Prevention of calcification of bioprosthetic heart valve cusp and aortic wall with ethanol and aluminum chloride.

BACKGROUND: Calcification is frequently associated with device failure of bioprostheses fabricated from either glutaraldehyde pretreated porcine aortic valves or bovine pericardium. It was hypothesized that differential pretreatment with ethanol-aluminum chloride will prove safe and efficacious for inhibiting the calcification of both the porcine aortic valve bioprosthetic cusp and the aortic wall. METHODS: Glutaraldehyde-fixed porcine aortic valves were subjected to differential aluminum chloride (AlCl3) and ethanol pretreatment; aortic wall segments were treated exclusively with AlCl3 (0.1 moles/L) for 45 minutes, 6 hours, or 8 hours (groups 3A, B, and C, respectively), followed by valve cusp incubations in ethanol (80%, pH 7.4). Nontreated control bioprosthetic valves were either stent-mounted porcine aortic valve bioprostheses (Carpentier-Edwards, group 1) (Edwards, Santa Anna, CA) or St. Jude Toronto SPV valves (St. Jude Medical, St. Paul, MN) (group 2). Mitral valve replacements were carried out in juvenile sheep for 150 days. RESULTS: Calcium in cusps from group 3A was 2.84 +/- 0.62 mg calcium/g tissue versus control, 22.79 +/- 8.46 mg calcium/g tissue, p = 0.04. Valves pretreated with AlCl3 for 45 minutes, 6 hours, and 8 hours had significantly lower levels of calcium in the aortic wall compared to controls (40.38 +/- 5.66, 26.77 +/- 4.02, and 28.94 +/- 8.25 mg calcium/g tissue for groups 3A, 3B, and 3C, respectively, vs 95.47 +/- 17.14 mg calcium/g tissue for group 1, p < 0.001, and 133.42 +/- 3.96 mg calcium/g tissue for group 2, p < 0.001). CONCLUSIONS: Differentially applied ethanol and aluminum chloride pretreatment significantly inhibited calcification of both the glutaraldehyde-fixed porcine aortic valve bioprosthetic cusp and the aortic wall.

Transcription factor Egr-1 in calcific aortic valve disease.

BACKGROUND AND AIM OF THE STUDY: Previous immunohistochemistry studies have shown that the transcription factor, Egr-1, is increased in human atherosclerotic lesions but is absent from the normal adjacent aortic wall. The hypothesis was investigated that Egr-1 is also increased in calcified heart valve cusps because of the unique presence in these tissues of proteins known to be regulated by Egr-1, such as tenascin C (TN-C). METHODS: Non-calcified and calcified human aortic valves were obtained at autopsy or from cardiac surgery. Egr-1 immunohistochemical studies were performed. The effects of Egr-1 on cellular proliferation and on mechanisms of calcification were also investigated using sheep aortic valve interstitial cell (SAVIC) cultures. Signal transduction pathways involving Egr-1 were studied with specific inhibitors. RESULTS: Immunohistochemical studies revealed that calcific aortic stenosis cusps contained a significantly higher level of Egr-1 in the spindle-shaped interstitial cells of calcified human aortic valves, but not white blood cells. By comparison, Egr-1 was detected at very low levels in the interstitial cells of non-calcified human aortic valve cusps. SAVIC cultivated on denatured versus native collagen substrates demonstrated a marked increase in Egr-1 levels (by Western blotting), and an absence of calcification in these cultures, compared to SAVIC grown on native collagen which calcified severely with little Egr-1 expression. Parallel increases in TN-C and osteopontin (OPN), both of which are proteins associated with heart valve calcification, were observed (by Western blotting) in SAVIC grown on denatured collagen. Furthermore, a protein kinase-C (PKC) inhibitor blocked the up-regulation of Egr-1 and TN-C, implicating PKC-dependent signaling control of Egr-1 and TN-C up-regulation. CONCLUSION: Egr-1 is up-regulated in human calcific aortic stenosis cusps compared to non-calcified normal cusps. Egr-1 up-regulation involves a PKC-dependent signaling pathway. TN-C and OPN appear to be co-regulated with Egr-1. Furthermore, in SAVIC cultures on denatured collagen, Egr-1 up-regulation was associated with inhibition of calcification. Taken together, these results suggest that complex Egr-1 mechanisms may be operative in calcific aortic stenosis.