Opposing effects of bisphosphonates and advanced glycation end-products on osteoblastic cells.
Patients with long-standing Diabetes mellitus can develop osteopenia and osteoporosis. We have previously shown that advanced glycation endproducts reduce the bone-forming activity of osteoblasts. Bisphosphonates are used for the treatment of various bone disorders, since they reduce osteoclastic function and survival, and stimulate osteoblastic bone-forming capacity. In this work we have investigated whether bisphosphonates are able to revert advanced glycation endproducts-induced deleterious effects in osteoblasts. MC3T3E1 and UMR106 osteoblastic cells were incubated with control or advanced glycation endproducts-modified bovine serum albumin, in the presence or absence of different doses of the bisphosphonates Alendronate, Pamidronate or Zoledronate. After 24-72 h of culture, we evaluated their effects on cell proliferation and apoptosis, type-1 collagen production, alkaline and neutral phosphatase activity, and intracellular reactive oxygen species production. Advanced glycation endproducts significantly decreased osteoblast proliferation, alkaline phosphatase activity and type 1 collagen production, while increasing osteoblastic apoptosis and reactive oxygen species production. These effects were completely reverted by low doses (10(-8) M) of bisphosphonates. High doses of bisphosphonates (10(-4)-10(-5) M) were toxic for osteoblasts. Nifedipine (L-type calcium channel blocker) did not affect the advanced glycation endproducts-induced decrease in osteoblastic proliferation, although it blocked the reversion of this effect by 10(-8) M Alendronate. Both advanced glycation endproducts and Alendronate inhibited the activity of intracellular neutral phosphatases. In conclusion, we show that bisphosphonates revert the deleterious actions of advanced glycation endproducts on osteoblastic cells, and that these effects of bisphosphonates depend on: (a) Ca(2+) influx through L-type voltage-sensitive channels, and (b) blockage of advanced glycation endproducts-induced reactive oxygen species generation.
Gangoiti
María Virginia
MV
Facultad de Ciencias Exactas, Universidad Nacional de La Plata, Argentina.
Cortizo
Ana María
AM
Arnol
Verónica
V
Felice
Juan Ignacio
JI
McCarthy
Antonio Desmond
AD
D016475:3T3 Cells; D019386:Alendronate / Q000008:administration & dosage / Q000494:pharmacology; D000818:Animals; D050071:Bone Density Conservation Agents / Q000008:administration & dosage / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D048909:Diabetes Complications / Q000503:physiopathology; D004164:Diphosphonates / Q000008:administration & dosage / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D017127:Glycation End Products, Advanced / Q000009:adverse effects*; D007093:Imidazoles / Q000008:administration & dosage / Q000494:pharmacology; D051379:Mice; D010006:Osteoblasts / Q000187:drug effects* / Q000378:metabolism; D010024:Osteoporosis / Q000209:etiology / Q000503:physiopathology; D000077268:Pamidronate; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000009:adverse effects*; D013997:Time Factors; D000077211:Zoledronic Acid
D050071:Bone Density Conservation Agents; D020746:Calcium Channels, L-Type; D004164:Diphosphonates; D017127:Glycation End Products, Advanced; D007093:Imidazoles; D017382:Reactive Oxygen Species; C081628:advanced glycation end products-bovine serum albumin; D012710:Serum Albumin, Bovine; D000077211:Zoledronic Acid; D000077268:Pamidronate; D002118:Calcium; D019386:Alendronate