A purified human platelet pellet lysate rich in neurotrophic factors and antioxidants repairs and protects corneal endothelial cells from oxidative stress

Widyaningrum, Rifa; Burnouf, Thierry; Nebie, Ouada; Delila, Liling; Wang, Tsung Jen

doi:10.1016/j.biopha.2021.112046

Cited by 20 publications

(17 citation statements)

References 74 publications

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“…), antiinflammatory cytokines (e.g., TGF-β), antioxidants (e.g., glutathione reductase, superoxide dismutase, and catalase), and other proteins with neuroprotective activities (e.g., interalpha trypsin inhibitor, ceruloplasmin, and optineurin) 34 In addition, the activity of HPPL on damaged corneal endothelium cells highlighted a protective effect against oxidative stress and promotion of cell−cell adherens junction and cadherin binding related to cell−cell adhesion relevant to wound healing. 78 We, therefore, hypothesize that such important proteins are also present in EVs found in platelet lysates, thereby explaining in part their functional activities. Other functional biomolecules potentially present in our isolated EVs include miRNA but further investigations about their role in neuroprotection are needed.…”

Section: Discussionmentioning

confidence: 95%

“…Our previous proteomics studies to understand the neuroprotective activity of HPPL in traumatic brain injury have identified its remarkable richness in neurotrophins (e.g., PDGF, VEGF, BDNF, etc. ), anti-inflammatory cytokines (e.g., TGF-β), antioxidants (e.g., glutathione reductase, superoxide dismutase, and catalase), and other proteins with neuroprotective activities (e.g., inter-alpha trypsin inhibitor, ceruloplasmin, and optineurin) In addition, the activity of HPPL on damaged corneal endothelium cells highlighted a protective effect against oxidative stress and promotion of cell–cell adherens junction and cadherin binding related to cell–cell adhesion relevant to wound healing . We, therefore, hypothesize that such important proteins are also present in EVs found in platelet lysates, thereby explaining in part their functional activities.…”

Section: Discussionmentioning

confidence: 99%

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Characterization and Chromatographic Isolation of Platelet Extracellular Vesicles from Human Platelet Lysates for Applications in Neuroregenerative Medicine

Nyam-Erdene

Nebie

Delila

et al. 2021

ACS Biomater. Sci. Eng.

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Human platelet lysates (HPLs) made from clinical-grade platelet concentrates are currently evaluated in the preclinical models of Parkinson’s disease, Alzheimer’s disease, traumatic brain injury, and others, as a new polyvalent neuroprotective biotherapy of the central nervous system. However, the presence and content of extracellular vesicles (EVs) in HPLs and their potential contribution to the neuroprotective and neurorestorative activities of HPLs are still unknown. We, therefore, characterized the EVs present in four different HPL preparations and after purification by size-exclusion chromatography. We then tested the effect of the isolated EVs on neuronal cell repair. We identified that all four HPLs contained a high and similar amount of EVs (1011 to 1012/mL) with a mean size ranging from ca. 50 to 300 nm and a negative zeta potential as determined by nanoparticle tracking analysis and dynamic light scattering. Western blot analysis revealed that the EVs present in HPLs expressed the clusters of differentiation 41 (CD41) and 61 (CD61) characteristic of platelets. These EVs were efficiently isolated from HPL proteins by Sepharose CL-2B size-exclusion column chromatography as confirmed by total protein determination and protein profile by sodium dodecyl sulfate polyacrylamide gel electrophoresis, with 73–85% recovery and maintenance of their size, negative zeta potential, and CD41 and CD61 expression. Interestingly, the EVs purified from the four HPLs exhibited a differential capacity to promote cell growth and migration in a wound-healing assay using SH-SY5Y neuronal cells, and one EV preparation stimulated network formation in primary neuronal cultures. These data indicated that the EVs present in HPLs have different neuroregenerative capacities and that some EV preparations may have interesting applications as a stand-alone therapy for usage in neuroregenerative medicine.

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 99%

Characterization and Chromatographic Isolation of Platelet Extracellular Vesicles from Human Platelet Lysates for Applications in Neuroregenerative Medicine

Nyam-Erdene

Nebie

Delila

et al. 2021

ACS Biomater. Sci. Eng.

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“…We determined the total protein concentration and growth factor content and performed proteomics analysis by LC‐MS/MS as described before 13 , 56 and in Supporting Information S1 .…”

Section: Methodsmentioning

confidence: 99%

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

Delila

Nebie

et al. 2022

Bioengineering & Transla Med

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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.

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“…A dual-media approach with alternating stabilisation (low-mitogenic) and proliferation (high mitogenic) conditions has been shown to be effective [25,44,45]. More recently, human platelet lysate (HPL) [46][47][48][49], small molecules [50,51] and mesenchymal stem cell derived conditioned media [52,53] have been reported to be helpful.…”

Section: Corneal Endothelial Cell Culture Protocolmentioning

confidence: 99%

Cell therapy in corneal endothelial disease

Wong

2022

Current Opinion in Ophthalmology

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Purpose of reviewEndothelial keratoplasty is the current gold standard for treating corneal endothelial diseases, achieving excellent visual outcomes and rapid rehabilitation. There are, however, severe limitations to donor tissue supply and uneven access to surgical teams and facilities across the globe. Cell therapy is an exciting approach that has shown promising early results. Herein, we review the latest developments in cell therapy for corneal endothelial disease.Recent findingsWe highlight the work of several groups that have reported successful functional outcomes of cell therapy in animal models, with the utilization of human embryonic stem cells, human-induced pluripotent stem cells and cadaveric human corneal endothelial cells (CECs) to generate populations of CECs for intracameral injection. The use of corneal endothelial progenitors, viability of cryopreserved cells and efficacy of simple noncultured cells, in treating corneal decompensation is of particular interest. Further additions to the collective understanding of CEC physiology, and the process of cultivating and administering effective cell therapy are reviewed as well.SummaryThe latest developments in cell therapy for corneal endothelial disease are presented. The continuous growth in this field gives rise to the hope that a viable solution to the large numbers of corneal blind around the world will one day be reality.

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A purified human platelet pellet lysate rich in neurotrophic factors and antioxidants repairs and protects corneal endothelial cells from oxidative stress

Cited by 20 publications

References 74 publications

Characterization and Chromatographic Isolation of Platelet Extracellular Vesicles from Human Platelet Lysates for Applications in Neuroregenerative Medicine

Characterization and Chromatographic Isolation of Platelet Extracellular Vesicles from Human Platelet Lysates for Applications in Neuroregenerative Medicine

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

Cell therapy in corneal endothelial disease

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