Tenocyte Containing Bioscaffolds And Treatment Using The Same

Abstract

The present invention relates to methods for preparing bioscaffolds useful in the repair of tears. More specifically, the invention relates to a method of treating rotator cuff tear in a mammalian subject in need thereof comprising the steps of: (i) selectively expanding tenocytes in vitro in culture medium comprising insulin or a functional derivative and a glucocorticoid or a glucocorticoid-like molecule to produce a culture of expanded tenocytes; (ii) seeding a bioscaffold with said expanded tenocytes to produce a tenocyte seeded bioscaffold; and (iii) implanting said tenocyte seeded bioscaffold proximal to the rotator cuff tear. The present invention also relates to a bioscaffold comprising cells, wherein more than 80% of said cells are tenocytes.

What is claimed: 
     
         1 . A tenocyte seeded bioscaffold for the treatment of rotator cuff tear comprising:
 (i) an amount of selectively expanded tenocyte cells that express one or more genes encoding the following: type I collagen, type III collagen, EphA4, scleraxis, Six1, COMP and Cbfa1;   (ii) a bioscaffold being a membrane, a fleece, a thread or mixtures thereof, wherein said bioscaffold comprises a type I/III collagen matrix or a small intestinal submucosa;   wherein said selectively expanded tenocyte cells have been in vitro cultured on said bioscaffold for sufficient time to adhere the tenocytes before use.   
     
     
         2 . The tenocyte seeded bioscaffold of  claim 1 , wherein the step of in vitro culturing of the tenocyte cells comprises:
 (i) culturing isolated tenocyte cells in medium comprising:
 (a) at least 10% v/v serum; 
 (b) insulin; and 
 (c) betamethasone to produce said culture of expanded tenocytes; 
   (ii) seeding the bioscaffold with said expanded tenocytes; and   (iii) culturing the bioscaffold and expended tenocytes for no more than five days to produce the tenocyte seeded bioscaffold.   
     
     
         3 . The tenocyte seeded bioscaffold of  claim 1 , wherein said tenocyte cells are isolated from a mammalian animal selected from the group consisting of a sheep, a cow, a pig and a human. 
     
     
         4 . The tenocyte seeded bioscaffold of  claim 1 , wherein said tenocyte cells are isolated from a human. 
     
     
         5 . The tenocyte seeded bioscaffold of  claim 1 , wherein said tenocyte cells are autologous. 
     
     
         6 . The tenocyte seeded bioscaffold of  claim 2 , wherein said medium comprises about 0.0006% w/v insulin and about 0.0002% w/v betamethasone. 
     
     
         7 . The tenocyte seeded bioscaffold of  claim 1 , wherein said selectively expanded tenocyte cells comprises at least 80% tenocyte cells. 
     
     
         8 . The tenocyte seeded bioscaffold of  claim 1 , wherein the step of seeding a bioscaffold with said expanded tenocytes to produce a tenocyte seeded bioscaffold is carried out at a cell density of 0.05 to 5 times the physiological cell density of native tendon. 
     
     
         9 . The tenocyte seeded bioscaffold of  claim 8 , wherein the cell density is at least 1×10 6  to 1×10 7  cells per cm 2 . 
     
     
         10 . The tenocyte seeded bioscaffold of  claim 8 , wherein the cell density is 4×10 6  cells per cm 2 . 
     
     
         11 . The tenocyte seeded bioscaffold of  claim 2 , wherein said culture medium comprises at least 15% v/v serum.