Abstract

Background - We evaluated neurotrophin (NF) levels and their impact on in vitro cell wound healing in eye drops from differently prepared blood sources (cord blood [CB], and peripheral blood [PB]) in the same donor, to avoid intrasubject biological variability.
Materials and methods - Twenty healthy adult donor PB samples, and twenty CB samples acquired at the time of delivery were processed to obtain serum (S), platelet-rich plasma (PRP), platelet-poor plasma (PPP), and S retrieved from PRP after activation with Ca-gluconate (PRP-R). The levels of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), glial-derived neurotrophic factor (GDNF), fibroblast growth factor (FGF), and epidermal growth factor (EGF) were assessed with a Luminex xMAP (Luminex Corporation), and by using multikine kits from R&D system, and were statistically analysed in the eight different preparations. The impact of S, PRP, PPP, PRP-R from both sources on a cell line responding to NF supplementation (MIO-M1, UCL Institute of Ophthalmology, London, UK) was tested with a scratch wound assay, and analysed by IncuCyte S3 equipment.
Results - All the preparations from CB showed higher NF levels, except for BDNF where no difference was found as compared to PB. PRP showed higher NF levels with respect to S, PPP and PRP-R in this decreasing order. Younger donors in PB contributed with higher NF levels. The scratch assay showed different cell migration results, with a complete wound closure only recorded with the supplementation of CB-S, and a progressive reduction by using PRP, PRP-R, and PPP from both sources.
Discussion - Protocols of preparation and choice of blood source determine different NF levels in the final products. The therapeutic use of a natural neurotrophin pool from blood sources might have a clinical impact in several different settings. Efforts are needed to standardise the manufacturing and the product content in order to establish and modulate the posology of the final supplementation.

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Authors

Sabrina Valente - DIMES, Alma Mater Studiorum University of Bologna, Bologna, Italy; Center for Applied Biomedical Research (CRBA), Alma Mater Studiorum, University of Bologna, Bologna, Italy

Nico Curti - DIMES, Alma Mater Studiorum University of Bologna, Bologna, Italy

Enrico Giampieri - DIMES, Alma Mater Studiorum University of Bologna, Bologna, Italy

Vanda Randi - Emilia Romagna Cord Blood Bank-Transfusion Service, IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy

Chiara Donadei - DIMES, Alma Mater Studiorum University of Bologna, Bologna, Italy

Marina Buzzi - Emilia Romagna Cord Blood Bank-Transfusion Service, IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy

Piera Versura - Center for Applied Biomedical Research (CRBA), Alma Mater Studiorum, University of Bologna, Bologna, Italy; Ophthalmology Unit, DIMES, Alma Mater Studiorum University of Bologna, IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy

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