Osteogenic devices

Claim 2

An osteogenic device for implantation in a mammal comprising osteogenic protein dispersed within a biocompatible, in vivo biodegradable matrix defining pores of a dimension sufficient to permit influx, proliferation and differentiation of migratory progenitor cells from the body of said mammal, the improvement wherein said osteogenic protein is produced by expression of recombinant DNA in a host cell, and comprises a pair of polypeptide chains, each of which has an amino acid sequence sufficiently duplicative of the sequence of COP-5 or COP-7 such that said pair of polypeptide chains, when disulfide bonded to produce a dimeric species, has a conformation capable of inducing endochondral bone formation in association with said matrix when implanted in a mammal.

Claim 5

The device of claim 1 or 2 wherein the sequence comprises: ##STR25##

Claim 6

The device of claim 1 or 2 wherein the sequence comprises: ##STR26##

Claim 7

The device of claim 1 or 2 wherein the sequence comprises: ##STR27##

Claim 8

The device of claim 1 or 2 wherein the sequence comprises: ##STR28##

Claim 9

The device of claim 1 or 2 wherein the sequence comprises: ##STR29##

Claim 10

The device of claim 1 or 2 wherein the sequence comprises: ##STR30##

Claim 11

The device of claim 1 or 2 wherein the sequence comprises: ##STR31##

Claim 12

The device of claim 1 or 2 wherein the sequence comprises: ##STR32##

Claim 13

The device of claim 1 or 2 wherein the sequence comprises: ##STR33##

Claim 14

The device of claim 1 or 2 wherein the sequence comprises: ##STR34##

Claim 15

The device of claim 1 or 2 wherein the sequence comprises: ##STR35##

Claim 16

The device of claim 1 or 2 wherein the sequence comprises: ##STR36##

Claim 17

An osteogenic device for implantation in a mammal, said device comprising osteogenic protein dispersed within a biocompatible, in vivo biodegradable matrix defining pores of a dimension sufficient to permit influx, proliferation and differentiation of migratory progenitor cells from the body of said mammal, the improvement wherein said osteogenic protein comprises substantially pure protein capable of inducing endochondral bone formation in said mammal, said osteogenic protein having a half maximum bone inducing activity of at least about 0.8 to 1.0 ng per mg of said matrix and comprising the amino acid sequence: V--P--K--P--C--C--A--P--T.

Claim 18

An osteogenic device for implantation in a mammal, said device comprising osteogenic protein dispersed within a biocompatible, in vivo biodegradable matrix defining pores of a dimension sufficient to permit influx, proliferation and differentiation of migratory progenitor cells from the body of said mammal, the improvement wherein said osteogenic protein comprises substantially pure protein capable of inducing endochondral bone formation in said mammal, said osteogenic protein having a half maximum bone inducing activity of at least about 0.8 to 1.0 ng per mg of said matrix and comprising the amino acid sequence: V--P--K--P--C--C--A--P--T.

Claim 19

The device of claim 1, 2, or 15 wherein said matrix comprises close-packed particulate matter having a particle size within the range of 70-850 μm.

Claim 20

The device of claim 1, 2, or 15 wherein said particulate matter has a particle size within the range of 70-420 μm.

Claim 21

The device of claim 1, 2, or 15 wherein said matrix comprises demineralized, protein-extracted, particulate, allogenic bone.

Claim 22

The device of claim 1, 2, or 15 wherein said matrix comprises a material selected from the group consisting of collagen, hydroxyapalite, tricalcium phosphate, polymers comprising lactic acid monomer units, polymers comprising glycolic acid monomer units, demineralized, guanidine-extracted allogenic bone, and mixtures thereof.

Claim 23

The device of claim 1, 2, or 15 wherein said matrix is shaped to span a non-union fracture in said mammal.

Claim 24

The device of claim 1, 2, or 15 disposed within the marrow cavity of allogenic bone.

Claim 25

The device of claim 1, 2, or 15 wherein said matrix comprises demineralized, protein extracted, particulate, deglycosylated xenogeneic bone.

Claim 26

The device of claim 22 wherein said matrix is treated with a protease.

Claim 27

The device of claim 15 wherein said osteogenic protein is unglycosylated.

Claim 28

The device of claim 24 wherein said osteogenic protein has a apparent molecular weight of about 27 kD when oxidized as determined by comparison to molecular weight standards in SDS-polyacrylamide gel electrophoresis.

Claim 29

The device of claim 15 wherein said osteogenic protein is glycosylated.

Claim 30

The device of claim 26 wherein said osteogenic protein has a molecular weight of about 30 kD when oxidized as determined by comparison to molecular weight standards in SDS-polyacrylamide gel electrophoresis.

Claim 31

The device of claim 15 wherein said osteogenic protein comprises a pair of polypeptide chains.

Claim 32

The device of claim 28 wherein one chain of said pair of polypeptide chains has a molecular weight of about 14 kD and the other has an apparent molecular weight of about 16 kD, both as determined after reduction by comparison to molecular weight standards in SDS-polyacrylamide gel electrophoresis.

Claim 33

The device of claim 28 wherein one chain of said pair of polypeptide chains has a molecular weight of about 16 kD and the other has an apparent molecular weight of about 18 kD, both as determined after reduction by comparison to molecular weight standards in SDS-polyacrylamide gel electrophoresis.

Claim 34

The device of claim 15 wherein said osteogenic protein has the approximate amino acid composition set forth below: ______________________________________ Amino acid Rel. no. Amino acid Rel. no. residue res./molec. residue res./molec. ______________________________________ Aspartic acid/ 22 Tyrosine 11 Asparagine Valine 14 Glutamic acid/ 24 Methionine 3 Glutamine Cysteine 16 Serine 24 Isoleucine 15 Glycine 29 Leucine 15 Histidine 5 Proline 14 Arginine 13 Phenylalanine 7 Threonine 11 Tryptophan ND Alanine 18 Lysine 12 ______________________________________

Claim 35

A method of inducing local cartilage or bone formation in a mammal comprising the step of implanting the device of claim 1, 2, or 15 in said mammal at a locus accessible to migratory progenitor cells of said mammal.

Claim 36

A method of inducing endochondral bone formation in a mammal comprising the step of implanting the device of claim 1 or 15 in said mammal at a locus accessible to migratory progenitor cells of said mammal.

Claim 37

A method of inducing endochondral bone formation in a non-union fracture in a mammal comprising the step of implanting in the fracture in said mammal the device of claim 20.

Claim 38

The device of claim 1 or 2 wherein the sequence comprises: ##STR37## wherein X 1 =(K or R); X 2 =(H, R, or K); X 3 =(P, S, E or Q); X 4 =(Y, K or F); X 5 =(D, S or E); X 6 =(R, S, K or A); X 7 =(V, L or I); X 8 =(N, Q, D or S); X 9 =(D, E or N); X 10 =(I or V); X 11 =(I or V); X 12 =(A or S); X 13 =(P, E, L or K); X 14 =(Y or F); X 15 =(H or D); X 16 =(F, Y or N); X 17 =(H, E or S); X 18 =(E or A); X 19 =(P, A or Q); X 20 =(F or Y); X 21 =(L, M or I); X 22 =(A, P or T); X 23 =(D, E or K); X 24 =(H or S)p; X 25 =(L, M or F); X 26 =(N or K); X 27 =(S, A or P); X 28 =(T or A); X 29 =(I, V, or T); X.sub. 30 =(V, I or L); X 31 =(T or S); X 32 =(L or I); X 33 =(N, H or R); X 34 =(S, A, F or N); X 35 =(V or I); X 36 =(P or S); X 37 =(G or E); X 38 =(K, Q, T or S); X 39 =(I or V); X 40 =(K or E); X 41 =(A, P or S); X 42 =(V or A); X 43 =(T or E); X 44 =(E, Q or K); X 45 =(L or M); X 46 =(S N or D); X 47 =(A, S or P); X 48 =(I, L or V); X 49 =(S or A); X 50 =(M, I or V); X 51 =(Y or F); X 52 =(L, F or Y); X 53 =(D or N); X 54 =(E, D or N); X 55 =(N or Q); X 56 =(E, D, S or K); X 57 =(N or K); X 58 =(V or I); X 59 =(K or R); X 60 =(N, K or H); X 61 =(Q, E, R or P); X 62 =(D, E or N); X 63 =(V or T); X 64 =(E, D or R); X 65 =(G, A, S or E); X 66 =(G or H); and X 67 =(R or H).

Claim 39

The device of claim 1 or 2 wherein the sequence comprises: ##STR38## wherein X 1 =(Y, K or F); X 2 =(D, S, or E); X 3 =(R, S, K or A); X 4 =(Y, L or I); X 5 =(N, Q, D or S); X 6 =(D, E or N); X 7 l =(I or V); X 8 =(I or V); X 9 =(A or S); X 10 =(P, E, L or K); X 11 =(Y or F); X 12 =(H or D); X 13 =(F, Y or N); X 14 =(H, E or S); X 15 =(E or A); X 16 =(P, A or Q); X 17 =(F or Y); X 18 =(L, M or I); X 19 =(A, P or T); X 20 =(D, E or K); X 21 =(H or S); X 22 =(L, M or F); X 22 =(N or K); X 27 =(S, A or P); X 25 =(T or A); X 26 =(I, V, or T); X 27 =(V, I or L); X 28 =(T or S); X 29 =(L or I); X 30 =(N, H or R); X 31 =(S, A, F or N); X 32 =(V or I); X 33 =(P or S); X 34 =(G or E); X 35 =(K, Q, T or S); X 36 =(I or V); X 37 =(K or E); X 38 =(A, P or S); X 39 =(V or A); X 40 =(T or E); X 41 =(E, Q or K); X 42 =(L or M); X 43 =(S N or D); X 44 =(A, S or P); X 45 =(I, L or V); X 46 =(S or A); X 47 =(M, I or V); X 48 =(Y or F); X 49 =(L, F or Y); X 50 =(D or N); X 51 =(E, D or N); X 52 =(N or Q); X 53 =(E, D, S or K); X 54 =(N or K); X 55 =(V or I); X 56 =(K or R); X 57 =(N, K or H); X 58 =(Q, E, R or P); X 59 =(D, E or N); X 60 =(V or T); X 61 =(E, D or R); X 62 =(G, A, S or E); X 62 =(G or H); and X 64 =(R or H).

Claim 40

The device of claim 1 or 2 wherein the sequence comprises: ##STR39##