INVESTIGATION OF ENDOTHELIAL CONTROL OF BONE BLOOD FLOW: DEVELOPMENT OF A RABBIT TIBIA EX VIVO PERFUSION MODEL

Abstract

An ex vivo rabbit tibia perfusion model was developed to investigate the vascular reactivity and vascular endothelial function of bone tissue. The tibiae of 36 adult New Zealand rabbits were isolated and perfused through the cannulated nutrient artery at a constant flow rate in an organ chamber. The increase in perfusion pressure caused by graded doses of norepinephrine produced a dose response curve, which was first recorded under control condition, and then during perfusion with acetylcholine (ACh), N G -monomethyl-L-arginine acetate (L-NMMA), and/or L-arginine (nitric oxide precursor) infusion. The optimal flow rate for rabbit tibia was 0.3 to 0.4 ml/min, depending on the weight of the tibia. The perfusion baseline and the norepinephrine dose-response curve (NEDRC) remained stable for at least eight hours. Acetylcholine (1 × 10-5 M ) infusion attenuated the NEDRC (33 ± 5%) for 1.5 hours. Thereafter, the vasorelaxant effect of ACh diminished due to depletion of nitric oxide, and ultimately caused an increase in the NEDRC due to a vasoconstrictive action. The rebound vasoconstriction did not occur in the presence of constant L-arginine infusion, which maintained nitric oxide production, and ACh continued to attenuate the NEDRC (36 ± 4.5%) for at least eight hours. L-NMMA, which inhibits the synthesis of nitric oxide, increased the vasoconstrictive effect of norepinephrine significantly (155 ± 16%). The rabbit tibia ex vivo model seems ideal for the study of bone vascular bed activity, due to its stable baseline, reliable NEDRC and cost efficiency. We have further demonstrated that the intraosseous vascular endothelium of rabbit tibia produces nitric oxide as one method of regulating bone perfusion.