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Self-seeding Heart Valve Replacement Solutions Based on Self-assembling Peptide-functionalized Acellular Pericardium
Monica Dettin, Roberta Danesin, Sabrina Facciolo, Annj Zamuner, Laura Iop, Michele Spina,
Gino Gerosa.
University of Padua, Padua, Italy.

OBJECTIVE: Current mechanical and biological prostheses for heart valve replacement offer good haemodynamic performance, but have significant limitations, such as lack of viability and inability of somatic adaptation, exposing already treated patients to high risk re-interventions. The goal of this project is the formulation of novel self-seeding valve substitutes with improved bioactive and biomechanical properties, by functionalizing decellularized scaffolds with self-assembling peptide (SAP) hydrogels.
METHODS: EAbuK SAPs were synthesized using Fmoc chemistry and solid phase approach. Hydrogels structure determination was carried out by Atomic Force Microscopy and Scanning Electron Microscopy analyses. Decellularized bovine pericardium was incubated with peptide solution in water for 24 hrs at 37°C, followed by PBS. Entrapment and distribution within interfibrillar spaces were visualized by means of fluorescence microscopy and quantified by HPLC after pericardium treatment with rhodamine-labelled peptides. Scaffold modifications in thickness, area and water content were evaluated before and after SAPs gelation. Biofunctionalized samples were submitted to in vitro tissue engineering experiments with human bone marrow- mesenchymal stem cells and macaque aortic valve interstitial cells.
RESULTS: SAPs showed a tendency to spontaneously adopt β-sheet conformation in monovalent cation solutions. Fluorescence microscope analyses showed widespread incorporation of SAPs in pericardium scaffolds. Interestingly no peptide release was revealed by HPLC in solution after extensive washing of SAPs-treated samples. An increase in thickness, mass and water content, concomitant to a reduction in area, were demonstrated in functionalized pericardium. Both seeded cells were able to adhere: further analyses on their differentiation are under study.
CONCLUSIONS: In this preliminary study, we demonstrated the penetration and retention of SAPs into decellularized pericardium samples. The evaluation of peptide-induced modifications of samples' area and thickness are in agreement with water enrichment. Filler properties of SAPs can be used to design a new class of easy-to-prepare, self-seeding heart valve substitutes. In addition to the already demonstrated capacity of SAPs to increase cell adhesion and growth, possible drug or protein delivery, as well as covalent functionalization with bioactive molecules make these peptides very interesting in cardiovascular tissue engineering applications.

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