title: Effects of heat-treatment on plasma rich in growth factors-derived autologous eye drop.
abstract: We have developed and characterized a new type of plasma rich in growth factors (PRGF) derived eye-drop therapy for patients suffering from autoimmune diseases. To determine the concentration of several growth factors, proteins, immunoglobulins and complement activity of the heat-inactivated eye-drop and to study its biological effects on cell proliferation and migration of different ocular surface cells, blood from healthy donors was collected, centrifuged and PRGF was prepared avoiding the buffy coat. The half volume of the obtained plasma supernatant from each donor was heat-inactivated at 56 °C for 1 h (heat-inactivated PRGF). The concentration of several proteins involved on corneal wound healing, immunoglubolins G, M and E and functional integrity of the complement system assayed by CH50 test were determined. The proliferative and migratory potential of inactivated and non-inactivated PRGF eye drops were assayed on corneal epithelial cells (HCE), keratocytes (HK) and conjunctival fibroblasts (HConF). Heat-inactivated PRGF preserves the content of most of the proteins and morphogens involved in its wound healing effects while reduces drastically the content of IgE and complement activity. Heat-inactivated PRGF eye drops increased proliferation and migration potential of ocular surface cells with regard to PRGF showing significant differences on proliferation and migration rate of HCE and HConF respectively. In summary, heat-inactivation of PRGF eye drops completely reduced complement activity and deceased significantly the presence of IgE, maintaining the biological activity of PRGF on ocular surface cells.
authors.affiliation: Biotechnology Institute (BTI), Vitoria, Spain. Electronic address: eduardoanitua@eduardoanitua.com.
authors.forename: E
authors.initials: E
authors.lastname: Anitua
authors.affiliation: Biotechnology Institute (BTI), Vitoria, Spain.
authors.forename: F
authors.initials: F
authors.lastname: Muruzabal
authors.affiliation: Biotechnology Institute (BTI), Vitoria, Spain.
authors.forename: M
authors.initials: M
authors.lastname: De la Fuente
authors.affiliation: Fundación de Investigación Oftalmológica, Instituto Oftalmológico Fernández-Vega, Oviedo, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Merayo-Lloves
authors.affiliation: Biotechnology Institute (BTI), Vitoria, Spain. Electronic address: gorka.orive@bti-implant.es.
authors.forename: G
authors.initials: G
authors.lastname: Orive
mesh_terms: D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003315:Cornea / Q000187:drug effects / Q000473:pathology; D003316:Corneal Diseases / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D006358:Hot Temperature; D006801:Humans; D009883:Ophthalmic Solutions / Q000494:pharmacology*; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology*; D053657:Platelet-Rich Plasma / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D014945:Wound Healing / Q000187:drug effects*
chemical_list: D009883:Ophthalmic Solutions; D010982:Platelet-Derived Growth Factor; D053773:Transforming Growth Factor beta1
keywords: EGF; ELISA; FBS; Fab fragment; Fc fragment; HCE; HConF; HK; IGF-I; IgE; IgG; IgM; Inact-PRGF; PBS; PDGF; PETIA; PRGF; PRP; TGF-β1; VEGF; autoimmune diseases; constant fragment of immunoglogulins; enzyme-linked immunosorbent assay; epithelial growth factor; fetal bovine serum; heat-inactivation; heat-treatment; human conjunctival fibroblasts; human corneal epithelial cells; human keratocytes; immunoglobulin E; immunoglobulin G; immunoglobulin M; inactivated PRGF; insulin-like growth factor I; ocular surface; particle-enhanced turbidimetric immunoassay; phosphate buffered saline; plasma rich in growth factors; platelet-derived growth factor; platelet-rich plasma; transforming growth factor-beta 1; variable fragment of immunoglogulins; vascular endothelial growth factor; wound healing