title: Anabolic potential of bone mineral in human periosteal fibroblasts using steroid markers of healing.
abstract: 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 5 alpha-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 (2 ml x 2) 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<0.001; one way ANOVA). Combined incubations of B and M resulted in similar yields. F inhibited DHT yields with both radiolabelled substrates by 2-3-fold (n=6; p<0.001) which was overcome by a combined incubation of F+B to values similar to those of controls (p<0.01). Documented pro-anabolic effects of minocycline were applicable as a standard for confirmation of responses to B. Significant increases in yields of DHT in response to B and M with both substrates indicate their anabolic potential in periosteal fibroblasts with implications for wound healing.
authors: Formerly at King's College London Dental Institute, Periodontology, King's College Dental Hospital, Denmark Hill, London SE5 9RW, UK.
authors: A
authors: A
authors: Suchak
authors: M
authors: M
authors: Soory
mesh_terms: D000328:Adult; D015113:Androstane-3,17-diol / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001842:Bone and Bones / Q000378:metabolism*; D002417:Cattle; D013196:Dihydrotestosterone / Q000378:metabolism; D004347:Drug Interactions; D005260:Female; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism*; D018120:Finasteride / Q000494:pharmacology; D006801:Humans; D008297:Male; D051379:Mice; D008875:Middle Aged; D008903:Minerals / Q000494:pharmacology*; D008911:Minocycline / Q000494:pharmacology; D010521:Periosteum / Q000166:cytology*; D013256:Steroids / Q000378:metabolism*; D013739:Testosterone / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects*
chemical_list: D015415:Biomarkers; D008903:Minerals; D013256:Steroids; D013196:Dihydrotestosterone; D015113:Androstane-3,17-diol; D013739:Testosterone; D018120:Finasteride; D008911:Minocycline