title: Purinergic transmitters inhibit bone formation by cultured osteoblasts.
abstract: Adenosine triphosphate (ATP) and other purinoceptor agonists cause a transient rise in [Ca2+]i in cultured osteoblast-like cells and have a mitogenic effect, as does parathyroid hormone (PTH), and there is evidence that ATP and PTH can act synergistically on osteoblasts. The likelihood that nucleotides, acting through purinoceptors, are important local factors in bone remodeling is therefore considerable. However, their effect on bone formation is unknown. We recently developed a culture system in which appositional bone formation occurs only in narrow grooves cut in a substratum. We have used this as an assay to measure the effects of ATP (50 and 500 mumol/L), ATP gamma S (20 mumol/L), 2-MeSATP (2 and 20 mumol/L), uridine triphosphate (UTP) (0.2, 2, and 20 mumol/L), adenosine (20 mumol/L), bovine PTH (0.25 and 0.5 IU/mL), rat PTH1-34 (10(-8) and 10(-7) mol/L), and rat PTHrP1-40 (10(-9) and 10(-8) mol/L) on bone formation by rat calvarial osteoblasts. The culture medium was renewed 3 times/week (every 2 or 3 days), and the number of bone loci and length and area of Alizarin red-stained mineralized bone formed in the grooves of each specimen in 16-29 days were measured. Compared with controls, ATP gamma S, 2-MeSATP, and ATP reduced the amount of bone formed in a 2-3 week culture period. Adenosine had no effect, and UTP either had no effect or at 2 mumol/L stimulated bone formation. PTH and PTHrP completely abolished bone formation in 4 week cultures. Our findings are consistent with evidence for more than one P2 purinoceptor subtype in bone, and show for the first time that the effect of ATP on appositional bone formation by osteoblasts in vitro is, like PTH and PTHrP, inhibitory.
authors.affiliation: Department of Anatomy and Developmental Biology, University College London, UK. s.jones@ucl.ac.uk
authors.forename: S J
authors.initials: SJ
authors.lastname: Jones
authors.forename: C
authors.initials: C
authors.lastname: Gray
authors.forename: A
authors.initials: A
authors.lastname: Boyde
authors.forename: G
authors.initials: G
authors.lastname: Burnstock
mesh_terms: D000241:Adenosine / Q000494:pharmacology; D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D001846:Bone Development / Q000187:drug effects*; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D004357:Drug Synergism; D010006:Osteoblasts / Q000187:drug effects* / Q000378:metabolism; D010281:Parathyroid Hormone / Q000494:pharmacology*; D044162:Parathyroid Hormone-Related Protein; D011506:Proteins / Q000494:pharmacology; D058913:Purinergic Agonists; D051381:Rats; D017207:Rats, Sprague-Dawley; D013329:Structure-Activity Relationship; D013873:Thionucleotides / Q000494:pharmacology; D014544:Uridine Triphosphate / Q000494:pharmacology
chemical_list: D010281:Parathyroid Hormone; D044162:Parathyroid Hormone-Related Protein; D011506:Proteins; D058913:Purinergic Agonists; D013873:Thionucleotides; C030812:adenosine 5'-O-(2-thiodiphosphate); D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000241:Adenosine; D002118:Calcium; D014544:Uridine Triphosphate