Patent Publication Number: US-2003225378-A1

Title: Mixing device for surgical sealants, and method thereof

Description:
RELATED APPLICATIONS  
     [0001] This application claims benefit of U.S. provisional application serial No. 60/375,683 filed on Apr. 26, 2002. 
    
    
     
       BACKGROUND OF THE INVENTION  
       [0002] Fluid and/or gaseous leaks can result from surgeries involving vascular, pulmonary, thoracic, spinal, meningeal, neural, hepatic, lymphatic, digestive, oncological, gynecological and renal tissues. The current standard of care involves the use of hemostats such as thrombin, gelatin and fibrin glue for diffuse bleeding, or the placement of drains until wound resolution.  
       [0003] Tissue sealants are topically applied fluid or gel media used to repair defects such as perforations or tears in tissues such as, for example, the lungs, vessels, the dura mater, and intestines. Tissue sealants are generally made ready for immediate use in surgery by first mixing together two or more components of the tissue sealant just prior to their application to tissue, followed by rapid application of the sealant to the tissue before the sealant cures and is no longer suitable for application to a tissue surface.  
       [0004] Typically, the preparation of the sealant occurs during the surgical procedure. Usually, one component of a sealant is provided in a powder form and must be dissolved in a diluent prior to mixing with the other sealant components. The powder is typically packaged in a vial. The seal of the vial must be broken by the user with a needle or other device and the diluent is injected into the vial to dissolve the powder. This process can make the sealant difficult to use, is fraught with delays, and is prone to error. Moreover, once mixed, the sealant begins to rapidly cure and if not used immediately, the sealant cures and is no longer useful as a sealant. There is, therefore, a need for devices and methods for preparing tissue sealants that are easy to mix, do not elicit severe adverse host reactions, and have a predictable cure rate.  
       SUMMARY OF THE INVENTION  
       [0005] The invention provides devices and methods for mixing elements of a composition. According to the invention, a device comprises three separate housings, which may be chambers. A device of the invention further comprises at least one mixing element for passing elements of a composition reciprocally between the housings. In use, a first element of a composition is placed in a first housing, a second element of the composition is placed in a second housing, and the two elements are mixed by reciprocal transfer through a third housing. The third housing is typically placed between the first and second housings to allow reciprocal passage of the elements between the housings for mixing. The result is a composition that is a mixture of the elements. In certain embodiments, the mixture contains more than two components. In that case, multiple components are placed into the housings. However, it is preferred that reactive components are sequestered until mixing is desired.  
       [0006] A device of the invention is useful for mixing components of a composition, such as a tissue sealant for sealing fluid or gas leaks from tissues, for example, the lungs, vessels, intestines or neural tissues. In one embodiment, a device of the invention comprises a first chamber comprising a first lumen for holding a first component. The device further comprises a second chamber comprising a second lumen for holding a second component. The first and second lumens may be in fluid communication. In a preferred embodiment a device of the invention comprises a third chamber comprising a third lumen, which may be in fluid communication with the other two lumens. Another feature of a device of the invention is a transferor or mixing element for reciprocal transfer of at least one of the first and second components between at least two of the first, second, and third chambers.  
       [0007] A third housing of a device of the invention may hold a third component. In one embodiment, a device of the invention further comprises a removable barrier interposed between at least two of the housings. The removable barrier may comprise a valve interposed between at least two of the housings. In one embodiment, the valve is a stop-cock.  
       [0008] In another embodiment, a soluble plug or an insoluble breakable membrane is used to divide one or more of the first, second, and third housings. Optionally, the first housing is in fluid communication with the second housing. In a further feature, the second housing is in fluid communication with the third housing. Optionally, the first housing is in fluid communication with the third housing.  
       [0009] A mixing element of a device according to the invention is selected from a plunger, a piston, a bladder, a pump, a propeller, an intraluminal ball or any combination thereof. In a particular embodiment according to the invention, the transferor or mixing element is a first plunger joined to the first housing and a second plunger joined to the third housing for reciprocally transferring at least one of the first and second components between the first housing and the third housing through the second housing. In another feature, at least two of the housings are detachable.  
       [0010] A preferred device of the invention comprises a slideably mounted member. The slideably mounted member contains a bore, the bore forming the second housing. Optionally, the second housing is further divided into a plurality of housings.  
       [0011] In one embodiment according to the invention, at least two of the housings are syringes. In another embodiment, the removable barrier is a soluble plug. In yet another embodiment, the removable barrier is an insoluble breakable membrane. In yet another embodiment, one or more housings include an obstruction to create turbulent flow to enhance mixing efficiency.  
       [0012] Another aspect of the invention relates to a system for preparing a tissue sealant with a predetermined cure rate. In one embodiment according to the invention, a system comprises a first chamber including a lumen, and a first mixture held in the first chamber. The first mixture, in one embodiment, includes an adhesive matrix and a cross-linker. The system also includes a second chamber. According to one embodiment, the second chamber is detachably fitted to the first chamber. In one embodiment, a second component is held in the second chamber for adjusting the pH of the first mixture held in the first chamber. In another embodiment according to the invention, the first mixture from the first chamber is mixed with the second component from the second chamber to form a second mixture and the second component adjusts the pH of the second mixture to form a sealant having the predetermined cure rate.  
       [0013] In one embodiment according to the invention, the second component held in the second chamber is a buffer. The buffer is selected from the group consisting of an ion exchange resin, organic acids, inorganic acids, acid exchange resins, organic bases, inorganic bases and alkaline exchange resins. Optionally, the buffer is selected from citric acid, phosphate, succinate, acidic salts, and morpholinoethanesulfonic acid. As another example, in another embodiment according to the invention, the pH of the first mixture is in the range of about 7-13. In this embodiment, the second component of the system lowers the pH of the second mixture to control the cure rate of the second mixture. In yet another embodiment, the pH of the first mixture is in the range of about 1-7. In this embodiment, the second component raises the pH of the second mixture to control the cure rate of the second mixture.  
       [0014] In another embodiment, the first mixture includes, for example, a carbodiimide crosslinker. In another embodiment, the lumen of the second chamber is coated with the second component.  
       [0015] Another aspect of the invention relates to a method for preparing a tissue sealant with a predetermined cure rate. In one embodiment according to the invention, the method includes fitting a first chamber holding a first mixture to a second chamber. In one embodiment, the mixture in the first chamber includes an adhesive matrix and a cross linker at about pH 7-13. In one embodiment, the method includes introducing the first mixture from the first chamber into the second chamber holding a buffer. The buffer in the second chamber adjusts the pH of the first mixture to a predetermined pH level. The method further includes the step of mixing the first mixture and the buffer in the second chamber to form a sealant having a predetermined cure rate.  
       [0016] These and other objects, along with advantages and features of the present invention herein disclosed, will become apparent through reference to the following description, the accompanying drawings, and the claims. Furthermore, it is to be understood that the features of the various embodiments described herein are not mutually exclusive and can exist in various combinations and permutations.  
     
    
    
     BRIEF DESCRIPTION OF THE FIGURES  
     [0017] The foregoing and other objects, features and advantages of the present invention disclosed herein, as well as the invention itself, will be more fully understood from the following description of preferred embodiments and claims when read together with the accompanying drawings. The drawings are not necessarily to scale, emphasis instead generally being placed upon illustrating the principles of the invention.  
     [0018]FIG. 1 is a schematic side view of a mixing device according to an illustrative embodiment of the invention.  
     [0019] FIGS.  2 - 4  are schematic side views of other embodiments of a mixing device according to an illustrative embodiment of the invention.  
     [0020]FIG. 5 is a schematic side view of a three chamber mixing device including a side port according to an illustrative embodiment of the invention.  
     [0021]FIG. 6 is a schematic side view of a three chamber mixing device according to an illustrative embodiment of the invention.  
     [0022] FIGS.  7 A- 7 C depict a method for mixing the components of a sealant in a mixing device according to an illustrative embodiment of the invention.  
     [0023]FIG. 8 is a schematic side view of a device including a fourth chamber formed by a barrier according to an illustrative embodiment of the invention.  
     [0024]FIGS. 9A and 9B are schematic side views of a mixing device including a rotationally mounted bore according to an illustrative embodiment of the invention.  
     [0025]FIG. 10A is a schematic side view of a mixing device including a slideable member according to an illustrative embodiment of the invention.  
     [0026]FIG. 10B is a schematic side section view of a mixing device including a slideable member in the closed position according to an illustrative embodiment of the invention.  
     [0027]FIG. 10C is a schematic side view of a mixing device including a slideable member according to an illustrative embodiment of the invention.  
     [0028]FIG. 10D is a schematic top view of one embodiment of a casing member illustrated in FIGS. 10A, 10B, and  10 C.  
     [0029]FIG. 10E is a schematic cross section of the casing member illustrated in FIG. 10D taken along line  10 E- 10 E of FIG. 10D.  
     [0030]FIG. 10F is a schematic end view of one embodiment of the casing member illustrated in FIG. 10D.  
     [0031]FIG. 10G is a schematic end view of one embodiment of the casing member illustrated in FIG. 10D.  
     [0032]FIG. 10H is a schematic cross section of the casing member illustrated in FIG. 10D taken along line  10 H- 10 H of FIG. 10D.  
     [0033]FIG. 10I is an isometric schematic depiction of one embodiment of a luer lock fitting that can be used in association with the casing member illustrated in FIG. 10D.  
     [0034]FIG. 10J is a schematic side view of one embodiment of the luer lock fitting illustrated in FIG. 10I.  
     [0035]FIG. 10K is a schematic cross section of the luer lock fitting illustrated in FIG. 10J taken along line  10 K- 10 K of FIG. 10J.  
     [0036]FIG. 10L is a schematic front view of a slideable member according to an illustrative embodiment of the invention.  
     [0037]FIG. 10M is a schematic side section view of the slideable member illustrated in FIG. 10L taken along line  10 M- 10 M of FIG. 10L.  
     [0038]FIG. 10N is a schematic side view of the slideable member illustrated in FIG. 10L.  
     [0039]FIG. 10O is a schematic top view of the slideable member illustrated in FIG. 10L.  
     [0040]FIG. 10P is a schematic side section view of a mixing device including a slideable member in an open position according to an illustrative embodiment of the invention.  
     [0041]FIGS. 11A and 11B are schematic side views of a mixing device including a plurality of chambers separated by a stationary tab according to an illustrative embodiment of the invention.  
     [0042]FIG. 12 is a schematic side view of a mixing device including a mixing chamber according to an illustrative embodiment of the invention. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION  
     [0043] In general, the invention provides a mixing device for mixing components of a composition, for example, a tissue sealant, and methods of use. In one illustrative embodiment, the sealant includes a mixture of albumin, a lipid, water, an enzyme, a surfactant and a cross-linker, for example, as described in U.S. application Ser. No. 09/747,293, the entirety of which is incorporated by reference herein. All of the embodiments of the device and method of the invention described herein have in common a series of chambers and mixing elements, e.g., transferors that allow for the controlled, rapid mixing of components of the tissue sealant.  
     [0044] Referring now to FIG. 1, in the illustrative embodiment, the device  2  according to the invention is a closed system including a first housing  10 , a second housing  20 , and a third housing  30  all in series. Each housing can be a chamber, with each chamber comprising a lumen  27 . In the illustrative embodiment, the lumen  27  of chamber  10  is in direct fluid communication with the lumen  27 ′ of the second chamber  20 , and the lumen  27 ′ of the second chamber  20  is in direct fluid communication with the lumen  27 ″ of the third chamber  30 . According to one feature of this embodiment, the contents of the first chamber  10  and the contents of the second chamber  20  are reciprocally transferable between the first chamber  10  and the third chamber  30  through the second chamber  20 , i.e., the contents may be moved back and forth between one chamber to another. In the illustrative embodiment, the second chamber  20  is a closed chamber except for end  22  and end  24  of the second chamber  20 . The ends  22 ,  24  are in fluid communication with the first chamber  10  and the third chamber  30 , respectively. Embodiments of the chambers include, for example, a tube, a cylinder, a bladder, or a syringe. In one exemplary embodiment according to the invention, the device  2  includes at least one mixing element, i.e., transferor  11  such as a pump or a plunger, for transferring the contents of one chamber to another chamber. According to one feature, the mixing device  2  includes more than one mixing element, for example, a first transferor  11  that is operatively joined to the first chamber  10 , and a second transferor  36  that is operatively joined to the third chamber  30 . According to another feature of the invention, the device  2  includes at least one removable barrier interposed between at least two of the chambers. The barrier can be a valve  12   a  positioned, for example, between the first chamber  10  and the second chamber  20 , and optionally a second barrier, e.g., a valve  12   b  positioned, for example, between the second chamber  20  and the third chamber  30 . The valves  12   a,    12   b  enable fluid communication between the lumens  27 ,  27 ′,  27 ″ of the chambers to be selective. The barrier can also be a gas, an insoluble breakable membrane, or a soluble plug (later described).  
     [0045] FIGS.  2 - 4  depict a mixing device  2  including a plurality of housings or chambers according to an illustrative embodiment of the invention. In the illustrative embodiment, the device  2  includes a first chamber  10  and a second chamber  20  (FIG. 2), a first chamber  10 , a second chamber  20 , and a third chamber  30  (FIG. 3), a first chamber  10 , a second chamber  20 , a third chamber  30 , and a fourth chamber  90  (FIG. 4). In other embodiments, the device  2  includes more than four chambers. According to one embodiment of the invention, any of the chambers may be detached from the device  2 . Alternatively, the one or more chambers are fixed to the device  2 , and are thus not detachable from the device  2 .  
     [0046] The housings, for example, the first chamber  10 , second chamber  20 , and third chamber  30  may be manufactured from materials such as metal, metal alloys, ceramics, glass, or plastics, such as polyvinylchloride or polyethylene, preferentially, polypropylene or polycarbonate. In a particular embodiment according to the invention, illustrated in FIG. 1, the first chamber  10  and the third chamber  30  are each a hypodermic-like syringe.  
     [0047] According to the illustrative embodiment, a valve  12  is operatively positioned between contiguous housings or chambers. For example, the illustrative embodiment of the device shown in FIG. 4 includes three valves  12 . A first valve  12   a  is positioned between the first chamber  10  and the second chamber  20 . A second valve  12   b  is positioned between the second chamber  20  and the third chamber  30 . A third valve  12   c  is positioned between the third chamber  30  and the fourth chamber  90 . Valves positioned between the chambers allow selective fluid communication between the lumens of the chambers.  
     [0048] In one embodiment, the device  2  includes three barrier-free chambers. At least two of the chambers hold a different sealant component. The device further includes a mixing element. The three chambers are assembled to permit fluid communication between the chambers and mixing of the components just prior to use.  
     [0049] Referring again to FIG. 1, according to the illustrative embodiment, the device  2  includes a first end  4  and a second end  6 . In a particular embodiment, the second end  6  is opposite to the first end  4  along a longitudinal axis indicated by arrow  3  between the first end  4  and the second end  6 . The first housing  10  is positioned at the first end  4  of the device  2 . The first valve  12   a  is positioned between the first housing  10  and the second housing  20  on the end  4   a  of the first housing  10  that is opposite to the first end  4  of the device  2 . In one illustrative embodiment, the first valve  12   a  is a two way stopcock operatively joined to the first housing  10  and to the first end  22  of the second housing  20 . In the same embodiment, the second valve  12   b  is also a two-way stopcock that is operatively joined to the second end  24  of the second housing  20  and the first end  32  of the third housing  30 .  
     [0050]FIG. 5 depicts a three housing device according to the invention including a three-way valve. In the illustrative three chamber embodiment, one or more of the valves  12  are a three-way valve, for example, including a side port  13 . With reference to FIG. 6, the device includes a second valve  12   b,  for example, a stopcock, shown in the closed position. The three-way valve  12   a  enables the contents of the first housing, i.e. the first chamber  10  and the contents of the second housing, i.e. the second chamber  20  to be reciprocally transferred between the first chamber  10  and the second chamber  20  with the use of the transferor  11  by allowing air to bleed from the otherwise closed system. A three-way valve also adds an additional port  13  for the introduction of another component to modify the sealant or adhesive. In one non-limiting example, as described in greater detail below, if a longer cure time of the sealant is required, the pH could be adjusted by adding a buffer component via the additional port  13  in the three-way valve. Thus, according to the invention the device  2  may have a plurality of valves of a variety of types, e.g., two-way and three-way valves.  
     [0051] FIGS.  7 A- 7 D depict a method for mixing a tissue sealant according to an illustrative embodiment of the invention. In one aspect, the invention provides a method for mixing the components of a tissue sealant in a mixing device  2 . The mixing device  2  includes one or more housings which hold one or more of the tissue sealant components. The housings can be chambers including lumens, where each lumen can hold one or more tissue sealing components. In one embodiment, for example, a first component  40  such as an albumin solution, described in U.S. Ser. No. 09/747,293 and incorporated by reference herein, is held in the first chamber  10  of the mixing device  2 . In the illustrative embodiment shown in FIG. 7A, the first chamber  10  is a syringe. The first valve  12   a  is operatively joined to the first chamber  10  and is in a closed position as illustrated in FIG. 7A. Referring now to FIG. 7B, for example, the first valve  12   a  is moved from the closed position (FIG. 7A) to an opened position shown in FIG. 7B. When the valve is in the open position, the first component  40  and a second component  50 , for example a diluent, such as water, held in the second chamber  20  begin to mix in the lumen  27  of the first chamber  10  and the lumen  27 ′ of the second chamber  20  forming a first mixture  45 . According to the illustrative embodiment, the first mixture  45  is rapidly reciprocally transferred between the second chamber  20  and the first chamber  10  by a mixing element, e.g., a transferor  11 . The transferor  11  can be a plunger, a piston, a bladder, an intraluminal ball, a propeller or a pump operatively joined to the first chamber  10 . To enable the first mixture  45  to be reciprocally transferred between the second chamber  20  and the first chamber  10 , an escape for air, i.e. a vent, must be provided. For example, in one embodiment a three-way valve located between the first chamber  10 , and the second chamber  20  (not shown) may provide the vent. Alternatively, if a vent is not provided, the first mixture  45  can be further mixed by aspirating the mixture into the first chamber  10  as illustrated in FIG. 7B.  
     [0052] Referring now to FIG. 7C, the second valve  12   b  is moved from a closed position (FIG. 7A) to an opened position. The first mixture  45  contained in the first chamber  10  and the second chamber  20  can then be pushed through the second valve  12   b  and into the third chamber  30  by the transferor  11 , for example a plunger  11 , or aspirated by the transferor  36 , for example, the plunger  36  operatively joined to the third chamber  30 . In the lumen  27 ″ of the third chamber  30 , the first mixture  45  is contacted with a third component  60 , for example, a cross-linker, such as, 1-ethyl-3-(3dimethylaminopropyl) carbodiimide HCl (EDC) to form a second mixture  65 . The second mixture  65  may be reciprocally transferred by the plunger  11  and the plunger  36  between the third chamber  30  and the second chamber  20  or the first chamber  10  to further mix by shearing the second mixture  65 , i.e., to mix the first component  40 , the second component  50  and the third component  60 , to form the tissue sealant. More specifically, referring to FIG. 7C, for example, plunger  11  is depressed manually to transfer the components within the first chamber  10  into the second chamber  20  and third chamber  30 . Simultaneously, plunger  36  of the third chamber  30  is pushed out of the chamber  30 . The plunger  36  is then depressed to reciprocally transfer the contents of the third chamber  30  between the third chamber  30  and the first chamber  10  through the second chamber  20  while plunger  11  of the first chamber  10  is simultaneously pushed out of chamber  10  (not shown). The number of reciprocal transfers between the first chamber  10 , the second chamber  20 , and the third chamber  30  depends on the number of transfers needed to thoroughly mix the contents of the chambers. In a particular embodiment, fifteen (15) transfers are needed to mix the components of the chambers to make the tissue sealant. After mixing, the tissue sealant is ready to be applied to a tissue defect such as a perforated lung, vessel, neural tissue or intestine. The disclosed number of chambers, tissue sealant components, and transfers are exemplary and are not intended to be limiting.  
     [0053] In another exemplary embodiment of this aspect of the invention, the device  2  further includes a fourth housing or chamber  90 . In one embodiment, the fourth chamber  90 , illustrated in FIG. 8, is formed by a barrier  72  such as a plug made from a soluble material. Alternatively the fourth chamber  90  may be formed by a barrier  72  such as an insoluble breakable membrane, for example, that will break upon application of a force to allow sealant components to mix. The barrier  72 , such as the soluble plug or insoluble breakable membrane is positioned in a chamber, for example, the third chamber  30  to divide the third chamber  30  into two chambers, i.e., the third chamber  30 , and the fourth chamber  90 . If a soluble plug is used to form the fourth chamber  90 , two powdered components of the sealant may be separated. If an insoluble breakable membrane is used to form the fourth chamber  90 , a liquid may be separated from either another liquid or a powder. If a plug is used to divide a chamber, the size of the plug  72  depends on the material used to make the plug  72  and the dimensions of the chamber. In one embodiment of this aspect of the invention, the plug  72  is made from about 0.5 to 6 mg, preferably about 2 mg to 4 mg of a lyophilized protein, e.g., albumin, e.g., bovine serum albumin (BSA). Other materials used to make the plug  72  include, but are not limited to, salts and inert polymers like starch and lipids.  
     [0054] If an insoluble breakable membrane  72  is used to divide a housing or chamber, the insoluble breakable membrane  72  may be made from polymeric materials such as polyethylene, particularly low molecular weight polyethylene, polypropylene, polycarbonate, polybutadiene, polybutadiene-nitrile, polystyrene, polyester, polyamide, polyisoprene, rubber, latex, nitrile, chloroprene, polyvinyl chloride, viton made by DuPont Dow Elastomers L.L.C. (Wilmington, Del.), or foil. The material used to make the barrier  72  should not interfere with the function of the tissue sealant. The number of housings or chambers that may be made by one or more barriers  72  or by the addition of one or more valves  12  is not limited to the specific embodiments disclosed and includes any number of housings or chambers as the circumstances require. In another embodiment, the barrier interposed between the first chamber  10  and the second chamber  20 , or the second chamber  20  and the third chamber  30  can be a soluble plug or an insoluble breakable membrane.  
     [0055] In another embodiment, the fourth chamber  90  holds, for example, a component  80 , for example, the cross linker or an enzyme. Alternatively, the fourth chamber  90  holds the diluent or the sealant matrix. Any of the chambers may hold any one or more of the components of the tissue sealant.  
     [0056] According to one embodiment, one or more of the first chamber  10 , second chamber  20 , third chamber  30  and fourth chamber  90  may hold one or more drugs, for example, antibiotics, anti-inflammatory drugs, growth factors, or other pharmaceutical or bioactive compounds.  
     [0057] Referring now to FIG. 9A, in another embodiment, the device  2  has a valve, i.e. a stop-cock, the stop-cock comprising a rotationally mounted bore  26  and a casing member  28 . The rotationally mounted bore  26 , for example, a tube, is rotationally mounted in the casing member  28 , the bore  26  forming the second chamber  20 . The casing member  28  includes ends  22   a  and  24   a  which are in fluid communication with the first chamber  10  and the third chamber  30  respectively. The second chamber  20  is a closed chamber except for ends  22  and  24 . The ends  22  and  24  are optionally in fluid communication with the first chamber  10  and the third chamber  30  respectively via bores  26   a  and  26   b  in the casing member  28 . The bores  26   a  and  26   b  extend from ends  22  and  24  to ends  22   a  and  24   a.  In one embodiment, the second chamber  20  is detachably connected to the first chamber  10 , and the third chamber  30 , for example, by luer lock fittings that are attached to or integral with the casing member  28 . The rotationally mounted bore  26  defines a lumen  27 ′ wherein the lumen  27 ′ of the rotationally mounted bore  26  can contain a component of the tissue sealant, for example, component  50 . A seal  29  is located between the casing member  28  and the rotationally mounted bore  26  to prevent leakage of the component  50 . In various embodiments, the seal  29  can be accomplished, for example, by a pressure fitting, or by one or more o-rings. The rotationally mounted bore  26  can rotate relative to the casing member  28  to move the second chamber  20  from the closed position shown in FIG. 9A, to the open position shown in FIG. 9B. In the open position illustrated in FIG. 9B, the lumen  27 ′ of the second chamber  20  is in fluid communication with the first chamber  10  and the third chamber  30  via bores  26   a  and  26   b  in the casing member  28 . When the rotationally mounted bore  26  is rotated so that the second chamber  20  is in the open position, the transferor  11  and the transferor  36 , for example, the plunger  11  and the plunger  36  can be used to reciprocally transfer the first component  40 , the second component  50 , and the third component  60  between the first chamber  10 , the second chamber  20  and third chamber  30 . Mixing is accomplished by reciprocally transferring the first component  40 , the second component  50  and the third component  60  between the first chamber  10 , the second chamber  20 , and the third chamber  30  as described hereinabove. In one embodiment, one or more powders may be placed in the second chamber  20 , and sealant matrix and diluent, for example, may be placed in the first chamber  10  and the third chamber  30  respectively. Alternatively, any of the sealant components can be placed in any chamber. After reciprocally transferring the contents of the housings or chambers to mix the sealant components, the tissue sealant is ready to be applied to a tissue defect.  
     [0058] In this aspect of the invention, the rotationally mounted bore  26  and the casing member  28  can be manufactured from, for example, one or more metals, a metal alloy, ceramic, glass, or plastics, such as polyvinylchloride, or polyethylene, preferentially, polypropylene or polycarbonate. Moreover, the diameter and length of the second chamber  20  can be optimized for peak mixing performance. In a particular embodiment, the diameter of the second chamber  20  is between 0.05 and 0.3 inches. In another embodiment, the diameter of the second chamber  20  is between 0.100 and 0.200 inches. In yet another embodiment, the diameter of the second chamber  20  is between 0.125 inches and 0.175 inches. In yet another embodiment, the length of the second chamber  20  is between 0.1 and 3 inches. In yet another embodiment, the length of the second chamber  20  is between 0.3 and 1.5 inches. In yet another embodiment, the length of the second chamber  20  is between 0.5 and 1.0 inches. In another embodiment according to the invention, the second chamber may be divided into two or more chambers by adding one or more barriers such as inert plugs or insoluble breakable membranes as described above (not shown), each chamber containing a component of the tissue sealant.  
     [0059] Referring now to FIG. 10A, in another embodiment, the device  2  has a slideable member  29  that can slide relative to a casing member  28  along a first axis. In operation, the slideable member  29  and the casing member  28  act together to form a valve. The slideable member  29  contains a cylindrical bore  62  disposed along a longitudinal axis that is perpendicular to the first axis. The bore  62  forms the second chamber  20 . The bore  62  extends from a first end  22  of the second chamber  20 , to a second end  24  of the second chamber  20 . Referring to FIGS.  10 B, and  10 D- 10 H, the casing member  28  further includes a channel  64  which is longitudinally disposed along the first axis. The bore  62  can be aligned with the first chamber  10  and third chamber  30  by slideably moving the slideable member  29  in the chamber  64 . The slideable member  29  and the casing member  28  can be made from any materials with appropriate mechanical strength that are also compatible with the sealant or adhesive. Specifically, materials such as, but not limited to, polycarbonate, polyethylene, nylon and polypropylene can be used.  
     [0060] Referring to FIGS.  10 D- 10 H, one embodiment of the casing member  28  is shown in greater detail. The casing member  28  has a first end  22   a  and a second end  24   a,  which are in fluid communication with the first chamber  10  and the third chamber  30  respectively. Referring to FIGS.  10 I- 10 K, the casing member  28  is connected to the first chamber  10  and the third chamber  30 , for example, by luer lock fittings  31 . The luer lock fittings  31  allow the casing member  28  to be connected to and detached from the first chamber  10 , and the third chamber  30 . Alternatively, the casing member  28  can be connected to the first chamber  10  and the third chamber  30  by a snap fit, or threading, for example. The first end  22   a  and the second end  24   a  of the casing member  28  are also in fluid communication with the channel  64  of the casing member  28  via bores  62   b  and  62   c.  In one embodiment illustrated in FIG. 10E, the top portion of the casing member  28  has a slot  66  which receives a pin  67 . In use (later described), the pin  67  can prevent the slideable member  29  from sliding in the channel  64  of the casing member  28 .  
     [0061] Referring now to FIGS.  10 L- 10 O, the slideable member  29  is depicted in greater detail. The slideable member  29  is snuggly received in the channel  64  of the casing member  28 . The bore  62  of the slideable member  29 , i.e. the second chamber  20 , can contain a component  50  of the tissue sealant. One or more seals  122  may be positioned between the casing member  28  and the slideable member  29  in a first circular slot  123  in the slideable member  29  to prevent leakage of the component  50 . The seals can be created, for example, but without limitation, through the use of a press fit, or through the use of o-rings. A second set of seals  125  located in a second circular slot  126  in the slideable member  29  further prevents leakage of the first component  40  and the third component  60  when the slideable member  29  is in the closed position illustrated in FIG. 10B (later described). The slideable member  29  may also contain a second bore  68  which can receive the pin  67  when the second chamber  20  is in the closed position illustrated in FIG. 10B.  
     [0062] In use, the slideable member  29  can be moved from a closed position as shown in FIGS. 10A and 10B, to an open position shown in FIGS. 10C and 10P, by sliding the slideable member  29  along the channel  64  in the casing member  28 . In the closed position as shown in FIG. 10B, the pin  67  can lock the slideable member  29 , such that the slideable member  29  is prevented from sliding in the channel  64 . To lock the slideable member  29 , the pin  67  is inserted into the slot  66  of the casing member  28  and also into the bore  68  of the slideable member  29 , which prevents movement of the slideable member  29  along the first axis. To enable movement of the slideable member  29  in the channel  64  of the casing member  28 , the pin  67  is removed from the bore  68  of the slideable member  29  and the slot  66  of the casing member  28 . When the slideable member  29  is actuated to slide in the channel  64  of the casing member  28 , the bore  62  of the slideable member  29  may be aligned with the bores  62   b  and  62   c  and ends  22   a  and  24   a  of the casing member  28 . The transferor  11  and the transferor  36 , respectfully, can then be used to reciprocally transfer the first component  40 , the second component  50 , and the third component  60  between the housings or chambers as described hereinabove.  
     [0063] In one embodiment, for example, powders are placed in the second chamber  20 , and a sealant matrix and diluent are placed in the first chamber  10  and the third chamber  30 , respectively. Any of the tissue sealant components may be placed in any chamber. Mixing of the sealant components is accomplished by reciprocally transferring the first component  40 , the second component  50 , and the third component  60  between the first chamber  10 , the second chamber  20 , and the third chamber  30  as described hereinabove.  
     [0064] In this aspect of the invention, the diameter and length of the second chamber  20  can be optimized to achieve peak mixing performance. In a particular embodiment, the diameter of the second chamber  20  is between 0.05 and 0.3 inches. In another embodiment, the diameter of the second chamber  20  is between 0.100 and 0.200 inches. In another embodiment, the diameter of the second chamber  20  is between 0.125 inches, and 0.175 inches. In yet another embodiment, the length of the second chamber  20  is between 0.1 and 3 inches. In yet another embodiment, the length of the second chamber  20  is between 0.3 and 1.5 inches. In yet another embodiment, the length of the second chamber  20  is between 0.5 and 1.0 inches. In another embodiment according to the invention, the second chamber may be divided into two or more chambers by adding one or more barriers  72 , for example inert plugs or insoluble breakable membranes into the second chamber  20  as described above (not shown).  
     [0065] In another embodiment according to the invention, illustrated in FIGS. 11A and 11B, the second housing, i.e. the second chamber  20  described above with respect to FIG.  10 A can be further divided into a plurality of chambers by a stationary tab  121 . For example, the stationary tab  121  can divide the second chamber  20  into a second chamber  141  and a third chamber  143 . The stationary tab  121  is an elongated member that is positioned along the first axis anywhere in the slideable member  29 . The stationary tab  121  includes a throughhole  145  that is aligned with ends  22   a  and  24   a  of the casing member  28 . As shown in FIG. 11A, when the slideable member  29  is in the closed position, the tab  121  separates the active components, which are contained in the second chamber  141  and the third chamber  143 . When the slideable member  129  is depressed into the channel  64  of the casing member  28  as shown in FIG. 11B, such that the slideable member  29  is in the open position, the chambers  10 ,  141 ,  143 , and  30  align permitting the chamber  141  and the chamber  143  to communicate via the throughhole  145  in the stationary tab  121 . When the chambers are aligned as shown in FIG. 11B, the components of the chambers can be reciprocally transferred between chambers  10 ,  141 ,  143 , and  30  through use of the transferor  11  and the transferor  36  as described hereinabove. After reciprocally transferring the contents of the chambers, and mixing together all of the sealant components, the tissue sealant is ready to be applied to a tissue defect. The number of chambers that may be made by one or more stationary tabs is not limited to the specific embodiments disclosed and includes any number of chambers as the circumstances require.  
     [0066] In another embodiment, according to the invention, one or more of the chambers further holds a visualant such as a soluble dye to be used as an indicator of adequate mixing of the components to form a homogeneous mixture. In yet another embodiment, the dye in one chamber may be a different color than the dye in another chamber, for example, a blue dye in one chamber, and a yellow dye in another chamber. When the contents of the chambers are mixed together, such as the yellow and blue dye, yet a third color may appear, such as a green color, when the contents are thoroughly mixed.  
     [0067] In another embodiment, according to the invention, one or more of the chambers further contains an obstruction to create turbulent flow, thus enhancing the mixing efficiency of the sealant. Examples of obstructions include, but are not limited to, static mixers, restrictors, propellers, and stationary meshes.  
     [0068] The components of the tissue sealant may assume a variety of physical forms. For example, one component may be a powder, another a gel, and another a fluid. Alternatively, two or more components may be a fluid or a gel. The powder, gel, or fluid may be held in the first chamber  10 , the second chamber  20 , the third chamber  30  or any other chamber, for example, the fourth chamber  90 . In a particular embodiment according to the invention, the component  40  in the first chamber  10  is a gel, the component  50  in the second chamber  20  is water, the component  60  in the third chamber  30  is a powder, and the component  80  in the fourth chamber  90  is a powder.  
     [0069] In another aspect, the invention comprises a device for mixing an adhesive, such as albumin, and EDC in a weakly active state, such as at an elevated pH. Referring now to FIG. 12, the device of this aspect of the invention includes a first chamber  10  connected to a second chamber  20 . The chambers  10  and  20  can optionally be detached. The first chamber  10  holds a first component  40 , such as, for example, albumin mixed with a cross-linker, for example, a carbodiimide cross-linker (EDC) to form a first mixture. The first component  40  may be alkaline at a pH at about 7-13, or more preferably at a pH at about 7-11, or most preferably at a pH at about 7-9. At this pH the albumin-EDC mixture is a semi-stable matrix that will not cure for a few minutes to a few hours. EDC reactivity is a function of pH and diminishes as pH rises. Thus, the higher the pH, the longer the time before the sealant cures. The first chamber  10  includes a transferor, such as a plunger  11 . In a particular embodiment according to the invention, the first chamber  10  is a syringe comprising a barrel and a plunger.  
     [0070] The second chamber  20  is a tube, cylinder, or static mixer holding a second component  50 . In this embodiment, the second component  50  is an acidic buffer, which lowers the pH of the first mixture. Non limiting examples of acidic buffers  50  of this invention include organic acids, inorganic acids, acid exchange resins, and ion exchange resins. More specifically, morpholinoethanesulfonic acid (MES) is an example of a buffer  50  that can be used in accordance with the invention. Other buffers  50  that can be used in accordance with the invention include citric acid, phosphate, succinates and other acidic salts. In another embodiment, the second component  50  may comprise an acid resin, such as Dowex® weakly acidic exchange resin. Optionally a first valve  12  may operatively join the first chamber  10  and the second chamber  20  (not shown).  
     [0071] According to this embodiment of the invention, the albumin and carbodiimide at a pH in the range of 7-13, i.e., the first component  40 , are introduced into the first chamber  10 . At a pH in the range of 7-13, the first mixture begins to cure at a low rate. The first chamber  10  is then joined to the second chamber  20  such that the first chamber  10  and the second chamber  20  are in fluid communication. Alternatively, the first chamber  10  and the second chamber  20  are joined before the albumin and carbodiimide mixture are added to the first chamber  10 . The valve  12  optionally separates the first chamber  10  and the second chamber  20  (not shown).  
     [0072] The first mixture is introduced by the action of the plunger  11  from the first chamber  10  into the second chamber  20 . In the second chamber  20 , the first mixture is mixed with the acidic buffer  50 . The buffer  50  is selected to cause the mixture resulting from the combination of the first mixture and the buffer  50  (hereinafter the second mixture) to reach a pH lower than the pH of the first mixture. For example, the pH of the first mixture may be lowered to a pH in the range of 1-7, preferably 3-7, and most preferably 5.5-6.5. The first mixture can be rapidly cured by passing it through the second chamber  20  containing the acidic buffer  50 . The second mixture comprised of the first mixture mixed with the buffer  50  will cure more rapidly due to a lower pH. Accordingly, the buffer  50  that is selected is the buffer that provides the desired pH and thus the desired cure rate. As used in the context of the invention described herein, cure is defined as the amount of time necessary to produce a non-fluid, cross-linked matrix. As the first mixture passes through the acidic buffer  50  which may be coated on the inner surface of the lumen  27 ′ of the second chamber  20 , the pH of the first mixture will be reduced thereby shortening the cure time of the sealant. Other factors will affect the cure rate, including but not limited to, the length and diameter of the second chamber  20 , the concentration of the acidic buffer  50 , the solubility of the buffer  50 , and the stability of the sealant cross-linker matrix  40 . Once the first mixture is mixed with the buffer  50 , the resulting second mixture is ready to be applied to tissue as a tissue sealant.  
     [0073] In an alternative embodiment of this aspect of the invention, referring still to FIG. 12, the device includes a first chamber  10  and a second chamber  20 . In this embodiment the first chamber  20  holds a first component  40 , such as an adhesive, for example, albumin. The first chamber  10  includes a transfer means, such as a plunger  11 . In a particular embodiment according to the invention, the first chamber  10  is a syringe having a barrel and a plunger. The second chamber  20  is a tube, cylinder, or static mixer and holds the second component  50 . According to this embodiment of the invention, the second component is a cross-linker, for example, EDC. Optionally a first valve  12  may operatively join the first chamber  20  and the second chamber  30  (not shown).  
     [0074] According to this embodiment of the invention, the first chamber  10  and the second chamber  20  are joined after the albumin  40  is added to the first chamber  10 . Alternatively, the first chamber  10  and the second chamber  20  are joined and after the first and second chambers are joined, albumin  40  is added to the first chamber  10 .  
     [0075] According to one feature of this embodiment, the first component  40  is introduced from the first chamber  10  into the second chamber  20 . In the second chamber  20 , the first component  40  is mixed with the crosslinker  50 . Once the first component  40  is mixed with the crosslinker  50 , the resulting mixture is ready to be applied to tissue as a tissue sealant.  
     [0076] In another embodiment of the invention, the pH of the first component  40  may be acidic at about a pH of 1-7, more preferably at about a pH of 3-7 and most preferably at about a pH of 5-7. In this embodiment of the invention, the second component  50  is an alkaline buffer that adjusts the pH of the first component  40 . Non limiting examples of alkaline buffers of this invention include organic bases, inorganic bases, alkaline exchange resins, or ion exchange resins.  
     [0077] When the first component  40  and the second component  50  are mixed as described above, the alkaline buffer  50  causes the mixture resulting from the combination of the first component  40  and the buffer  50  to reach a pH higher than the pH of the first component  40 . For example, the pH may be raised to a pH in the range of 7-13, preferably 7-11, and most preferably 7-9. The first component  40  can be rapidly cured by passing it through the second chamber  20  containing the alkaline buffer. The mixture of the first component  40  and the buffer  50  will cure more rapidly due to a raised pH. Accordingly, the buffer  50  that is selected is the buffer that provides the desired pH and thus the desired cure rate. As the first component  40  passes through the alkaline buffer  50  which may be coated on the inner surface of the lumen  27 ′ of the second chamber, the pH of the first component  40  will be raised thereby shortening the cure time of the sealant. Other factors will affect the cure rate, including the concentration of the alkaline buffer  50 , the solubility of the buffer  50 , and the stability of the sealant crosslinker matrix  40 . Once the first component  40  is mixed with the buffer  50 , the resulting mixture is ready to be applied to tissue as a tissue sealant.  
     [0078] Other embodiments incorporating the concepts disclosed herein may be used without departing from the spirit and scope of the invention. The described embodiments are to be considered in all respects as only illustrative and not restrictive.