Compositions Derived from Placenta and Methods of Producing the Same

Abstract

Compositions comprising unseparated amnion/chorion derived from the placenta and methods of preparing and using those compositions are provided. Washing or preservation of placental tissue according to the methods of the disclosure may allow for one or more benefits such as more efficient removal of blood remnants, retention of wound healing and tissue regeneration components, better handling characteristics, increased absorption potential, or improved healing capacity. The present invention also includes methods of healing a wound of the skin, eye, nerve, tendon, or dura comprising applying the compositions of the invention to the wound.

1 - 68 . (canceled) 
     
     
         69 . A dehydrated composition comprising a separated amnion and an intermediate layer, wherein the intermediate layer is substantially intact, and wherein the dehydrated composition is substantially free of blood remnants. 
     
     
         70 . The dehydrated composition of  claim 69 , wherein the composition is dehydrated by lyophilization, dehydrated by heat-drying, dehydrated by oven-drying, dehydrated by air-drying, or dehydrated by chemical dehydration. 
     
     
         71 . The dehydrated composition of  claim 69 , wherein the composition has a dry mass of less than about 1 mg/cm 2  to about 3 mg/cm 2 . 
     
     
         72 . The dehydrated composition of  claim 69 , wherein the composition has a hemoglobin content of less than about 13 ng/mg of dry mass of the composition. 
     
     
         73 . The dehydrated composition of  claim 69 , wherein the composition has a hemoglobin content of dry mass of the composition that is reduced in an amount greater than about 50%, 60%, 70%, 80%, 90%, or 95% compared to an unprocessed dried placental tissue. 
     
     
         74 . The dehydrated composition of  claim 69 , wherein the composition has a mean thickness of less than 10 μm to about 60 μm. 
     
     
         75 . The dehydrated composition of  claim 69 , wherein the composition comprises one or more additional sheets comprising a perforated or unperforated unseparated amnion/chorion sheet. 
     
     
         76 . A dehydrated composition comprising a separated amnion and an intermediate layer, wherein the intermediate layer is substantially intact, wherein the dehydrated composition is substantially free of blood remnants, and wherein the composition comprises one or more additional sheets comprising a separated chorion sheet. 
     
     
         77 . The dehydrated composition of  claim 76 , wherein the composition comprises nucleated cells, collagens, fibronectins, fibroblasts, growth factors, cytokines, proteoglycans, glycoproteins, tissue inhibitors of metalloproteinases, and/or hyaluronic acid. 
     
     
         78 . The dehydrated composition of  claim 76 , wherein the composition has a hemoglobin content of less than about 13 ng/mg of dry mass of the composition. 
     
     
         79 . The dehydrated composition of  claim 76 , wherein the composition has a mean thickness of about 10 μm to about 700 μm. 
     
     
         80 . The method of  claim 76 , wherein the separated amnion is detached from the placental disc and umbilical cord. 
     
     
         81 . A dehydrated composition comprising a separated chorion and an intermediate layer, wherein the intermediate layer is substantially intact, and wherein the dehydrated composition is substantially free of blood remnants. 
     
     
         82 . The dehydrated composition of  claim 81 , wherein the composition comprises one or more additional sheets comprising a perforated or unperforated unseparated amnion/chorion sheet. 
     
     
         83 . The dehydrated composition of  claim 81 , wherein the composition comprises one or more additional sheets comprising a separated amnion sheet. 
     
     
         84 . A method of preparing a dehydrated composition comprising an intermediate layer comprising:
 (a) obtaining an unseparated placental tissue dissected away from the placental disc;   (b) (i) separating the chorion from the amnion and intermediate layer to obtain a separated amnion sheet comprising amnion and an intermediate layer, wherein the intermediate layer is substantially intact on the separated amnion and appears by visual inspection to cover at least about 60% of the separated amnion; or   (ii) separating the amnion from the chorion and intermediate layer to obtain a separated chorion sheet comprising chorion and an intermediate layer, wherein the intermediate layer is substantially intact on the separated chorion and appears by visual inspection to cover at least about 60% of the separated chorion;   (c) performing at least one washing step comprising washing the separated amnion sheet or separated chorion sheet with at least one suitable washing medium to remove substantially all blood remnants; and   (d) dehydrating the separated amnion sheet or separated chorion sheet, wherein the dehydrating comprises lyophilizing, heat-drying, oven-drying, air-drying, or chemically dehydrating the sheet.   
     
     
         85 . The method of  claim 84 , wherein the method comprises (i) layering a separated chorion sheet on the separated amnion sheet; or (ii) layering a separated amnion sheet on the separated chorion sheet prior to dehydrating. 
     
     
         86 . The method of  claim 84 , wherein the unseparated placental tissue is from a human donor, and wherein the method comprises screening the human donor for one or more of HIV-1, HIV-2, HBV, HCV, HTLV-I, HTLV-II, CMV, or syphilis. 
     
     
         87 . A method of treating a wound comprising applying to the wound a dehydrated composition comprising an amnion, an intermediate layer, and a chorion, wherein the composition is substantially free of blood remnants. 
     
     
         88 . A method of treating a wound comprising applying to the wound a dehydrated composition comprising an amnion, an intermediate layer, and a chorion, wherein the composition is substantially free of blood remnants, and wherein the intermediate layer has a mean thickness of from about 5 μm to about 200 μm. 
     
     
         89 . The method of  claim 88 , wherein the dehydrated composition has
 (a) a dry mass of about 1 mg/cm 2  to about 25 mg/cm 2 ; and/or   (b) a total protein content of about 0.1 mg/cm 2  to about 2.0 mg/cm 2 .   
     
     
         90 . The method of  claim 88 , wherein the composition has a water absorption potential of about 1 mg absorbed/cm 2  to about 40 mg absorbed/cm 2 . 
     
     
         91 . The method of  claim 88 , wherein the composition comprises (i) the chorion layered on the amnion and intermediate layer; or (ii) the amnion layered on the chorion and intermediate layer. 
     
     
         92 . The method of  claim 88 , wherein the composition is an unseparated amnion/chorion sheet. 
     
     
         93 . The method of  claim 88 , wherein the amnion, intermediate layer, and/or chorion is detached from the placental disc and umbilical cord. 
     
     
         94 . The method of  claim 88 , wherein the composition comprises nucleated cells, collagens, fibronectins, fibroblasts, growth factors, cytokines, proteoglycans, glycoproteins, tissue inhibitors of metalloproteinases, and/or hyaluronic acid. 
     
     
         95 . A method of treating a wound comprising applying to the wound the dehydrated composition of  claim 76 . 
     
     
         96 . A method of treating a wound comprising applying to the wound the dehydrated composition of  claim 69 .