Artificial dermis and method of preparation

Abstract

Artificial dermis ( 1 ) obtained from plasma with platelets ( 2 ) and human fibroblasts. The plasma with platelets ( 2 ) is obtained from the fractionating of total blood ( 4 ) from the patient ( 8 ) by light centrifugation, and the human fibroblasts ( 3 ) from a skin biopsy ( 5 ). Clotting is obtained by adding calcium. This artificial dermis ( 1 ) provides for the rapid growth of the keratinocytes ( 6 ) seeded on its surface to build an artificial skin ( 7 ) which can easily be transplanted. Large areas of artificial dermis ( 1 ) are obtained from a small skin biopsy ( 5 ) and minimal quantities of plasma with platelets ( 2 ), which being enriched with cytokines and platelet growth factors, strengthens the proliferation of the cells seeded, both inside and on the surface. The artificial skin ( 7 ) obtained can be used to treat major burn treatments, chronic skin ulcers, etc., or be used, by employing genetically altered cells, as a vehicle for gene therapy.

1 . Artificial dermis generated by the use of a matrix obtained by clotting human plasma, wherein the plasma contains platelets and dermal cells are embedded in it.  
   
   
       2 . Artificial dermis according to  claim 1 , wherein the plasma with platelets undergoes viral inactivation.  
   
   
       3 . Artificial dermis according to  claim 1 , wherein a biocompatible polymer is added.  
   
   
       4 . Artificial dermis according to  claim 1 , wherein the dermal cells are fibroplasts.  
   
   
       5 . Artificial dermis according to  claim 1 , wherein the dermal cells are mesenchyme stem cells or endothelial cells.  
   
   
       6 . A method of using an artificial dermis according to  claim 1 , further comprising cultivation of human keratinocytes on the surface to form an artificial skin suitable for transplantation.  
   
   
       7 . A method of using an artificial dermis according to  claim 1 , further comprising the cultivation of other epithelial cells (oral, urogenital . . . ) on its surface to make up an artificial internal epithelia (oral mucosa, bladder mucosa . . . ) suitable for transplantation.  
   
   
       8 . Method of obtaining an artificial dermis, the method including the following steps; obtaining plasma with platelets, by low-speed centrifugation of total blood from a donor and its subsequent freezing, culturing dermal cells from a skin biopsy using known methods, clotting the previously de-frosted plasma with platelets, by adding calcium salts, to obtain a fibrin matrix, embedding dermal cells inside the fibrin.  
   
   
       9 . Method of obtaining an artificial dermis according to  claim 8 , wherein thrombin is added during the clotting.  
   
   
       10 . Method of obtaining an artificial dermis according to  claim 9 , wherein the thrombin is added in a concentration of 0.2 UI/ml.  
   
   
       11 . Method of obtaining an artificial dermis according to  claim 8 , wherein calcium salts consists of 1% Cl 2 Ca in a solution of 1 ml for every 3 to 6 ml of plasma with platelets.