title: The use of tissue-nonspecific alkaline phosphatase (TNAP) and PHOSPHO1 inhibitors to affect mineralization by cultured cells.
abstract: Here, we describe methods to evaluate the ability of small molecules inhibitors of TNAP and PHOSPHO1 in preventing mineralization of primary cultures of murine vascular smooth muscle cells. The procedures are also applicable to primary cultures of calvarial osteoblasts. These cell-based assays are used to complement kinetic testing during structure-activity relationship studies aimed at improving scaffolds in the generation of pharmaceuticals for the treatment for medial vascular calcification.
authors.affiliation: Sanford Children's Health Research Center, Sanford-Burnham Medical Research Institute, La Jolla, CA, USA.
authors.forename: Tina
authors.initials: T
authors.lastname: Kiffer-Moreira
authors.forename: Sonoko
authors.initials: S
authors.lastname: Narisawa
mesh_terms: D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors*; D000818:Animals; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D057166:High-Throughput Screening Assays; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000201:enzymology*; D032389:Myocytes, Smooth Muscle / Q000201:enzymology* / Q000378:metabolism; D010006:Osteoblasts / Q000187:drug effects / Q000201:enzymology; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors*; D012886:Skull / Q000166:cytology; D013329:Structure-Activity Relationship; D061205:Vascular Calcification / Q000201:enzymology*
chemical_list: D004791:Enzyme Inhibitors; C552022:PHOSPHO1 protein, mouse; C510878:ALPL protein, mouse; D000469:Alkaline Phosphatase; D010744:Phosphoric Monoester Hydrolases