Acellular biologic composition and method of manufacture
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 preparing a biological composition for use comprises the steps of:
collecting, recovering and processing bone marrow from a cadaver donor;
mechanically separating by spinning or tumbling cellular and non-cellular components of bone marrow from cadaverous bone using a plurality of cycles, wherein the first cycle has a decanted fluid of unwanted cells and debris which is discarded, thereafter subsequent cycles are performed, each subsequent cycle creating a decanted fluid which is retained containing the cellular and non-cellular components, the retained decanted fluid is combined in a collection jar prior to centrifugation and passed through a blood filter to remove any bone or spicules or clumps from the cells and the non-whole cell components;
concentrating the separated cellular and non-cellular components by centrifugation;
separation of cellular and non-cellular components by density gradient centrifugation;
collecting non-cellular fractions or non-cellular components or combinations in a fluid above an interface containing whole cells;
washing the non-cellular fractions or non-cellular components or combinations thereof to create a mixture;
quantifying non-whole cell fraction from the fluid above the interface has a concentration exceeding zero;
suspending the mixture in a polyampholyte cryoprotectant;
freezing the mixture.
2. The method of preparing the biological composition for use of claim 1 further comprises the steps of:
thawing the mixture; and
implanting the thawed mixture with a bone blend being intermixed by packing, injection or any other suitable means into a patient.
3. The method of preparing the biological composition for use of claim 2 wherein the step of thawing the mixture occurs at a temperature of 37 degrees C. for 2 to 3 minutes in a warm water bath.
4. The method of preparing the biological composition for use of claim 2 further comprises the step of:
diluting the thawed mixture in saline without spinning.
5. The method of preparing the biological composition for use of claim 3 further comprises the step of:
diluting the thawed mixture in saline without spinning.