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Francisco Jose Muruzabal, Maria De la Fuente, Jesus Merayo-Lloves, Gorka Orive, Eduardo Anitua; Comparing the biological outcomes of autologous serum versus plasma rich in growth factors (PRGF-Endoret) eye drops on ocular surface wound healing.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):4689.
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To evaluate and compare the biological and regeneration responses of keratocytes and conjunctival fibroblasts to either autologous serum eye drops or Plasma rich in growth factors (PRGF-Endoret) eye drops.
Blood from three healthy donors was collected. The blood sample from each donor was processed according to the following methods to obtain the corresponding 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 the complete supernatant fraction was collected and diluted to 20% by addition of 0.9% sterile saline serum. Plasma rich in growth factors (PRGF) eye drop was obtained using the Endoret (PRGF) kit in Ophthalmology (BTI Biotechnology Institute, S.L., Vitoria, Spain). Briefly, blood was collected into 9 mL tubes containing sodium citrate as anticoagulant, was centrifuged at 580g for 8 minutes, whole plasma column was drawn off avoiding the buffy coat, activated with calcium chloride, incubated at 37 C degrees for one hour, and finally, the released supernatants were collected by aspiration, filtered and aliquoted and stored at - 80 C degrees until use. PDGF-AB, VEGF, EGF, FGFb and TGF-β1 were quantified in each type of blood-derivative eye-drops. Primary human cells including keratocytes and conjunctival fibroblasts were used to perform the “in vitro” investigations. The potential of PRGF and AS in promoting wound healing was evaluated by means of proliferation and migration assays. Fibroblast cells 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 differentiation was evaluated.
PRGF eye drops showed significant higher levels of all growth factors analyzed with regard to AS. Furthermore, PRGF eye drops enhanced significantly the biological outcomes of both keratocytes and conjunctival fibroblasts, and reduced TGF-β1-induced myofibroblast differentiation in contrast to autologous serum eye drops (AS).
These results suggest that cellular response of keratocytes and conjunctival fibroblasts to PRGF eye drops is significantly better than to AS. PRGF may improve the treatment of ocular surface wound healing minimizing the scar formation.
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