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.

中文翻译：

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去除细胞外囊泡的病毒安全纳米过滤人血小板裂解物在帕金森病和创伤性脑损伤模型中的神经保护活性


人血小板裂解物（HPL）的脑给药是一种潜在的新兴生物疗法，用于治疗中枢神经系统的神经退行性疾病和创伤性疾病。 HPL 由混合血小板浓缩物制备，从而增加了病毒风险，生产过程应采用强有力的病毒减少处理。我们评估了使用亲水性再生纤维素中空纤维的 19 ± 2 nm 病毒去除纳滤工艺对神经保护性热处理 HPL (HPPL) 的性能。加标实验表明，使用免疫定量聚合酶链式反应测定，小鼠模拟颗粒的 20–22 nm 无包膜微小病毒的去除率为 >5.30 log。纳米过滤 HPPL (NHPPL) 含有一系列神经营养因子，如 HPPL。细胞外囊泡 (EV) 去除率为 >2 log，与促血栓形成磷脂酰丝氨酸和促凝血活性的表达降低相关。 LC-MS/MS 蛋白质组学表明，大约。 NHPPL 中仍存在 80% 的 HPPL 蛋白，包括神经营养因子、细胞因子和抗氧化剂，而与某些感染和癌症相关途径、促凝血和 EV 相关的蛋白质则被去除。 NHPPL 在帕金森病 (PD) 隆德人中脑多巴胺能神经元模型中保持完整的神经保护活性，刺激 SH-SY5Y 神经元细胞的分化，并在创伤性脑损伤 (TBI) 小鼠模型中鼻内给药后显示出保留的抗炎功能。因此，HPL 纳滤是可行的，可降低病毒、血栓形成和促凝血风险，并保留 PD 和 TBI 神经元临床前模型中的神经保护和抗炎特性。