Bone induction system and methods
An osteogenic composition suitable for use in bone and soft tissue reparative procedures is provided for herein. Methods for preparing osteogenic composition from blood and bone marrow are also provided for. The osteogenic compositions contain therapeutically effective concentrations of one or more osteogenic agents such as BMP-2, TGF-β, PDGF and VEGF.
1. A method of preparing an osteogenic composition comprising:
obtaining a biological sample, said biological sample comprising whole blood, bone marrow aspirate, or a combination of whole blood and bone marrow aspirate, from which cells and one or more osteogenic agents are derived;
at least partially lysing the biological sample to produce a lysate, said lysate comprising solid material and one or more osteogenic agents, wherein lysing the biological sample comprises acidifying the biological sample by contacting the biological sample with an amount of a hydrochloric acid solution, thereby producing an acidified lysate;
neutralizing the acidified lysate;
separating the lysate into a first fraction and a second fraction, said first fraction comprising the one or more osteogenic agents, and said second fraction comprising the solid material; and
collecting and concentrating the first fraction, thereby obtaining an osteogenic composition enriched for the one or more osteogenic agents, including one or more of BMP-2, VEGF, PDGF and TGF-β.
2. The method of claim 1 , wherein the bone marrow aspirate being passed through a three-way valve system one or more times prior to lysing the biological sample.
3. The method of claim 1 , wherein lysing the biological sample further comprises subjecting the biological sample to one or more freeze/thaw cycles.
4. The method of claim 1 , wherein neutralizing the acidified lysate comprises contacting the lysate with an alkaline composition.
5. The method of claim 1 , wherein neutralizing the acidified lysate comprises contacting the lysate with a phosphate buffer.
6. The method of claim 1 , wherein separating the lysate comprises applying the lysate to a tangential flow hollow fiber membrane filtration system, wherein the filtrate comprises the first fraction and the retentate comprises the second fraction.
7. The method of claim 1 , wherein separating the lysate comprises centrifuging the lysate so that the supernatant comprises the first fraction and the pellet comprises the second fraction.
8. The method of claim 1 , wherein concentrating the first fraction comprises applying the first fraction to a hollow fiber membrane system, wherein a molecular weight cut-off of the membrane is about 10-20 kD.
9. The method of claim 1 , further comprising combining the osteogenic composition with a pharmaceutical carrier suitable for bone or soft tissue reparative therapies.
10. The method of claim 9 , wherein the pharmaceutical carrier is in the form of a block, paste, cement, powder, granule, putty, liquid, gel, or a combination thereof, and wherein the pharmaceutical carrier comprises a ceramic, a polymer, a metal, allograft bone particles, autograft bone particles, demineralized bone matrix particles, or a mixture thereof.