title: In Vivo Confocal Microscopy 1 Year after Autologous Cultured Limbal Stem Cell Grafts.
abstract: OBJECTIVE
  To correlate clinical, impression cytologic, and in vivo confocal microscopy findings on the corneal surface after cultured limbal stem cell transplantation.
  
  
  METHODS
  Prospective, interventional, noncomparative, masked case series.
  
  
  METHODS
  Thirteen patients with limbal stem cell deficiency after unilateral (9 eyes) or bilateral (2 eyes) chemical burn, liquid nitrogen injury (1 eye), or herpes simplex virus infection (1 eye).
  
  
  METHODS
  Limbal cells were harvested from healthy or less affected eyes, cultured on 3T3 cells and fibrin glue, and transplanted to the patient's injured eye. Patients underwent clinical examination and impression cytologic examination of the central cornea before and 1 year after intervention. In vivo confocal microscopy scans were obtained in all corneal quadrants after 1 year. The interexamination agreement was established by calculation of the Cohen's κ coefficient.
  
  
  METHODS
  Results of surgery were assessed considering clinical signs (successful: restoration of transparent, avascular, and stable corneal epithelium without neovascularization in central corneal surface; partially successful: recurrence of superficial neovascularization; failed: recurrent epithelial defects, pannus, and inflammation), phenotype of cells covering the corneal surface (conjunctivalized corneal surface: cytokeratin 12 [cK12]-negative and mucin 1 [MUC1]-positive cells; mixed epithelium: cK12-positive and MUC1-positive cells; corneal epithelium: cK12-positive and MUC1-negative cells), and cell morphologic features (corneal epithelium: multilayered polygonal and flat cells with hyperreflective nuclei; conjunctival epithelium: stratified cuboidal or polygonal cells, hyperreflective cytoplasm, and barely defined borders; epithelial transition: transition of epithelial cells from the cornea to the conjunctiva over the corneal surface).
  
  
  RESULTS
  We found a moderate to substantial degree of concordance between confocal microscopy and clinical evaluation (κ = 0.768) and between confocal microscopy and impression cytologic analysis (κ = 0.629). Confocal microscopy showed that 46.2% of patients exhibited corneal epithelium in the central and peripheral cornea, 30.8% showed an irregular mixed corneal and conjunctival epithelium, and 23.0% showed conjunctival epithelium. Palisades of Vogt were absent in all (100.0%) patients, and the cornea-conjunctiva epithelial transition localized approximately 1 mm internally on the cornea.
  
  
  CONCLUSIONS
  Confocal microscopy provides objective measures of the corneal epithelium and may significantly improve the evaluation of outcomes after cultured limbal stem cell graft.
authors.affiliation: Eye Clinic, Department of Neurological and Movement Sciences, University Hospital, Verona, Italy. Electronic address: emilio.pedrotti@univr.it.
authors.forename: Emilio
authors.initials: E
authors.lastname: Pedrotti
authors.affiliation: Eye Clinic, Department of Neurological and Movement Sciences, University Hospital, Verona, Italy.
authors.forename: Mattia
authors.initials: M
authors.lastname: Passilongo
authors.affiliation: Eye Clinic, Department of Neurological and Movement Sciences, University Hospital, Verona, Italy; The Veneto Eye Bank Foundation, Venice, Italy.
authors.forename: Adriano
authors.initials: A
authors.lastname: Fasolo
authors.affiliation: Ophthalmology Clinic, Department of Medicine and Ageing Sciences, University "G. D'Annunzio" of Chieti and Pescara, Chieti, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Nubile
authors.affiliation: Eye Clinic, Department of Neurological and Movement Sciences, University Hospital, Verona, Italy.
authors.forename: Graziella
authors.initials: G
authors.lastname: Parisi
authors.affiliation: Eye Clinic, Department of Neurological and Movement Sciences, University Hospital, Verona, Italy; Ophthalmology Clinic, Department of Medicine and Ageing Sciences, University "G. D'Annunzio" of Chieti and Pescara, Chieti, Italy.
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Mastropasqua
authors.affiliation: Eye Clinic, Department of Neurological and Movement Sciences, University Hospital, Verona, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Ficial
authors.affiliation: The Veneto Eye Bank Foundation, Venice, Italy.
authors.forename: Marina
authors.initials: M
authors.lastname: Bertolin
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Enzo
authors.initials: E
authors.lastname: Di Iorio
authors.affiliation: The Veneto Eye Bank Foundation, Venice, Italy.
authors.forename: Diego
authors.initials: D
authors.lastname: Ponzin
authors.affiliation: Eye Clinic, Department of Neurological and Movement Sciences, University Hospital, Verona, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Marchini
mesh_terms: D016475:3T3 Cells / Q000166:cytology; D000328:Adult; D000368:Aged; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002478:Cells, Cultured; D018920:Coculture Techniques; D003316:Corneal Diseases / Q000378:metabolism / Q000473:pathology* / Q000628:therapy*; D019573:Epithelium, Corneal / Q000637:transplantation; D005260:Female; D006801:Humans; D053551:Keratin-12 / Q000378:metabolism; D016850:Limbus Corneae / Q000166:cytology* / Q000378:metabolism; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal; D008875:Middle Aged; D018396:Mucin-1 / Q000378:metabolism; D011446:Prospective Studies; D033581:Stem Cell Transplantation; D014182:Transplantation, Autologous
chemical_list: D015415:Biomarkers; D053551:Keratin-12; C060192:MUC1 protein, human; D018396:Mucin-1