April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Silk Fibroin Membranes as a Carrier for the Treatment of Corneal Lesions.
Author Affiliations & Notes
  • Alvaro Meana
    Superficie Ocular, Fundacion de Investigacion Oftalmologica, Oviedo, Spain
    Universidad de Oviedo, Oviedo, Spain
  • Jose L Cenis
    Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario, Murcia, Spain
  • Salvador D Aznar-Cervantes
    Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario, Murcia, Spain
  • Manuel Chacón
    Superficie Ocular, Fundacion de Investigacion Oftalmologica, Oviedo, Spain
    Universidad de Oviedo, Oviedo, Spain
  • Natalia Vázquez
    Superficie Ocular, Fundacion de Investigacion Oftalmologica, Oviedo, Spain
    Universidad de Oviedo, Oviedo, Spain
  • David Cereijo
    Fundacion Prodintec, Gijon, Spain
  • Jose Antonio Rodríguez-Cortés
    Fundacion Prodintec, Gijon, Spain
  • David Álvarez
    Fundacion Prodintec, Gijon, Spain
  • Jesus Merayo-Lloves
    Superficie Ocular, Fundacion de Investigacion Oftalmologica, Oviedo, Spain
    Universidad de Oviedo, Oviedo, Spain
  • Footnotes
    Commercial Relationships Alvaro Meana, None; Jose Cenis, None; Salvador Aznar-Cervantes, None; Manuel Chacón, None; Natalia Vázquez, None; David Cereijo, None; Jose Antonio Rodríguez-Cortés, None; David Álvarez, None; Jesus Merayo-Lloves, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 1450. doi:
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      Alvaro Meana, Jose L Cenis, Salvador D Aznar-Cervantes, Manuel Chacón, Natalia Vázquez, David Cereijo, Jose Antonio Rodríguez-Cortés, David Álvarez, Jesus Merayo-Lloves; Silk Fibroin Membranes as a Carrier for the Treatment of Corneal Lesions.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1450.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: We analyze the viability of silk fibroin (SF) membranes as a carrier of corneal cells and their potential use as a treatment of corneal lesions.

Methods: Cocoons of Bombyx mori were degummed (boiling in 0.02N Na2CO3) and SF was dissolved in 9.3M LiBr and dialyzed against distilled water into dialysis bags (3,500Da MWCO) for 3 days. Then, 580µL of the resultant 5% w/v SF solution was cast upon plastic Petri dish 5,8cm in diameter to give a 10 microns thickness film. Once dried at room temperature, the water-annealing was performed by placing the SF films in a water-filled desiccator in vacuum conditions for 24h. Cells were obtained from normal New Zealand white rabbits and normal human corneoscleral rims obtained during surgery after 8mm trephination of the graft. Limbal stem cells were obtained from explants of 2-3mm in diameter of the limbal region. Stromal and endothelial cells were obtained by digesting the stromal tissue and the dissected endothelium with Tripsin/EDTA and Collagenase I respectively. Cells were cultured in different culture media onto SF membranes using a handling device designed by Prodintec Foundation for our laboratory. Corneal cells growing on the SF membranes were examined by phase contrast microscopy, scanning electron microscopy (SEM) and immunocytochemistry for antibodies: Cytokeratin High Molecular Weight and p.63, Vimentin and ZO-1. Limbal stem cell deficiency (LSCD) and Descemet Membrane and Endothelial Keratoplasty (DMEK) are currently being performed in order to test the capabilities of SF membranes in vivo.

Results: Stromal and limbal stem cells showed a high proliferation capacity in human and rabbit primary cell cultures and were able to attach and grow onto SF membranes while maintaining their morphology and cellular markers (Vimentin and Cytokeratin) while showing a highly population of p.63 positive cells in the limbal stem cell cultures. Rabbit endothelial cells were effectively cultured on SF membrane while maintaining a positive stain of ZO-1, however, human endothelial cells were unable to attach and proliferate in vitro on this type of membrane.

Conclusions: SF membranes could be improved and studied for their use as a carrier for the treatment of corneal diseases.

Keywords: 482 cornea: epithelium • 481 cornea: endothelium  
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