[In vitro study of the effect of bisphosphonates on mineralization induced by a composite material: poly 2(hydroxyethyl) methacrylate coupled with alkaline phosphatase].
We have immobilized the mineralizing agent alkaline phosphatase (AlkP) in a hydrophilic polymer (poly 2(hydroxyéthyl) methacrylate) (pHEMA) in a copolymerization technique. Histochemical study on polymer sections revealed that AlkP has retained its biological activity. The image analysis of sections using a tessellation method showed a lognormal distribution of the area of the tiles surrounding AlkP particles thus confirming a homogeneous distribution of the enzyme in the polymer. Pellets of pHEMA-AlkP were incubated with a synthetic body fluid containing organic phosphates (beta-glycerophosphate). Mineral deposits with a rounded shape (calcospherites) were obtained in about 17 days. We have investigated the effects of three bisphosphonates (etidronate, alendronate and tiludronate) on this system. Bisphosphonates at a concentration of 10(-2) M totally inhibited AlkP in solution at a concentration of 10(-4) mg/ml. Inhibition has been reported being due to the chelation of a metal cofactor (Zn2+). Etidronate and alendronate appeared to inhibit the calcospherite deposition onto the pHEMA-AlkP material in a similar way. Both bisphosphonates possess three sites for mineral complexion. On the other hand, tiludronate having only two sites was associated with a reduced inhibitory effect on mineralization. When used in microgravity conditions, mineralization was impaired with etidronate and larger crystals were obtained with tiludronate. However, these effects were obtained in non-physiological conditions (a 20 degrees C temperature was used during the STS80 flight of the space shuttle). The pHEMA-AlkP material provides an interesting method to study the effects of pharmacological compounds and environmental factors on the bone and cartilage mineralization process.
LHEA-Laboratoire d'Histologie-Embryologie, Faculté de Médecine, Angers, France.
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Barbier
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Chappard
D000469:Alkaline Phosphatase; D004164:Diphosphonates; D011102:Polyhydroxyethyl Methacrylate
D004164:Diphosphonates; D011102:Polyhydroxyethyl Methacrylate; D000469:Alkaline Phosphatase