Autologous bone graft substitute composition

A method of preparation of an autologous bone graft substitute composition, comprising the steps of:

• (1) mixing:
• a) autologous blood;
• b) one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13, and combinations thereof, in a range of from 0.002 mg per ml to 1 mg per ml of autologous blood, wherein the one or more analogs are protein dimers having 7 cysteines and an inter disulfide bridge at the 4th cysteine and lack affinity for naturally occurring BMP antagonist, Noggin and N-terminal amino acid introduction;
• and
• c) a compression resistant matrix selected from the group consisting of a bone allograft, bone autograft, hydroxyapatite (HA), tri-calcium phosphate (TCP), and combinations thereof;
• (2) incubating components of step (1) for a period sufficient to form a coagulum gel comprising a fibrin-meshwork reinforced with the compression resistant matrix, the coagulum gel containing the one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13, whereby the coagulum gel and the compression resistant matrix provide a sustained release of the one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13.

A method of inducing or promoting bone growth by treatment of a bone with an autologous bone graft substitute composition comprising:

• (i) autologous blood;
• (ii) one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13, and combinations thereof, in a range of from 0.002 mg per ml to 1 mg per ml of autologous blood, wherein the one or more analogs are protein dimers having 7 cysteines and an inter disulfide bridge at the 4th cysteine and lack affinity for naturally occurring BMP antagonist, Noggin and N-terminal amino acid introduction;
• and
• (iii) a compression resistant matrix selected from the group consisting of a bone autograft, bone allograft, hydroxyapatite, tri-calcium phosphate, and combinations thereof;
• wherein the autologous blood forms a coagulum gel comprising a fibrin-meshwork reinforced with the compression resistant matrix, the coagulum gel containing the one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13, whereby the coagulum gel provides a sustained release of the one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13, whereby the autologous bone graft substitute composition induces ectopic bone formation.

An autologous bone graft substitute composition for inducing new bone formation, promoting bone growth and treating of bone defect, wherein the composition comprises:

• (i) autologous blood;
• (ii) one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13, and combinations thereof, in a range of from 0.002 mg per ml to 1 mg per ml of autologous blood, wherein the one or more analogs are protein dimers having 7 cysteines and an inter disulfide bridge at the 4th cysteine and lack affinity for naturally occurring BMP antagonist, Noggin and N-terminal amino acid introduction;
• (iii) a compression resistant matrix selected from the group consisting of a bone autograft, bone allograft, hydroxyapatite, tri-calcium phosphate, and combinations thereof;
• and
• (iv) a blood clotting agent in a range from above 15 mM to 50 mM per ml of autologous blood;
• the blood clotting agent being selected from the group consisting of:
• pharmacologically acceptable calcium, strontium or magnesium salts in ionic solution or nanoparticles and Ca++microsphere conjugates;
• wherein the autologous blood forms a coagulum gel comprising the one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13 and the autologous blood clotting agent, said coagulum gel having a structure and rheological properties that provide a sustained release of the one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13.

A method of preparation of an autologous bone graft substitute composition, comprising the steps of:

• (1) mixing:
• a) autologous blood;
• b) one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, BMP-13, and combinations thereof, in a range of from 0.002 mg per ml to 1 mg per ml of autologous blood, wherein the one or more analogs are protein dimers having 7 cysteines and an inter disulfide bridge at the 4th cysteine and lack affinity for naturally occurring BMP antagonist, Noggin and N-terminal amino acid introduction;
• c) a compression resistant matrix selected from the group consisting of a bone autograft, bone allograft, hydroxyapatite, tri-calcium phosphate, and combinations thereof;
• and
• d) a blood clotting agent in a range from above 15 mM to 50 mM per ml of autologous blood;
• the blood clotting agent being selected from the group consisting of:
• pharmacologically acceptable calcium, strontium or magnesium salts in ionic solution or nanoparticles and Ca++microsphere conjugates;
• (2) incubating components of step (1) for a period sufficient to form a coagulum gel, said coagulum gel having a structure and rheological properties resulting in a sustained release of the one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13.

A method of inducing or promoting bone growth by treatment of a bone with an autologous bone graft substitute composition comprising:

• (i) autologous blood;
• (ii) one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, BMP-13, and combinations thereof, in a range of from 0.002 mg per ml to 1 mg per ml of autologous blood, wherein the one or more analogs are protein dimers having 7 cysteines and an inter disulfide bridge at the 4th cysteine and lack affinity for naturally occurring BMP antagonist, Noggin and N-terminal amino acid introduction;
• (iii) a compression resistant matrix selected from the group consisting of a bone autograft, bone allograft, hydroxyapatite, tri-calcium phosphate, and combinations thereof;
• and
• (iv) a blood clotting agent in a range from above 15 mM to 50 mM per ml of autologous blood;
• the blood clotting agent being selected from the group consisting of:
• pharmacologically acceptable calcium, strontium or magnesium salts in ionic solution or nanoparticles and Ca++microsphere conjugates;
• wherein the autologous blood forms a coagulum gel comprising the one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13 and the autologous blood clotting agent, said coagulum gel having a structure and rheological properties that provide a sustained release of the one or more analogs of BMP-6, BMP-2, BMP-7, BMP-4, BMP-5, BMP-8, BMP-9, BMP-12, and BMP-13, whereby the autologous bone graft substitute composition induces ectopic bone formation.