Comparative effects of anti-inflammatory corticosteroids in human bone-derived osteoblast-like cells

Abstract

While effects of inhaled corticosteroids on serum markers of bone metabolism in normal and asthmatic subjects have been reported, there are little data on the direct effects of these corticosteroids on end-organs such as bone. The results presented here compare the effects of budesonide and its epimers (22S- and 22R-budesonide), fluticasone and dexamethasone on growth and differentiation of cultured human bone cells. Osteoblast-like cells were cultured from human foetal bone chips grown to confluence and used at first subculture. At concentrations of 10(-11)-10(-7) M each corticosteroid (CS) caused a dose-dependent decrease in [3H]thymidine incorporation into deoxyribonucleic acid (DNA), median effective concentration (EC50): fluticasone (0.06 nM) >22R (0.26 nM) >22S (0.4 nM) >budesonide (0.47 nM) >dexamethasone (1.5 nM). Each CS resulted in a dose-dependent increase in alkaline phosphatase activity, EC50: fluticasone (0.14 nM) >22R (0.2 nM)=22S (0.2 nM) >budesonide (0.4 nM) >dexamethasone (1.6 nM). The 1,25 dihydroxyvitamin D3 (1,25(OH)2D3)-stimulated osteocalcin production was decreased in the presence of each CS, EC50: fluticasone (0.02 nM) >22S (0.1 nM) >22R (0.2 nM) >budesonide (1.0 nM) >dexamethasone (1.8 nM). In human bone cells the potencies of fluticasone and budesonide in relation to dexamethasone are not dissimilar to those derived from human lymphocytes in vitro.

Effects of β2 Agonists, Corticosteroids, and Novel Therapies on Rhinovirus-Induced Cytokine Release and Rhinovirus Replication in Primary Airway Fibroblasts

“…Fluticasone is a more potent corticosteroid in inhibiting inflammation [22], and our data reflect this fact. It is also interesting to note that, although IL-8 is released more abundantly than IL-6 [23], this selective inhibition suggests that IL-8 may at least in part be steroid insensitive, and other studies have also demonstrated that certain cytokines are steroid insensitive [24, 25].…”

Clinical, cellular, microscopic, and ultrastructural studies of a case of fibrogenesis imperfecta ossium

“…Similar to the effects seen with BzATP, the patient’s cells responded in a paradoxical manner when treated with 1,25-dihydroxyvitamin D. Usually with increasing concentrations of 1,25-dihydroxyvitamin D, osteoblast cell proliferation decreases 6,10 and this was seen in the control biopsy cells. The patient cells, however, significantly increased proliferation in culture with a concentration of 10 −9  mol·L −1 1,25-dihydroxyvitamin D concentration( P <0.001; Figure 3c).…”

“…Cells were plated into 96-well plates at 1×10 5 cells per well and incubated at 37 °C for 24 h. The medium was then replaced with serum-reduced OptiMEM media and further incubated for 24 h, before the addition of treatments (benzoylbenzoyl-ATP (BzATP) or 1,25-dihydroxyvitamin D in vehicle) for another 24 h. Tritiated thymidine (final concentration 20 nCi·μL −1 ) was added to each well and incubated for 4 h. Cell proliferation was indirectly determined by the incorporation of tritiated (methyl,1′2- 3 H) thymidine, as previously described. 6 Scintillation vials were read for 5 min in a liquid beta-scintillation counter (Packard Tri-Carb 1990CA, Packard Instrument Company, Downers Grove, IL, USA). A bicinchoninic acid (BCA) assay was also carried out for cell proliferation measuring the amount of protein per well, in accordance with the manufacturer’s instructions.…”

“…Alkaline phosphatase was determined as previously described, 6 using a modification of the method of Lowry 7 that uses p -nitrophenyl phosphate as the substrate of alkaline phosphatase. Plates were read at 405 nm to detect the alkaline phosphatase product ( p -nitrophenolate).…”

Science and physicianly practice: Are they compatible?

“…If there is systemic absorption of inhaled corticosteroids, it is possible that there are unwanted effects on bone, because osteoporosis is a major long‐term side‐effect with oral corticosteroids. We investigated the effects of fluticasone and budesonide on human osteoblast‐like cells in vitro 16 and found that the rank order of potency was fluticasone > budesonide, reflecting the same order as for suppression of the hypothalamic–pituitary–adrenal axis 14 and for clinical efficacy 17 . These findings suggested that the more potent inhaled corticosteroids, such as fluticasone, are likely to be more potent in terms of unwanted effects if there is systemic absorption.…”