Neuroprotective activity of a virus‐safe nanofiltered human platelet lysate depleted of extracellular vesicles in Parkinson's disease and traumatic brain injury models

Abstract

Brain administration of human platelet lysates (HPL) is a potential emerging biotherapy of neurodegenerative and traumatic diseases of the central nervous system. HPLs being prepared from pooled platelet concentrates, thereby increasing viral risks, manufacturing processes should incorporate robust virus‐reduction treatments. We evaluated a 19 ± 2‐nm virus removal nanofiltration process using hydrophilic regenerated cellulose hollow fibers on the properties of a neuroprotective heat‐treated HPL (HPPL). Spiking experiments demonstrated >5.30 log removal of 20–22‐nm non‐enveloped minute virus of mice‐mock particles using an immuno‐quantitative polymerase chain reaction assay. The nanofiltered HPPL (NHPPL) contained a range of neurotrophic factors like HPPL. There was >2 log removal of extracellular vesicles (EVs), associated with decreased expression of pro‐thrombogenic phosphatidylserine and procoagulant activity. LC‐MS/MS proteomics showed that ca. 80% of HPPL proteins, including neurotrophins, cytokines, and antioxidants, were still found in NHPPL, whereas proteins associated with some infections and cancer‐associated pathways, pro‐coagulation and EVs, were removed. NHPPL maintained intact neuroprotective activity in Lund human mesencephalic dopaminergic neuron model of Parkinson's disease (PD), stimulated the differentiation of SH‐SY5Y neuronal cells and showed preserved anti‐inflammatory function upon intranasal administration in a mouse model of traumatic brain injury (TBI). Therefore, nanofiltration of HPL is feasible, lowers the viral, prothrombotic and procoagulant risks, and preserves the neuroprotective and anti‐inflammatory properties in neuronal pre‐clinical models of PD and TBI.

Proteomics of human platelet lysates and insight from animal studies on platelet protein diffusion to hippocampus upon intranasal administration

“…Nanofiltration impact is not dramatic, making it a valuable process to ensure HPPL viral safety without significantly altering protein composition. 19 Even though our data show significant differences in the proteomes of HPLs depending on their preparation methods, we must acknowledge that variations within the same type of HPL could be linked to the methods how the PCs are collected, the proteomic methods used, and the size of the HPL pool. These factors need to be considered to contextualize the extent of the observed differences among different HPLs and within each HPL category.…”

“…MHPPL were then filtered through 0.001-m 2 planova-20N (19-nm pore size; Asahi Kasei, Tokyo, Japan), a filter dedicated to the removal of blood-borne viruses, to form NHPPL. 19 To minimize individual differences in the proteome of platelet concentrates, we pooled the HPLs from three different donors.…”

“… 24,25 For instance, HPLs can be prepared through direct freeze-thawing of PCs suspended in plasma or from isolated platelets. 14,26,27 Some undergo calcium salt-triggered seroconversion, 26 while others are heat-treated 14,28 or subjected to virus-reduction treatments, 19,26,29 each with its potential effects on protein content and function. Given these variabilities, conducting comprehensive proteomic analyses using advanced techniques of label-free and tandem mass tag (TMT)-labeled mass spectrometry (MS) can be of a significant scientific value to objectively guide the translational applications of HPLs.…”

Emergency Treatment and Photoacoustic Assessment of Spinal Cord Injury Using Reversible Dual‐Signal Transform‐Based Selenium Antioxidant

“…Measurement of Cellular ROS: In order to verify the ROS scavenging capacity of Se@BDP-DOH via the flow cytometry experiments and immunocytochemistry experiments, SH-SY5Y cells were selected in this study because it is often used as a cell model for the study of nervous system diseases. [36,46] Cells were placed into 96-well cell culture plates and incubated with dihydroethidium (DHE, S0063, Beyotime,) for 0.5 h at the temperature of 37°C in an atmosphere of 5% CO 2 . Later, the OD value was detected by a 96-well plate reader (Thermo, USA) for every 20 min at 300 nm.…”

Innovative applications of platelet derivatives in light of information presented during the 2022 virtual congress of the International Society of Blood Transfusion (ISBT); selected issues

“…A 5% solution of IHPPL after a week of incubation with microglia and neurons showed no toxicity towards them and did not induce inflammatory activity (as marked by no expression of inflammatory factors -tumor necrosis factor; TNF and anti-cyclooxygenase, COX-2). With support of research on neuron division, there is hope for implementation of lysates to clinical procedures for management of the disorders of the nervous system [6,9,10].…”