Soft tissue xenografts

Abstract

A method for preparing a soft tissue xenograft includes the steps of removing at least a portion of a soft tissue from a non-human animal to provide a xenograft; washing the xenograft in saline and alcohol: subjecting the xenograft to cellular disruption treatment; and either digesting the xenograft with a glycosidase or glycosidase digestion followed by treatment for sialylation. A soft tissue xenograft for implantation into a human includes a portion of a soft tissue from a non-human animal, wherein the portion has extracellular matrix and substantially only dead cells. The matrix and dead cells have substantially no surface α-galactosyl moieties and have sialic acid molecules linked to at least a portion of surface carbohydrate moieties. Each of the xenografts of the invention has substantially the same mechanical properties as a corresponding native soft tissue.

1 - 29 . (Canceled)  
     
     
         30 . A method of preparing a soft tissue xenograft for implantation into a human, which comprises: 
 a. removing at least a portion of a soft tissue from a non-human animal to provide a xenograft;    b. washing the xenograft in water and alcohol;    c. subjecting the xenograft to a cellular disruption treatment; and    d. treating the xenograft with a glycosidase to remove a plurality of first surface carbohydrate moieties, whereby the xenograft has substantially the same mechanical properties as a corresponding portion of a native soft tissue, where the glycosidase has a concentration in a range of about 1 mU/ml to about 1000 U/ml.    
     
     
         31 . The method of  claim 30 , wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint, and optionally the removing step comprises removing the medial or lateral meniscus having a superior principal surface and an inferior principal surface, the outer portions of the principal surfaces being joined by an outer lateral surface, and the inner portions of the principal surfaces being joined by an inner lateral surface;    b) a ligament or tendon, and optionally the removing step comprises removing with the portion a first block of bone attached to a first end of the portion, and further optionally the removing step comprises removing with the portion a second block of bone attached to a second end of the portion opposite the first end;    c) an articular cartilage, and optionally the removing step comprises removing with the portion a layer of subchondral bone; and    d) a combination thereof.    
     
     
         32 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of  claim 31 .  
     
     
         33 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of  claim 30 .  
     
     
         34 . A soft tissue xenograft for implantation into a human comprising a portion of a soft tissue from a non-human animal, wherein the portion has substantially no surface carbohydrate moieties which are susceptible to glycosidase digestion, whereby the portion has substantially the same mechanical properties as a corresponding portion of a native soft tissue; wherein the portion of the soft tissue has extracellular components and substantially all of the cells in the tissue are dead, the extracellular components and the dead cells having substantially no surface α-galactosyl moieties, wherein the xenograft has been treated with a glycosidase having a concentration in a range of about 1 mU/ml to about 1000 U/ml to remove a plurality of first surface carbohydrate moieties.  
     
     
         35 . The soft tissue xenograft of  claim 34  wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint;  
 b) an articular cartilage from a joint;  
 c) a ligament or tendon; and  
 d) a combination thereof.  
 
     
     
         36 . A method of preparing a soft tissue xenograft for implantation into a human, which comprises: 
 a. removing at least a portion of a soft tissue from a non-human animal to provide a xenograft;    b. washing the xenograft in water and alcohol; and    c. subjecting the xenograft to a cellular disruption treatment;    whereby the xenograft has substantially the same mechanical properties as a corresponding portion of a native soft tissue.    
     
     
         37 . The method of  claim 36 , wherein (i) the cellular disruption treatment comprises freeze/thaw cycling or exposure to gamma radiation; and (ii) further comprises the steps of: 
 subsequent to step (c), treating a plurality of second surface carbohydrate moieties on the xenograft with a plurality of capping molecules to cap at least a portion of the second surface carbohydrate moieties, whereby the xenograft is substantially non-immunogenic.    
     
     
         38 . The method of  claim 37 , wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint;    b) a ligament or tendon;    c) an articular cartilage; and    d) a combination thereof.    
     
     
         39 . The method of  claim 36 , wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint;    b) a ligament or tendon;    c) an articular cartilage; and    d) a combination thereof.    
     
     
         40 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of  claim 36 .  
     
     
         41 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of  claim 37 .  
     
     
         42 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of  claim 38 .  
     
     
         43 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of  claim 39 .  
     
     
         44 . A method of preparing a soft tissue xenograft for implantation into a human, which comprises: 
 a. removing at least a portion of a soft tissue from a non-human animal to provide a xenograft;    b. washing the xenograft in water and alcohol;    c. subjecting the xenograft to a cellular disruption treatment;    d. treating the xenograft with a glycosidase to remove a plurality of first surface carbohydrate moieties; and    e. treating a plurality of second surface carbohydrate moieties on the xenograft with a plurality of sialic acid molecules to cap at least a portion of the second surface carbohydratemoieties, whereby the xenograft is substantially non-immunogenic and has substantially the same mechanical properties as a corresponding portion of a native soft tissue.    
     
     
         45 . The method of  claim 44 , wherein (i) the cellular disruption treatment comprises freeze/thaw cycling or exposure to gamma radiation; and (ii) the capping step comprises treating the second surface carbohydrate moieties on the xenograft with the sialic acid molecules having a concentration in a range of about 0.01 mM to about 100 mM.  
     
     
         46 . The method of  claim 45 , wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint;    b) a ligament or tendon;    c) an articular cartilage; and    d) a combination thereof.    
     
     
         47 . The method of  claim 44 , wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint;    b) a ligament or tendon;    c) an articular cartilage; and    d) a combination thereof.    
     
     
         48 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of  claim 44 .  
     
     
         49 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of  claim 45 .  
     
     
         50 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of  claim 46 .  
     
     
         51 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of  claim 47 .  
     
     
         52 . A soft tissue xenograft for implantation into a human comprising a portion of a soft tissue from a non-human animal, wherein the portion has extracellular components and substantially all of the cells in the tissue are dead, the extracellular components and the dead cells having substantially no surface α-galactosyl moieties and having a plurality of sialic acid molecules linked to at least a portion of a plurality of surface carbohydrate moieties on the xenograft, whereby the portion of the soft tissue is substantially non-immunogenic and has substantially the same mechanical properties as a corresponding portion of a native soft tissue.  
     
     
         53 . The soft tissue xenograft of  claim 52  wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint;  
 b) an articular cartilage from a joint;  
 c) a ligament or tendon; and  
 d) a combination thereof.  
 
     
     
         54 . A method of preparing a soft tissue xenograft for implantation into a human, which comprises the steps of: 
 a) removing at least a portion of a soft tissue from a non-human animal to provide a xenograft;    b) washing the xenograft in water and alcohol;    c) subjecting the xenograft to a cellular disruption treatment; and    d) treating a plurality of surface carbohydrate moieties on the xenograft with a plurality of capping molecules to cap at least a portion of the surface carbohydrate moieties, whereby the xenograft is substantially non-immunogenic and has substantially the same mechanical properties as a corresponding portion of a native soft tissue.    
     
     
         55 . The method of  claim 54 , wherein (i) the cellular disruption treatment comprises freeze/thaw cycling or exposure to gamma radiation; and (ii) further comprises a step selected from the group consisting of: 
 a) prior to step c, piercing the xenograft;    b) following step c, treating the xenograft with a second enzyme;    c) following step c, treating the xenograft with one or more agents selected from the group consisting of anticalcification agents, antithrombotic agents, antibiotics, and growth factors;    d) following step c, sterilizing the xenograft;    e) following step c, treating the xenograft with polyethylene glycol;    f) following step c, exposing the xenograft to a crosslinking agent in a vapor form; and    g) combinations thereof.    
     
     
         56 . The method of  claim 54 , wherein (i) the cellular disruption treatment comprises freeze/thaw cycling or exposure to gamma radiation; and (ii) the capping step comprises treating the surface carbohydrate moieties on the xenograft with sialic acid molecules having a concentration in a range of about 0.01 mM to about 100 mM.  
     
     
         57 . The method of  claim 54 , wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint;    b) a ligament or tendon;    c) an articular cartilage; and    d) a combination thereof.    
     
     
         58 . The method of  claim 55 , wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint;    b) a ligament or tendon;    c) an articular cartilage; and    d) a combination thereof.    
     
     
         59 . The method of  claim 56 , wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint;    b) a ligament or tendon;    c) an articular cartilage; and    d) a combination thereof.    
     
     
         60 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of any one of claims  54 .  
     
     
         61 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of any one of claims  55 .  
     
     
         62 . An article of manufacture comprising a substantially non-immunogenic soft tissue xenograft for implantation into a human, produced by the method of any one of claims  56 .  
     
     
         63 . A soft tissue xenograft for implantation into a human comprising a portion of a soft tissue from a non-human animal, wherein the portion has extracellular components and substantially all of the cells in the tissue are dead, the extracellular components and the dead cells having a plurality of capping molecules linked to at least a portion of a plurality of surface carbohydrate moieties on the xenograft, whereby the portion of the soft tissue is substantially non-immunogenic and has substantially the same mechanical properties as a corresponding portion of a native soft tissue.  
     
     
         64 . The soft tissue xenograft of  claim 63  wherein the soft tissue is selected from the group consisting of: 
 a) a medial or lateral meniscus from a knee joint, and optionally the medial or lateral meniscus comprises a superior principal surface and an inferior principal surface, the outer portions of the principal surfaces being joined by an outer lateral surface, and the inner portions of the principal surfaces being joined by an inner lateral surface;  
 b) an articular cartilage from a joint, and optionally the portion has attached a layer of subchondral bone;  
 c) a ligament or tendon, and optionally the portion has a first block of bone attached to a first end of the portion, and further optionally the portion has a second block of bone attached to a second end of the portion opposite the first end; and  
 d) a combination thereof.