Comparison of cytotoxic effects of bisphosphonates in vitro and in vivo.
We compared the cytotoxic effects of alendronate (ALN) and incadronate (YM175) on isolated rabbit osteoclasts in vitro and on rats in vivo. In the in vitro experiment, each bisphosphonate was added to the culture of isolated osteoclasts at the final concentration of 3 x 10(-5), 3 x 10(-4), or 3 x 10(-3) M, and the amount of creatine phosphokinase (CPK) released into the medium was taken as an index of cytotoxicity at 5, 10, and 24 hours after the treatment. Also viability of osteoclasts, measured in terms of trypan blue exclusion, was assessed at 24 hours after the treatment. In YM175-treated groups, CPK activity in the medium increased in a concentration-dependent manner with time, and phase-contrast microscopic observation revealed damaged cell membranes and nuclear deterioration in YM175-treated osteoclasts. As a result, the viability of the osteoclasts was decreased at the concentrations of 3 x 10(-4) and 3 x 10(-3) M. However, in the ALN-treated groups, neither CPK activity nor viability of isolated osteoclasts changed significantly compared with control levels even at 3 x 10(-3) M for up to 24 hours. In the in vivo experiment, each bisphosphonate was administered separately to normal rats (7 weeks old, Sprague-Dawley) by intravenous injection at 1, 5, or 25 nmol/kg. Two days after the injection, the animals were euthanized, and the plasma Ca concentration and total CPK activity were measured. In YM175-injected rats, the CPK activity increased at 25 mmol/kg, and a slight decrease in the plasma Ca level was seen at this dose. In contrast, in ALN-injected rats, CPK activity did not increase even at 5 or 25 mmol/kg, and the plasma Ca level did decrease significantly compared with controls. Isozyme analysis revealed that, not only was CPK activity increased in the BB type in YM175-injected rats, it was also increased in the MB and MM types. In conclusion, alendronate, unlike YM175, does not have any cytotoxic effects on osteoclasts either in vitro or in vivo.
Ito
M
M
Pharmacological Research Department, TEIJIN Institute for Bio-Medical Research, Tokyo, Japan.
Chokki
M
M
Ogino
Y
Y
Satomi
Y
Y
Azuma
Y
Y
Ohta
T
T
Kiyoki
M
M
D019386:Alendronate / Q000633:toxicity*; D000818:Animals; D002118:Calcium / Q000097:blood; D002462:Cell Membrane / Q000187:drug effects / Q000473:pathology; D002470:Cell Survival / Q000187:drug effects; D003402:Creatine Kinase / Q000378:metabolism; D004164:Diphosphonates / Q000633:toxicity*; D007275:Injections, Intravenous; D007527:Isoenzymes; D008297:Male; D010010:Osteoclasts / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D011817:Rabbits; D051381:Rats; D017207:Rats, Sprague-Dawley; D014343:Trypan Blue / Q000378:metabolism
D004164:Diphosphonates; D007527:Isoenzymes; C071542:cimadronate; D003402:Creatine Kinase; D014343:Trypan Blue; D002118:Calcium; D019386:Alendronate