Methods and compositions for culturing a biological tooth

Abstract

Tooth tissues include the pulp mesenchyme that forms the dentin and an epithelium that is responsible for enamel formation. Cells from these tissues were obtained from porcine third molars and were seeded onto a biodegradable scaffold composed of a polyglycolic acid-polylactic acid copolymer. Cell polymer constructs were then surgically implanted into the omentum of athymic nude rats so that the constructs would have a blood supply and these tissues were allowed to develop inside the rats. Histological analysis of 7.5 week-old implants revealed a dentin-like collagenous matrix containing hydroxyapatite mineral surrounding a core of mesenchymal cells that appeared analogous to pulp tissue. Infrequently, columnar epithelial cells were observed as a single layer on the outside of the dentin-like matrix similar to the actual arrangement of ameloblasts over dentin during early tooth development. Developing tooth tissues derived from such cell polymer constructs could eventually be surgically implanted into the gum of an edentulous recipient where the construct would receive a blood supply and develop to maturity, providing the recipient with a biological tooth replacement.

1 . A method of generating tooth tissue comprising: applying tooth germ cells onto a biodegradable polymer scaffold and allowing the tooth germ cells to develop into a tooth.  
     
     
         2 . The method of  claim 1 , further comprising: 
 forming a tooth mold containing the biodegradable polymer scaffold.    
     
     
         3 . The method of  claim 1 , wherein the biodegradable polymer scaffold has a shape of a tooth.  
     
     
         4 . The method of  claim 3 , wherein the biodegradable polymer scaffold has a shape of a human tooth.  
     
     
         5 . The method of  claim 1 , wherein the tooth germ cells are mammalian.  
     
     
         6 . The method of  claim 1 , wherein the tooth germ cells are porcine.  
     
     
         7 . The method of  claim 1 , wherein the tooth germ cells comprise one or more of cells selected from the group consisting of cells from an enamel organ, cells from a pulp organ and tissue cultured cells derived from a tooth tissue.  
     
     
         8 . The method of  claim 1 , wherein the tooth germ cells are applied to the biodegradable polymer scaffold with between about 20 to 50 million cells per square centimeter of scaffold.  
     
     
         9 . The method of  claim 1 , wherein the biodegradable polymer scaffold is selected from the group consisting of poly(lactide), poly(glycolide) and poly(L-lactide-co-glycolide).  
     
     
         10 . The method of  claim 1 , wherein the biodegradable polymer scaffold is coated with collagen prior to said applying.  
     
     
         11 . The method of  claim 1 , further comprising: 
 implanting the scaffold into an omentum of a host mammal.    
     
     
         12 . The method of  claim 11 , wherein the host mammal is a rat.  
     
     
         13 . A method of generating tooth tissue comprising: 
 forming a biodegradable polymer scaffold;    applying tooth germ cells onto the biodegradable polymer scaffold; and    implanting the scaffold to which tooth germ cells have been applied into a host animal.    
     
     
         14 . The method of  claim 13 , further comprising: 
 preparing a tooth mold, wherein the biodegradable polymer scaffold is formed in the tooth mold.    
     
     
         15 . The method of  claim 13 , wherein the biodegradable polymer scaffold is in a shape of a tooth.  
     
     
         16 . The method of  claim 15 , wherein the biodegradable polymer scaffold is in a shape of a human tooth.  
     
     
         17 . The method of  claim 13 , wherein the tooth germ cells are mammalian.  
     
     
         18 . The method of  claim 13 , wherein the tooth germ cells comprise one or more of cells selected from the group consisting of cells from an enamel organ, cells from a pulp organ and tissue cultured cells derived from a tooth tissue.  
     
     
         19 . The method of  claim 13 , wherein the tooth germ cells are applied to the biodegradable polymer scaffold with between about 20 to 50 million cells per square centimeter of scaffold.  
     
     
         20 . The method of  claim 13 , wherein the biodegradable polymer scaffold is selected from the group consisting of poly(lactide), poly(glycolide) and poly(L-lactide-co-glycolide).  
     
     
         21 . The method of  claim 13 , wherein after applying, the tooth germ cells are allowed to attach to the scaffold for at least one hour prior to said implanting.  
     
     
         22 . The method of  claim 13 , wherein the biodegradable polymer scaffold is coated with collagen prior to said applying.  
     
     
         23 . The method of  claim 13 , wherein said implanting is into an omentum of the host mammal.  
     
     
         24 . A tooth produced from tooth germ cells and a biodegradable polymer scaffold.  
     
     
         25 . The tooth of  claim 24 , wherein the tooth is in a shape of a human tooth.  
     
     
         26 . The tooth of  claim 24 , wherein the tooth germ cells comprise one or more of cells selected from the group consisting of cells from an enamel organ, cells from a pulp organ and tissue cultured cells derived from a tooth tissue.  
     
     
         27 . The tooth of  claim 24 , wherein the biodegradable polymer scaffold is selected from the group consisting of poly(lactide), poly(glycolide) and poly(L-lactide-co-glycolide).  
     
     
         28 . The method of  claim 1 , wherein said tooth germ cells are human.  
     
     
         29 . The method of  claim 1 , further comprising: 
 implanting said tooth into the gum of a host mammal.    
     
     
         30 . The method of  claim 13 , wherein said implanting is into a mouth of the host mammal.