Effect of Microtopographical Cues on Human Keratocyte Orientation and Gene Expression

Abstract

Purpose: To determine if microtopographical cues can influence the orientation and extracellular matrix production of human keratocytes in vitro.

Materials and Methods: Human keratocytes were cultured on grooved and ungrooved polycaprolactone films for up to 3 weeks. The cell morphology was examined using ordinary light microscopy, reflective microscopy, and scanning electron microscopy. The cells gene expression was examined using a GEArray Q series gene expression kit.

Results: Cells initially appeared to orientate in the direction of the grooves. Cells cultured on ungrooved films exhibited random orientations. For longer culture periods on grooved membranes, a second cell layer formed on top of the initial layer at an angle orientation to the initial layer. Analysis of mRNA showed that several genes involved in the production of integrins and matrix metalloproteinases were either up-regulated or down-regulated in the presence of the grooves.

Conclusions: The introduction of microtopographical cues has been shown to influence the orientation of keratocytes and alter their gene expression. This pilot study reveals some important findings that can be used in the development of bioengineered corneas.

KEYWORDS::

• Bioengineered cornea
• Collagen fibrils
• Grooved-polycaprolactone
• Keratocytes
• Microtopographical cues

ACKNOWLEDGMENT

We would like to acknowledge Prof. A. Curtis in Glasgow University for providing us with the polycaprolactone for this project; Mr. P McDonell and Mr. S Shah from Birmingham and Midland Eye Centre for technical assistance. KT acknowledges The Priory of the Order of St.John Research Fellowship in Ophthalmology and North Staffordshire Medical Institute for the funding.

Declaration of interest: The authors report no conflict of interest. The authors alone are responsible for content and writing of the paper.