Apoptosis of Corneal Stromal Cells Induced by Telopeptides: An In vitro Study
Vimalin Jeyalatha
L&T Microbiology Research Centre, Vision Research Foundation, Sankara Nethralaya 41, College Road, Chennai, 600 006, India and Birla Institute of Technology and Science (BITS), Pilani-333 031, Rajasthan, India
Jambulingam Malathi *
L&T Microbiology Research Centre, Vision Research Foundation, Sankara Nethralaya 41, College Road, Chennai, 600 006, India
Prema Padmanabhan
Department of Cornea and Refractive Surgery, Medical Research Foundation, Sankara Nethralaya 41, College Road, Chennai, 600 006, India
Hajib N. Madhavan
L&T Microbiology Research Centre, Vision Research Foundation, Sankara Nethralaya 41, College Road, Chennai, 600 006, India
*Author to whom correspondence should be addressed.
Abstract
Background: Uncontrolled collagen degradation is the characteristics of keratoconus. Degradation of type 1 collagen releases telopeptides. The role of telopeptides on the apoptosis of corneal stromal cells have not been studied so far.
Methods: Human primary corneal stromal cells was cultivated and treated with varying concentrations of synthetic telopeptides (3.012µg, 6.125µg, 12.25µg, 12.25µg, 23.5µg, 47µg and 94µg) and incubated for 24 hours, 48 hours and 72 hours. MTT and TUNEL assay was performed following incubation. The difference between the number of viable cells present in the treated and untreated cells were considered for the analysis.
Results: Primary corneal stromal cells treated with varying concentrations of synthetic telopeptide at 24 hours and 48 hours had no morphological or apoptotic changes, the viability remained 100%. The percentage viability was altered after 72 hours of incubation with the synthetic telopeptide. 47µg/ml, 94µg/ml concentrations of telopeptide showed considerable decrease in the cell viability (p<0.05, t test).
Conclusion: The current study aims to evaluate the effect of telopeptides on primary corneal stromal cells in vitro and results revealed that the synthetic telopeptide had apoptotic effect. Similar phenomenon can be exhibited in keratoconus stromal cells.
Keywords: Keratoconus, keratocytes, apoptosis, telopeptides, collagen