title: Corneal reconstruction with tissue-engineered cell sheets composed of human immature dental pulp stem cells.
abstract: OBJECTIVE
  To determine the outcome of the use of a tissue-engineered cell sheet composed of human undifferentiated immature dental pulp stem cells (hIDPSC) for ocular surface reconstruction in an animal model of total limbal stem cell deficiency (LSCD).
  
  
  METHODS
  LSCD was induced by the application of 0.5 M NaOH to the right eye of rabbits for 25 seconds (mild chemical burn [MCB]) and for 45 seconds (severe chemical burn [SCB]). After 1 month, a superficial keratectomy was performed to remove the fibrovascular pannus that covered the animals' burned corneas. A tissue-engineered hIDPSC sheet was transplanted onto the corneal bed and then covered with deepithelialized human amniotic membrane (AM). In the respective control groups, the denuded cornea was covered with AM only. After 3 months, a detailed analysis of the rabbit eyes was performed with regard to clinical aspect, histology, electron microscopy, and immunohistochemistry.
  
  
  RESULTS
  Corneal transparency of the rabbit eyes that underwent hIDPSC transplantation was improved throughout the follow-up, while the control corneas developed total conjunctivalization and opacification. Rabbits from the MCB group showed clearer corneas with less neovascularization. The clinical data were confirmed by histologic analysis that showed healthy uniform corneal epithelium, especially in the MCB group. The presence of hIDPSC was detected using an anti-hIDPSC antibody. The corneal tissue also showed positive immunostaining with anti-human antibodies. In the control corneas, none of these antigens were detected.
  
  
  CONCLUSIONS
  Overall, these data showed that transplantation of a tissue-engineered hIDPSC sheet was successful for the reconstruction of corneal epithelium in an animal model of LSCD.
authors.affiliation: Centro Avançado de Superfície Ocular and Setor de Córnea e Doenças Externas, Instituto da Visão, Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: José Alvaro Pereira
authors.initials: JA
authors.lastname: Gomes
authors.forename: Bábyla
authors.initials: B
authors.lastname: Geraldes Monteiro
authors.forename: Gustavo Barreto
authors.initials: GB
authors.lastname: Melo
authors.forename: Ricardo Luiz
authors.initials: RL
authors.lastname: Smith
authors.forename: Marcelo
authors.initials: M
authors.lastname: Cavenaghi Pereira da Silva
authors.forename: Nelson Foresto
authors.initials: NF
authors.lastname: Lizier
authors.forename: Alexandre
authors.initials: A
authors.lastname: Kerkis
authors.forename: Humberto
authors.initials: H
authors.lastname: Cerruti
authors.forename: Irina
authors.initials: I
authors.lastname: Kerkis
mesh_terms: D000818:Animals; D002057:Burns, Chemical / Q000473:pathology / Q000601:surgery; D018929:Cell Culture Techniques; D003316:Corneal Diseases / Q000601:surgery*; D003782:Dental Pulp / Q000166:cytology*; D004195:Disease Models, Animal; D005126:Eye Burns / Q000139:chemically induced / Q000473:pathology / Q000601:surgery; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D016850:Limbus Corneae / Q000601:surgery* / Q000648:ultrastructure; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D046529:Microscopy, Electron, Transmission; D013508:Ophthalmologic Surgical Procedures; D011817:Rabbits; D019651:Plastic Surgery Procedures; D023822:Tissue Engineering; D014183:Transplantation, Heterologous