Osteogenic devices

Abstract

Disclosed are (1) osteogenic devices comprising a matrix containing substantially pure natural-sourced mammalian osteogenic protein; (2) DNA and amino acid sequences for novel polypeptide chains useful as subunits of dimeric osteogenic proteins; (3) vectors carrying sequences encoding these novel polypeptide chains and host cells transfected with these vectors; (4) methods of producing these polypeptide chains using recombinant DNA technology; (5) antibodies specific for these novel polypeptide chains; (6) osteogenic devices comprising these recombinantly produced proteins in association with an appropriate carrier matrix; and (7) methods of using the osteogenic devices to mimic the natural course of endochondral bone formation in mammals.

1 - 133 . (canceled)  
     
     
         134 . An osteogenic protein comprising a polypeptide chain capable of inducing endochondral bone formation in association with a matrix when implanted in a mammal, wherein the polypeptide chain is further characterized as having cysteine residues in the same relative positions as the cysteine skeleton sequence:  
       
         
           
                 
                 
               
                     
                 
                   (SEQ ID NO:31) 
                     
                 
                 
                 
                 
               
                     
                     1        10        20        30       40 
                     
                 
                     
                     XXXXXXXXXXXXXXXXXXXXXXXXCXXXCXXXXXXCXXXX 
                 
                     
                     
                 
                     
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                   XXXXXXXXXXXXXXXXXXXXCCXXXXXXXXXXXXXXXXXX 
                 
                     
                     
                 
                     
                            90    97 
                 
                     
                       XXXXXXXXXXXXXCXCX, 
                 
                     
                     
                 
             
                
                
               
            
             
                
                
                
                
                
                
                
                
                
               
            
           
         
         or an allelic variant or mutant protein thereof that has an altered conserved C-terminal cysteine skeleton, wherein the protein or the allelic variant or mutant protein is capable of forming a dimeric species having a conformation capable of inducing bone formation in a mammal, and wherein each X represents any amino acid.  
       
     
     
         135 . An osteogenic protein comprising a polypeptide chain capable of inducing endochondral bone formation in association with a matrix when implanted in a mammal, wherein the polypeptide chain is further characterized as having cysteine residues in the same relative positions as the cysteine skeleton sequence depicted in amino acid residues 335 to 431 of SEQ ID NO:1:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   1     10        20       30        40 
                     
                 
                     
                      XXXXXXXXXXXXXXXXXXXXXXXXCXXXCXXXXX 
                 
                     
                     
                 
                     
                               50        60        70 
                 
                     
                      XXXXXXXXXXXXXXXXXXXXXXXXXXCCXXXXXX 
                 
                     
                     
                 
                     
                           80       90       100 
                 
                     
                      XXXXXXXXXXXXXXXXXXXXXXXXXCXCX, 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
               
            
           
         
         or an allelic variant or mutant protein thereof that has an altered conserved C-terminal cysteine skeleton, wherein the normal protein or allelic variant or mutant protein is capable of forming a dimeric species having a conformation capable of inducing bone formation in a mammal, and wherein each X represents any amino acid.  
       
     
     
         136 . An osteogenic protein comprising a polypeptide chain capable of inducing endochondral bone formation in association with a matrix when implanted in a mammal, wherein the polypeptide chain comprises an amino acid sequence encoded by a nucleic acid that hybridizes selectively to a nucleic acid encoding amino acid residues 335 to 431 of SEQ ID NO: 1 in 40% formamide, 5×SSPE, 5× Denhardt's Solution, and 0.1% SDS at 37° C. overnight, and washing in 0.1×SSPE, 0.1% SDS at 50°, wherein the polypeptide chain is further characterized as having a mutation that alters the conserved C-terminal cysteine skeleton, and further wherein the mutant polypeptide chain is capable of forming a dimeric species having a conformation capable of inducing endochondral bone formation in a mammal.  
     
     
         137 . The osteogenic protein of  claim 134 , wherein the protein is dimeric.  
     
     
         138 . The osteogenic protein of  claim 134 , wherein the polypeptide is glycosylated.  
     
     
         139 . The osteogenic protein of  claim 134 , wherein the polypeptide is unglycosylated.  
     
     
         140 . An osteogenic device for implantation in a mammal, the device comprising: 
 a biocompatible, in vivo biodegradable matrix defining pores of a dimension sufficient to permit influx, differentiation, and proliferation of migratory progenitor cells from the body of the mammal, and    an osteogenic protein of  claim 134 .    
     
     
         141 . The osteogenic device of  claim 140 , wherein the matrix comprises collagen and at least one material selected from the group consisting of polymers comprising lactic acid monomer units, polymers comprising glycolic acid monomer units, bone, hydroxyapatite, calcium phosphate, muscle, and tissue.  
     
     
         142 . A method of inducing endochondral bone formation in a mammal comprising the step of implanting the device of  claim 140  in the mammal at a locus accessible to migratory progenitor cells of the mammal.  
     
     
         143 . A method of inducing local cartilage formation in a mammal comprising the step of implanting the device of  claim 140  in the mammal at a locus accessible to migratory progenitor cells of the mammal.