Generation and activity of equine osteoclasts in vitro: effects of the bisphosphonate pamidronate (APD).
Equine osteoclast-like cells (OCLs) were generated from the bone marrow (BM) of two ponies and one horse in the presence of RANKL, the receptor activator of NF kappa B ligand and macrophage colony-stimulating factor (M-CSF). The phenotype of these cells was confirmed by demonstration of characteristics typical of osteoclasts (OCs) including: the expression of tartrate-resistant acid phosphatase (TRAP), the vitronectin receptor (VNR) and the calcitonin receptor (CTR), the demonstration of responsiveness to calcitonin (CT) and the ability to form resorption lacunae on ivory slices and calcium phosphate films. The bisphosphonate pamidronate (APD) dose-dependently inhibited resorption of calcium phosphate films by equine OCLs with an IC(50) of 5.8 x 10(-7) M in one horse. APD also dose-dependently inhibited the number of OCLs present in BM cultures after 7 days. However, this effect is most likely attributable to increased OCL death rather than decreased OCL formation. Paradoxically, ADP appeared to cause an early, transient, increase in OCL formation in BM cultures, however, this effect was reversed after 7 days. These preliminary in vitro data support the potential use of APD in clinical conditions characterised by increased bone turnover such as osteomyelitis, osteitis, septic osteoarthritis, navicular disease, cystic bone lesions and immobilisation-induced osteoporosis and provide useful information for future pharmacokinetic studies and clinical trials in vivo.
Gray
A W
AW
Equine Orthopaedic Research Group, University of Cambridge, Department of Clinical Veterinary Medicine, Madingley Road, Cambridge, UK.
Davies
M E
ME
Jeffcott
L B
LB
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D001853:Bone Marrow; D001862:Bone Resorption; D002130:Calcium Phosphates / Q000378:metabolism; D016923:Cell Death; D002478:Cells, Cultured; D004164:Diphosphonates / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation; D006736:Horses; D020128:Inhibitory Concentration 50; D007457:Iodine Radioisotopes; D010010:Osteoclasts / Q000166:cytology* / Q000187:drug effects*; D010012:Osteogenesis / Q000187:drug effects; D000077268:Pamidronate; D019030:Receptors, Vitronectin / Q000378:metabolism; D013194:Staining and Labeling; D013997:Time Factors
D002130:Calcium Phosphates; D004164:Diphosphonates; D007457:Iodine Radioisotopes; D019030:Receptors, Vitronectin; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; D000135:Acid Phosphatase; C485829:calcium phosphate, dibasic, anhydrous; D000077268:Pamidronate