Stabilized, sterilized collagen scaffolds with active adjuncts attached

Abstract

Bioimplants and methods of making the bioimplants are provided. The bioimplants comprise biological tissues having conjugated thereto adjunct molecules. The biological tissues are sterilized with a chemical sterilizing agent, such as a water soluble carbodiimide. The processes of making the bioimplants include a process in which an adjunct molecule is conjugated to a biological tissue during the sterilization process.

1 . A bioimplant comprising a chemically sterilized biological tissue and at least one adjunct, wherein the adjunct is covalently conjugated to the biological tissue. 
     
     
         2 . The bioimplant of  claim 1 , wherein the chemically sterilized biological tissue is sterilized with a carbodiimide, such as EDC, optionally in the presence of an alkanol, such as a C 2 -C 4  alkanol, especially isopropanol. 
     
     
         3 . The bioimplant of  claim 1 , wherein the chemically sterilized biological tissue is crosslinked with a carbodiimide, optionally in the presence of a bifunctional crosslinking agent. 
     
     
         4 . The bioimplant of  claim 1 , wherein the biological tissue comprises native tissue, processed tissue in native form, processed tissue in non-native form, a composite or a complex composite. 
     
     
         5 . The bioimplant of  claim 4 , wherein: (1) said native tissue comprises bone, tendon, ligament, dermis, fascia, pericardium, and combinations thereof, including bone-connective tissue combinations, such as bone-tendon combinations and bone-ligament-bone combinations; (2) said processed tissue in native form comprises crosslinked tissue, decellularized crushed bone fragments, decellularized collagen or other decellularized and/or defatted bone, tendon, ligament, fascia or bone-connective tissue combinations, such as bone-ligament-bone or bone-tendon combinations; (3) said processed tissue in non-native form comprises solubilized or purified collagen from connective tissue, gelatin from mammals or fish or demineralized bone; (4) said composites comprise combinations of native tissues, processed tissues in native form and/or processed tissues in non-native form, such as pericardium with gelatin, bone with gelatin, purified collagen with gelatin or demineralized bone with solubilized or purified collagen; and (5) said complex composite comprises native tissue, processed tissue in native form, processed tissue in non-native form or a composite of native tissue, processed tissue in native form and/or processed tissue in non-native form with a biocompatible material such as a hydrogel, an alginate and/or chitosan. 
     
     
         6 . The bioimplant of  claim 1 , wherein the adjunct is a protein, a small peptide, a ribonucleic acid, a deoxyribonucleic acid, a polysaccharide, glycosaminoglycan (GAG) or an antibiotic. 
     
     
         7 . The bioimplant of  claim 6 , wherein the adjunct comprises: (1) one or more proteoglycans, glycosaminoglycans, growth factors, including any member of the transforming growth factor (TGF) superfamily and proteoglycans; (2) a deoxyribonucleic acid; (3) ribonucleic acid, such as a small interfering RNA or microRNA; (4) an antibiotic selected from aminoglycosides, the amphenicols, the ansamycins, the β-lactams, the lincosamides, the macrolides, the polypeptide antibiotics, the tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, the nitroftirans, the quinolones, the sulfonamides, the sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibernol. 
     
     
         8 . The bioimplant of  claim 1 , wherein the adjunct comprises: (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (4) a ribonucleic acid such as a small interfering RNA (siRNA) or a microRNA; or (5) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibemol. 
     
     
         9 . The bioimplant of  claim 1 , wherein the biological tissue comprises collagen, purified collagen or solubilized collagen. 
     
     
         10 . The bioimplant of  claim 1  in the form of a suture, a sheet, an implantable valve, an implantable sponge or an implantable paste. 
     
     
         11 . A process of making the bioimplant of  claim 1 , comprising:
 (a) contacting a biological tissue with an adjunct to form a combination; and   (b) contacting the combination with a chemical sterilizing agent to form the bioimplant.   
     
     
         12 . The process of  claim 11 , wherein the sterilizing agent is a carbodiimide, such as EDC, optionally in the presence of an alkanol, such as a C 2 -C 4  alkanol, especially isopropanol. 
     
     
         13 . The process of  claim 11 , wherein the wherein the biological tissue comprises native tissue, processed tissue in native form, processed tissue in non-native form, a composite or a complex composite. 
     
     
         14 . The process of  claim 11 , wherein: (1) said native tissue comprises bone, tendon, ligament, dermis, fascia, pericardium, said native tissue comprises bone, tendon, ligament, dermis, fascia, pericardium, and combinations thereof, including bone-connective tissue combinations, such as bone-tendon combinations and bone-ligament-bone combinations; (2) said processed tissue in native form comprises crosslinked tissue, decellularized crushed bone fragments, decellularized collagen or other decellularized and/or defatted bone, tendon, ligament, fascia or bone-connective tissue combinations, such as bone-ligament-bone or bone-tendon combinations; (3) said processed tissue in non-native form comprises solubilized or purified collagen from connective tissue, gelatin from mammals or fish or demineralized bone; (4) said composites comprise combinations of native tissues, processed tissues in native form and/or processed tissues in non-native form, such as pericardium with gelatin, bone with gelatin, purified collagen with gelatin or demineralized bone with solubilized or purified collagen; and (5) said complex composite comprises native tissue, processed tissue in native form, processed tissue in non-native form or a composite of native tissue, processed tissue in native form and/or processed tissue in non-native form with a biocompatible material such as a hydrogel, an alginate and/or chitosan. 
     
     
         15 . The process of  claim 11 , wherein the adjunct is a protein, a small peptide, a ribonucleic acid, a deoxyribonucleic acid, a polysaccharide, glycosaminoglycan (GAG) or an antibiotic. 
     
     
         16 . The process of  claim 15 , wherein the adjunct comprises: (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (4) ribonucleic acid such as a small interfering RNA (siRNA) or a microRNA; or (5) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibernol. 
     
     
         17 . The process of  claim 11 , wherein the adjunct is (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (3) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibemol. 
     
     
         18 . The process of  claim 11 , wherein the biological tissue comprises collagen, purified collagen or solubilized collagen. 
     
     
         19 . The process of  claim 11 , further comprising shaping or forming the biological tissue into the form of a suture, a sheet, an implantable valve, an implantable sponge or an implantable paste. 
     
     
         20 . A bioimplant made by a process comprising contacting a biological tissue with an adjunct molecule in the presence of a sterilizing agent. 
     
     
         21 . A process of making a sterilized biological implant, comprising:
 (a) contacting a starting tissue with an adjunct to form an intermediate;   (b) freezing the intermediate product of (a) to produce a frozen intermediate;   (c) lyophilizing the frozen intermediate from (b) to produce a lyophilized intermediate; and   (d) contacting the lyophilized intermediate with a sterilizing solution comprising a carbodiimide sterilizing agent to produce the biological implant.   
     
     
         22 . The process of  claim 21 , wherein said sterilizing agent is a carbodiimide, such as EDC, optionally in the presence of an alkanol, such as a C 2 -C 4  alkanol, especially isopropanol. 
     
     
         23 . The process of  claim 21 , wherein the starting tissue is a native tissue, a processed tissue in native form or a composite tissue. 
     
     
         24 . The process of  claim 23 , wherein the starting tissue is: (1) a native tissue comprising bone, tendon, ligament, dermis, fascia, pericardium, and combinations thereof, including bone-connective tissue combinations, such as bone-tendon combinations and bone-ligament-bone combinations; (2) said processed tissue in native form comprises crosslinked tissue, decellularized crushed bone fragments, decellularized collagen or other decellularized and/or defatted bone, tendon, ligament, fascia or bone-connective tissue combinations, such as bone-ligament-bone or bone-tendon combinations; or (3) a composite comprising combinations of native tissues, processed tissues in native form and/or processed tissues in non-native form, such as pericardium with gelatin, bone with gelatin, purified collagen with gelatin or demineralized bone with solubilized or purified collagen. 
     
     
         25 . The process of  claim 21 , wherein the adjunct is a protein, a small peptide, a ribonucleic acid, a deoxyribonucleic acid, a polysaccharide, glycosaminoglycan (GAG) or an antibiotic. 
     
     
         26 . The process of  claim 25 , wherein the adjunct comprises: (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (4) ribonucleic acid such as a small interfering RNA (siRNA) or a microRNA; or (5) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibernol. 
     
     
         27 . The process of  claim 21 , wherein the adjunct is (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibemol. 
     
     
         28 . The process of  claim 21 , wherein the biological tissue comprises collagen, purified collagen or solubilized collagen. 
     
     
         29 . The process of  claim 21 , further comprising shaping or forming the biological tissue into the form of a suture, a sheet, an implantable valve, an implantable sponge or an implantable paste. 
     
     
         30 . A bioimplant produced by the process of  claim 21 . 
     
     
         31 . A process of making a sterilized biological implant, comprising:
 (a) preparing a composition comprising a starting tissue;   (b) freezing the composition from (a) to form a frozen composition;   (c) lyophilizing the frozen composition from (b) to form a lyophilized composition; and   (d) contacting the lyophilized composition from (c) with a sterilizing solution comprising a sterilizing agent and an adjunct to produce the biological implant.   
     
     
         32 . The process of  claim 31 , wherein said sterilizing agent is a carbodiimide, such as EDC, optionally in the presence of an alkanol, such as a C 2 -C 4  alkanol, especially isopropanol. 
     
     
         33 . The process of  claim 31 , wherein the starting tissue is a processed tissue in native form or a complex composite. 
     
     
         34 . The process of  claim 33 , wherein the starting tissue is: (1) a processed tissue in native form comprising crosslinked tissue, decellularized crushed bone fragments, decellularized collagen or other decellularized and/or defatted bone, tendon, ligament, fascia or bone-connective tissue combinations, such as bone-ligament-bone or bone-tendon combinations; or (2) complex composite comprising native tissue, processed tissue in native form, processed tissue in non-native form or a composite of native tissue, processed tissue in native form and/or processed tissue in non-native form with a biocompatible material such as a hydrogel, an alginate and/or chitosan. 
     
     
         35 . The process of  claim 31 , wherein the adjunct is a protein, a small peptide, a ribonucleic acid, a deoxyribonucleic acid, a polysaccharide, glycosaminoglycan (GAG) or an antibiotic. 
     
     
         36 . The process of  claim 35 , wherein the adjunct comprises: (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (4) ribonucleic acid such as a small interfering RNA (siRNA) or a microRNA; or (5) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibernol. 
     
     
         37 . The process of  claim 31 , wherein the adjunct is (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (3) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibernol. 
     
     
         38 . The process of  claim 31 , wherein the biological tissue comprises collagen, purified collagen or solubilized collagen. 
     
     
         39 . The process of  claim 31 , further comprising shaping or forming the biological tissue into the form of a suture, a sheet, an implantable valve, an implantable sponge or an implantable paste. 
     
     
         40 . A biological implant produced by the process of  claim 31 . 
     
     
         41 . A process of making a sterilized biological implant, comprising:
 (a) contacting a starting tissue with a crosslinking agent to at least partially crosslink the starting tissue to produce a crosslinked tissue; and   (b) contacting the crosslinked tissue from (a) with a sterilizing solution comprising a sterilizing agent and an adjunct to produce the biological implant.   
     
     
         42 . The process of  claim 41 , wherein said sterilizing agent is a carbodiimide, such as EDC, optionally in the presence of an alkanol, such as a C 2 -C 4  alkanol, especially isopropanol. 
     
     
         43 . The process of  claim 41 , wherein the starting tissue is a native tissue or a processed tissue in native form. 
     
     
         44 . The process of  claim 43 , wherein the starting tissue is: (1) a native tissue comprising bone, tendon, ligament, dermis, fascia, pericardium, or combinations thereof, including bone-connective tissue combinations, such as bone-tendon combinations and bone-ligament-bone combinations; or (2) a processed tissue in native form comprising crosslinked tissue, decellularized crushed bone fragments, decellularized collagen or other decellularized and/or defatted bone, tendon, ligament, fascia or bone-connective tissue combinations, such as bone-ligament-bone or bone-tendon combinations. 
     
     
         45 . The process of  claim 41 , wherein the adjunct is a protein, a small peptide, a ribonucleic acid, a deoxyribonucleic acid, a polysaccharide, glycosaminoglycan (GAG) or an antibiotic. 
     
     
         46 . The process of  claim 45 , wherein the adjunct comprises: (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (4) ribonucleic acid such as a small interfering RNA (siRNA) or a microRNA; or (5) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibernol. 
     
     
         47 . The process of  claim 41 , wherein the adjunct is (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (4) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibemol. 
     
     
         48 . The process of  claim 41 , wherein the biological tissue comprises collagen, purified collagen or solubilized collagen. 
     
     
         49 . The process of  claim 41 , further comprising shaping or forming the biological tissue into the form of a suture, a sheet, an implantable valve, an implantable sponge or an implantable paste. 
     
     
         50 . A bioimplant produced by the process of  claim 41 . 
     
     
         51 . A process of making a sterilized biological implant, comprising:
 (a) contacting a starting tissue with a crosslinking agent and an adjunct to at least partially crosslink the starting tissue to produce a crosslinked tissue adjunct-conjugated tissue; and   (b) contacting the crosslinked tissue from (a) with a sterilizing solution comprising a sterilizing agent to produce the biological implant.   
     
     
         52 . The process of  claim 51 , wherein said sterilizing agent is a carbodiimide, such as EDC, optionally in the presence of an alkanol, such as a C 2 -C 4  alkanol, especially isopropanol. 
     
     
         53 . The process of  claim 51 , wherein the starting tissue is a processed tissue in non-native form. 
     
     
         54 . The process of  claim 53 , wherein the starting tissue is a processed tissue in non-native form comprising solubilized or purified collagen from connective tissue, gelatin from mammals or fish or demineralized bone. 
     
     
         55 . The process of  claim 51 , wherein the adjunct is a protein, a small peptide, a ribonucleic acid, a deoxyribonucleic acid, a polysaccharide, glycosaminoglycan (GAG) or an antibiotic. 
     
     
         56 . The process of  claim 55 , wherein the adjunct comprises: (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (4) ribonucleic acid such as a small interfering RNA (siRNA) or a microRNA; or (5) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, #lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibernol. 
     
     
         57 . The process of  claim 51 , wherein the adjunct is (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (3) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibernol. 
     
     
         58 . The process of  claim 51 , wherein the biological tissue comprises collagen, purified collagen or solubilized collagen. 
     
     
         59 . The process of  claim 51 , further comprising shaping or forming the biological tissue into the form of a suture, a sheet, an implantable valve, an implantable sponge or an implantable paste. 
     
     
         60 . A bioimplant produced by the process of  claim 51 . 
     
     
         61 . A process of making a sterilized biological implant, comprising:
 (a) contacting a starting tissue with a sterilizing solution comprising a sterilizing agent to produce a sterilized intermediate;   (b) contacting the sterilized intermediate from (a) with an adjunct and a conjugating agent to produce an implant; and   (c) optionally subjecting the implant to another sterilization step; whereby a sterilized biological implant is produced.   
     
     
         62 . The process of  claim 61 , wherein said sterilizing agent is a carbodiimide, such as EDC, optionally in the presence of an alkanol, such as a C 2 -C 4  alkanol, especially isopropanol. 
     
     
         63 . The process of  claim 61 , wherein the starting tissue is a processed tissue in non-native form. 
     
     
         64 . The process of  claim 63 , wherein the starting tissue is: (1) a native tissue, such as bone, tendon, ligament, dermis, fascia, pericardium, and combinations thereof, including bone-connective tissue combinations, such as bone-tendon combinations and bone-ligament-bone combinations; (2) a processed tissue in non-native form, such as solubilized or purified collagen from connective tissue, gelatin from mammals or fish or demineralized bone; or (3) combinations of native tissues, processed tissues in native form and/or processed tissues in non-native form, such as pericardium with gelatin, bone with gelatin, purified collagen with gelatin or demineralized bone with solubilized or purified collagen. 
     
     
         65 . The process of  claim 61 , wherein the adjunct is a protein, a small peptide, a ribonucleic acid, a deoxyribonucleic acid, a polysaccharide, glycosaminoglycan (GAG) or an antibiotic. 
     
     
         66 . The process of  claim 65 , wherein the adjunct comprises: (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (4) ribonucleic acid such as a small interfering RNA (siRNA) or a microRNA; or (5) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibernol. 
     
     
         67 . The process of  claim 61 , wherein the adjunct is (1) one or more proteoglycans or glycosaminoglycans, (2) one or more proteins, such as: (a) any member of the Transforming Growth Factor (TGF) superfamily, such as BMP-2, BMP-4 and BMP-7, transforming growth factor-β (TGF-β); (b) platelet derived growth factor (PDGF); (c) fibroblast growth factor (FGF); (d) insulin-like growth factors (IGF); (e) cartilage-derived growth factors (CDGF); (3) a deoxyribonucleic acid selected from genes, gene fragments and antisense DNA; (4) an antibiotic, such as one or more aminoglycosides, amphenicols, ansamycins, β-lactams, lincosamides, macrolides, polypeptide antibiotics, tetracyclines, cycloserine, mupirocin, tuberin, 2,4-diaminopyrimidines, nitrofurans, quinolones, sulfonamides, sulfones, clofoctol, hexedine, methenamine, nitroxoline, taurolidine and xibemol. 
     
     
         68 . The process of  claim 61 , wherein the biological tissue comprises collagen, purified collagen or solubilized collagen. 
     
     
         69 . The process of  claim 61 , further comprising shaping or forming the biological tissue into the form of a suture, a sheet, an implantable valve, an implantable sponge or an implantable paste. 
     
     
         70 . A bioimplant produced by the process of  claim 61 . 
     
     
         71 . A bioimplant of  claim 1 , wherein the adjunct retains at least some of its native activity after it has been conjugated to the biological tissue. 
     
     
         72 . A bioimplant of  claim 1 , wherein the adjunct is adapted to be released in vivo and the adjunct, once release in vivo possesses at least some of its native activity.