Bone induction system and methods
View Patent ↗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 cells and one or more osteogenic agents, wherein the biological sample comprises whole blood;
contacting said biological sample with 0.15 N HCl, thereby lysing the biological sample to produce a lysate, said lysate comprising solid material and one or more osteogenic agents;
neutralizing said lysate to a pH of from about 4 to 10 by contacting said lysate with an alkaline composition;
separating the lysate into a first fraction and a second fraction, said first fraction substantially comprising the one or more osteogenic agents, and said second fraction substantially comprising the solid material, wherein the solid material of said second fraction including cellular debris and/or precipitated proteins; and
collecting said first fraction, thereby obtaining an osteogenic composition having one or more osteogenic agents.
2. The method of claim 1 , wherein neutralizing the acidified lysate comprises contacting the lysate with a phosphate buffer.
3. The method of claim 1 , wherein separating the lysate comprises applying the lysate to a tangential flow hollow fiber membrane filtration system such that the filtrate substantially comprises the first fraction and the retentate substantially comprises the second fraction.
4. The method of claim 1 , further comprising concentrating the first fraction.
5. The method of claim 4 , wherein concentrating the first fraction comprises applying the first fraction to a hollow fiber membrane system having a porous membrane.
6. The method of claim 5 , wherein a molecular weight cut off of the porous membrane is about 10-20 kDa.
7. The method of claim 1 , further comprising contacting the first fraction with a pharmaceutical carrier suitable for use in bone repair surgeries.
8. The method of claim 1 , wherein the first fraction is an osteogenic composition enriched for osteogenic factors, including one or more of BMP-2, VEGF, PDGF and TGF-beta, which is applied to a pharmaceutical carrier suitable for use in bone and soft tissue reparative therapies.
9. The method of claim 8 , wherein the pharmaceutical carrier is in the form of a block, paste, cement, powder, granule, putty, liquid, gel, or a combination thereof, which is comprised of at least one of a ceramic, a polymer, a metal, allograft bone, autograft bone, demineralized bone matrix, or a mixture thereof.
10. An osteogenic composition prepared by a process comprising:
obtaining a biological sample, said biological sample comprising cells and one or more osteogenic agents, wherein the biological sample comprises whole blood;
contacting said biological sample with 0.15 N HCl, thereby lysing the biological sample to produce a lysate, said lysate comprising solid material and one or more osteogenic agents;
neutralizing said lysate to a pH of from about 4 to 10 by contacting said lysate with an alkaline composition;
separating the lysate into a first fraction and a second fraction, said first fraction substantially comprising the one or more osteogenic agents, and said second fraction substantially comprising the solid material; and
collecting said first fraction, thereby obtaining an osteogenic composition having one or more osteogenic agents, and admixing the first fraction with a pharmaceutical carrier suitable for use in bone repair surgeries, wherein the pharmaceutical carrier is in the form of a block, paste, cement, powder, granule, putty, liquid, gel, or a combination thereof, which is comprised of at least one of a ceramic, a polymer, a metal, allograft bone, autograft bone, demineralized bone matrix, or a mixture thereof.
11. The composition of claim 10 , wherein neutralizing the acidified lysate comprises contacting the lysate with a phosphate buffer.
12. The composition of claim 10 , wherein separating the lysate comprises applying the lysate to a tangential flow hollow fiber membrane filtration system such that the filtrate substantially comprises the first fraction and the retentate substantially comprises the second fraction.
13. The composition of claim 10 , further comprising concentrating the first fraction.
14. The composition of claim 10 , wherein a concentration of the first fraction is formed by applying the first fraction to a hollow fiber membrane system having a porous membrane.
15. The composition of claim 14 , wherein molecular weight cut off of the porous membrane is about 10-20 kDa.
16. The composition of claim 10 , wherein the first fraction is an osteogenic composition enriched for osteogenic factors, including one or more of BMP-2, VEGF, PDGF and TGF-beta, which is applied to the pharmaceutical carrier suitable for use in bone and soft tissue reparative therapies.