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Novel TRI™ Heart Valve Biomaterial Resists Calcification and Elicits Reduced Inflammatory Response
Hobey Tam, Narendra Vyavahare.
Clemson University, Clemson, SC, USA.

OBJECTIVE: We have developed a novel, proprietary fabrication method (TRI™) that utilizes carbodiimide crosslinking chemistry, neomycin trisulfate, and pentagalloyl glucose to stabilize extracellular matrix components in soft tissues to resist (1) calcification and (2) structural degradation. In this study, we demonstrate that porcine aortic valve leaflets treated with TRI™ resist calcification in vivo and may be more biocompatible than glutaraldehyde (GLUT) leaflets, the current market gold standard.
METHODS: Freshly harvested porcine aortic heart valve leaflets were treated with either 0.6% GLUT or TRI™. Differential scanning calorimetry (DSC) and enzymatic degradation was used to analyze samples for tissue stability. Treated leaflets were subcutaneously implanted into juvenile rats for 90 days to assess in vivo biocompatibility. Pre and post implanted samples were analyzed for mineralization via inductively coupled plasma mass spectrometry (ICP) and Alizarin red staining. Hematoxylin and Eosin staining (H&E) was utilized to evaluate fibrous capsule thickness, macrophage activity, and foreign body giant cell formation. Lectin histochemistry was used to detect Galα epitope left in the tissue. Immunohistochemistry (IHC) was used to detect macrophages and T-cells in explants.
REUSLTS:
Table 1: Tissue stability assessment.

Collagenase Stability (% wt. loss)Elastase Stability (% wt. loss)GAGase Stability (% GAGs lost)Td by DSC (oC)
GLUT1.52±1.0716.8±2.157.3±4.989.7±0.9
TRI0.42±0.424.6±1.9*9.5±0.4*90.1±1.2
*Indicates significant difference (p<0.05) from GLUT. H&E on GLUT explants revealed chronic persistence of macrophages, formation of foreign body giant cells, and moderate fibrous capsule thickness. Dark, black nodules were found throughout the GLUT explants that are not clear as to if they are calcification nodules or if it is phagocytic/immunogenic activity. IHC is being performed to determine if these are host macrophages/T-cells infiltrating the implant. TRI™ elicited little inflammatory response with minimal fibrous capsule formation and no morphological changes within the implant. No Galα was detected in either GLUT or TRI™ leaflets.
CONCLUSIONS: Our data suggest that TRI™ is a more stable biomaterial, effectively resists calcification, and elicits a markedly reduced inflammation response from the body thus TRI™ may be a more suitable biomaterial than GLUT for bioprosthetic heart valve fabrication.
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