Coated biological composition

Abstract

A coated biological composition has a mixture of biologic material and a volume of a liquid protectant. The mixture of biologic material has non-whole cellular components or whole cells or combinations of the non-whole cellular components and whole cells, wherein the mixture is compatible with biologic function. The volume of a liquid protectant is intermixed with the mixture of biologic material, wherein the liquid protectant forms a coating externally enveloping each of the non-whole cellular components, if any, and each of the whole cells, if any, of the mixture of biologic material, to form the coated biological composition. The coated biological composition is frozen and thereafter thawed and then frozen a second time for storage or frozen at least once and thawed and stored under refrigeration above freezing, or frozen and thawed and then concentrated by drying, or while frozen without thawing lyophilized for ambient or room temperature storage.

What is claimed is: 
     
         1 . A coated biological composition comprising:
 a mixture of biologic material having non-whole cellular components including vesicular components and active and inactive components of biological activity, cell fragments, cellular excretions, cellular derivatives, and extracellular components, or whole cells or combinations of the non-whole cellular components and whole cells, wherein the mixture is compatible with biologic function;   a volume of a liquid protectant intermixed with the mixture of biologic material; and   wherein the liquid protectant forms a coating externally enveloping each of the non-whole cellular components, if any, and each of the whole cells, if any, of the mixture of biologic material, to form the coated biological composition and wherein the coated biological composition is frozen and thereafter thawed and then frozen a second time for storage or frozen at least once and thawed and stored under refrigeration above freezing, or frozen and thawed and then concentrated by drying, or while frozen without thawing lyophilized for ambient or room temperature storage.   
     
     
         2 . The coated biological composition of  claim 1  wherein the thawed coated biological composition re-frozen for storage prior to use. 
     
     
         3 . The coated biological composition of  claim 1  wherein the thawed coated biological composition is refrigerated above freezing for storage prior to use. 
     
     
         4 . The coated biological composition of  claim 1  wherein the thawed coated composition is processed to be stored at room temperature for storage. 
     
     
         5 . The coated biological composition of  claim 1  wherein the thawed coated biological composition is concentrated by drying prior to being stored at room temperature to form a dried coated biological composition. 
     
     
         6 . The coated biological composition of  claim 5  wherein the coated biological composition is freeze dried or hypothermic dehydrated. 
     
     
         7 . The coated biological composition of  claim 6  wherein the dried coated biological composition is micronized into particles of 1000 microns or less. 
     
     
         8 . The coated biological composition of  claim 6  wherein the dried coated biological composition is micronized into particles of 400 microns or less. 
     
     
         9 . The coated biological composition of  claim 1  wherein the liquid protectant is a polyampholyte protectant or polyampholyte cryoprotectant. 
     
     
         10 . The coated biological composition of  claim 1  wherein the liquid protectant is a polyampholyte protectant or polyampholyte cryoprotectant, or a glycerol-based protectant, or dimethyl sulfoxide (DMSO) or glycols or trehalose or other molecular hydrogel preservatives or sucrose or dextrose based protectant. 
     
     
         11 . The coated biological composition of  claim 1  wherein the liquid protectant is dimethyl sulfoxide (DMSO). 
     
     
         12 . The coated biological composition of  claim 5  wherein the dried coated biological composition has a moisture content of 5% or less. 
     
     
         13 . The coated biological composition of  claim 12  wherein an initial volume of the liquid protectant when reduced to a solid for drying or freeze-drying yields 5% or less of the initial volume. 
     
     
         14 . The coated biological composition of  claim 13  wherein the dried coated biological composition when reconstituted in a liquid is suitable for direct implantation. 
     
     
         15 . The coated biological composition of  claim 1  further comprises a volume of cartilage particles, the particles intermixed with the biological material and coated with the liquid protectant. 
     
     
         16 . The coated biological composition of  claim 1  further comprises a volume of nucleus pulposus particles. 
     
     
         17 . The coated biological composition of  claim 1  wherein the coating deters attachment to other cells for a predetermine time. 
     
     
         18 . The coated biological composition of  claim 1  wherein the coating buffers inflammation. 
     
     
         19 . The coated biological composition of  claim 1  wherein the liquid protectant is a polyampholyte protectant of a polyamine polymer compound. 
     
     
         20 . The coated biological composition of  claim 1  wherein the coating retards or reduces premature differentiation of the whole cells of the mixture. 
     
     
         21 . The coated biological composition of  claim 1  wherein the coating sustains regenerative potential and biologic function of the mixture during preservation and implantation. 
     
     
         22 . The coated biological composition of  claim 1  wherein the coating is configured to be metabolized after implantation after a predetermined time. 
     
     
         23 . The coated biological composition of  claim 22  wherein the predetermined time is three or more days. 
     
     
         24 . The coated biological composition of  claim 23  wherein the predetermined time is up to six days. 
     
     
         25 . The coated biological composition of  claim 1  wherein the coating forms a spherical shell about each whole cell. 
     
     
         26 . The coated biological composition of  claim 19  wherein the polyampholyte protectant is a cryoprotectant. 
     
     
         27 . The coated biological composition of  claim 26  wherein the polyampholyte protectant forms a strong hydrophilic characteristic of the coating to protect the cell membrane external of the whole cells. 
     
     
         28 . The coated biological composition of  claim 27  wherein the protectant when frozen as a cryoprotectant interrupts crack propagation externally protecting the cell membrane. 
     
     
         29 . The coated biological composition of  claim 1  wherein the mixture is mechanically selected allogeneic biologic material derived from bone marrow. 
     
     
         30 . The coated biological composition of  claim 1  further comprises bone particles, the bone particles being added to the mixture and coated with liquid protectant. 
     
     
         31 . The coated biological composition of  claim 30  wherein the bone particles include a mixture of cortical bone particles and cancellous bone particles. 
     
     
         32 . The coated biological composition of  claim 29  wherein the mixture of mechanically selected material derived from bone marrow further includes a select number of non-whole cell fractions including one or more of exosomes, transcriptosomes, proteasomes, membrane rafts, lipid rafts. 
     
     
         33 . The coated biological composition of  claim 32  wherein the combination of non-whole cell components with a select number of the non-whole cell fractions sustains pluripotency in both graft or host cells or combinations thereof. 
     
     
         34 . The coated biological composition of  claim 33  wherein the select number of the non-whole cell fractions sustains pluripotency in graft or host cells or combinations thereof includes differentiated committed cells and non-differentiated and non-committed cells. 
     
     
         35 . The coated biological composition of  claim 29  wherein the biological composition is predisposed to demonstrate or support elaboration of active volume or spatial geometry consistent in morphology with that of endogenous bone. 
     
     
         36 . The coated biological composition of  claim 1  wherein the biological composition extends regenerative resonance that compliments or mimics tissue complexity. 
     
     
         37 . The coated biological composition of  claim 1  wherein the mixture is treated in the protectant prior to preservation or cryopreservation or freeze drying. 
     
     
         38 . The coated biological composition of  claim 37  wherein the protectant creates a physical or electrical or chemical gradient or combination thereof for tissue regeneration. 
     
     
         39 . The coated biological composition of  claim 38  wherein the gradient has a physical characteristic of modulus or topography such as charge density, field shape or cryo- or chemo-taxic tendencies. 
     
     
         40 . The coated biological composition of  claim 38  wherein the gradient has a chemical characteristic of spatially changing compositions of density or species of functional molecules, wherein the molecules can offer a fixed catalytic function as a co-factor. 
     
     
         41 . The coated biological composition of  claim 38  wherein the gradient has an electrical characteristic of charge based or pH based or electron affinities that confer metastability in biologic potential. 
     
     
         42 . The coated biological composition of  claim 29  wherein the bone marrow which is derived from a cadaver has separation-enhanced non-whole cell fractions vitality including one or more of the following: separating the fractions from cells heightens their vitality, reversing “arrest” of donors, accentuating responsive molecular coupling, matrix guarding in neutralizing inflammation or providing a basis for metabolic satience by balancing stimulus for repair. 
     
     
         43 . The coated biological composition of  claim 1  wherein the liquid protectant is a cryoprotectant polyampholyte of carboxylated polylysine. 
     
     
         44 . The coated biological composition of  claim 36  wherein the regenerative resonance occurs in the presence or absence of a refractory response. 
     
     
         45 . The coated biological composition of  claim 43  wherein the cryopreservation occurs at a temperature that is sub-freezing. 
     
     
         46 . The coated biological composition of  claim 45  wherein the cryopreservation temperature is from 0 degrees C. to −200 degrees C. 
     
     
         47 . The coated biological composition of  claim 1  wherein the mixture creates a physical or electrical or chemical gradient or combination thereof for tissue regeneration. 
     
     
         48 . The coated biological composition of  claim 47  wherein the gradient has a physical characteristic such as modulus or topography. 
     
     
         49 . The coated biological composition of  claim 47  wherein the gradient has a chemical characteristic such as spatially changing compositions of density or species of functional molecules. 
     
     
         50 . The coated biological composition of  claim 47  wherein the gradient has an electrical characteristic such as charge based or pH based. 
     
     
         51 . The coated biological composition of  claim 1  can be organelle fragments. 
     
     
         52 . The coated biological composition of  claim 1  wherein active and inactive components of biological activity can be extants of the human metabolome. 
     
     
         53 . The coated biological composition of  claim 43  wherein the composition is maintained at ambient temperature prior to freeze drying. 
     
     
         54 . The coated biological composition of  claim 1  wherein the mixture of biological material is intermixed with a liquid protectant of a polyampholyte protectant for direct implantation wherein said protectant is a 1-50 w/w % aqueous solution of at least one polyamine polymer compound comprised of at least one polymer of units having side-chain amino groups, said at least one polymer of units being selected from a group consisting of ε-poly-L-lysine, α-poly-L-lysine, poly-arginine, allylamine polymer and partially methoxy-carbonylated allylamine polymer; and 50-99 mol % of amino groups, other than those forming amino-acid-to-amino-acid linkages, of said at least one polymer compound is blocked with carboxylic anhydride to form pendant moieties, each of which is linked to main chain of the polymer via an amide linkage and essentially has a not-blocked carboxylic group. 
     
     
         55 . The coated biological composition according to  claim 54 , wherein said liquid protectant is obtained by dissolving the at least one polyamine polymer compound in a physiological solution. 
     
     
         56 . The coated biological composition according to  claim 55 , wherein the physiological solution is a saline, Dulbecco-modified eagle MEM culture medium (DMEM), or a culture medium for cells or tissues. 
     
     
         57 . The coated biological composition according to  claim 54 , wherein said at least one polymer compound is ε-poly-L-lysine having number-average molecular weight in a range of 1000-20,000. 
     
     
         58 . The coated biological composition according to  claim 54 , wherein remaining side-chain amino groups or remaining side-chain and terminal amino groups of the at least one polymer compound are not blocked by covalent bonding. 
     
     
         59 . A method of making a coated biological composition comprises the steps of:
 collecting, recovering and processing bone marrow from a cadaver donor;   mechanically separating cellular and non-cellular components of bone marrow from cadaverous bone;   concentrating by centrifugation and filtering;   separation by density gradient centrifugation;   collecting non-cellular fractions or non-cellular components or combinations thereof of predetermined density;   washing the non-cellular fractions or non-cellular components or combinations thereof to create a mixture;   quantifying non-whole cell fraction concentration exceeds zero;   suspending to a predetermined concentration in a liquid protectant;   freezing the mixture at a predetermined controlled rate; and   thereafter thawing and then freezing a second time for storage or frozen at least once thawed and stored under refrigeration above freezing, or frozen and thawed and then concentrated by drying, or while frozen without thawing lyophilized for ambient or room temperature storage.   
     
     
         60 . A method of preparing the mixture for use made according to the method of  claim 59  wherein the liquid protectant is a polyampholyte protectant or polyampholyte cryoprotectant, or a glycerol-based protectant, or dimethyl sulfoxide (DMSO) or glycols or trehalose or other molecular hydrogel preservatives or sucrose or dextrose based protectant. 
     
     
         61 . The method of preparing the mixture for use made according to the method of  claim 59  further comprises the step of adding a volume of cartilage particles, the particles intermixed with the biological material and coated with the liquid protectant. 
     
     
         62 . The method of preparing the mixture for use made according to the method of  claim 59  further comprises the step of adding a volume of nucleus pulposus particles, the particles intermixed with the biological material and coated with the liquid protectant. 
     
     
         63 . The method of preparing the mixture for use made according to the method of  claim 59  further comprises the step of adding a volume of bone particles, the particles intermixed with the biological material and coated with the liquid protectant. 
     
     
         64 . The method of preparing the mixture for use made according to the method of  claim 59  by the steps of:
 diluting the mixture in saline without spinning; and 
 implanting the diluted mixture by packing, injection or any other suitable means into a patient. 
 
     
     
         65 . The method of preparing the mixture for use made according to the method of  claim 64  wherein the step of diluting the mixture includes warming the mixture at a temperature of 37 degrees C. for 2 to 3 minutes in a warm water bath.