Estudo comparativo da eficácia do etanol e do ácido L-glutâmico na prevenção da calcificação das cúspides e parede aórtica porcina: estudo experimental em ratos

INTRODUCTION: The glutataldehyde (GDA) treated pigs cusps are one of most employed tissues in bioprosthesis, but is late post-implant calcification is main cause of its failure. BACKGROUND: This study aims at comparing and analyzing two methods (ethanol 80% and l-glutamic acid 0.8%) to prevent calcification in pig cusps and aortic wall implanted subcutaneously in rats, the cusps and aortic wall segments of the control were in glutaraldehyde (GDA), during a 15, 30 and 60 days period after the implant. MATERIAL AND METHODS: We used 45 young rats, distributed in 3 groups of 15 rats each, which in turn were subdivided in 3 subgroups of 5 rats each, in which we implanted one cusp and one aortic wall segment in 2 subcutaneous pouches for each rat. We called each group as follows: GDA (control group), E80% (the group whose structures were previously prepared with ethanol 80%) and GA 0.8% (group previously prepared with L-glutamic acid 0.8%); in those groups we measured calcium and performed a microscopic analysis seeking for any calcification, its location and intensity; inflammatory infiltrate, location and type, during a 15, 30, and 60-day period after the implant. RESULTS: Calcium was found in the aortic cusp in the E80% group (1.30±0.21 mg calcium/mg tissue) at day 15, (1.05±0.22 mg calcium/mg tissue) at day 30, and (0.53±0.42 mg calcium/mg tissue) at day 60; in the GA 0.8% group (12.17±0.66 mg calcium/mg tissue) at day 15, (15.31±2.82 mg calcium/mg tissue) at day 30, and (34.24±16.28 mg calcium/mg tissue) at day 60; and in the control group, GDA at day 15 (12.44±2.26 mg calcium/mg tissue), at day 30 (13.44±3.34 mg calcium/mg tissue), and at day 60 (50.85±8.71 µg calcium/mg tissue). As for the calcium measured in the aortic wall, in the E80% group we found (4.62±0.68 µg calcium/mg tissue) at day 15, (9.47±2.59 µg calcium/mg tissue) at day 30, and (23.56±7.75 µg calcium/mg tissue) at day 60; in the GA 0.8% group at day 15 (4.31±0.85 µg calcium/mg tissue), at day 30 (7.69±1.48 µg calcium/mg tissue), and at day 60 (20.50±1.22 µg calcium/mg tissue); and in the control group (GDA) at day 15 (7.34±1.32 µg calcium/mg tissue), at day 30 (9.28±0.76 µg calcium/mg tissue), at day 60 (27.60±1.08 µg calcium/mg tissue). Microscopic evaluation of the aortic cusp, showed a progressive calcification in those fixed with GDA. Such process was found partially in the GA 0.8% group, and totally absent in the E80% group. As for the assessment of the aortic wall segments, we also observed progressive calcification, which was not inhibited by the treatment with either GA 0.8% or E80%. CONCLUSIONS: We concluded that a pre-treatment with ethanol at 80% inhibited calcification in pig aortic cusps, however it was not as effective on the aortic wall. However, L-glutamic acid at 0.8% did show that it minimizes calcification in the aortic wall. Further studies are required, to evidence if the anti-calcifying action of ethanol 80% is kept if the pig aortic bioprostheses are implanted in the circulatory system.