Abstract
PurposeTo examine the effect of prolactin (PRL) on human corneal stromal fibroblasts (CSFs), derived from healthy individuals and from keratoconus (KC) patients, in vitro, specifically assessing physiological and elevated PRL concentrations as apparent during pregnancy.MethodsEye bank corneas of 3 female and 3 male healthy individuals as well as the corneal buttons of 3 female and 3 male KC patients were utilized for this study. The endothelium of the cornea was removed with sterile surgical scalpels, the probes were washed repeatedly with Dulbecco’s PBS and corneoscleral rims were trimmed off. Subsequently the corneal stroma was digested with collagenase type I and the harvested CSFs were cultured. We then examined (1) cell proliferation, (2) cell viability and (3) cytokine release of CSFs upon exposure to prolactin in vitro.ResultsWith respect to viability and proliferation our experiments did not show significant differences between CSFs exposed to different PRL concentrations. Our data show a significantly lower IL-8 concentration in normal CSFs exposed to 10ng/ml PRL compared to 0ng/ml and 1000ng/ml at 5 hours post exposition. Moreover, we can report significantly lower secretion of IL-8, IL-6, HGF, VEGF and FGFb in KC CSFs compared to normal CSFs, independent of PRL exposure, as determined by cytokine ELISA.ConclusionOur data in part points towards corneal cytokine secretion as a possible link between altered stromal PRL concentrations and KC progression. However, in our small dataset a significant influence of PRL concentration on cytokine secretion can only be described for IL-8 in normal CSFs. Further our results contribute to existing reports on the importance of cytokines in KC development, with an emphasis on significantly lower cytokine secretion in KC CSFs compared to normal controls.
Highlights
Keratoconus (KC) is a progressive condition, in which the cornea acquires an ectatic cone-like shape, which is caused by thinning of the corneal stroma [1]
With respect to viability and proliferation our experiments did not show significant differences between corneal stromal fibroblasts (CSFs) exposed to different PRL concentrations
Our data show a significantly lower IL-8 concentration in normal CSFs exposed to 10ng/ml PRL compared to 0ng/ ml and 1000ng/ml at 5 hours post exposition
Summary
Keratoconus (KC) is a progressive condition, in which the cornea acquires an ectatic cone-like shape, which is caused by thinning of the corneal stroma [1]. Even though KC has been described as a non-inflammatory disease for a long time, recent studies have reported a significant role of cytokines in KC development, suggesting the involvement of inflammatory mechanisms [3, 4]. Keratocytes are the housekeepers of the corneal stroma, contributing to corneal homeostasis [5]. Quiescent, these cells are activated in pathological processes and, hold a pivotal position in the KC trajectory [6]. Hormonal imbalances have been connected to KC development: for instance, changes in T4 concentrations, like apparent in thyroid gland dysfunction, have been shown to alter the expression of collagens within the cornea [7]
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