title: Does platelet-rich plasma freeze-thawing influence growth factor release and their effects on chondrocytes and synoviocytes?
abstract: PRP cryopreservation remains a controversial point. Our purpose was to investigate the effect of freezing/thawing on PRP molecule release, and its effects on the metabolism of chondrocytes and synoviocytes. PRP was prepared from 10 volunteers, and a half volume underwent one freezing/thawing cycle. IL-1β, HGF, PDGF AB/BB, TGF-β1, and VEGF were assayed 1 hour and 7 days after activation. Culture media of chondrocytes and synoviocytes were supplemented with fresh or frozen PRP, and, at 7 days, proliferation, gene expression, and secreted proteins levels were evaluated. Results showed that in the freeze-thawed PRP the immediate and delayed molecule releases were similar or slightly lower than those in fresh PRP. TGF-β1 and PDGF AB/BB concentrations were significantly reduced after freezing both at 1 hour and at 7 days, whereas HGF concentration was significantly lower in frozen PRP at 7 days. In fresh PRP IL-1β and HGF concentrations underwent a significant further increase after 7 days. Similar gene expression was found in chondrocytes cultured with both PRPs, whereas in synoviocytes HGF gene expression was higher in frozen PRP. PRP cryopreservation is a safe procedure, which sufficiently preserves PRP quality and its ability to induce proliferation and the production of ECM components in chondrocytes and synoviocytes.
authors.affiliation: Nano-Biotechnology Laboratory, Rizzoli Orthopaedic Institute, Via di Barbiano 1, 40136 Bologna, Italy.
authors.forename: Alice
authors.identifier: 0000-0002-2683-3743
authors.initials: A
authors.lastname: Roffi
authors.affiliation: II Clinic-Biomechanics Laboratory and Nano-Biotechnology Laboratory, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy.
authors.forename: Giuseppe
authors.identifier: 0000-0002-8136-0977
authors.initials: G
authors.lastname: Filardo
authors.affiliation: Laboratory of Immunorheumatology and Tissue Regeneration/RAMSES, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy.
authors.forename: Elisa
authors.identifier: 0000-0001-5678-6506
authors.initials: E
authors.lastname: Assirelli
authors.affiliation: Laboratory RAMSES, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy.
authors.forename: Carola
authors.initials: C
authors.lastname: Cavallo
authors.affiliation: Immunohematology and Transfusion Medicine and Cell and Musculoskeletal Tissue Bank, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy.
authors.forename: Annarita
authors.initials: A
authors.lastname: Cenacchi
authors.affiliation: Laboratory of Immunorheumatology and Tissue Regeneration Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy ; Department of Medical and Surgical Science, University of Bologna, Via Giuseppe Massarenti 9, 40138 Bologna, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Facchini
authors.affiliation: Laboratory of Immunorheumatology and Tissue Regeneration/RAMSES, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy.
authors.forename: Brunella
authors.initials: B
authors.lastname: Grigolo
authors.affiliation: II Clinic-Biomechanics Laboratory and Nano-Biotechnology Laboratory, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy.
authors.forename: Elizaveta
authors.initials: E
authors.lastname: Kon
authors.affiliation: Laboratory of Immunorheumatology and Tissue Regeneration Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy ; Department of Medical and Surgical Science, University of Bologna, Via Giuseppe Massarenti 9, 40138 Bologna, Italy.
authors.forename: Erminia
authors.initials: E
authors.lastname: Mariani
authors.affiliation: Clinical Pathology Unit, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy.
authors.forename: Loredana
authors.initials: L
authors.lastname: Pratelli
authors.affiliation: Laboratory of Immunorheumatology and Tissue Regeneration/RAMSES, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy.
authors.forename: Lia
authors.identifier: 0000-0003-3031-170X
authors.initials: L
authors.lastname: Pulsatelli
authors.affiliation: II Clinic-Biomechanics Laboratory and Nano-Biotechnology Laboratory, Rizzoli Orthopaedic Institute, Via di Barbiano 1/10, 40136 Bologna, Italy.
authors.forename: Maurilio
authors.initials: M
authors.lastname: Marcacci
mesh_terms: D000328:Adult; D049109:Cell Proliferation / Q000187:drug effects; D019902:Chondrocytes / Q000187:drug effects*; D015925:Cryopreservation; D003470:Culture Media; D005615:Freezing; D006801:Humans; D008297:Male; D010982:Platelet-Derived Growth Factor / Q000187:drug effects / Q000378:metabolism*; D053657:Platelet-Rich Plasma / Q000737:chemistry*; D020574:Proto-Oncogene Proteins c-sis / Q000187:drug effects / Q000378:metabolism; D013582:Synovial Fluid / Q000166:cytology / Q000187:drug effects*
chemical_list: D003470:Culture Media; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; C095477:platelet-derived growth factor AB