Effects of metallic ions and diphosphonates on inhibition of pericardial bioprosthetic tissue calcification and associated alkaline phosphatase activity.
This study focused on the association of extrinsic alkaline phosphatase (AP) activity with both early and advanced calcification of glutaraldehyde-pretreated bovine pericardial bioprosthetic (GPBP) tissue, and the inhibition of both calcification and AP activity by pre-incubation in diphosphonates (sodium-ethanehydroxydiphosphonate [NaEHDP], aminopropanehydroxydiphosphonate [APD]) and metallic salts (FeCl3, Ga(NO3)3, AICI3). GPBP specimens were implanted subcutaneously in 3 wk old male rats after pre-incubation. Following explantation of the tissue at 72 h and 21 d, calcification was assessed morphologically by light microscopy and chemically by atomic adsorption spectroscopy for calcium content and by molybdate complexation for phosphorus. AP activity was characterized by enzymatic hydrolysis of paranitrophenyl phosphate and by histochemical studies. In both control and pretreated groups, AP levels were greater in 72 h explants than 21 d retrievals, which demonstrated extensive calcification in control explants. All pre-incubations that resulted in inhibition of calcification after 21 d, except for APD, were associated with 72 h AP content which was lower than control specimens. The typical time of initiation of calcification was 72 h, as determined by previous studies with this model system. Covalently bound APD inhibited calcification. Increased AP activity in the APD group may be due to the toxicity of this agent with resultant acute inflammation, or other incompletely understood effects of diphosphonates on calcification and AP. Furthermore, EHDP and Ga3+ incubations were also associated with decreased GPBP AP at 72 h compared to control, but were not effective for inhibiting calcification after 21 d. We concluded that inhibition of peak GPBP AP activity is not necessarily associated with the prevention of GPBP mineralization.
Department of Pediatrics, University of Michigan Medical School, Ann Arbor 48109-0576.
D
D
Hirsch
F J
FJ
Schoen
R J
RJ
Levy
D000469:Alkaline Phosphatase / Q000378:metabolism*; D000818:Animals; D001705:Bioprosthesis; D002114:Calcinosis / Q000517:prevention & control*; D002417:Cattle; D004164:Diphosphonates / Q000494:pharmacology*; D012968:Etidronic Acid / Q000494:pharmacology*; D005976:Glutaral; D006868:Hydrolysis; D008297:Male; D008670:Metals / Q000494:pharmacology*; D008854:Microscopy, Electron; D000077268:Pamidronate; D010496:Pericardium / Q000187:drug effects* / Q000201:enzymology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013054:Spectrophotometry, Atomic
D004164:Diphosphonates; D008670:Metals; D000469:Alkaline Phosphatase; D012968:Etidronic Acid; D000077268:Pamidronate; D005976:Glutaral