rich Fibrin on Central and Peripheral Nervous System Neurons-Implications for Biomaterial Applicability

Ivo Lambrichts; Esther Wolfs; Annelies Bronckaers; Pascal Gervois; Tim Vangansewinkel

doi:10.3390/ijms241814314

The Effect of Leukocyte- and Platelet-Rich Fibrin on Central and Peripheral Nervous System Neurons-Implications for Biomaterial Applicability

Int J Mol Sci. 2023 Sep 20;24(18):14314. doi: 10.3390/ijms241814314.

Authors

Ivo Lambrichts¹, Esther Wolfs¹, Annelies Bronckaers¹, Pascal Gervois¹, Tim Vangansewinkel^{1

2}

Affiliations

¹ Cardio and Organ Systems, Biomedical Research Institute, UHasselt-Hasselt University, 3590 Diepenbeek, Belgium.
² VIB, Center for Brain & Disease Research, Laboratory of Neurobiology, 3000 Leuven, Belgium.

Abstract

Leukocyte- and Platelet-Rich Fibrin (L-PRF) is a second-generation platelet concentrate that is prepared directly from the patient's own blood. It is widely used in the field of regenerative medicine, and to better understand its clinical applicability we aimed to further explore the biological properties and effects of L-PRF on cells from the central and peripheral nervous system. To this end, L-PRF was prepared from healthy human donors, and confocal, transmission, and scanning electron microscopy as well as secretome analysis were performed on these clots. In addition, functional assays were completed to determine the effect of L-PRF on neural stem cells (NSCs), primary cortical neurons (pCNs), and peripheral dorsal root ganglion (DRG) neurons. We observed that L-PRF consists of a dense but porous fibrin network, containing leukocytes and aggregates of activated platelets that are distributed throughout the clot. Antibody array and ELISA confirmed that it is a reservoir for a plethora of growth factors. Key molecules that are known to have an effect on neuronal cell functions such as brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), vascular endothelial growth factor (VEGF), and platelet-derived growth factor (PDGF) were slowly released over time from the clots. Next, we found that the L-PRF secretome had no significant effect on the proliferative and metabolic activity of NSCs, but it did act as a chemoattractant and improved the migration of these CNS-derived stem cells. More importantly, L-PRF growth factors had a detrimental effect on the survival of pCNs, and consequently, also interfered with their neurite outgrowth. In contrast, we found a positive effect on peripheral DRG neurons, and L-PRF growth factors improved their survival and significantly stimulated the outgrowth and branching of their neurites. Taken together, our study demonstrates the positive effects of the L-PRF secretome on peripheral neurons and supports its use in regenerative medicine but care should be taken when using it for CNS applications.

Keywords: leukocyte- and platelet-rich fibrin; neural stem cells; neuritogenesis; neurotoxicity; primary cortical neurons; sensory neurons.

MeSH terms

Biocompatible Materials*
Humans
Leukocytes
Neurons
Peripheral Nervous System
Platelet-Rich Fibrin*
Vascular Endothelial Growth Factor A

Substances

Biocompatible Materials
Vascular Endothelial Growth Factor A

Grants and funding

This research was supported by grants to I.L. from the Research Foundation Flanders (‘Fonds Wetenschappelijk Onderzoek Vlaanderen—FWO’, G0C1916N), and T.V. is a postdoc fellow also funded by the FWO (12Z2620N).