Conditioned compositions for tissue restoration

Abstract

The invention provides a composition conditioned for the remodeling, restoration, repair, or replacement of tissue within a host. The composition is conditioned by culturing cells on the matrix and/or by exposing the cultured cells or matrix to one or more stressors.

1 . A composition, comprising: 
 a decellularized matrix, the matrix conditioned by cells cultured on the matrix in vitro whereby the decellularized conditioned matrix has enhanced restorative, remodeling, replacement or repair properties when placed in contact with a tissue in need of restoration, remodeling, replacement or repair.    
     
     
         2 . The composition of  claim 1 , wherein the matrix comprises a naturally occurring extracellular matrix (ECM).  
     
     
         3 . The composition of  claim 2  wherein the ECM comprises at least a portion of tunica submucosa.  
     
     
         4 . The composition of  claim 2  wherein the ECM comprises at least a portion of epithelial basement membrane.  
     
     
         5 . The composition of  claim 2  wherein the ECM comprises at least a portion of tunica propria.  
     
     
         6 . The composition of  claim 2  wherein at least a portion of the ECM comprises dermis.  
     
     
         7 . The composition of  claim 2  wherein at least a portion of the ECM comprises subcutaneous tissue.  
     
     
         8 . The composition of  claim 2  wherein at least a portion of the ECM comprises pancreatic connective tissue.  
     
     
         9 . The composition of  claim 2  wherein at least a portion of the ECM comprises tissue harvested from the stomach.  
     
     
         10 . The composition of  claim 2  wherein at least a portion of the ECM comprises tissue harvested from the intestine.  
     
     
         11 . The composition of  claim 2  wherein at least a portion of the ECM comprises tissue harvested from the urinary bladder.  
     
     
         12 . The composition of  claim 2  wherein at least a portion of the ECM comprises tissue harvested from the skin.  
     
     
         13 . The composition of  claim 2  wherein at least a portion of the ECM comprises tissue harvested from a tissue comprising a mucous membrane.  
     
     
         14 . The composition of  claim 1  wherein the matrix comprises a structural or functional component of a naturally occurring ECM.  
     
     
         15 . The composition of  claim 1  wherein the matrix comprises a degradation product of a structural or functional component of a naturally occurring ECM.  
     
     
         16 . The composition of  claim 1  wherein the matrix conditioning cells are selected from the group consisting of a primary, secondary, and immortalized cell population.  
     
     
         17 . The composition of  claim 1  wherein the matrix conditioning cells are selected from the group consisting of fibroblasts, keratinocytes, astroglial cells, epithelial cells, endothelial cells, glial cells, neural cells, cells of the blood and precursors thereof, and hepatocyte and precursor cells thereof.  
     
     
         18 . The composition of  claim 1  wherein the matrix conditioning cells express a biologically active molecule of interest.  
     
     
         19 . The composition of  claim 18  wherein the biologically active molecule of interest comprises a protein.  
     
     
         20 . The composition of  claim 19  wherein the protein is selected from the group consisting of angiogenin, angiopoietin-1, Del-1, acidic-fibroblast growth factor (aFGF), basic-fibroblast growth factor (bFGF), granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), hepatocyte growth factor (HGF), Interleukin-8 (IL-8), leptin, placental growth factor (PlGF), platelet-derived endothelial cell growth factor (PD-ECGF), platelet-derived growth factor-BB (PDGF-BB), pleiotrophin (PTN), proliferin, transforming growth factor-alpha (TGF-alpha), transforming growth factor-beta (TGF-beta), tumor necrosis factor-alpha (TNF-alpha), vascular endothelial growth factor (VEGF), bone morphogenetic protein-2 (BMP-2) and stromal derived growth factor-1.  
     
     
         21 . The composition of  claim 19  wherein the protein is a growth factor.  
     
     
         22 . The composition of  claim 21  wherein the growth factor is an angiogenic growth factor.  
     
     
         23 . The composition of  claim 21  wherein the growth factor is an osteogenic growth factor.  
     
     
         24 . The composition of  claim 1  wherein the matrix conditioning cells are genetically-modified cells.  
     
     
         25 . The composition of  claim 24  wherein the genetically-modified cells are transfected cells comprising an exogenous nucleic acid and which express a protein of interest.  
     
     
         26 . The composition of  claim 24  wherein the genetically-modified cells are transfected with a nucleic acid that encodes a protein of interest.  
     
     
         27 . The composition of  claim 24  wherein the genetically-modified cells express a biologically active molecule of interest.  
     
     
         28 . The composition of  claim 27  wherein the biologically active molecule of interest is a VEGF protein, a bFGF protein, a BMP protein, or a stromal derived growth factor-1 protein.  
     
     
         29 . The composition of  claim 27  wherein the biologically active molecule of interest is a recombinant protein.  
     
     
         30 . The composition of  claim 2  wherein the ECM is harvested from a tissue of a vertebrate.  
     
     
         31 . The composition of  claim 1  wherein the tissue in need of restoration, remodeling, replacement or repair comprises a tissue selected from the group consisting of bone, skin, heart, a tissue of the urogenital tract, a tissue of the gastrointestinal tract, nervous tissue, joint tissue and connective tissue.  
     
     
         32 . The composition of  claim 1  wherein the matrix conditioning cells are exposed to at least one stressor.  
     
     
         33 . The composition of  claim 32  wherein the at least one stressor comprises hypoxia.  
     
     
         34 . The composition of  claim 32  wherein the at least one stressor comprises hypercarbia.  
     
     
         35 . The composition of  claim 32  wherein the at least one stressor comprises an electrical current.  
     
     
         36 . The composition of  claim 1  wherein the matrix is exposed to at least one stressor.  
     
     
         37 . The composition of  claim 36  wherein the stressor comprises mechanical loading.  
     
     
         38 . A composition, comprising: 
 a matrix conditioned by cells cultured on the matrix in vitro and by exposure to at least one stressor, whereby the conditioned matrix has enhanced restorative, remodeling, replacement or repair properties when placed in contact with a tissue in need of restoration, remodeling, replacement or repair.    
     
     
         39 . The composition of  claim 38  wherein the matrix conditioning cells are selected from the group consisting of fibroblasts, keratinocytes, astroglial cells, epithelial cells, endothelial cells, glial cells, neural cells, cells of the blood and precursors thereof, and hepatocyte and precursor cells thereof.  
     
     
         40 . The composition of  claim 38  wherein the matrix conditioning cells are autologous cells.  
     
     
         41 . The composition of  claim 38  wherein the at least one stressor comprises hypoxia.  
     
     
         42 . The composition of  claim 38  wherein the at least one stressor comprises hypercarbia.  
     
     
         43 . The composition of  claim 38  wherein the at least one stressor comprises an electrical current.  
     
     
         44 . The composition of  claim 38  wherein the at least one stressor comprises mechanical loading.  
     
     
         45 . The composition of  claim 38  wherein the matrix conditioning cells express a biologically active molecule of interest.  
     
     
         46 . The composition of  claim 45  wherein the biologically active molecule of interest comprises a protein.  
     
     
         47 . The composition of  claim 38  wherein the matrix conditioning cells are genetically-modified cells.  
     
     
         48 . The composition of  claim 47  wherein the genetically-modified cells express a biologically active molecule of interest.  
     
     
         49 . The composition of  claim 48  wherein the biologically active molecule of interest is a protein.  
     
     
         50 . The composition of  claim 49  wherein the protein is a recombinant protein.  
     
     
         51 . A method for inducing restoration, remodeling, replacement or repair of a tissue in a mammal, the method comprising: 
 providing a decellularized matrix;    conditioning the matrix by culturing cells on the matrix in vitro; and    decellularizing the matrix wherein the decellularized conditioned matrix induces restoration, remodeling, replacement or repair of the tissue.    
     
     
         52 . The method of  claim 51  further comprising the step of exposing the matrix conditioning cells to at least one stressor.  
     
     
         53 . The method of  claim 51  further comprising the step of exposing the matrix to at least one stressor.  
     
     
         54 . The method of  claim 51  wherein the tissue comprises a tissue selected from the group consisting of blood vessel tissue, heart tissue and bone tissue.  
     
     
         55 . The method of  claim 51  wherein the matrix conditioning cells express a biologically active molecule of interest.  
     
     
         56 . The method of  claim 55  wherein the biologically active molecule of interest comprises a growth factor.  
     
     
         57 . The method of  claim 55  wherein the biologically active molecule of interest comprises a VEGF protein, a stromal derived growth factor-1 protein, or a bone morphogenic protein.  
     
     
         58 . The method of  claim 51  wherein the matrix conditioning cells are genetically-modified cells.  
     
     
         59 . The method of  claim 58  wherein the genetically-modified cells express a biologically active molecule of interest.  
     
     
         60 . The method of  claim 59  wherein the biologically active molecule of interest is a recombinant protein.  
     
     
         61 . The method of  claim 51  wherein the matrix is harvested from a tissue of a vertebrate.  
     
     
         62 . A composition for tissue replacement, repair, restoration, or remodeling, comprising: 
 a decellularized matrix comprising a structural or functional component of a naturally occurring ECM, a degradation product of a structural or functional component of a naturally occurring ECM, or a combination thereof, wherein the matrix is conditioned by culturing cells on the matrix in vitro.    
     
     
         63 . The composition of  claim 62  wherein the matrix conditioning cells are exposed to at least one stressor.  
     
     
         64 . The composition of  claim 62  wherein the matrix is exposed to at least one stressor.  
     
     
         65 . A composition, comprising: 
 a matrix, the matrix conditioned for tissue restoration, remodeling, replacement or repair by exposure to at least one stressor.    
     
     
         66 . The composition of  claim 65 , wherein the stressor is electric current.