Biomaterials and implants for enhanced cartilage formation, and methods for making and using them

Abstract

The invention provides products of manufacture, e.g., biomaterials and implants, for cartilage maintenance and/or formation in-vivo, in-vitro, and ex-vivo, using nanotechnology, e.g., using nanotube, nanowire, nanopillar and/or nanodepots configured on surface structures of the products of manufacture.

1 . A product of manufacture comprising:
 wherein optionally the product of manufacture is a cell-, cartilage- and/or bone growth-enhancing or cell differentiation-enhancing product of manufacture, or a bone- or cartilage-maintaining and/or bone or cartilage growth-enhancing product of manufacture, or an implant,   (a) nanostructures comprising a nanotube, nanowire, nanopore, nanoribbon and/or a nanopillar surface configuration on a Ti and/or Ti-comprising alloy, or on a Ti-coated or Ti alloy-coated surface, or on a TiO 2  and/or TiO 2  alloy surface or coating, wherein the Ti and/or Ti-comprising alloy or the TiO 2  and/or TiO 2  alloy surface or coating, or the Ti-coated or Ti alloy-coated surface, comprises one or more surfaces (or a subsurface or a partial surface) of the product of manufacture,   wherein optionally the nanostructures (nanotubes, nanowires, nanopores, nanoribbons and/or nanopillars) comprise a metal and/or a metal alloy comprising a Ti, a Zr, a Hf, a Nb, a Ta, a Mo and/or a W, or an oxide of a Ti, a Zr, a Hf, a Nb, a Ta, a Mo and/or a W,   wherein optionally the nanostructures (nanotubes, nanowires, nanopores, nanoribbons and/or nanopillars) are formed directly and/or indirectly on and/or attached to a Ti surface and/or a Ti-coated surface, or Ti oxide surface and/or a Ti oxide-coated surface,   wherein optionally the nanotubes have a diameter dimension in the range of between about 30 to 600 nm outside diameter, or between about 50 to 400 nm diameter, or between about 70 to 200 nm diameter, and/or optionally a height dimension in the range of between about 30 to 10,000 nm, and/or optionally between about 200 to 2,000 nm thickness, or between about 200 to 500 nm thickness,   wherein optionally the Ti surface and/or Ti-coated surface, or Ti oxide surface and/or a Ti oxide-coated surface, comprises: the surface of a wire or microwire; the surface of a springy and/or a hairy wire or microwire; the surface of a mesh or mesh screen; the surface of an implant; a “pre-patterned” and/or a “pre-etched” surface made by machining or mask patterning and/or etching of the surface of the product of manufacture structure,   wherein optionally the three-dimensional Ti wire or microwire is between about 10 to 100 μm in diameter and/or the Ti wire or microwire is a springy and compliant wire or microwire,   wherein optionally the material used for the three-dimensional springy, coil, wire, or mesh screen scaffold comprises at least one of a metal or an alloy selected from the group consisting of Ti, Zr, Hf, Nb, Ta, Mo and W, or an alloy or an oxide or a mixture thereof, or stainless steel, or a Co—Cr—Ni—Mo alloy (commonly known as MP35N alloy),   wherein the surface of the springy wire scaffold contains vertically configured nanotube or nanopore arrays with about 30 to 600 nm diameter, preferably 70 to 200 nm diameter, and about 200 to 2,000 nm thickness, and preferably 200 to 500 nm thickness,   wherein optionally the Ti or Ti oxide alloy or Ti or Ti oxide on the Ti-coated, or Ti oxide-coated or Ti alloy-coated surface is between about 100 to 2000 μm thick; and   wherein optionally the product of manufacture structure of (a) comprises (i) oxides of alloys comprising Ti or a Ti oxide or a TiO 2  by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more in weight %, or (ii) oxides of alloys comprising Zr, Hf, Nb, Ta, Mo, W, by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more weight %, or (ii) a ceramic, a polymer, a plastic, a Si-comprising composition, a Au-comprising composition, a Pd-comprising composition, a Pt-comprising composition, or a stainless steel;   (b) the product of manufacture of (a), and further comprising a chondrocyte, a stem cell, a totipotent cell, a multipotent progenitor cell and/or a pluripotent cell, wherein the chondrocyte functionality, as indicated by the degree of extracellular matrix formation, is increased by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% or 90% or more, as compared with the identical material but without the TiO 2  nanotube or nanopillar surface configuration;   (c) the product of manufacture of (a) or (b), further comprising a chondrocyte, a colony-forming unit-fibroblast (CFU-F), a marrow stromal cell or mesenchymal stem cell (MSC), a stem cell, a totipotent cell, a multipotent progenitor cell and/or a pluripotent cell, wherein optionally the cell is implanted in, seeded in or placed in the product of manufacture in-vivo, in-vitro, and/or ex-vivo;   (d) the product of manufacture of (b) or (c), wherein the stem cell is a mesenchymal stem cell (MSC), an adult stem cell, an induced pluripotent stem cell (abbreviated as iPS cell or iPSC) and/or an embryonic stem cell;   (e) the product of manufacture of any of (b) to (d), wherein the chondrocyte is an autologous chondrocyte, a hypertrophic chondrocyte, or a human chondrocyte;   (f) the product of manufacture of any of (a) to (d), further comprising on the surface of the product of manufacture a nano-depot, a microcavity and/or a macrocavity comprising a cell, a drug and/or a biological agent,   wherein optionally the nanotube or a nanopillar, or microcavity and/or a macrocavity, acts as a depot or storage area comprising a cell, a drug and/or a biological agent,   wherein optionally the microcavity has an entrance dimension of between about 1 to 100 micrometer, or a macrocavity having an entrance dimension of between about 100 to 1,000 micrometer; or   (g) the product of manufacture of any of (a) to (f), having a structure as illustrated in any one of  FIGS. 16 to 29 .   
     
     
         2 . The product of manufacture of  claim 1 , wherein the product of manufacture comprises (a) a thin coating of a metal, a metal oxide, and/or an alloy at least about 1, 2, 3, 4, 5, 10, 15 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm or 100 nm or more nm in thickness at the surface, and/or (b) at least a portion of the surface underneath comprises a vertically aligned and adhering nanotube, nanoribbon, nanowire and/or nanopillar array structure or structures, and/or a plurality of recessed nanopore structures. 
     
     
         3 . The product of manufacture of  claim 1 , wherein the entrance dimension of the nano-depot, nanotube and/or nanopore is reduced (constricted or impeded) by a selective deposition of a metal or an alloy, a metal oxide and/or alloy oxide, and/or another compound, to induce a partial bottlenecking (constricting) configuration to slow down or impede the release rate of a compound or a substance stored in the nano-depot, nanotube and/or nanopore,
 wherein optionally the compound or substance comprises a drug and/or a biological agent stored in the nano-depot, nanotube and/or nanopore,   wherein optionally the slowing down or impeding of the release rate of the compound or a substance stored in the nano-depot, nanotube and/or nanopore is at least by a factor of 2 or 3 or slower, or at least by a factor of about 10 or slower, than the case of non-bottlenecked (non-constricted) structure,   wherein optionally the other compound used to partially bottleneck (constrict or impeded) the nano-depot and/or nanopore comprises a nitride, a fluoride, a carbide and/or a polymer material,   wherein optionally the product of manufacture surface has a multiplex and/or a duplex distribution of nanostructure structures with different dimensions such that the product of manufacture comprises both one or more nano-depot, nanotube and/or nanopore structures having bottle-necked (constricted or impeded) pore structures together with nano-depot, nanotube and/or nanopore structures which do not have the bottleneck diameter (constricted or impeded opening) reductions,   wherein optionally the relative area fraction of bottle necked (constricted or impeded opening) nano-depot, nanotube and/or nanopore structures in the product of manufacture is in the range of about 2% to 50% of the total available surface area of the product of manufacture, or in the range of about 2% to 50% of the total available surface area available for stimulating cell growth, cartilage growth and/or bone deposition.   
     
     
         4 . The product of manufacture of  claim 1 , wherein the product of manufacture further comprises a chemical, a drug and/or a biological agent,
 and optionally the chemical, drug and/or biological agent comprises a small molecule, a growth factor, a collagen, a protein, a biomolecule, a gene, a nucleic acid, an RNA or a DNA, a nucleic acid expression vector, an antibiotic, a hormone, a therapeutic drug, a functional particle, a liposome, or a magnetic, metallic, ceramic or a polymer particle; or, a differentiation-inducing chemical, drug and/or biomolecule,   and optionally the chemical, drug and/or biological agent is attached to or coated on the product of manufacture, or is stored in a nanopore, nanodepot and/or nanotube, or the chemical, drug and/or biological agent is attached to, coated on or stored between nanostructures comprising a plurality of nanopillars, nanotubes, nanowires and/or nanoribbons,   and optionally the chemical, drug and/or biological agent comprises (are) a fibroblast growth factor (FGF), an epidermal growth factor (EGF), a vascular endothelial growth factor (VEGF), a transforming growth factor beta-1 (TGF-β1) or a transforming growth factor beta-2 (TGF-β2), a bone morphogenic protein (BMP), an agent that stimulates chondrocyte growth, maintenance and/or differentiation, a chemical or biomolecule osteogenic-inducing agent, a fibroblast growth factor and/or a vascular endothelial growth factor, a bisphosphonate, a chemical agent that suppresses the bone loss by suppressing osteoclasts (the type of bone cell that breaks down bone tissue),   wherein optionally the chemical, drug and/or biological agent are positioned on the side of an implant surface intended for cartilage growth and comprise (are) chondrogenic inducing agents, and/or a chemical or a biomolecule-comprising agent that stimulates chondrocyte growth, maintenance and/or differentiation;   and optionally a biological agent positioned on another or opposite side of the implant surface is intended for bone growth and optionally comprises a chemical, drug and/or biological agent that stimulates or maintains bone growth; or   (c) the product of manufacture of (b), wherein the bone morphogenic protein (BMP) is (or comprises) bone morphogenetic protein 2 (BMP-2), bone morphogenetic protein 3 (BMP-3), bone morphogenetic protein 4 (BMP-5), bone morphogenetic protein 5 (BMP-5), bone morphogenetic protein 6 (BMP-6), bone morphogenetic protein 7 (BMP-7), bone morphogenetic protein 8 (BMP-8a), bone morphogenetic protein 10 (BMP-10), bone morphogenetic protein 15 (BMP-15).   
     
     
         5 . The product of manufacture of any of  claim 4 , wherein the functional particles comprise magnetic oxide particles or metallic particles utilized for remotely actuated RF heating and creation of temperature gradient for accelerated or switch-on, or switch-off release of the chemical, drug and/or biological agent stored in the nanodepot space. 
     
     
         6 . A method of fabricating a chondrocyte attachment-enhancing and/or chondrocyte growth-enhancing product of manufacture comprising a nanotube, nanowire, nanopore and/or nanopillar configuration comprising:
 (a) use of anodization, formation and selective phase removal of a two-phase mask layer using diblock copolymer layer, spinodally decomposing alloy layer, or two-phased alloy film, followed by selective etching of a biomaterial surface to produce a nanotube or nanopillar surface configuration on a surface of the product of manufacture;   (b) spot-welding, or induction melting-bonding, or electron-beam (“e-beam”) bonding, or laser bonding, or braze-bonding, a plurality of nanotubes or nanowires onto a TiO 2  base on a surface of the product of manufacture, wherein the base comprises: a Ti, Zr, Hf, Nb, Ta, Mo or W; or a Ti alloy or oxide, a TiO 2 , an Au or Au oxide, a Pt or Pt oxide, a Pd or a Pd oxide; a mixture comprising an alloy or an oxide of one or more of Ti, Au, Pt or Pd; or, an oxide of an alloy comprising Zr, Hf, Nb, Ta, Mo or W;   (c) fabricating on the base surface of the product of manufacture a nanotube, nanopore and/or nanopillar configuration, wherein the base comprises a bulk metal or alloy, deposited thin film or deposited thick layer selected from: a Ti, Zr, Hf, Nb, Ta, Mo or W; or a Ti alloy comprising Ti, Al, and V, or Ti alloy one or more of Ti, Au, Pt or Pd; or, an alloy comprising Zr, Hf, Nb, Ta, Mo or W;   (d) the method of any of (a) to (c), wherein the product of manufacture comprises a surface having a duplex distribution of the nanostructure dimensions such that a nanopore or nano-depot has an intentionally bottle-necked or constricted pore structure or opening, wherein optionally the bottle-necked or constricted pore structure or opening results in a slower release of a stored agent or composition, and optionally the agent or composition comprises a drug and/or a biological agent, and optionally nanostructure with bottle-necked or constricted pore structures or openings are mixed and distributed together with regular (non-bottle-necked or non-constricted pore structures or openings) nanotubes or nanopores (which do not have the bottleneck diameter reduction), and optionally the relative area fraction of the bottle necked agent or composition release region is in the range of about 2% to 50% of the total available surface area of the product of manufacture, or in the range of about 2% to 50% of the total available surface area available for cartilage and/or bone growth or attachments, wherein optionally the product of manufacture comprises a configuration as illustrated in  FIG. 13 ; or   (e) the method of any of (a) to (d), wherein the product of manufacture comprises a product of manufacture composition of  claim 1 , or the product of manufacture has a structure as illustrated in any one of  FIGS. 5 ,  13  and  16  to  29 .   
     
     
         7 . A method of fabricating a product of manufacture comprising a macroscale added-on scaffold structure for 3-dimensional cartilage construction using protruding springy wires, mesh screens, vertical pillar array columns, comprising
 (i) (a) attaching or forming a plurality of space-containing and/or springy protruding surface scaffold structures for three-dimensional chondrocyte assembly and cartilage growth,   wherein optionally space-containing and/or springy protruding surface scaffold structures are attached or formed on a surface of the product of manufacture by spot-welding, or induction melting-bonding, or electron-beam (“e-beam”) bonding, or laser bonding, or braze-bonding of a plurality of wires coils, mesh screens onto a surface of the product of manufacture,   wherein optionally the material used for the three-dimensional springy, coil, wire, or mesh screen scaffold comprises a metal or an alloy or an oxide thereof selected from Ti, Zr, Hf, Nb, Ta, Mo or W, or an alloy or an oxide comprising at least one of these elements, or a stainless steel, or a Co- or Cr-comprising alloy, or a Co—Cr—Ni—Mo alloy,   wherein optionally the wire diameter in the range of between about 10 to 100 um,   wherein optionally the wires in the attached three-dimensional scaffold have a surface structure of either nanotubes or nanopores having diameter in the range of between about 30 to 600 nm, or between about 70 to 200 nm,   wherein optionally the wires in the attached three-dimensional scaffold have a thickness of between about 300 to 400 nm, between about 200 to 500 nm, to between about 100 to 600 nm,   wherein the material used for the three-dimensional springy, coil, wire, or mesh screen scaffold comprises a metal or alloy selected from Ti, Zr, Hf, Nb, Ta, Mo or W, or alloys containing at least one of these elements, or stainless steel, or Co—Cr—Ni—Mo alloy (commonly known as MP35N alloy),   wherein the surface of the springy wire scaffold contains vertically configured nanotube or nanopore arrays with about 30 to 600 nm diameter, preferably 70 to 200 nm diameter, and about 200 to 2,000 nm thickness, and preferably 200 to 500 nm thickness,   wherein a base material onto which the springy three-dimensional metal scaffold is attached comprises a bulk metal or alloy, deposited thin film or deposited thick layer selected from: a Ti, Zr, Hf, Nb, Ta, Mo or W; or a Ti alloy comprising Ti, Al, and V, or Ti alloy one or more of Ti, Au, Pt or Pd; or, an alloy comprising Zr, Hf, Nb, Ta, Mo or W,   (ii) the method of (i), further comprising introducing a chondrocyte-growth-enhancing nanostructure on the surface of the three-dimensional scaffold wire or pillar surface by use of anodization, formation and selective phase removal of a two-phase mask layer using diblock copolymer layer, spinodally decomposing alloy layer, or two-phased alloy film, followed by selective etching of a biomaterial surface to produce a nanotube or nanopillar surface configuration; or   (iii) the method of (i) or (ii), wherein the product of manufacture comprises a composition of  claim 1 , or the product of manufacture has a structure as illustrated in any one of  FIGS. 5 ,  13  and  16  to  29 .   
     
     
         8 - 10 . (canceled) 
     
     
         11 . A patch bone implant piece comprising a product of manufacture of  claim 1 , wherein optionally the product of manufacture serves a dual purpose of comprising at least one exposed surface that enhances a chondrocyte growth and cartilage formation while comprising another surface (e.g., an opposing surface or a bottom surface) facing the existing bone to induce a strong osseo-integration. 
     
     
         12 . (canceled) 
     
     
         13 . An implant, or a bone implant, or a patch implant comprising a product of manufacture of  claim 1 , optionally comprising Ti or TiO 2 . 
     
     
         14 - 15 . (canceled) 
     
     
         16 . A chondrocyte cell culture substrate for enhanced or new chondrocyte, stem cell and/or extracellular matrix production comprising the product of manufacture  claim 1 . 
     
     
         17 . A method for making an implant comprising a cell, comprising:
 (a) (i) providing a chondrocyte, a colony-forming unit-fibroblast (CFU-F), a marrow stromal cell (MSC), a stem cell, a totipotent cell, a multipotent progenitor cell and/or a pluripotent cell;   (ii) providing a product of manufacture of  claim 1 , wherein optionally the product of manufacture comprises a TiO 2 -comprising nanotube, nanowire and/or nanopore; and   (iii) adding the cell of (a) to the product of manufacture of (b) under cell culture conditions;   (b) the method of (a), wherein the stem cell is a mesenchymal stem cell, an adult stem cell, an induced pluripotent stem cell (abbreviated as iPS cell or iPSC) and/or an embryonic stem cell;   (c) the method of (a), wherein the product of manufacture is fabricated as a bone implant, or a patch implant, or patch bone implant piece;   (d) the method of any of (a) to (c), wherein the cell culture conditions comprise use of cell growth and/or cell differentiation factors;   (e) the method of (d), wherein the cell growth and/or cell differentiation factors comprise a drug or chemical or biological agent that promotes the growth, maintenance and/or regeneration of a cell;   (f) the method of (e) wherein the drug or chemical or biological agent that promotes the growth, maintenance and/or regeneration of a cell promotes the differentiation growth, maintenance and/or regeneration of a chondrocyte, a stem cell, a totipotent cell, a multipotent progenitor cell and/or a pluripotent cell;   (g) the method of (d), (e) or (f), wherein the cell growth, cell differentiation factor or biological agent comprises a chondrogenic agent or a bone morphogenic protein (BMP) or an agent, drug or chemical that stimulates chondrocyte growth, maintenance and/or differentiation, or a fibroblast growth factor and/or a vascular endothelial growth factor,   and optionally the chondrogenic agent is placed on an implant surface region intended for cartilage growth, and optionally bone morphogenic protein (BMP) is placed on an implant surface region intended for bone growth, and optionally fibroblast growth factor and/or vascular endothelial growth factor are placed on an implant surface region intended for bone growth for osseointegration attachment to the existing bone structure;   (h) the method of (g), wherein the bone morphogenic protein (BMP) is (or comprises) a bone morphogenetic protein 2 (BMP-2), a bone morphogenetic protein 3 (BMP-3), a bone morphogenetic protein 4 (BMP-5), a bone morphogenetic protein 5 (BMP-5), a bone morphogenetic protein 6 (BMP-6), a bone morphogenetic protein 7 (BMP-7), a bone morphogenetic protein 8 (BMP-8a), a bone morphogenetic protein 10 (BMP-10), a bone morphogenetic protein 15 (BMP-15);   (i) the method of (g), wherein the wherein the drug or chemical or biological agent comprises a fibroblast growth factor (FGF), an epidermal growth factor (EGF), a vascular endothelial growth factor (VEGF), a transforming growth factor beta-1 (TGF-β1) or a transforming growth factor beta-2 (TGF-β2), a bone morphogenic protein (BMP) (e.g., an agent that stimulates chondrocyte growth, maintenance and/or differentiation), fibroblast growth factors and/or vascular endothelial growth factors   (j) the method of any of (d) to (i), wherein cell growth, cell differentiation factor or biological agent comprises a recombinant protein, or an autologous protein, or a human protein;   (k) the method of any of (a) to (i), wherein the cell culture conditions comprise a chondrogenic-inducing media;   (l) the method of (k), wherein the chondrogenic-inducing media comprises one, several of all of: a serum-free DMEM, an ascorbate, a dexamethasone, L-proline, sodium pyruvate, ITS-plus, an antibiotic and/or a recombinant protein; or   (m) the method of any of (a) to (l), wherein the cell is implanted in, seeded in or placed in the implant in-vivo, in-vitro, and/or ex-vivo.