Three-dimensional porous hybrid scaffold and manufacture thereof

Abstract

The present invention refers to a three-dimensional porous hybrid scaffold for tissue engineering and methods of its manufacture and use.

1 . A three-dimensional hybrid scaffold for tissue engineering comprising:
 a first layer made of a decellularized biological material;   a second porous layer connected to the surface of said first layer, wherein said second layer is a porous bioadhesive; and   a third porous layer connected to the surface of said second layer which is located opposite the surface to which said first layer is connected, wherein said third layer is a three-dimensional porous polymer scaffold.   
     
     
         2 . The hybrid scaffold according to  claim 1 , wherein said third layer is an electrospun three-dimensional porous scaffold. 
     
     
         3 . The hybrid scaffold according to  claim 2 , wherein said electrospun scaffold is obtained by
 providing an electrospinning apparatus;   forming crystals from a molecule or group of molecules, which are in vapor phase comprised in the surrounding atmosphere, at the surface of a collector of said electrospinning apparatus, wherein the reaction chamber of said electrospinning apparatus has a temperature which allows formation of crystals at said surface of said collector;   electrospinning a solution comprising at least one polymer dissolved therein around the crystals;   continuing the formation of crystals and the electrospinning simultaneously; and   removing the crystals by sublimation.   
     
     
         4 . The hybrid scaffold according to  claim 3 , wherein said collector of said electrospinning apparatus is composed of said first and second layer and thus said electrospun scaffold is electrospun directly onto the surface of said second layer. 
     
     
         5 . The hybrid scaffold according to  claim 1 , wherein said biological material is selected from the group consisting of small intestine, liver, pancreas, urinary bladder, stomach, bladder, vascular system, bile duct, alimentary canal, respiratory tract, kidney, spleen, heart, heart valve, bone, skin or fragments or parts thereof. 
     
     
         6 . The hybrid scaffold according to  claim 1 , wherein said biological material is esophageal mucosa. 
     
     
         7 . The hybrid scaffold according to  claim 1 , wherein said biological material is derived from an individual which is selected from the group consisting of mammal, reptiles and insects. 
     
     
         8 . The hybrid scaffold according to  claim 7 , wherein said mammal is selected from the group consisting of porcine, bovine, ovine, rabbit, monkey and human. 
     
     
         9 . The hybrid scaffold according to  claim 1 , wherein said bioadhesive is selected from the group consisting of fibrin glue, polyvinylpyrolidone, polyvinylpyrolidone/vinyl acetate copolymers, polyethylene glycol, platelated gel, chitosan or gelatin-resorcin-formaldehyde. 
     
     
         10 . The hybrid scaffold according to  claim 9 , wherein said bioadhesive is fibrin glue. 
     
     
         11 . The hybrid scaffold according to  claim 1 , wherein the polymer for said three-dimensional porous polymer scaffold is a biodegradable and/or biocompatible polymer. 
     
     
         12 . The hybrid scaffold according to  claim 1 , wherein said second porous layer and said third layer have a pore size which is between about 10 nm to about 500 μm. 
     
     
         13 . The hybrid scaffold according to  claim 1 , wherein the thickness of said bioadhesive layer is about 10 to 1000 μm before said third layer is connected to said bioadhesive second layer. 
     
     
         14 . A method of manufacturing a three-dimensional hybrid scaffold wherein the three-dimensional hybrid scaffold comprises:
 a first layer made of a decellularized biological material;   a second porous layer connected to the surface of said first layer, wherein said second layer is a porous bioadhesive; and   a third porous layer connected to the surface of said second layer which is located opposite the surface to which said first layer is connected, wherein said third layer is a three-dimensional porous polymer scaffold,   the method comprising:   chilling a first layer of decellularized biological material to 0° C. or below;   applying a second layer to the surface of said chilled first layer, wherein said second layer is an aqueous solution of a bioadhesive, wherein said second layer is applied prior to formation of ice crystals on said chilled first layer; and   applying a third layer to a side of said second layer which is not the side facing said first layer to form a three-dimensional porous scaffold on the second layer,
 wherein the applying of the third layer occurs in the presence of an electric field and in an environment having (i) a humidity ranging from about 40% to about 80% and (ii) a temperature which is different from temperature of the side of said second layer facing the chilled first layer to create a temperature gradient; 
 wherein said third layer is applied in the presence of ice crystals formed on said second layer; and 
 wherein the applying of the third layer occurs in at least two cycles having an interval in between each cycle. 
   
     
     
         15 . The method according to  claim 14 , wherein said third layer is comprised of an electrospun scaffold. 
     
     
         16 . (canceled) 
     
     
         17 . The method according to  claim 14 , wherein said second layer is applied to said chilled first layer using a method selected from the group consisting of spraying, electrospraying, electrospinning, spin coating, dip coating, casting and brushing. 
     
     
         18 . (canceled) 
     
     
         19 . The method according to  claim 14 , further comprising controlling the degradation time of said second layer by controlling a pore size of said second layer which is achieved by increasing or decreasing the temperature of the chilled first layer. 
     
     
         20 . The method according to  claim 14 , wherein said bioadhesive is selected from the group consisting of fibrin glue, polyvinylpyrolidone, chitosan and gelatin-resorcin-formaldehyde. 
     
     
         21 . The method according to  claim 20 , wherein said bioadhesive is fibrin glue. 
     
     
         22 . A medicament comprising a three-dimensional hybrid scaffold according to  claim 1  for autologous, allogenic, xenogenic transplantation of tissue. 
     
     
         23 . The medicament according to  claim 22 , wherein said tissue is selected from the group consisting of small intestine, liver, pancreas, urinary bladder, stomach, bladder, vascular system, bile duct, alimentary canal, respiratory tract, kidney, spleen, heart, heart valve, bone, skin or fragments or parts thereof. 
     
     
         24 . (canceled) 
     
     
         25 . The method according to  claim 14 , further comprising removing the ice crystals and forming pores in place of the ice crystals; wherein the pores have a pore size ranging from about 50 μm to about 500 μm. 
     
     
         26 . The method according to  claim 14 , wherein the second layer has a thickness ranging from about 10 p.m to about 1000 p.m. 
     
     
         27 . The method according to  claim 14 , wherein the interval comprises 5 minutes.