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J. Merayo-Lloves, C. Alonso-Ron, G. Ferrara, M. Chacon; Platelet Rich Growth Factor (PRGF) Promotes Corneal Human Epithelium Wound Healing in vitro. Invest. Ophthalmol. Vis. Sci. 2010;51(13):372.
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To asses the potential therapeutic benefits of Platelet Rich Growth Factor (PRGF) in restoring corneal epithelial wound in a human corneal epithelium wound healing in vitro model.
PRGF was obtained by selective centrifugation (8 minutes, 580g) of 9 mL of blood acoording with BTI Protocol (BTI, Vitoria, Spain). The fraction rich in platelets (2 & 3) was activated with 50 µL/mL calcium chloride.SV40 immortalized human corneal epithelial cells (HCE; Araki-Sasaki K. et al., 1995) were grown in 24-wells plates in DMEM:F-12 medium supplemented with 7.5% FBS, 10 ng/mL EGF, 0.5 µg/mL insulin, 1% DMSO and penicillin/streptomycin mixture (100 units-µg/mL) [Complete Feeding Medium, CFM] until confluence.Wounds were produced using a pipette tip centered in each well by a cylindrical peace of methacrylate with a hole in the middle. The wounds were photographed every 4 or 12 h and their area was measured using the Image J software (NIH).Estimated Rate of Healing (ERH) was determined by the slope obtained after fitting the decrease of wound area time course to a line function.The effects on wound healing of different concentrations (20 to 100%) of PRGF and Human Serum were tested in wounded human epithelium cell cultured in DMEM:F-12 supplemented with penicillin/streptomycin, in CFM or in DMEM:F-12 supplemented with penicillin/streptomycin (Basic Medium, BM).
Epithelium wound did not heal in BM wells (negative control). Addition of PRGF promoted the clousure of the experimental wound (ERH -1.47 ± 0.2) with significative differences with human serum. The maximal wound rate was reached with CFM (ERH = -3.95 0.09), ( positive control).
PRGF promotes corneal epithelium wound healing in vitro. This findings address the possibility of using this trophic factor for the treatment of corneal epithelium injuries in human patients and open the possibility of adding specific growth factors to the autologous formulation.
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