Fluid concentrator, autologous concentrated body fluids, and uses thereof

Abstract

The present invention provides devices and methods for concentrating a fluid and for treating a patient with the concentrated fluid. The concentrator apparatus includes a main housing ( 12 ) defining a separation chamber ( 14 ), a filter housing ( 48 ) containing a filter ( 46 ) comprising a filter element, a piping ( 44 ) for moving concentrated fluid from the separation chamber to the filter, and ports ( 32 ) for pressurizing the concentrated fluid past the filter element of the filter. The present invention also provides a variety of uses of concentrated body fluids, including autologous concentrated body fluid. The concentrated fluids can be used for example in surgical applications, including graft applications such as allograft, xenograft and autograft applications.

1 . A fluid concentrator comprising:
 a main housing defining a separation chamber for holding a biological fluid;   a filter housing connected to the separation chamber;   a filter held by the filter housing;   and piping for moving a fraction of the biological fluid from the separation chamber to the filter;   wherein the separation chamber comprises an adjustably positionable first port in fluid communication with the piping   
     
     
         2 . The fluid concentrator of  claim 1 , further comprising one or more ports for pressuring the fraction of the fluid past the filter. 
     
     
         3 . The fluid concentrator of  claim 2 , further comprising a pump moveably mountable relative to the one or more ports for pressuring the fraction of the fluid past the filter. 
     
     
         4 . The fluid concentrator of  claim 3 , wherein the pump is a syringe or a vacuum pump comprising a reservoir. 
     
     
         5 . The fluid concentrator of  claim 1 , further comprising at least one port for applying vacuum pressure across the filter. 
     
     
         6 . The fluid concentrator of  claim 1 , wherein the separation chamber comprises a second port to introduce the biological fluid into the separation chamber. 
     
     
         7 . The fluid concentrator of  claim 1 , wherein the separation chamber defines a longitudinal orientation during centrifugation, and wherein the filter housing and filter define a longitudinal flow direction of the extracted fraction during filtration, with the longitudinal flow direction corresponding to the longitudinal orientation of the separation chamber. 
     
     
         8 . The fluid concentrator of  claim 7 , further comprising a pump movably mountable relative to one or more parts for pressuring a separated fraction of the third part of the filter and a concentration detector positioned along the longitudinal flow direction of the filter. 
     
     
         9 . The fluid concentrator of  claim 8 , wherein the concentration detector is positioned between the filter and the pump. 
     
     
         10 . The fluid concentrator of  claim 8 , wherein the concentration detector comprises a light scattering flow cell, absorbance cell, or spectrophotometric device. 
     
     
         11 . The fluid concentrator of  claim 1 , wherein the separation chamber comprises a filter. 
     
     
         12 . The fluid concentrator of  claim 11 , wherein the filter comprises an affinity membrane, affinity support, affinity column, packed bed matrix, beads, particles, or growth matrix. 
     
     
         13 . The fluid concentrator of  claim 12 , wherein the filter comprises one or more ligands. 
     
     
         14 . The fluid concentrator of  claim 13 , wherein ligand is a receptor or antibody. 
     
     
         15 . The fluid concentrator of  claim 13 , wherein the one or more ligands bind one or more growth factors, one or more differentiation factors, one or more chemotactic factors, one or more adhesion molecules, or a combination thereof. 
     
     
         16 . The fluid concentrator of  claim 12 , wherein the growth matrix comprises a network of one or more nanofibers; a nanofibrillar structure; glass, silicon, or plastic comprising an etched or micropatterned surface; glass, silicon, or plastic surface comprising macropores or nanopores; a polymer scaffold, an extruded scaffold, a woven or net textile, a rod, a screw, a wire, a mesh, or a cage. 
     
     
         17 . The fluid concentrator of  claim 1 , wherein the biological fluid is a body fluid. 
     
     
         18 . The fluid concentrator of  claim 17 , wherein the body fluid comprises blood, plasma, serum, urine, saliva, mucus, cerebrospinal fluid, lymphatic fluid, seminal fluid, amniotic fluid, or vitreous fluid. 
     
     
         19 . The fluid concentrator of  claim 17 , wherein the body fluid comprises tissue. 
     
     
         20 . The fluid concentrator of  claim 19 , wherein the tissue comprises bone, bone marrow, muscle tissue, brain, heart, liver, lung, stomach, small intestine, large intestine, colon, uterus ovary, testis, cartilage, soft tissue, skin, subcutaneous tissue, or breast tissue. 
     
     
         21 . A method of producing a concentrated biological fluid, comprising:
 separation a biological fluid in the fluid concentrator of  claim 1 ;   withdrawing a first layer of the separated biological fluid from the separation chamber of  claim 1 ; and   filtering the withdrawn layer by passing the layer through the filter of  claim 1 .   
     
     
         22 . The method of  claim 21 , wherein the biological fluid wherein the biological fluid is a body fluid. 
     
     
         23 . The method of  claim 22 , wherein the body fluid comprises blood, plasma, serum, urine, saliva, mucus, cerebrospinal fluid, lymphatic fluid, seminal fluid, amniotic fluid, or vitreous fluid. 
     
     
         24 . The method of  claim 22 , wherein the body fluid comprises tissue. 
     
     
         25 . The method of  claim 24 , wherein the tissue comprises bone, bone marrow, muscle tissue, brain, heart, liver, lung, stomach, small intestine, large intestine, colon, uterus ovary, testis, cartilage, soft tissue, skin, subcutaneous tissue, or breast tissue. 
     
     
         26 . The method of  claim 22 , wherein the biological fluid is blood. 
     
     
         27 . The method of  claim 26 , wherein withdrawn layer is protein poor plasma (PPP) or protein rich plasma (PRP). 
     
     
         28 . The method of  claim 22 , wherein the physiological ratio of one or more components in the withdrawn layer are substantially the same as the physiological ratio of the same one or more components in said biological fluid. 
     
     
         29 . The method of  claim 22 , wherein one or more components in the withdrawn layer are concentrated about:
 a) 1:10,   b) 1:20,   c) 1:30,   d) 1:40,   e) 1:50,   f) 1:100,   g) 1:200,   h) 1:500, or   i) 1:1000   
       compared to the concentration of the same one or more components in said biological fluid. 
     
     
         30 . The method of  claim 22 , wherein one or more components of the withdrawn layer are concentrated:
 a) from about 1:20 to about 1:50,   b) from about 1:20 to about 1:100,   c) from about 1:50 to about 1:100,   d) from about 1:50 to about 1:200,   e) from about 1:100 to about 1:200,   f) from about 1:200 to about 1:500,   g) from about 1:200 to about 1:1000, or   h) from about 1:500 to about 1:1000   
       compared to the concentration of the same one or more components in said biological fluid. 
     
     
         31 . A method of reconstituting a graft material, comprising reconstituting the graft material in a concentrated biological fluid produced by the method of  claim 21 . 
     
     
         32 . The method of  claim 31 , wherein the graft material is autogenic, allogenic, or xenogenic. 
     
     
         33 . The method of  claim 32 , wherein the graft material comprises tendon, ligament, bone, cartilage, or skin. 
     
     
         34 . The method of  claim 31 , wherein the concentrated biological fluid is autologous. 
     
     
         35 . The method of  claim 31 , wherein the concentrated biological fluid is PRP or PPP. 
     
     
         36 . An implantable graft material comprising a polymeric coating, the polymeric coating comprising one or more reactive functional groups that can react with or covalently bind one or more growth factors, differentiation factors, chemotactic factors, adhesion molecules, or a combination thereof. 
     
     
         37 . An implantable graft comprising a coating agent, the coating agent comprising a nonpolymeric core molecule having attached thereto, directly or indirectly, one or more substituents comprising negatively charged groups or reactive species that interact with positively charged moieties of one or more growth factors, differentiation factors, chemotactic factors, adhesion molecules, or a combination thereof. 
     
     
         38 . The graft of  claim 36 , wherein the polymeric coating provides for the controlled release of said one or more growth factors, differentiation factors, chemotactic factors, adhesion molecules, or combination thereof. 
     
     
         39 . The graft of  claim 37 , wherein the coating agent provides for the controlled release of said one or more growth factors, differentiation factors, chemotactic factors, adhesion molecules, or combination thereof. 
     
     
         40 . The graft of  claim 36 , wherein the polymeric coating comprises nanofibers or microfibers. 
     
     
         41 . The graft of  claim 38 , wherein the rate of release of said one or more growth factors, differentiation factors, chemotactic factors, adhesion molecules, or combination thereof is determined by the rate of degradation or dissolution of one or more polymers, copolymers, or combination thereof comprising the polymeric coating. 
     
     
         42 . A method of concentrating a protein from a biological fluid, comprising:
 separating the biological fluid in the fluid concentrator of  claim 1 ;   withdrawing a fraction of the separated biological fluid comprising the protein from the separation chamber of  claim 1 ; and   filtering the withdrawn fraction of the separated biological by passing the layer through the filter of  claim 1 .   
     
     
         43 . The method of  claim 42 , wherein the protein comprises a growth factor, differentiation factor, chemotactic factor, or adhesion molecule. 
     
     
         44 . The method of  claim 42 , wherein the biological fluid comprises blood, tissue, PRP, or PPP. 
     
     
         45 . The method of  claim 42 , wherein the filter comprises an affinity membrane, affinity support, or affinity column. 
     
     
         46 . The method of  claim 45 , wherein the filter comprises one or more ligands that specifically bind the protein. 
     
     
         47 . The method of  claim 46 , wherein the ligand is a receptor or antibody. 
     
     
         48 . The method of  claim 42 , wherein the separation chamber comprises a density gradient. 
     
     
         49 . A method of promoting bone healing, comprising administering a concentrated biological fluid produced by the method of  claim 21  to the bone. 
     
     
         50 . The method of  claim 49 , wherein the concentrated biological fluid is mixed with a bone void filler. 
     
     
         51 . The method of  claim 50 , wherein the bone void filler comprises demineralized bone matrix. 
     
     
         52 . A method of promoting cartilage healing, comprising administering a concentrated biological fluid produced by the method of  claim 21  to damaged, wounded, or severed cartilage. 
     
     
         53 . The method of  claim 52 , wherein the concentrated biological fluid is mixed with an artificial polymer or collagen-based cartilage replacement product. 
     
     
         54 . A method of promoting cell or tissue regeneration following surgery or injury, comprising administering a concentrated biological fluid produced by the method of  claim 21  to the surgical site or injury. 
     
     
         55 . The method of  claim 54 , wherein the tissue is soft tissue. 
     
     
         56 . A method of promoting wound healing, comprising administering a concentrated biological fluid produced by the method of  claim 21  to the wound. 
     
     
         57 . The method of  claim 56 , wherein the wound is a surgical incision, abrasion, ulcer, or burn. 
     
     
         58 . A method of treating an orthopedic disorder in a patient, comprising administering to the patient at the site of the orthopedic disorder a concentrated biological fluid produced by the method of  claim 21 . 
     
     
         59 . The method of  claim 58 , wherein the concentrated biological fluid is mixed with bone void filler. 
     
     
         60 . The method of  claim 59 , wherein the bone void filler comprises demineralized bone matrix. 
     
     
         61 . The method of  claim 58 , wherein the orthopedic disorder comprises spinal fusion, spinal defect, bone trauma, bone cyst, bone tumor, bone fracture, filling of osseous defect, joint augmentation, sinus augmentation, ridge preservation, joint revision, or posterolateral fusion procedure. 
     
     
         62 . The method of  claim 21 , wherein the concentrated biological fluid comprises one or more growth factors, one or more different ration factors, one or more chemotactic factors, one or more adhesion molecules, stem cells, or a combination thereof. 
     
     
         63 . The method of  claim 21 , wherein the concentrated biological fluid is PRP, PPP, or PRP+PPP. 
     
     
         64 . The method of  claim 21 , wherein the withdrawn layer comprises stem cells and the biological fluid comprises bone marrow or cord blood. 
     
     
         65 . The method of  claim 21 , wherein the biological fluid has been separated by density gradient centrifugation, the biological fluid comprises seminal fluid, and the withdrawn layer comprises X-genotype sperm but not Y-genotype sperm or Y-genotype sperm but not X-genotype sperm. 
     
     
         66 . The method of  claim 21 , wherein the biological fluid has been separated by pH gradient centrifugation, the biological fluid comprises seminal fluid, and the withdrawn layer comprises X-genotype sperm but not Y-genotype sperm or Y-genotype sperm but not X-genotype sperm. 
     
     
         67 . The fluid concentrator of  claim 1 , wherein the separation chamber comprises a density gradient. 
     
     
         68 . A surgical glue or sealant comprising a concentrated biological fluid produced by the method of  claim 21 . 
     
     
         69 . A method of culturing cells or tissue, comprising
 separating a biological fluid in the fluid concentrator of  claim 1 ;   withdrawing a first layer of cells in the separated biological fluid from the separation chamber of  claim 1 ; and   passing the withdrawn cell layer through the filter of  claim 1 , wherein the filter is a growth matrix, thereby seeding the growth matrix with a concentrated fraction of cells.   
     
     
         70 . The method of  claim 21 , wherein separating comprises centrifugation or gravitational weight separation. 
     
     
         71 . The method of  claim 69 , further comprising culturing the growth matrix comprising the concentrated cells under conditions that promote proliferation and/or differentiation of the cells. 
     
     
         72 . The method of  claim 69 , wherein the biological fluid comprises blood or disrupted tissue. 
     
     
         73 . The method of  claim 69 , wherein the growth matrix comprises one or more growth factors, differentiation factors, chemotactic factors, adhesion molecules, stem cells, or a combination thereof. 
     
     
         74 . The method of  claim 69 , wherein the growth matrix comprises a network of one or more nanofibers; a nanofibrillar structure; glass, silicon, or plastic comprising an etched or micropatterned surface; glass, silicon, or plastic surface comprising macropores or nanopores; a polymer scaffold, an extruded scaffold, a woven or net textile, a rod, a screw, a wire, a mesh, or a cage. 
     
     
         75 . The method of  claim 21 , wherein the concentrated biological fluid is autologous.