Anabolic potential of bone mineral in human periosteal fibroblasts using steroid markers of healing

• Published in: Steroids 2013-05, Vol.78 (5), p.462-467
• Main Authors: Suchak, A., Soory, M.
• Format: Article
• Language: English
• Subjects: Adult; Anabolic effects; Androstane-3,17-diol - metabolism; Animals; Biomarkers - metabolism; Bone and Bones - metabolism; Bone mineral; Cattle; Dihydrotestosterone - metabolism; Drug Interactions; Female; Fibroblasts - drug effects; Fibroblasts - metabolism; Finasteride - pharmacology; Humans; Male; Mice; Middle Aged; Minerals - pharmacology; Minocycline; Minocycline - pharmacology; Periosteal fibroblasts; Periosteum - cytology; Steroid markers; Steroids - metabolism; Testosterone - metabolism; Wound Healing - drug effects
• ISSN: 0039-128X; 1878-5867
• DOI: 10.1016/j.steroids.2013.02.002

► Deproteinized bone mineral has specific anabolic potential in periosteal fibroblasts. ► Specific AR mediated actions in response to finasteride are validated by minocycline. ► AR mediated specificity is relevant to actions of deproteinized bone matrix in vitro. ► Results are interpreted in the context of androgen metabolites as markers of healing. ► Utilization of two androgen substrates validates similar responses to testing agents. A deproteinized natural cancellous bone mineral (B) was studied in a cell culture model for its anabolic potential using two radiolabelled steroid substrates, 14C-testosterone (14C-T) and 14C-4-androstenedione (14C-4-A) independently; in the presence or absence of the anti-androgen finasteride (F) and minocycline (M). Culture medium was assayed for the biologically active metabolite 5alpha-dihydrotestosterone (DHT) a marker of regenerative potential and wound healing. Confluent monolayer cultures of human periosteal fibroblasts were incubated in Eagle’s minimum essential medium with each of the substrates 14C-T and 14C-4-A. Incubations were performed with previously established optimal concentrations of B5 (milligrams/ml), M25 (μg/ml) and F5 (μg/ml) alone and in combination (n=6) for 24h. The eluent was solvent extracted with ethyl acetate (2mlx2) and subjected to TLC in a benzene/acetone solvent system (4:1 v/v) for separation of metabolites; they were quantified using a radioisotope scanner. The yield of DHT was increased over controls in response to B and M with both substrates 14C-T and 14C-4-A by 1.7, 1.8-fold and 1.7, 1.6-fold respectively (n=6; p