Abstract
Purpose:
Previous studies have demonstrated that androgen treatments significantly influence gene expression in human meibomian gland epithelial (HMG) cells and conjunctival epithelial cells in vitro and in animal models. Androgen insufficiency is known to be a significant risk factor for the development of meibomian gland dysfunction (MGD). In this study we sought to determine the effect of two androgens, testosterone (TT) and dehydroepiandrosterone (DHEA) on HMG cell proliferation and lipid production.
Methods:
Immortalized HMG cells were cultured in serum free (KSFM) or serum containing media (DMEM/F12 containing 10% serum) in the absence or presence of TT at concentrations ranging from 1 nM to 1µM or DHEA at concentrations ranging from 12.5 µM to 100 µM. After 24h incubation, cell proliferation was evaluated using a live/dead viability assay kit. Lipid production in HMG cells was determined following staining with lipid dye Nile red and quantification of fluorescence measured on a Tecan Spectra Fluor Plus spectrophotometer.
Results:
Testosterone increased HMG cell proliferation in a dose dependent manner, with overall 19% (±6%) increase at 1 nM of TT, 29% (±11%) increase at 10 nM TT, and 37% (±17%) increase at 100 nM TT after treatment for 24 h. Increased lipid productions from 37% (±19%) up to 62% (± 26%) were observed in HMG cells after 24 h treatments of TT at concentrations ranging from 1 nM to 1µM. HMG cell proliferation increased by 33% (±13%) and 44% (±16%) after 24 h incubation with 12.5 and 25 µM of DHEA (p<0.05), respectively. Differentiated cells showed a 46% (±5%) and 43% (±1%) increase in lipid production when treated with DHEA at 12.5 and 50 µM respectively for 24 h.
Conclusions:
Androgens increased cell proliferation and lipid production in HMG cells up to 24h. However, the long term effect of androgen on cell proliferation and lipid production remains to be determined. These findings may be useful in the development of androgen based topical therapy for the treatment of MGD.
Keywords: 654 proliferation •
583 lipids