Preservation of biomaterial properties and methods of storing

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.

What is claimed is: 
     
         1 . A composition comprising a biomaterial placed in a solution that includes at least one agent that reduces or prevents a loss of biomaterial properties, 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 an enzyme inhibitor of a matrix metalloproteinase having a concentration ranging from 1.0 nM to 1 mM.   
     
     
         2 . The composition of  claim 1 , wherein the biomaterial properties comprise extracellular matrix integrity, cell viability, or a combination thereof. 
     
     
         3 . The composition of  claim 2 , wherein extracellular matrix integrity comprises extracellular matrix permeability, extracellular matrix water content, extracellular matrix glycosaminoglycan content, or any combination thereof. 
     
     
         4 . The composition of  claim 1 , wherein the biomaterial comprises cartilage. 
     
     
         5 . The composition of  claim 1 , wherein the biomaterial comprises chondrocytes in an extracellular matrix. 
     
     
         6 . The composition of  claim 1 , wherein the solution does not include fetal bovine serum. 
     
     
         7 . The composition of  claim 1 , wherein the animal product-free solution is an extracellular-type solution that is isotonic. 
     
     
         8 . The composition of  claim 1 , wherein the animal product-free solution is an intracellular-type solution that is isotonic. 
     
     
         9 . The composition of  claim 1 , wherein the enzyme inhibitor minimizes an enzymatic activity to reduce or prevent the loss of biomaterial properties, wherein the biomaterial properties include extracellular matrix integrity. 
     
     
         10 . The composition of  claim 1 , wherein the matrix metalloproteinase comprises one or more members selected from the group consisting of MMP 1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP10, MMP 11, MMP12, MMP13, MMP14, MMP15, MMP16, MMP17, MMP 19, MMP 20, MMP 21, MMP 23A, MMP23B, MMP24, MMP25, MMP26, MMP27, and MMP28. 
     
     
         11 . The composition of  claim 1 , wherein the enzyme inhibitor is selected from the group consisting of doxycycline, TIMPs, a compound that up-regulates endogenous TIMPs, PCK3145, BB-2516, and BB-94. 
     
     
         12 . A method for storing a biomaterial comprising preparing the composition of  claim 1 , wherein preparing the composition of  claim 1  includes placing the biomaterial in a solution that includes at least one agent that reduces or prevents a loss of biomaterial properties. 
     
     
         13 . The method of  claim 12 , further comprising storing the biomaterial placed in the solution at a temperature ranging from −25° C. to +35° C. 
     
     
         14 . A composition comprising an animal product-free solution, wherein
 the solution includes at least one matrix metalloproteinase inhibitor at concentrations ranging from 1.0 nM to 1000 μM,   the animal product-free solution is an intracellular-type solution that does not include a cell culture media, and   the matrix metalloproteinase inhibitor reduces or inhibits enzymatic activity of at least one of MMP1, MMP2, MMP3, MMP7, MMP8, MMP9, MMP10, MMP11, MMP12, MMP13, MMP 14, MMP15, MMP16, MMP17, MMP19, MMP20, MMP21, MMP23A, MMP23B, MMP24, MMP25, MMP26, MMP27, MMP28 or any combination thereof.   
     
     
         15 . The composition of  claim 14 , wherein the at least one matrix metalloproteinase inhibitor reduces or inhibits enzymatic activity of at least two of MMP1, MMP8, MMP9, and MMP13. 
     
     
         16 . The composition of  claim 14 , wherein the at least one matrix metalloproteinase inhibitor reduces or inhibits enzymatic activity of at least three of MMP1, MMP8, MMP9, and MMP13. 
     
     
         17 . The composition of  claim 14 , wherein the at least one matrix metalloproteinase inhibitor is selected from the group consisting of doxycycline, TIMPs, a compound that up-regulates endogenous TIMPs, PCK3145, BB-2516, and BB-94. 
     
     
         18 . The composition of  claim 14 , wherein doxycycline is the at least one matrix metalloproteinase inhibitor and is present at a concentration of from 1.0 nM to 1000 μM. 
     
     
         19 . The composition of  claim 18 , wherein the intracellular-type solution further comprises a nutrient cocktail that includes at least one of the following components: D-glucose, glycine, L-arginine hydrochloride, L-cystine hydrochloride, L-glutamine, L-histidine hydrochloride, L-isoleucine, L-leucine, L-lysine hydrochloride, L-methionine, L-phenylalanine, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, choline, D-calcium pantothenate, folic acid, niacinamide, pyridoxine, riboflavin, thiamine, inositol, any salt thereof, or any combination thereof. 
     
     
         20 . A method comprising storing a biomaterial at hypothermic temperatures in the composition of  claim 14 , wherein
 the composition includes at least one additive that promotes retention of extracellular matrix integrity and cell viability, and   the at least one additive comprises an enzyme inhibitor, an amino acid, a plurality of amino acids, a sugar, a plurality of sugars, a lipid, a plurality of lipids, a vitamin, a plurality of vitamins, or any combination thereof.