title: Serum from plasma rich in growth factors regenerates rabbit corneas by promoting cell proliferation, migration, differentiation, adhesion and limbal stemness.
abstract: OBJECTIVE
  To evaluate the regenerating potential and the mechanisms through which the autologous serum derived from plasma rich in growth factors (s-PRGF) favours corneal wound healing in vitro and in vivo.
  
  
  METHODS
  We compared the effect of various concentrations of s-PRGF versus fetal bovine serum (FBS) and control treatment in rabbit primary corneal epithelial and stromal cells and wounded rabbit corneas. Cell proliferation was measured using an enzymatic colorimetric assay. In vitro and in vivo wound-healing progression was assessed by image-analysis software. Migration and invasion were evaluated using transfilter assays. Histological structure was analysed in stained sections. Protein expression was evaluated by immunohistochemistry.
  
  
  RESULTS
  s-PRGF promoted the robust proliferation of epithelial cultures at any concentration, similar to FBS. Likewise, s-PRGF and FBS produced similar re-epithelialization rates in in vitro wound-healing assays. In vivo, s-PRGF treatment accelerated corneal wound healing in comparison with control treatment. This difference was significant only for 100% s-PRGF treatment in our healthy rabbit model. Histological analysis confirmed normal epithelialization in all cases. Immunohistochemistry showed a higher expression of cytokeratins 3/76 and 15, zonula occludens-1 and alpha-smooth muscle actin proteins as a function of s-PRGF concentration. Notably, keratocyte density in the anterior third of the stroma increased with increase in s-PRGF concentration, suggesting an in vivo chemotactic effect of s-PRGF on keratocytes that was further confirmed in vitro.
  
  
  CONCLUSIONS
  s-PRGF promotes proliferation and migration and influences limbal stemness, adhesion and fibrosis during corneal healing.
authors.affiliation: Department of Cell Biology and Histology, School of Medicine and Nursing, BioCruces Health Research Institute, University of the Basque Country, Begiker, Leioa, Spain.|Department of Ophthalmology, BioCruces Health Research Institute, University Hospital of Cruces, Begiker, Barakaldo, Spain.
authors.forename: Jaime
authors.initials: J
authors.lastname: Etxebarria
authors.affiliation: Department of Cell Biology and Histology, School of Medicine and Nursing, BioCruces Health Research Institute, University of the Basque Country, Begiker, Leioa, Spain.
authors.forename: Sara
authors.initials: S
authors.lastname: Sanz-Lázaro
authors.affiliation: Department of Cell Biology and Histology, School of Medicine and Nursing, BioCruces Health Research Institute, University of the Basque Country, Begiker, Leioa, Spain.
authors.forename: Raquel
authors.initials: R
authors.lastname: Hernáez-Moya
authors.affiliation: Department of Cell Biology and Histology, School of Medicine and Nursing, BioCruces Health Research Institute, University of the Basque Country, Begiker, Leioa, Spain.|R & D Department, Instituto Clínico-Quirúrgico de Oftalmología, Bilbao, Spain.
authors.forename: Vanesa
authors.initials: V
authors.lastname: Freire
authors.affiliation: R & D Department, Instituto Clínico-Quirúrgico de Oftalmología, Bilbao, Spain.|Department of Ophthalmology, School of Medicine and Nursing, BioCruces Health Research Institute, University of the Basque Country, Begiker, Leioa, Spain.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Durán
authors.affiliation: Department of Cell Biology and Histology, School of Medicine and Nursing, BioCruces Health Research Institute, University of the Basque Country, Begiker, Leioa, Spain.
authors.forename: María-Celia
authors.initials: MC
authors.lastname: Morales
authors.affiliation: Department of Cell Biology and Histology, School of Medicine and Nursing, BioCruces Health Research Institute, University of the Basque Country, Begiker, Leioa, Spain.
authors.forename: Noelia
authors.initials: N
authors.lastname: Andollo
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003315:Cornea / Q000473:pathology*; D065306:Corneal Injuries / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D019573:Epithelium, Corneal / Q000473:pathology*; D005260:Female; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D009883:Ophthalmic Solutions; D053657:Platelet-Rich Plasma; D011817:Rabbits; D062326:Re-Epithelialization / Q000502:physiology; D044967:Serum
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D009883:Ophthalmic Solutions
keywords: cornea regeneration; s-PRGF; serum eye drops; wound healing