Non-glycerol stabilized bone graft
A demineralized bone matrix (DBM) or other matrix composition is provided that has been stabilized by lowering the pH of the composition, reducing the water content, adding water substitutes, and/or increasing the amount of deuterated water present in the composition in order to reduce the activity of endogenous degrading enzymes such as proteases. A hydrated form of a stabilized DBM composition may be stable up to a year at room temperature at acidic pH. The acidified DBM compositions may be further stabilized by the addition of a stabilizing agent such as deuterated water, water substitutes, polymers, protease inhibitors, glycerol or hydrogels.
1. A demineralized bone matrix composition comprising:
demineralized bone matrix; and
at least one non-glycercol stabilizing means;
wherein the composition retains at least 50% of its original osteoinductivity after one year at room temperature, and
wherein the non-glycerol stabilizing means is a non-glycerol polyol selected from the group consisting of polynivyl alcohols and polyethylene glycols.
2. The composition of claim 1 , wherein the composition does not include glycerol.
3. The composition of claim 1 , wherein the demineralized bone matrix is in the form selected from the group consisting fibers, plates, particles, threads, and gels.
4. The composition of claim 1 further comprising water.
5. The composition of claim 1 further comprising hyaluronic acid.
6. The composition of claim 1 , wherein the non-glyercol stabilizing means is a protease inhibitor or combination of protease inhibitors.
7. The composition of claim 6 , wherein the protease inhibitor is selected from the group consisting of aprotinin, 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF), amastatin-HCl, alpha1-antichymotrypsin, antithrombin III, alpha1-antitrypsin, 4-aminophenylmethane sulfonyl-fluoride (APMSF), arphamenine A, arphamenine B, E-64, bestatin, CA-074, CA-074-Me, calpain inhibitor I, calpain inhibitor II, cathepsin inhibitor, chymostatin, diisopropylfluorophosphate (DFP), dipeptidylpeptidase IV inhibitor, diprotin A, E-64c, E-64d, E-64, ebelactone A, ebelactone B, EGTA, elastatinal, foroxymithine, hirudin, leuhistin, leupeptin, alpha2-macroglobulin, phenylmethylsulfonyl fluoride (PMSF), pepstatin A, phebestin, 1,10-phenanthroline, phosphoramidon, chymostatin, benzamidine HCl, antipain, epsilon-aminocaproic acid, N-ethylmaleimide, trypsin inhibitor, 1-chloro-3-tosylamido-7-amino-2-heptanone (TLCK), 1-chloro-3-tosylamido-4-phenyl-2-butanone (TPCK), trypsin inhibitor, sodium EDTA, and combinations thereof.
8. The composition of claim 1 , wherein the pH of the composition is below 7.
9. The composition of claim 1 , wherein the pH of the composition is below 5.
10. The composition of claim 1 , wherein the pH of the composition is below 4.
11. The composition of claim 1 , wherein the pH of the composition is below 2.
12. The composition of claim 1 , wherein the pH of the composition is between approximately 3 and 4.
13. The composition of claim 1 , wherein the pH of the composition is between approximately 4 and 5.
14. The composition of claim 1 , wherein the composition retains at least 75% of its original osteoinductivity after 1 year at room temperature.
15. The composition of claim 1 , wherein the composition retains at least 90% of its original osteoinductivity after 1 year at room temperature.
16. The composition of claim 1 , wherein the composition retains at least 75% of its original osteoinductivity after 2 years at room temperature.
17. The composition of claim 1 , wherein the composition retains at least 90% of its original osteoinductivity after 2 years at room temperature.
18. The composition of claim 1 further comprising at least one exogenous osteoinductive or osteogenic agent.
19. The composition of claim 1 further comprising:
a non-glycerol carrier.
20. The composition of claim 19 , wherein the carrier is selected from the group consisting of hyaluronic acid, collagens, lipids, polymers, proteins, and water.
21. The composition of claim 19 , wherein the carrier is selected from the group consisting of hyaluronic acid, collagens, lipids, polymers, and water.
22. The composition of claim 19 , wherein the carrier is selected from the group consisting of deuterated water (D 2 O), protease inhibitors, non-glycerol polyols, sorbitol, and acids.
23. A demineralized bone matrix composition comprising:
an exogenous destabilizing agent.
24. The composition of claim 23 , wherein the exogenous destabilizing agent is a protease.
25. The composition of claim 23 , wherein the exogenous destabilizing agent is a tissue comprising a protease.