Material and method for engraftment of a composite biocompatible skin graft on the neodermis of artificial skin

Abstract

Autologous cultured keratinocytes are grown on a biosynthetic and biocompatible substratum following pre-seeding with autologous or allogenic dermal fibroblasts. The resultant composite material may then be applied on the neodermis of artificial skin which had been previously engrafted on the patient. The composite material, and specifically Composite Biocompatible Skin Graft (CBSG) material comprises autologous keratinocytes and allogenic or autologous dermal fibroblasts grown on Laserskin. A method for cultivating the CBSG includes the application of dermal fibroblasts onto the substratum as a feeder layer and then the inoculation of autologous keratinocytes on the resultant structure. A method for engraftment comprises first applying an artificial skin with a protective silicone membrane on a wound area, thereby allowing vascularization; and following vascularization, removing the silicone membrane and engrafting the CBSG material onto the vascularized artificial skin.

What is claimed is:  
     
         1 . A method for cultivation of graftable skin comprising: 
 growing a layer of dermal fibroblasts upon at least an upper side of a biosynthetic substratum of a derivative of benzyl esterified hyaluronic acid; and, after said dermal fibroblast layer becomes at least sub-confluent,    growing a layer of keratinocytes over said dermal fibroblasts upon said upper side of said substratum to form a composite skin graft material, said keratinocytes being harvested from a target donor patient.    
     
     
         2 . The method according to  claim 1  wherein said dermal fibroblasts are allogenic.  
     
     
         3 . The method according to  claim 1  wherein said dermal fibroblasts are autologous.  
     
     
         4 . A method for cultivation of graftable skin comprising: 
 growing a first layer of dermal fibroblasts upon a first basal side of a biosynthetic substratum of a derivative of benzyl esterified hyaluronic acid;    growing second layer of dermal fibroblasts upon a second upper side of said biosynthetic substratum; and    after said second dermal fibroblast layer becomes at least sub-confluent, growing a layer of keratinocytes over said dermal fibroblasts upon said upper side of said substratum to form a composite skin material, said keratinocytes having been harvested from a target donor patient.    
     
     
         5 . The method according to  claim 4  wherein said dermal fibroblasts are allogenic.  
     
     
         6 . The method according to  claim 4  wherein said dermal fibroblasts are autologous.  
     
     
         7 . A method for cultivation of graftable skin comprising: 
 growing a layer of keratinocytes upon an upper side of a substratum of a biosynthetic substratum of a derivative of benzyl esterified hyaluronic acid to form a composite skin graft material, said keratinocytes having been harvested from a target donor patient.    
     
     
         8 . A graftable skin material compromising a composite of: 
 a biosynthetic substratum of a derivative of benzyl esterified hyaluronic acid;    a layer of dermal fibroblasts upon at least an upper side of said biosynthetic substratum; and    a layer of keratinocytes over said dermal fibroblasts upon said upper side of said substratum, said keratinocytes having been harvested from a target donor patient.    
     
     
         9 . The material according to  claim 8  wherein said dermal fibroblasts are allogenic.  
     
     
         10 . The material according to  claim 8  wherein said dermal fibroblasts are autologous.  
     
     
         11 . A graftable skin material comprising a composite of: 
 a biosynthetic substratum of a derivative of benzyl esterified hyaluronic acid;    a first layer of dermal fibroblasts upon a first basal side of said biosynthetic substratum;    a second layer of dermal fibroblasts upon a second upper side of said biosynthetic substratum; and    a layer of keratinocytes over said dermal fibroblasts upon said upper side of said substratum, said keratinocytes having been harvested from a target donor patient.    
     
     
         12 . The material according to  claim 11  wherein said dermal fibroblasts are allogenic.  
     
     
         13 . The material according to  claim 11  wherein said dermal fibroblasts are autologous.  
     
     
         14 . A graftable skin material comprising: 
 a substratum of a biosynthetic substratum of a derivative of benzyl esterified hyaluronic acid; and    a layer of keratinocytes upon an upper side of said substratum, said keratinocytes having been harvested from a target donor patient.    
     
     
         15 . A method for grafting a graftable skin material comprising the steps of: 
 applying an artificial skin substrate upon a wound bed of a recipient patient; said artificial skin substrate comprising a layer of collagen-glycoaminoglycan on a basal side to be juxtaposed to said wound bed and a covering membrane of silicone on an opposing upper side;    allowing a sufficient time to form a vascularized wound bed under said collagen-glycoaminoglycan; thereupon    removing said silicone membrane; and thereupon    applying a basal side of a sheet of cultivated skin material over said collagen-glycoaminoglycan, said cultivated skin material comprising at least a layer of keratinocytes upon an upper side of a substratum, said keratinocytes being harvested from a target donor patient.    
     
     
         16 . The method according to  claim 15  wherein said cultivated skin material further comprises a layer of dermal fibroblasts upon at least an upper side of a biosynthetic substratum and wherein said layer of keratinocytes is over said dermal fibroblasts.  
     
     
         17 . The method according to  claim 16  wherein said cultivated skin material further comprises a layer of dermal fibroblasts upon said basal side of said biosynthetic substratum.