title: Implementation of a more physiological plasma rich in growth factor (PRGF) protocol: Anticoagulant removal and reduction in activator concentration.
abstract: Plasma rich in growth factors (PRGF) is a biological therapy that uses patient's own growth factors for promoting tissue regeneration. Given the current European regulatory framework in which anticoagulant solution in blood extraction tubes could be considered as a medicinal product, a new PRGF protocol has been developed. The actual protocol (PRGF-A) and the new one (PRGF-B) have been performed and compared under Good Laboratory Practices. PRGF-A protocol uses extraction tubes with 0.9 mL of trisodium citrate as anticoagulant and 50 μL of calcium chloride/mL PRGF to activate it. The PRGF-B reduces the amount of sodium citrate and calcium chloride to 0.4 mL and to 20 μL, respectively. Basic hematological parameters, platelet function, the scaffold obtaining process, growth factors content, and the biological effect were compared between both PRGF obtaining protocols.
  
  
  RESULTS
  PRGF-B protocol led to a statistically significant higher enrichment and recovery of platelets regarding to the PRGF-A. Hypotonic stress response by platelets was significantly better in the new protocol. A statistically significant decrease in the basal platelet activation status of PRGF-B compared to PRGF-A was also observed. The duration of the lag phase in the platelet aggregation assay was statistically lower for the PRGF-B protocol. Both the clotting and the clot retraction time were significantly reduced in the B protocol. A higher growth factor concentration was detected in the plasma obtained using the PRGF-B protocol. The new PRGF obtaining protocol, with a reduction in the amount of anticoagulant and activator, has even improved the actual one.
authors.affiliation: a BTI-Biotechnology Institute , Vitoria , Spain.|b Eduardo Anitua Foundation for Biomedical Research , Vitoria , Spain.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Anitua
authors.affiliation: a BTI-Biotechnology Institute , Vitoria , Spain.
authors.forename: Roberto
authors.initials: R
authors.lastname: Prado
authors.affiliation: a BTI-Biotechnology Institute , Vitoria , Spain.
authors.forename: María
authors.initials: M
authors.lastname: Troya
authors.affiliation: a BTI-Biotechnology Institute , Vitoria , Spain.
authors.forename: Mar
authors.initials: M
authors.lastname: Zalduendo
authors.affiliation: a BTI-Biotechnology Institute , Vitoria , Spain.
authors.forename: María
authors.initials: M
authors.lastname: de la Fuente
authors.affiliation: a BTI-Biotechnology Institute , Vitoria , Spain.
authors.forename: Ander
authors.initials: A
authors.lastname: Pino
authors.affiliation: a BTI-Biotechnology Institute , Vitoria , Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: Muruzabal
authors.affiliation: a BTI-Biotechnology Institute , Vitoria , Spain.|b Eduardo Anitua Foundation for Biomedical Research , Vitoria , Spain.
authors.forename: Gorka
authors.initials: G
authors.lastname: Orive
mesh_terms: D000328:Adult; D000925:Anticoagulants; D001772:Blood Cell Count; D001780:Blood Coagulation Tests; D001781:Blood Component Removal / Q000379:methods; D001792:Blood Platelets / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use; D008297:Male; D008875:Middle Aged; D009997:Osmotic Pressure; D015539:Platelet Activation; D010974:Platelet Aggregation; D053657:Platelet-Rich Plasma / Q000737:chemistry
chemical_list: D000925:Anticoagulants; D036341:Intercellular Signaling Peptides and Proteins
keywords: Fibrin; PRGF; growth factors; platelet-rich plasma; platelets