Postdoctoral Research Fellow University of Sydney Sydney, New South Wales, Australia
Abstract: Limbal stem cell deficiency (LSCD) is a major cause of blindness worldwide. The condition arises from the depletion or dysfunction of limbal epithelial stem cells (LESCs), a specialized stem cell population located in the limbus at the corneal edge. LSCD can arise from physical or chemical injuries, infections, or genetic causes, and its treatment poses significant challenges due to the loss of the cornea's natural immune privilege, heightening the risk of immune rejection. LESC transplantation is the most effective treatment for LSCD. However, achieving sufficient LESC transfer and precise placement in the eye are critical barriers to transplantation. We previously pioneered silicon hydrogel contact lenses (CLs) for LESC transplantation and demonstrated that CLs offer a practical substratum for adhesion, migration, and rapid expansion of LESCs. In this study, we aimed to assess the morphology, adhesiveness and proliferative capacity of LESCs on novel CSIRO's two distinct coatings, aminomalononitrile-30T (AMN-30T) and poly (ethylene glcol)-based cRGD. Human telomerase-immortalized limbal epithelial stem cells (T-LSCs) were seeded on coated and uncoated CLs for 2 weeks. The effectiveness of CLs in maintaining the T-LSCs morphology, adhesion, and cellular connections was evaluated using light and scanning electron microscopy (SEM). AMN-30T-coated CLs exhibited notable improvement in cell adhesion and proliferation compared to both cRGD-coated and control CLs, underscoring the impact of CL composition on cellular behaviour. While cRGD-coated CLs were less effective in promoting adhesion, cells seeded across all CL types were morphologically similar, typically uniform with evident cell-to-cell contacts and the development of multiple cell layers. SEM revealed extensive microvilli on the apical surface and cell projections that were suggestive of anchorage points on the CL surface. In conclusion, this study offers preliminary insights into the potential use of novel-coated CLs as therapeutic scaffolds for cultivating LESCs. Future studies employing cell-laden CLs in a murine model of LSCD will further validate their efficacy for LESC transplantation and ocular surface disease treatment.
Funding Source: This research was funded by the Medical Research Future Fund (MRFF) Stem Cell Therapies Mission Grant (Application ID: 20222757).
