Preservation of biomaterial properties and methods of storing
Inventors
Assignees
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Publication Number
US-11930809-B2
Publication Date
2024-03-19
Expiration Date
Abstract
Described herein are enhanced compositions and methods for storing biomaterials. In certain aspects, these biomaterials include natural and engineered eukaryotic tissues. The methods described herein include storing these biomaterials in such a manner that reduces or prevents the loss of biomaterial properties (e.g., extracellular matrix integrity, cell viability, or a combination thereof) occurring either during storage or after removal of the biomaterial from storage. In certain aspects, these biomaterials will be stored in animal product-free solutions containing an agent that prevents or reduces the loss of extracellular matrix integrity.
Core Innovation
The disclosure describes enhanced biomaterial storage that preserves cell viability and extracellular matrix integrity, where the extracellular matrix integrity includes extracellular matrix permeability. The problem addressed is loss of biomaterial properties during and after cold storage, including loss of extracellular matrix permeability and associated degradation signals that reduce chondrocyte viability and cartilage matrix preservation.
A composition is prepared by placing a biomaterial in a solution including at least one agent that reduces or prevents loss of biomaterial properties, with the solution being an animal product-free solution. The biomaterial comprises chondrocytes in an extracellular matrix or cartilage, and the extracellular matrix permeability is maintained by adjusting the solution to hypothermic temperature and then storing the biomaterial at hypothermic temperature.
The disclosure further emphasizes preservation under hypothermic conditions using an agent that includes doxycycline at a concentration ranging from 10 μM to 30 μM. Documented examples address extracellular-type solutions and intracellular-type preservation media under hypothermic storage, and correlate improved post-storage viability with retention of extracellular matrix permeability and related matrix features such as water content and glycosaminoglycans.
Claims Coverage
The partial claim set includes three independent claims. Across them, the core coverage centers on forming an animal product-free composition containing an agent that reduces or prevents loss of cell viability and extracellular matrix integrity, including extracellular matrix permeability, followed by hypothermic temperature adjustment and hypothermic storage, with chondrocytes in extracellular matrix or cartilage and doxycycline used at defined concentration ranges.
Animal product-free hypothermic composition preserving cell viability and extracellular matrix permeability
A method for storing a biomaterial comprising preparing a composition by placing a biomaterial in a solution that includes at least one agent that reduces or prevents a loss of biomaterial properties, the biomaterial properties comprising cell viability and extracellular matrix integrity, the extracellular matrix integrity including extracellular matrix permeability; after formation of the composition, adjusting the solution comprising the biomaterial to a hypothermic temperature; then reducing or preventing the loss of the cell viability and the extracellular matrix permeability by storing the biomaterial in the solution at a hypothermic temperature ranging from 0° C. to +5° C. for a storage period of hours to 1 month; wherein the solution is an animal product-free solution.
Doxycycline concentration for preserving chondrocytes in cartilage extracellular matrix under hypothermic storage
A method for storing a biomaterial comprising preparing a composition by placing a biomaterial in a solution that includes at least one agent that reduces or prevents a loss of biomaterial properties, the biomaterial properties comprising cell viability and extracellular matrix integrity, the extracellular matrix integrity including extracellular matrix permeability; after formation of the composition, adjusting a temperature of the solution comprising the biomaterial to a hypothermic temperature ranging from 0° C. to +5° C.; then reducing or preventing the loss of the cell viability and the extracellular matrix permeability by storing the biomaterial in the solution at a hypothermic temperature ranging from 0° C. to +5° C. for a storage period of hours to 1 month; wherein the solution is an animal product-free solution; the biomaterial comprises chondrocytes in an extracellular matrix or cartilage; and the at least one agent comprises doxycycline having a concentration ranging from 10 μM to 30 μM.
Two-step hypothermic temperature adjustment and storage to preserve extracellular matrix integrity
A method for storing a biomaterial comprising preparing a composition by placing a biomaterial in a solution that includes at least one agent that reduces or prevents a loss of biomaterial properties, the biomaterial properties comprising cell viability and extracellular matrix integrity, the extracellular matrix integrity including extracellular matrix permeability; after formation of the composition, adjusting a temperature of the solution comprising the biomaterial to a hypothermic temperature ranging from −25° C. to +10° C.; then reducing or preventing the loss of the cell viability and the extracellular matrix permeability by storing the biomaterial in the solution at a hypothermic temperature ranging from 0° C. to +5° C. for a storage period of hours to 1 month; wherein the solution is an animal product-free solution; the biomaterial comprises chondrocytes in an extracellular matrix or cartilage; and the at least one agent comprises doxycycline having a concentration ranging from 10 μM to 30 μM.
Across the independent claims, the inventive coverage is directed to storing chondrocytes in an extracellular matrix or cartilage by preparing an animal product-free solution composition containing an agent that reduces or prevents loss of cell viability and extracellular matrix integrity, including extracellular matrix permeability, while applying hypothermic temperature adjustment and hypothermic storage. The claims further specify doxycycline as the agent at 10 μM to 30 μM and define hypothermic temperature ranges for the adjustment step and storage step, including a two-step temperature scheme in one independent claim.
Stated Advantages
Reducing or preventing loss of cell viability during storage.
Reducing or preventing loss of extracellular matrix permeability during storage.
Documented Applications
Storage of biomaterial comprising chondrocytes in an extracellular matrix or cartilage.
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