Soluble bone marrow protein compositions, methods of making, and uses thereof
View Patent ↗Provided herein are soluble bioactive factor solutions, grafting scaffolds containing the bioactive factor solutions, and methods of making and using the same.
1. A method of making a soluble bone marrow protein composition, the method comprising:
harvesting bone marrow from a cadaver donor to obtain harvested bone marrow, where the harvested bone marrow contains bone marrow cells;
heating the harvested bone marrow to about 37° C. to about 60° C.;
lysing one or more of the bone marrow cells to obtain a bone marrow cell lysate, wherein lysing is carried out at about 37° C. to about 60° C., wherein lysing is carried out in a solution selected from the group consisting of: water, acetic acid, hydrochloric acid, tricholoroacetic acid, hydrofluoric acid, hydrocyanic acid, hydrogen sulfide; and
dehydrating at least a part of the bone marrow cell lysate to obtain a dehydrated soluble bone marrow protein composition.
2. The method of claim 1 , further comprising the step of fractionating the bone marrow cell lysate via centrifugation to obtain a soluble bone marrow protein fraction after performing the step of lysing the bone marrow cells.
3. The method of claim 1 , further comprising the step of washing the harvested bone marrow, wherein the step of washing occurs before the step of lysing.
4. The method of claim 1 , further comprising the step of washing the harvested bone marrow, wherein the step of washing the harvested bone marrow occurs simultaneously with the step of lysing.
5. The method of claim 1 , further comprising the step of adding a stabilization solution prior to the step of dehydrating.
6. The method of claim 1 , further comprising the step of filtering the bone marrow cell lysate.
7. The method of claim 6 , wherein filtering removes at least nucleic acids from the bone marrow cell lysate.
8. The method of claim 1 , further comprising the step of selectively filtering the bone marrow cells to obtain a desired cell population prior to or during the step of lysing the bone marrow cells.
9. The method of claim 8 , wherein the step of lysing further comprises lysing the desired cell population.
10. The method of claim 8 , wherein the desired cell population is mesenchymal stem cells.
11. The method of claim 1 , wherein an amount of the dehydrated soluble bone marrow protein composition is implanted in or otherwise administered to a subject in need thereof.
12. The method of claim 5 , wherein an amount of the dehydrated soluble bone marrow protein composition in stabilization solution is implanted in or otherwise administered to a subject in need thereof.
13. The method of claim 1 further comprising applying an amount of the dehydrated soluble bone marrow protein composition to a scaffold.
14. The method of claim 13 , wherein the scaffold is implanted in or otherwise administered to a subject in need thereof.
15. The method of claim 5 , further comprising applying an amount of the dehydrated soluble bone marrow protein composition in stabilization solution to a scaffold.
16. The method of claim 15 , wherein the scaffold is implanted in or otherwise administered to a subject in need thereof.
17. An implant comprising:
a soluble bone marrow protein composition, wherein the soluble bone marrow composition is made by a method according to claim 1 ; and
a non-bone scaffold, wherein one or more intracellular bioactive factors, cells, or one or more intracellular bioactive factors and cells of the soluble bone marrow protein composition are bound to the non-bone scaffold.
18. The implant of claim 17 , wherein the bioactive factor is a protein selected from the group consisting of: a bone morphogenetic protein; epidermal growth factor, an insulin-like growth factor, a fibroblast growth factor, vascular endothelial growth factor, otseoprotegerin, and osteopontin.
19. The implant of claim 17 , wherein the non-bone scaffold is a synthetic scaffold.