Cryogel 3d scaffolds and methods for producing thereof

Abstract

A method of producing a cryogel-based multicompartment 3D scaffold is herein disclosed. The method comprises the steps of: a) providing a first frozen polymeric layer on a refrigerated support kept at subzero temperature; b) providing subsequent polymeric layers to obtain a stack of polymeric layers by possibly modulating the subzero temperature of the refrigerated support; c) optionally incubating the final polymeric structure at subzero temperature; and d) placing the produced cryogel at a temperature above 0° C., wherein each subsequent layer i) is deposited on the previous one after freezing of this latter; ii) is deposited on the previous one before the complete polymerization of this latter; and iii) is deposited with a temperature higher than the freezing temperature of the previously deposited layer. Cryogel scaffolds obtained from said method are also disclosed.

1 - 15 . (canceled) 
     
     
         16 . A method of producing a multicompartment three-dimensional scaffold, said method comprising:
 c) raising the temperature of a stack of frozen polymeric layers from a temperature below 0° C. to a temperature above 0° C.,   wherein the stack of frozen polymeric layers comprises crosslinked polymers,   wherein following step (c), the stack comprises a crosslinked polymer matrix in one or more of the polymeric layers having pores with a mean pore size between 1 μm and 500 μm.   
     
     
         17 . The method of  claim 16 , comprising forming the frozen polymeric layers by depositing a second layer comprising a second precursor polymeric liquid on a first layer comprising a first precursor polymeric liquid, prior to complete polymerization of the first layer. 
     
     
         18 . The method of  claim 16 , comprising:
 a) depositing a first liquid comprising a first precursor of a first polymeric material to form a first frozen polymeric layer on a refrigerated support kept below 0° C. temperature, prior to step (c).   
     
     
         19 . The method of  claim 18 , comprising:
 b) repeatedly depositing a second liquid comprising a second precursor of a second polymeric material to form a plurality of subsequent polymeric layers on the refrigerated support at below 0° C.,   wherein the first frozen polymeric layer and said plurality of subsequent polymeric layers forming a stack of polymeric layers,   wherein step (b) further comprises polymerizing the first and/or second precursor,   wherein the first precursor and the second precursor are the same or different, and   wherein step (b) is performed after step (a) and prior to step (c).   
     
     
         20 . A multicompartment three-dimensional scaffold produced by the method of  claim 16 ,
 wherein the stack of layers that comprise the multicompartment three-dimensional scaffold are mechanically connected by a built-in intermediate layer, having a thickness smaller than those of its adjacent layers.   
     
     
         21 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the crosslinked polymer matrix comprises chitosan, alginate, cellulose, acrylate, gelatin, or collagen. 
     
     
         22 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the crosslinked polymer matrix comprises polysaccharides (e.g., cellulose, agarose, alginate, starch, chitosan and others), polypeptides (e.g., silk, collagen, gelatin and others), amelogenin or synthetic polymers such as polyurethanes, poly-olefins, polyethylene glycol (PEG), poly(glycolide) (PGA), poly-L-lactide (PLA), carboxymethylcellulose (CMC) or poly(lactide-co-glycolide) (PLGA). 
     
     
         23 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the crosslinked polymer matrix comprises hyaluronic acid, chondroitinsulfate, heparansulfate, heparine, or keratansulfate. 
     
     
         24 . The multicompartment three-dimensional scaffold of  claim 20 , wherein at least one compartment comprises bioactive compounds, cells, tissue fragments, or proteins or a combination thereof. 
     
     
         25 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the scaffold is suitable for tissue engineering and/or cell transplantation. 
     
     
         26 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the shape and size of the scaffold is adapted to fit into a defect site as defined from medical imaging. 
     
     
         27 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the scaffold is degradable. 
     
     
         28 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the scaffold is permanent. 
     
     
         29 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the scaffold comprises degradable and permanent parts. 
     
     
         30 . The multicompartment three-dimensional scaffold of  claims 20 , wherein part of the scaffold is porous, and part of the scaffold is non-porous. 
     
     
         31 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the scaffold is patterned on a medical device. 
     
     
         32 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the scaffold is flowable and injectable, optionally wherein the scaffold is sterilizable and biocompatible. 
     
     
         33 . The multicompartment three-dimensional scaffold of  claim 20 , wherein the scaffold's internal organization is close to the architecture of native tissues or organs. 
     
     
         34 . The method of  claim 19 , comprising a second liquid comprising encapsulated cells. 
     
     
         35 . A multi-component system comprising non-stoichiometric amounts of two or more reactants of a crosslinked 3D structure, wherein the two or more reactants can react to form a biomaterial-based ink that is capable of being printed and polymerizing at subzero temperatures to form a frozen crosslinked 3D structure.