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

Abstract

Three-dimensional solid contractile collagen scaffolds having improved biological properties and electromechanical performance are provided. The scaffolds can be used for cell transplantation, including the fixation of biocompatible reagents and adhesion molecules which control cell adhesion, apoptosis, survival and/or differentiation simultaneously. Grafting adhesion molecules to the collagen matrices renders the scaffold 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 method for grafting and optimizing the presentation of adhesion peptides or biological agents in the scaffold is provided.

1 . A three-dimensional solid contractile scaffold made of collagen, synthetic collagen or collagen derivative, wherein:
 the scaffold has a sponge structure obtained by cross-linking a preparation of collagen, synthetic collagen or collagen derivative using a DHT-mediated cross-linking method;   the collagen, synthetic collagen or collagen derivative is modified by covalent bonding with an adhesion molecule;   the scaffold allows differentiation of contractile cells and contractibility; and   the scaffold is free of any tumoral extract.   
     
     
         2 . The three-dimensional scaffold of  claim 1 , wherein the preparation is cryodried before cross-linking. 
     
     
         3 . The three-dimensional scaffold of  claim 1 , wherein the collagen, synthetic collagen or collagen derivative is associated with a natural or synthetic polymer. 
     
     
         4 . The three-dimensional scaffold according to  claim 1 , having a pore size of 30 to 200 micrometers. 
     
     
         5 . The three-dimensional solid contractile scaffold according to  claim 1 , wherein the adhesion molecules are added to the scaffold using an heterogeneous solid phase synthesis. 
     
     
         6 . The three-dimensional solid contractile scaffold according to  claim 1 , wherein the collagen is further chemically or enzymatically cross-linked. 
     
     
         7 . The three-dimensional solid contractile scaffold according to  claim 6 , wherein the collagen is chemically cross-linked using a cross-linking agent selected from genipin, nordihydroguaiaretic acid aglycone, geniposidic acid, epoxy compounds, dialdehyde starch, glutaraldehyde, formaldehyde, dimethyl suberimidate, carbodiimides, succinimidyls, diisocyanates, aryl azide, glyceraldehyde, cyanamide, diimides, dimethyladipate, ruterine and nordihydroguaiaretic acid. 
     
     
         8 . The three-dimensional scaffold according to  claim 1 , wherein the adhesion molecule is an RGD containing peptide. 
     
     
         9 . The three-dimensional scaffold according to  claim 1 , wherein the scaffold is biodegradable. 
     
     
         10 . The three-dimensional scaffold according to  claim 1 , further comprising an heterobifunctional reactant bound by one end to the collagen scaffold and by another end to the adhesion molecule. 
     
     
         11 . The three-dimensional scaffold of  claim 10 , wherein the heterobifunctional reactant has a length of 30 to 40 angstroms. 
     
     
         12 . The three-dimensional scaffold of  claim 10 , wherein the heterobifunctional reactant is sulfo-LC-SPD. 
     
     
         13 . The three-dimensional scaffold according to  claim 1 , having an electric threshold lower than 3 V/cm. 
     
     
         14 . The three-dimensional scaffold according to  claim 1 , further comprising an agent for controlling the adhesion, survival, proliferation, apoptosis and differentiation of cells, angiogenesis, ischemia, degradation, turnover and immunogenicity of extracellular matrix, inflammatory or immune response, cellular homing and myocardial function. 
     
     
         15 . The three-dimensional scaffold according to  claim 14 , wherein the agent is selected from the group consisting of collagen, fibrin or fibrinogen, a cytokine, a chemokine, eicosanoides, glycoproteins, glycosaminoglycans, keratane and chemotactic agents, a growth factor receptor, an enzyme, a hormone or its receptor, an angiogenic factor, a factor for promoting or inhibiting vascularization, a vaccine antigen, an antibody, a coagulation factor, a normalization protein, a transcription factor, a differentiation factor, a receptor, DNA, cDNA, DNA-aptamers, a toxin, a structural protein, an adhesion molecule, a drug, a therapeutic agent, a chemotherapeutic agent, an antibiotic, antifungals and antibacterials, antiviral agents, antimicrobials, anti-infectious agents and any fragment, variant or combinations thereof. 
     
     
         16 . The three-dimensional scaffold according to  claim 14 , wherein the agent is chemically bound, absorbed, adsorbed or free. 
     
     
         17 . The three-dimensional scaffold according to  claim 1 , further comprising a population of cells associated or not associated with the modified three-dimensional scaffold. 
     
     
         18 . The three-dimensional scaffold according to  claim 17 , wherein the associated cells of the population of cells are selected from the group consisting of contractile cells and cells with contractile capability upon differentiation. 
     
     
         19 . The three-dimensional scaffold according to  claim 18 , wherein the cells of the population of cells are selected from the group consisting of progenitors for contractile cells isolated from fetal, neonatal or adult tissue such as Sca1 cells, skeletal myoblasts or myocytes, cardiomyocites (fetal, neonatal or adult), cardiomyocyte or myocyte progenitors, human marrow mesenchymal cells, predifferentiated cells (using electric treatments, hypoxia, hypothermia or freezing, by stimulating or removal of some chemokines, chemical products, biological products), embryonic stem cells and genetically engineered cells. 
     
     
         20 . The three-dimensional scaffold according to  claim 17 , further comprising non-contractile cells such as endothelial cells and fibroblasts. 
     
     
         21 . A method for preparing a tissue that is either spontaneously contractile or contractile upon stimulation, comprising utilizing the three-dimensional scaffold according to  claim 1 . 
     
     
         22 . A method for producing a product selected from the group consisting of replacement heart tissue, heart patch, cardiac regeneration tissue, healing acceleration device, myocardial tissue, valvular tissue, device for controlling cardiac dilatation, device for promoting, remodeling or improving cardiac contractility, device for regenerating or treating myocardial condition or pathological myocardium, and device for cell therapy, comprising utilizing the three-dimensional scaffold according to  claim 1 . 
     
     
         23 . The method according to  claim 20 , wherein the scaffold is implanted in vivo free of a population of contractile cells. 
     
     
         24 . The method according to  claim 22 , wherein the scaffold is applied onto the ventricle, into the heart to replace part or the entirety of the myocardial thickness or into the pericardial cavity. 
     
     
         25 . The method according to  claim 22 , wherein the scaffold is fixed onto the myocardium. 
     
     
         26 . A method of using the three-dimensional scaffold according to  claim 1 , comprising implanting the scaffold into an ectopic site and then transferring the scaffold onto a heart area.