Nano-scale replication of the extracellular matrix underlying the anterior corneal epithelium of the primate

Christopher J Murphy, S. L. Goodman

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Purpose: To replicate the extracellular matrix underlying the anterior corneal epithelium with synthetic materials for anatomical study and (potentially) to improve epithelial cell behavior in corneal prosthetics. Methods: Freshly excised 5mm central corneal buttons from young adult Rhesus macaques were incubated in Dispase II or EDTA for variable time periods. The anterior corneal epithelium was completely removed using meticulous dissection to expose the underlying basement membrane, and the corneal button placed in a molding chamber. A methacrylate prepolymer was introduced into the chamber and allowed to polymerize. Tissue was then digested away and the resultant topographically inverse surface replica was imaged with high resolution SEM and compared to corneal tissue processed by conventional methods. The inverse replicas are then used as templates for preparing positive replicas of the matrix. Results: EDTA was superior to Dispase II by exposing cell-free matrix with minimal handling, and synthetic replica topography mimicked tissue morphology from the gross to the macromolecular scale (circa 50 nm). Conclusions: The nano-scale topography of the anterior corneal epithelial matrix may be fabricated with synthetic polymers, and hence recreate the 3-D structural-mechanical cellular environment of native tissue. Such biomimetic synthetic matrixes will be useful in examining surface topology effects on the anterior corneal epithelium, and may find application in improving cellular adhesion on corneal prosthetics. Support: NEI EY 10841-01, Whitaker Foundation. PrI: None.

Original languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number3
StatePublished - Feb 15 1996
Externally publishedYes

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Corneal Epithelium
Primates
Extracellular Matrix
Edetic Acid
Corneal Topography
Biomimetics
Methacrylates
Macaca mulatta
Basement Membrane
Dissection
Young Adult
Polymers
Epithelial Cells
dispase

ASJC Scopus subject areas

  • Ophthalmology

Cite this

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abstract = "Purpose: To replicate the extracellular matrix underlying the anterior corneal epithelium with synthetic materials for anatomical study and (potentially) to improve epithelial cell behavior in corneal prosthetics. Methods: Freshly excised 5mm central corneal buttons from young adult Rhesus macaques were incubated in Dispase II or EDTA for variable time periods. The anterior corneal epithelium was completely removed using meticulous dissection to expose the underlying basement membrane, and the corneal button placed in a molding chamber. A methacrylate prepolymer was introduced into the chamber and allowed to polymerize. Tissue was then digested away and the resultant topographically inverse surface replica was imaged with high resolution SEM and compared to corneal tissue processed by conventional methods. The inverse replicas are then used as templates for preparing positive replicas of the matrix. Results: EDTA was superior to Dispase II by exposing cell-free matrix with minimal handling, and synthetic replica topography mimicked tissue morphology from the gross to the macromolecular scale (circa 50 nm). Conclusions: The nano-scale topography of the anterior corneal epithelial matrix may be fabricated with synthetic polymers, and hence recreate the 3-D structural-mechanical cellular environment of native tissue. Such biomimetic synthetic matrixes will be useful in examining surface topology effects on the anterior corneal epithelium, and may find application in improving cellular adhesion on corneal prosthetics. Support: NEI EY 10841-01, Whitaker Foundation. PrI: None.",
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