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Effect of microtopographical cues on human keratocyte orientation and gene expression
Kong Y Then1, Ying Yang, Mark Ahearne
1Institute for Science & Technology in Medicine, Keele University, England, UK.
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.
