Development of stromal differentiation patterns in heterotypical models of artificial corneas generated by tissue engineering
We carried out a histological characterization analysis of the stromal layer of human heterotypic cornea substitutes generated with extra-corneal cells to determine their putative usefulness in tissue engineering. Human bioartificial corneas were generated using nanostructured fibrin-agarose biomate...
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Published in | Frontiers in bioengineering and biotechnology Vol. 11; p. 1124995 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Switzerland
Frontiers Media S.A
23.03.2023
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Subjects | |
Online Access | Get full text |
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Summary: | We carried out a histological characterization analysis of the stromal layer of human heterotypic cornea substitutes generated with extra-corneal cells to determine their putative usefulness in tissue engineering.
Human bioartificial corneas were generated using nanostructured fibrin-agarose biomaterials with corneal stromal cells immersed within. To generate heterotypical corneas, umbilical cord Wharton's jelly stem cells (HWJSC) were cultured on the surface of the stromal substitutes to obtain an epithelial-like layer. These bioartificial corneas were compared with control native human corneas and with orthotypical corneas generated with human corneal epithelial cells on top of the stromal substitute. Both the corneal stroma and the basement membrane were analyzed using histological, histochemical and immunohistochemical methods in samples kept in culture and grafted
for 12 months in the rabbit cornea.
Our results showed that the stroma of the bioartificial corneas kept
showed very low levels of fibrillar and non-fibrillar components of the tissue extracellular matrix. However,
implantation resulted in a significant increase of the contents of collagen, proteoglycans, decorin, keratocan and lumican in the corneal stroma, showing higher levels of maturation and spatial organization of these components. Heterotypical corneas grafted
for 12 months showed significantly higher contents of collagen fibers, proteoglycans and keratocan. When the basement membrane was analyzed, we found that all corneas grafted
showed intense PAS signal and higher contents of nidogen-1, although the levels found in human native corneas was not reached, and a rudimentary basement membrane was observed using transmission electron microscopy. At the epithelial level, HWJSC used to generate an epithelial-like layer in
corneas were mostly negative for p63, whereas orthotypical corneas and heterotypical corneas grafted
were positive.
These results support the possibility of generating bioengineered artificial corneas using non-corneal HWJSC. Although heterotypical corneas were not completely biomimetic to the native human corneas, especially
,
grafted corneas demonstrated to be highly biocompatible, and the animal cornea became properly differentiated at the stroma and basement membrane compartments. These findings open the door to the future clinical use of these bioartificial corneas. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Present addresses: Carmen Morales-Álvarez, GENYO, Centre for Genomics and Oncological Research: Pfizer, University of Granada, Andalusian Regional Government, PTS Granada, Granada, Spain; Department of Biochemistry and Molecular Biology III, Faculty of Medicine, University of Granada, Granada, Spain Reviewed by: Andri K. Riau, Singapore Eye Research Institute (SERI), Singapore These authors have contributed equally to this work Edited by: Mona Kamal Marei, Alexandria University, Egypt Damien Harkin, Queensland University of Technology, Australia This article was submitted to Tissue Engineering and Regenerative Medicine, a section of the journal Frontiers in Bioengineering and Biotechnology |
ISSN: | 2296-4185 2296-4185 |
DOI: | 10.3389/fbioe.2023.1124995 |