Publication:
Keratin-chitosan membranes as scaffold for tissue engineering of human cornea

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Vazquez-30-813-821-2015.pdf(10.75 MB)
Date
2015
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Authors
Vázquez, Natalia ; Chacón, Manuel ; Meana, Álvaro ; Menéndez-Menénde, Yolanda z ; Ferrero-Gutierrez, Amaia ; Cereijo-Martín, David ; Naveiras, Miguel ; Merayo-Lloves, Jesús
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Publisher
F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología
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DOI
DOI: 10.14670/HH-11-585
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info:eu-repo/semantics/article
Description
Abstract
Purpose: To study the attachment and growth of human corneal cells on keratin-chitosan membranes. The end goal is to develop a bioengineered cornea based on this material. Methods: Keratin-chitosan membranes were prepared as previously described by Tanabe et al., 2002. Briefly, 7.15 mg/cm2 of keratin dialysate was mixed with 10wt% chitosan solution and 20 wt% glycerol. The solution was cast into a silicone mold and dried at 50°C for 36 hours. Eyes were attained from a local eye bank after penetrant-keratoplastic surgery. Human epithelial, stromal and endothelial cells were obtained of the limbal, stromal and endothelial regions. Cells were cultured on keratin-chitosan membranes, as well as on plastic dishes as controls. When cultured cells reached confluence, they were fixed, incubated with primary antibodies (E-cadherin, cytokeratin high molecular weight (CK), vimentin and Na+/K+ ATPase) and visualized by indirect immunocytochemistry. Results: Epithelial, stromal and endothelial cells were able to attach and grow on keratin-chitosan membranes. All the cells maintained their morphology and cellular markers, both in the membrane and on the culture plate. Epithelial cells stained positively for CK and Ecadherin. A positive vimentin stain was observed in all stromal cells, while endothelial cells were positive forvimentin and Na+/K+ ATPase, but negative for Ecadherin. Conclusions: Keratin-chitosan membranes have been shown to be a good scaffold for culturing epithelial, stromal and endothelial corneal cells; therefore, future applications of keratin-chitosan membranes may be developed for reconstruction of the cornea.
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Keratin-chitosan scaffold , Human limbal epithelial cells , Human corneal stromal cells , Human corneal endothelial cells
Citation
Histology and Histopathology, vol. 30, nº 7, (2015)
URI
http://hdl.handle.net/10201/96581
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Histology and histopathology Vol.30, nº7 (2015)
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