A feline assay using osteoclasts generated in vitro from peripheral blood for screening anti-resorptive agents.
Musculo-skeletal diseases are a major cause of pain and suffering in cats and several conditions involve increased bone resorption by osteoclasts. However, little is known about the biology of these cells in the cat. In this study we established a method to generate feline osteoclasts from blood mononuclear cells stimulated by macrophage colony stimulating factor (M-CSF) and receptor activator of NF-kappaB ligand (RANKL). Cultured osteoclasts are multinucleated, express tartrate resistant acid phosphatase (TRAP), form F-actin rings and resorb bone. They express alpha(v)beta3 vitronectin receptor and osteoclast enzymes, cathepsin K and MMP9; the myeloid antigen, CD18, and the megakaryocyte/platelet integrin, CD41, are absent. This phenotype is typical of osteoclasts from other species. Three resorption inhibitors were examined for activity against feline osteoclasts. Calcitonin, bisphosphonate and RGD integrin inhibitory peptide all reduced bone resorption at doses similar to those efficacious in rabbit or human. We conclude that blood-derived osteoclast cultures are a suitable in vitro system for assessing the ability of drugs to inhibit bone resorption in domestic cats.
Muzylak
M
M
Bone and Mineral Centre, Department of Medicine, University College London, UK.
Flanagan
A M
AM
Ingham
K
K
Gunn
N
N
Price
J
J
Horton
M A
MA
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D001862:Bone Resorption / Q000188:drug therapy* / Q000378:metabolism / Q000662:veterinary*; D018821:CD18 Antigens / Q000378:metabolism; D002116:Calcitonin / Q000494:pharmacology; D002371:Cat Diseases / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D056657:Cathepsin K; D002403:Cathepsins / Q000378:metabolism; D002415:Cats; D018929:Cell Culture Techniques; D004164:Diphosphonates / Q000494:pharmacology; D019483:Disintegrins / Q000494:pharmacology; D007150:Immunohistochemistry / Q000662:veterinary; D007527:Isoenzymes / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D018613:Microscopy, Confocal / Q000662:veterinary; D010010:Osteoclasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D040201:Platelet Membrane Glycoprotein IIb; D019030:Receptors, Vitronectin / Q000378:metabolism; D000071681:Tartrate-Resistant Acid Phosphatase
D018821:CD18 Antigens; D004164:Diphosphonates; D019483:Disintegrins; D007527:Isoenzymes; D040201:Platelet Membrane Glycoprotein IIb; D019030:Receptors, Vitronectin; D002116:Calcitonin; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; D002403:Cathepsins; C534130:CTSK protein, human; D056657:Cathepsin K; D020780:Matrix Metalloproteinase 9