Compositions for regenerating tissue that has deteriorated, and methods for using such compositions

Abstract

A composition for promoting regeneration of tissue which has degenerated in a subject as a result of a disease or disorder and a method of using the composition is provided. The composition comprises a biodegradable acellular matrix, and passaged autologous fibroblasts substantially free of immunogenic proteins, e.g., culture medium serum-derived proteins, integrated within the matrix. The method of using the composition to promote regeneration of tissue involves placing the composition on a site of degenerated tissue in a subject so that the composition promotes tissue regeneration at the site. The composition and the method of its use have applications promoting regeneration of tissue (i) that has degenerated as a result of numerous diseases or disorders or (ii) that has a defect, including, but not limited to, defects of the oral mucosa, trauma to the oral mucosa (e.g., extraction of a tooth), periodontal disease, diabetes, cutaneous ulcers, venous stasis, scars of the skin, or wrinkles of the skin. Also provided is an injectable composition comprising any type of collagen and passaged autologous fibroblasts substantially free of immunogenic proteins, e.g., culture medium serum-derived proteins, for correcting defects in skin, such as wrinkles or scars, and for augmenting tissue in the subject, particularly facial tissue.

1 . A method for repairing tissue that has been damaged in a subject, wherein the method comprises: 
 a) providing a pharmaceutical composition comprising: (i) autologous passaged fibroblasts substantially free of proteins derived from xenogeneic serum of xenogeneic serum-containing culture medium; and (ii) a pharmaceutically acceptable carrier;    b) identifying a site in the subject of: (i) tissue degeneration due to a disease or disorder; or (ii) a tissue defect, wherein the tissue degeneration or the tissue defect in the subject comprises a defect of an oral mucosa, trauma to an oral mucosa, trauma to an oral bone, periodontal disease, diabetes, cutaneous ulcers, or venous stasis; and    c) injecting a therapeutically effective amount of the pharmaceutical composition subadjacent to the tissue degeneration or defect,    wherein the injecting results in repair of the tissue degeneration or the defect    
     
     
         2 . The method of  claim 1 , wherein the tissue degeneration or the tissue defect comprises periodontal disease.  
     
     
         3 . The method of  claim 2 , wherein the periodontal disease comprises periodontal degeneration.  
     
     
         4 . The method of  claim 2 , wherein the periodontal disease comprises gingivitis.  
     
     
         5 . The method of  claim 2 , wherein the periodontal disease comprises a non-healing wound of a palatal mucosa.  
     
     
         6 . The method of  claim 2 , wherein the periodontal disease comprises a non-healing wound of a gingival mucosa.  
     
     
         7 . The method of  claim 1 , wherein the tissue degeneration or the tissue defect comprises a defect of an oral mucosa.  
     
     
         8 . The method of  claim 7 , wherein the defect of an oral mucosa is caused by trauma.  
     
     
         9 . The method of  claim 7 , wherein the defect of an oral mucosa is caused by a dermatosis.  
     
     
         10 . The method of  claim 7 , wherein the defect of an oral mucosa is caused by aphthous stomatitis.  
     
     
         11 . The method of  claim 7 , wherein the defect of an oral mucosa is caused by an infection.  
     
     
         12 . The method of  claim 1 , wherein the tissue degeneration or the tissue defect comprises trauma to an oral mucosa.  
     
     
         13 . The method of  claim 12 , wherein the trauma to an oral mucosa is caused by extraction of a tooth.  
     
     
         14 . The method of  claim 1 , wherein the tissue degeneration or the tissue defect comprises trauma to an oral bone.  
     
     
         15 . The method of  claim 14 , wherein the trauma to an oral bone is caused by extraction of a tooth.  
     
     
         16 . The method of  claim 14 , wherein the oral bone is a maxillary bone.  
     
     
         17 . The method of  claim 14 , wherein the oral bone is a mandibular bone  
     
     
         18 . The method of  claim 1 , wherein the step of providing a pharmaceutical composition comprises: 
 a) taking a biopsy of dermis comprising fibroblasts from a subject;    b) separating fibroblasts from said biopsy so as to provide fibroblasts substantially free of extracellular matrix and non-fibroblast cells;    c) placing the fibroblasts in a culture medium comprising between 0.0% and about 20% serum in order to grow fibroblasts;    d) incubating the fibroblasts in a serum-free medium for at least 2 hours at between about 30° C. and about 37.5° C. to form passaged fibroblasts;    e) exposing the passaged fibroblasts to a proteolytic enzyme so as to suspend the passaged fibroblasts; and    f) adding a pharmaceutically acceptable carrier to the suspended passaged fibroblasts to form the pharmaceutical composition.    
     
     
         19 . The method of  claim 18 , wherein the biopsy is taken from gums or palate of the subject.  
     
     
         20 . The method of  claim 18 , wherein the biopsy is taken from skin of the subject.  
     
     
         21 . The method of  claim 18 , wherein the proteolytic enzyme is trypsin.