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Francisco Jose Muruzabal, Maria de la Fuente, Ana Cristina Riestra, Jesus Merayo-Lloves, Gorka Orive, Eduardo Anitua; Effects of autologous serum and plasma rich in growth factors (PRGF-Endoret) eye drops on ocular surface myofibroblast differentiation.. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):734.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate and compare the effect of autologous serum eye drops and Plasma rich in growth factors (PRGF-Endoret) eye drops in TGF-β1-induced myofibroblast differentiation of keratocytes and conjunctival fibroblasts.
Blood from healthy donors was collected and was processed according to the following methods to obtain both blood derivatives: Autologous serum (AS): blood was collected in tubes without anticoagulant, was incubated for 20 minutes at room temperature, centrifuged at 2,000 g for 10 minutes, after that supernatant fraction was collected and diluted to 20% with 0.9% sterile saline serum. Plasma rich in growth factors (PRGF) eye drop: blood was collected into 9 mL tubes, was centrifuged at 580g for 8 minutes, whole plasma column was drawn off avoiding the buffy coat, incubated at 37 0C for one hour, and the released supernatants were collected and filtered. Keratocytes and conjunctival fibroblasts were induced to myofibroblast differentiation after the treatment with 2.5 ng/ml of TGF-β1. The capability of PRGF and AS to prevent and inhibit TGF-β1-induced myodifferentiation was evaluated by the detection of some myofibroblast characteristic cytoskeletal proteins like smooth muscle actin (SMA), desmin or vimentin. PRGF and SA potential to induce fibroblast and myofibroblast proliferation or apoptosis (ki-67 or TUNNEL assay respectively) were also assayed by immunofluorescence staining.
The expression of the three fibrotic markers in both types of fibroblasts was higher after treatment with AS compared to PRGF. In particular, the expressions of α-SMA and desmin were significantly higher after culturing cells with AS compared to PRGF. In addition, results showed that PRGF induced the highest cell proliferation profile rate in TGF-β1-induced cultured cells. Fibroblast and myofibroblast cells shown ki67 positive staining after treatment with PRGF or AS, being significantly higher in those cells which were treated with PRGF regarding to SA. Apoptotic cells were not observed after treatment with PRGF or AS.
The present results suggest that PRGF exerts enhanced biological outcomes than AS. PRGF may improve the treatment of ocular surface wound healing minimizing the scar formation when compared to AS. Results obtained herein suggest that PRGF protects and reverses the myofibroblast phenotype while promotes cell proliferation.
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