Support with crosslinked marine collagen for tissue engineering and manufacture of biomaterials

Abstract

The invention relates to a method of in vitro testing of the efficacy of a potentially active substance comprising monitoring the effect of said potentially active substance on an artificial skin, comprising a composite product forming a collagen support comprising at least one porous collagen layer covered on at least one side with a collagen membrane component selected from the group consisting of a collagen membrane prepared by compression of a collagen sponge at a pressure of at least about 50 bar and of a collagen membrane comprising a collagen film prepared by drying a collagen gel separately from the porous collagen layer, thereby providing a reliable method for finding new potentially active substances.

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         42 . A support for tissue engineering comprising a porous matrix prepared from an aquatic collagen gel which has undergone a lyophilization step, and wherein the porous matrix is crosslinked by a chemical crosslinking agent.  
     
     
         43 . The support of  claim 42 , wherein the crosslinking agent is selected from diphenylphosphorylazide or DPPA, a carbodiimide, N-hydroxysuccinimide, and glutaraldehyde.  
     
     
         44 . The support of  claim 42 , wherein the aquatic collagen is coming from teleost fish skin teleost.  
     
     
         45 . The support of  claim 42 , wherein the aquatic collagen is coming from the skin of a flat fish.  
     
     
         46 . The support of  claim 42 , wherein the aquatic collagen is coming from the skin of a flat fish industrially fished selected from the sole, the dab, the turbot and the brill.  
     
     
         47 . The support of  claim 42 , wherein the porous matrix comprises an aquatic collagen admixed with chitosan or at least one glycosaminoglycan.  
     
     
         48 . The support of  claim 42 , wherein said porous matrix is covered on at least one side with an essentially compact collagen membrane selected from a collagen film prepared by drying a second collagen gel in air or in gaseous fluid, and from a compressed collagen sponge.  
     
     
         49 . The support of  claim 42 , wherein the porous matrix comprises living cells selected from the group consisting of normal living cells, genetically modified living cells and malignant living cells.  
     
     
         50 . The support of  claim 49 , wherein the living cells are selected from the group consisting of fibroblasts, keratinocytes, melanocytes, Langerhans' cells originating from the blood, endothelial cells originating from the blood, blood cells, macrophages, lymphocytes, adipocytes, sebocytes, chondrocytes, osteocytes, osteoblasts and Merkel's cells originating from the blood.  
     
     
         51 . The support of  claim 42 , wherein the porous matrix contains living fibroblasts selected from normal, genetically modified or malignant fibroblasts.  
     
     
         52 . The support of  claim 48 , wherein the essentially compact collagen membrane contains normal, genetically modified or malignant living cells selected from keratinocytes, melanocytes, Merkel's cells originating from the blood, Langerhans' cells originating from the blood, sebocyte cells originating from the blood, and nerve cells.  
     
     
         53 . The support of  claim 49 , wherein the living cells originate from human subjects.  
     
     
         54 . The support of  claim 51 , wherein the living cells originate from young subjects.  
     
     
         55 . The support of  claim 51 , wherein the living cells originate from old subjects.  
     
     
         56 . The support of  claim 48 , wherein the compression of the compressed collagen sponge is carried out at a pressure of at least about 50 bar (50.10 5  Pa) at a temperature of between 20° C. and 80° C.  
     
     
         57 . The support of  claim 48 , wherein the essentially compact membrane is prepared prior to combination with the porous layer by preparing first the compact collagen membrane and then by depositing it on said aquatic collagen gel before the combination of the collagen gel and of the compact collagen membrane is frozen and lyophilized.  
     
     
         58 . The support of  claim 42 , in the form of a biomaterial, a reconstructing conjunctive tissue, or a reconstructed skin.  
     
     
         59 . A method selected from a method for studying cutaneous ageing, and a method for testing the efficacy of actives ingredients with regard to slowing down or delaying cutaneous ageing comprising the use of a support as defined in  claim 42 , wherein the porous matrix comprises living cells selected from the group consisting of normal living cells, genetically modified living cells and malignant living cells.  
     
     
         60 . A method selected from a method for studying cutaneous ageing, and a method for testing the efficacy of actives ingredients with regard to slowing down or delaying cutaneous ageing comprising the use of a support as defined in  claim 48 , wherein the porous matrix comprises living cells selected from the group consisting of normal living cells, genetically modified living cells and malignant living cells.