Corneal reconstruction with tissue-engineered cell sheets composed of human immature dental pulp stem cells.
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.
Gomes
José Alvaro Pereira
JA
Centro Avançado de Superfície Ocular and Setor de Córnea e Doenças Externas, Instituto da Visão, Universidade Federal de São Paulo, São Paulo, Brazil.
Geraldes Monteiro
Bábyla
B
Melo
Gustavo Barreto
GB
Smith
Ricardo Luiz
RL
Cavenaghi Pereira da Silva
Marcelo
M
Lizier
Nelson Foresto
NF
Kerkis
Alexandre
A
Cerruti
Humberto
H
Kerkis
Irina
I
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