title: Quantification of corneal neovascularization after ex vivo limbal epithelial stem cell therapy.
abstract: OBJECTIVE
  To assess cultured limbal epithelial stem cell transplantation in patients with limbal stem cell deficiency by analyzing and quantifying corneal neovascularization.
  
  
  METHODS
  This retrospective, interventional case series included eight eyes with total limbal stem cell deficiency. Ex vivo limbal epithelial stem cells were cultured on human amniotic membrane using an animal-free culture method. The clinical parameters of limbal stem cell deficiency, impression cytology, and quantification of corneal neovascularization were evaluated before and after cultured limbal stem cell transplantation. The area of corneal neovascularization, vessel caliber (VC), and invasive area (IA) were analyzed before and after stem cell transplantation by image analysis software. Best-corrected visual acuity (BCVA), epithelial transparency, and impression cytology were also measured.
  
  
  RESULTS
  One year after surgery, successful cases showed a reduction (improvement) of all three parameters of corneal neovascularization [neovascular area (NA), VC, IA], while failed cases did not. NA decreased a mean of 32.31% (P=0.035), invasion area 29.37% (P=0.018) and VC 14.29% (P=0.072). BCVA improved in all eyes (mean follow-up, 76±21mo). Epithelial transparency improved significantly from 2.00±0.93 to 0.88±1.25 (P=0.014). Impression cytology showed that three cases failed after limbal epithelial stem cell therapy before 1y of follow-up.
  
  
  CONCLUSIONS
  This method of analyzing and monitoring surface vessels is useful for evaluating the epithelial status during follow-up, as successful cases showed a bigger reduction in corneal neovascularization parameters than failed cases. Using this method, successful cases could be differentiated from failed cases.
authors.affiliation: Department of Ophthalmology, Clínica Universidad de Navarra, Pamplona 31008, Spain.
authors.forename: Adriano
authors.initials: A
authors.lastname: Guarnieri
authors.affiliation: Department of Ophthalmology, Clínica Universidad de Navarra, Pamplona 31008, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Moreno-Montañés
authors.affiliation: Department of Ophthalmology, Clínica Universidad de Navarra, Pamplona 31008, Spain.
authors.forename: Belén
authors.initials: B
authors.lastname: Alfonso-Bartolozzi
authors.affiliation: Department of Ophthalmology, Clínica Universidad de Navarra, Pamplona 31008, Spain.
authors.forename: Alfonso L
authors.initials: AL
authors.lastname: Sabater
authors.affiliation: Department of Cell Therapy, Clínica Universidad de Navarra, Pamplona 31008, Spain.
authors.forename: María
authors.initials: M
authors.lastname: García-Guzmán
authors.affiliation: Department of Cell Therapy, Clínica Universidad de Navarra, Pamplona 31008, Spain.
authors.forename: Enrique J
authors.initials: EJ
authors.lastname: Andreu
authors.affiliation: Department of Cell Therapy, Clínica Universidad de Navarra, Pamplona 31008, Spain.
authors.forename: Felipe
authors.initials: F
authors.lastname: Prosper
keywords: corneal neovascularization; impression cytology; limbal stem cell; limbal stem cell deficiency; stem cell therapy