Bioenergetic bone
A biological composition has a mixture of mechanically selected allogeneic biologic material derived from bone marrow. The mixture has non-whole cellular components including vesicular components and active and inactive components of biological activity, cell fragments, cellular excretions, cellular derivatives, and extracellular components. The mixture including non-whole cell fractions including one or more of exosomes, transcriptosomes, proteasomes, membrane rafts, lipid rafts. The mixture is compatible with biologic function.
1. A method of making a biological composition, comprising:
(a) grinding a cut vertebral body from a human cadaver donor to produce a mixture comprising crushed vertebral bone and bone marrow;
(b) mechanically separating cellular and non-cellular components of the bone marrow from the vertebral bone in the mixture by tumbling and sieving the vertebral bone from a decanted fluid of the bone marrow comprising cellular components comprising whole cells, and non-whole cellular components comprising non-whole cell components;
(c) concentrating the decanted fluid by centrifugation;
(d) filtering the decanted fluid through a blood filter to form a mixture including the cellular and non-whole cellular components and remove any remaining vertebral bone;
(e) separating the cellular components and non-whole cellular components of the bone marrow directly from the mixture of step (d) by density gradient centrifugation, wherein the separation by centrifugation establishes a cell density gradient;
(f) collecting non-cellular fractions, non-cellular components, or combinations thereof of predetermined density from the gradient;
(g) washing the non-cellular fractions, non-cellular components, or combinations thereof to create a mixture;
(h) suspending the mixture of step (g) to a predetermined concentration in a polyampholyte cryoprotectant to form a coated biological composition;
(i) freezing the coated biological composition at a predetermined controlled rate; and
(j) packaging a bone blend having particles of 100 to 300 μm of demineralized cortical bone, mineralized cortical bone, and mineralized cancellous bone with the coated biological composition.
2. The method of claim 1 , wherein the coated biological composition is frozen in step (i) before packaging separately with the bone blend in step (i), and wherein the method further comprises:
(k) thawing the frozen coated biological composition;
(l) diluting the thawed coated biological composition in saline;
(m) mixing the coated biological composition with the bone blend; and
(n) implanting the diluted coated biological composition and the bone blend by packing, injection, or any other suitable means into a patient.
3. The method of claim 2 , wherein the step of thawing the frozen coated biological composition occurs at a temperature of 37 degrees C. for 2 to 3 minutes.
4. The method of claim 2 , wherein the thawed coated biological composition mixed with the bone blend is re-frozen after step (m) for storage prior to step (n).
5. The method of claim 2 , wherein the thawed coated biological composition mixed with the bone blend is refrigerated after step (m) for storage prior to step (n).
6. The method of claim 2 , wherein the thawed coated composition mixed with the bone blend is stored at room temperature after step (m) for storage prior to step (n).
7. The method of claim 1 , wherein the bone blend is intermixed with the coated biological composition prior to freezing in step (i).
8. The method of claim 7 , further comprising:
(k) thawing the frozen coated biological composition intermixed with the bone blend;
(l) diluting the thawed coated biological composition intermixed with the bone blend in saline; and
(m) implanting the diluted coated biological composition intermixed with the bone blend by packing, injection, or any other suitable means into a patient.
9. The method of claim 8 , wherein the step of thawing the frozen coated biological composition intermixed with the bone blend occurs at a temperature of 37 degrees C. for 2 to 3 minutes.
10. The method of claim 8 , wherein the thawed coated biological composition intermixed with the bone blend is re-frozen after step (l) for storage prior to step (m).
11. The method of claim 8 , wherein the thawed coated biological composition intermixed with the bone blend is refrigerated after step (l) for storage prior to step (m).
12. The method of claim 8 , wherein the thawed coated composition intermixed with the bone blend is stored at room temperature after step (l) for storage prior to step (m).