Collagen scaffold modified by covalent grafting of adhesion molecules, associated methods and use thereof for cardiovascular and thoracic cell therapy and contractile tissue engineering

摘要

Materials and methods for improving the biological properties and electromechanical performance of collagen scaffolds used for cell transplantation, including the fixation of biocompatible reagents and adhesion molecules which control cell adhesion, apoptosis, survival and/or differentiation simultaneously. The grafting of adhesion molecules to collagen matrices renders same suitable for use in vascular and cardiothoracic surgery/medicine, as well as in cell therapy for the heart and in artificial heart muscle engineering. Also, a simple method for grafting and optimising the presentation of adhesion peptides or biological agents when the scaffold is made from collagen and/or contains accessible thiol, amine or carboxyl groups.

主权项

1. A method for making a solid three-dimensional scaffold for cell therapy, tissue engineering and medico-surgical devices in the thoracic and surgical field, comprising:
providing a scaffold support comprising a cross-linked collagen matrix, the collagen matrix being a collagen sponge and the collagen being purified from collagen-containing tissue; further crosslinking the collagen matrix; and modifying the scaffold support by covalently binding adhesion molecules to the collagen, wherein the adhesion molecules are RGD-containing peptides, said binding comprising covalently and uni-directionally binding primary amine groups present in the adhesion molecules to primary amine groups present in the collagen utilizing the heterobifunctional coupling reagent sulfosuccinimidyl 6-(3′-[2-pyridyldithio]-propionamido)hexanoate (Sulfo-LC-SPDP), said binding being performed by reacting the collagen with Sulfo-LC-SPDP, wherein the Sulfo-LC-SPDP covalently binds to a primary amine group of the collagen; reacting the adhesion molecules with Sulfo-LC-SPDP, wherein the Sulfo-LC-SPDP covalently binds to a primary amine group of the adhesion molecule; and reacting the Sulfo-LC-SPDP covalently bound to the collagen with the Sulfo-LC-SPDP covalently bound to the adhesion molecule, the Sulfo-LC-SPDP coupling reagents covalently binding together to form a disulfide bond, wherein the RGD-modified collagen matrix allows survival and differentiation of implanted contractile or potential contractile cells and forms a spontaneously contractile three dimensional scaffold.