Effects of Mometasone Furoate on Human Keratinocytes and Fibroblasts in vitro

Abstract

The long-term treatment of inflammatory skin diseases with topical glucocorticoids is limited by their side effects such as skin atrophy, delayed wound healing and striae distensae. Mometasone furoate (MF) is a newly synthesized glucocorticoid with the advantage of increasing efficacy and reducing the number of adverse effects. The aim of our study was to compare the effects of MF and conventional fluorinated corticosteroids on a human keratinocyte cell line (HaCat) and human skin fibroblasts in vitro. Monolayer cultures of these cell lines were exposed to different concentrations of the active compounds for 5 days to analyze the influence on morphology and proliferation. Chemotaxis of HaCat cells and fibroblasts was studied in blind-well Boyden chambers using collagen type I and fibroblast-conditioned medium as a chemoattractant. Additionally, fibroblasts were used to investigate the contraction of collagen gels since lattice contraction appears to model the contraction of skin wounds. All glucocorticoids tested influenced fibroblast and keratinocyte proliferation in a dose-dependent manner, yet the effect was clearly more marked with fluorinated corticosteroids than with MF. Similar effects were obtained using the chemotaxis assay. At low concentrations (10^–9 M) MF exerted almost no influence, while the conventional fluorinated substances inhibited direct migration significantly. Contraction of collagen gels was inhibited completely by betamethasone valerate at high concentrations (10^–5– 10^–3 M), but only partially inhibited by MF at its highest concentration (10^–3 M). Although MF reveals high anti-inflammatory activity similar to that known for conventional fluorinated derivatives of corticosteroids, the study shows that MF has less effect in the tested in vitro systems. Therefore, it remains to be seen whether these data might indicate the possibility of a dissociation between the inflammatory activity and the inhibition of the biosynthetic capacities of fibroblasts and keratinocytes by modification of the steroidal structure of corticosteroids.

Fussel-15, a New Player in Wound Healing, Is Deregulated in Keloid and Localized Scleroderma

“…7 The lattices were prepared in ultra-low attachment 6-well plates (Corning Life Sciences, Lowell, MA), combining 1.5 mL of a 1.5ϫ concentrated medium containing FCS with 0.35 mL of collagen solution (3.85 mg/mL; BD Biosciences) and 0.25 mL of cell suspension (1 ϫ 10 6 cells/mL). This mixture polymerizes within 20 minutes at 37°C.…”

Test systems for the determination of glucocorticoid receptor ligand induced skin atrophy

“…Discrepancies observed between in vitro experiments might be due to indirect effects of GCs on fibroblasts by affecting the synthesis or actions of various factors produced by other cell types. Yet, more recent studies show anti-proliferative effects of GCs on primary human fibroblasts 20,26 and HaCaT cells, a human keratinocyte cell line, 20 only. Beside their anti-proliferative effects in cutaneous cells, GCs also affect collagen metabolism.…”

“…19 Skin atrophy can be modeled in vitro, in vivo and in humans. Classically, in vitro tests for skin atrophy assess proliferation of cutaneous cells, 20 or collagen synthesis in primary human fibroblasts. 21 Recently, epidermal thickness and collagen synthesis in three-dimensional full-thickness skin models (FTSM) were developed to determine GC ligand induced skin atrophy in vitro.…”

320 Activated hepatic stellate cells express keratinocyte growth factor in chronic liver disease

“…31 Human keratinocytes were obtained from CellSystems (St. Katharinen, Germany). Fibroblasts and keratinocytes were cultured on uncoated tissue culture flasks in Dulbecco's modified Eagle's medium supplemented with 10% heat-inactivated fetal calf serum, 100 IU/ml penicillin, and 100 g/ml streptomycin in a humidified atmosphere of 5% CO 2 in air.…”