Bone tissue engineering by ex vivo stem cells ongrowth into three-dimensional trabecular metal

Abstract

Adult autologous stem cells cultured on a porous, three-dimensional tissue scaffold-implant for bone regeneration by the use of a hyaluronan and/or dexamethasone to accelerate bone healing alone or in combination with recombinant growth factors or transfected osteogenic genes. The scaffold-implant may be machined into a custom-shaped three-dimensional cell culture system for support of cell growth, reservoir for peptides, recombinant growth factors, cytokines and antineoplastic drugs in the presence of a hyaluronan and/or dexamethasone alone or in combination with growth factors or transfected osteogenic genes, to be assembled ex vivo in a tissue incubator for implantation into bone tissue.

1 . A three-dimensional tissue scaffold-implant for supporting tissue on-growth, the scaffold-implant comprising: 
 a lattice having a matrix of interconnected pores which form surfaces in three dimensions;    an inert, bio-compatible material covering the surfaces; and    at least one of a hyaluronan, dexamethasone, protein, peptide, transcript factor, cytokine, therapeutic agent, chitosan, polymer, osteogenic gene and growth factor, covering the material.    
     
     
         2 . The tissue scaffold-implant of  claim 1 , wherein the inert, biocompatible material comprises a metal.  
     
     
         3 . The tissue scaffold-implant of  claim 2 , wherein the metal comprises tantalum.  
     
     
         4 . The tissue scaffold-implant of  claim 1 , wherein the inert, biocompatible material comprises a metal alloy.  
     
     
         5 . The tissue scaffold-implant of  claim 1 , further comprising living cells covering the biocompatible material, the cells selected from the group consisting of bone marrow cells, osteoblasts, mesenchymal stem cells, embryonic stem cells, gene transfected cells, endothelial cells and combinations thereof.  
     
     
         6 . The tissue scaffold-implant of  claim 1 , further comprising tissue grown over the bio-compatible material.  
     
     
         7 . The tissue scaffold-implant of  claim 6 , wherein the tissue comprises bone.  
     
     
         8 . A method of forming tissue, the method comprising: 
 providing a three-dimensional tissue scaffold comprising a lattice having a matrix of interconnected pores which form surfaces in three dimensions, and an inert, biocompatible material covering the surfaces;    covering the material covered surfaces of the scaffold with living cells; and    culturing the scaffold to grow tissue on and in the scaffold.    
     
     
         9 . The method of  claim 8 , wherein prior to the culturing step, further comprising the step of applying at least one of a hyaluronan, dexamethasone, protein, peptide, transcript factor, cytokine, therapeutic agent, chitosan, polymer, osteogenic gene and growth factor, to the material covered surfaces.  
     
     
         10 . The method of  claim 9 , wherein the applying step includes encapsulating the at least one of the hyaluronan, dexamethasone, protein, peptide, transcript factor, cytokines therapeutic agent, chitosan, polymer, osteogenic gene and growth factor.  
     
     
         11 . The method of  claim 10 , wherein the encapsulation is performed by complex sandwich conjugation with a polymeric material.  
     
     
         12 . The method of  claim 11 , wherein the culturing step is performed by placing the scaffold in a medium and incubating the medium and scaffold.  
     
     
         13 . The method of  claim 12 , further comprising the step of implanting the scaffold in a body of one of an animal and a human being.  
     
     
         14 . The method of  claim 9 , wherein the culturing step is performed by implanting the scaffold in the body of one of an animal and a human being.  
     
     
         15 . The method of  claim 10 , wherein the culturing step is performed by implanting the scaffold in the body of one of an animal and a human being.  
     
     
         16 . The method of  claim 8 , wherein the culturing step is performed by placing the scaffold in a medium and incubating the medium and scaffold.  
     
     
         17 . The method of  claim 16 , further comprising the step of implanting the scaffold in a body of one of an animal and a human being.  
     
     
         18 . The method of  claim 8 , wherein the culturing step is performed by implanting the scaffold in the body of one of an animal and a human being.  
     
     
         19 . The method of  claim 8 , wherein the covering step is performed by cell transplantation.  
     
     
         20 . The method of  claim 8 , wherein the inert, biocompatible metal comprises tantalum.  
     
     
         21 . The method of  claim 8 , wherein the living cells are selected from the group consisting of bone marrow cells, osteoblasts, mesenchymal stem cells, embryonic stem cells, gene transfected cells, endothelial cells and combinations thereof.  
     
     
         22 . The method of  claim 8 , wherein the tissue comprises bone.  
     
     
         23 . A method of making an implant for supporting tissue on-growth, the method comprising: 
 providing a three-dimensional tissue scaffold comprising a lattice having a matrix of interconnected pores which form surfaces in three dimensions, and an inert, biocompatible metal covering the surfaces; and    covering the metal covered surfaces of the scaffold with living cells.    
     
     
         24 . The method of  claim 23 , further comprising the step of applying at least one of a hyaluronan, dexamethasone, protein, peptide, transcript factor, cytokine, therapeutic agent, chitosan, polymer, osteogenic gene and growth factor, to the metal covered surfaces.  
     
     
         25 . The method of  claim 24 , wherein the applying step includes encapsulating at least one of the hyaluronan, dexamethasone, protein, peptide, transcript factor, cytokines, therapeutic agent, chitosan, polymer, osteogenic gene and growth factor.  
     
     
         26 . The method of  claim 25 , wherein the encapsulation is performed by complex sandwich conjugation with a polymeric material.  
     
     
         27 . The method of  claim 26 , further comprising the step of culturing the scaffold to grow tissue on and in the scaffold.  
     
     
         28 . The method of  claim 27 , wherein the culturing step is performed by placing the scaffold in a medium and incubating the medium and scaffold.  
     
     
         29 . The method of  claim 23 , further comprising the step of culturing the scaffold to grow tissue on and in the scaffold.  
     
     
         30 . The method of  claim 29 , wherein the culturing step is performed by placing the scaffold in a medium and incubating the medium and scaffold.  
     
     
         31 . The method of  claim 25 , wherein the culturing step is performed by implanting the scaffold in the body of one of an animal and a human being.  
     
     
         32 . The method of  claim 23 , wherein the covering step is performed by cell transplantation.  
     
     
         33 . The method of  claim 23 , wherein the inert, biocompatible material comprises tantalum.  
     
     
         34 . The method of  claim 23 , wherein the living cells are selected from the group consisting of bone marrow cells, osteoblasts, mesenchymal stem cells, embryonic stem cells, gene transfected cells, endothelial cells and combinations thereof.  
     
     
         35 . The method of  claim 34 , wherein cells are encapsulated in at least one of a hyaluronan and collagen, or chitosan with a polymeric material by complex sandwich conjugation.  
     
     
         36 . The method of  claim 28 , wherein the culturing method is performed in one of or any combination of a static, dynamic medium flow, pulsatile flow, microgravity and multidirectional gravity culturing environment.  
     
     
         37 . The method of  claim 23 , wherein the tissue comprises bone.