Aim This study aimed to test whether human platelet lysate (HPL) has neurotrophic ability for corneal nerve regeneration.
Methods We measured the neurotrophic factors in human peripheral serum (HPS) and two commercially available HPLs, UltraGRO and PLTMax. In vitro, we compared the growth rates, neuronal differentiation and immunostaining of neuron markers in mouse neuroblastoma cell line (Neuro-2a) and primary culture of mouse trigeminal ganglion cells that were cultivated in different concentrations of fetal bovine serum, HPS and HPL. In vivo, we created corneal wounds on Sprague Dawley rats with a rotating burr and evaluated the effects of topical HPL on wound healing and corneal nerve regeneration by in vivo confocal microscopy and corneal aesthesiometry.
Results HPLs had significantly higher concentrations of various neurotrophic factors compared with HPS (p<0.05). In Neuro-2a cells, 3% HPL was better at promoting neuronal growth and differentiation compared with HPS at the same concentration. HPL was also found to have superior neurotrophic effects compared with HPS in primary cultures of mouse trigeminal ganglion cells. In vivo, HPL-treated eyes had better corneal epithelial wound healing rate, nerve regeneration length and corneal touch threshold compared with eyes treated with artificial tears (p<0.05).
Conclusion HPL has significantly higher concentrations of neurotrophic factors compared with HPS. It showed not only in vitro but also in vivo corneal neurotrophic abilities. Our results suggest that HPL may have a potential role in the treatment of diseases related to corneal nerve damage or degeneration.
- platelet lysate
- corneal nerve regeneration
- neurotrophic keratopathy
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C-TH and H-SC are joint first authors.
Contributors W-LC conceived and planned the experiments and obtained institutional review board approvals. M-YC contributed to sample preparation and carried out the experiments. K-CH and F-RH contributed to the interpretation of the results. C-TH, H-SC and LWC took the lead in writing the manuscript. All authors provided critical feedback and helped shape the research, analysis and manuscript.
Funding This study was supported, in part, by the Department of Medical Research and the Advanced Ocular Surface and Corneal Nerve Regeneration Center at National Taiwan University Hospital in Taipei, Taiwan.
Competing interests None declared.
Patient consent for publication Not required.
Ethics approval The preparation of the human peripheral serum in this study was approved by the Institutional Review Board for Human Studies at the National Taiwan University Hospital (201510123RINB). The animal use protocol has been approved by the Institutional Animal Care and Use Committee of National Taiwan University College of Medicine and College of Public Health (IACUC-20180120), and followed the ARVO Statement for the Use of Animal in Ophthalmic and Vision Research.
Provenance and peer review Not commissioned; externally peer reviewed.
Data availability statement All data relevant to the study are included in the article or uploaded as supplementary information.
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