Chronic nonhealing ulcers are responsible for considerable morbidity, given the increasing prevalence of type II diabetes and other comorbid conditions that further worsen healing. This study introduced shelf-stable decellularized and lyopreserved human amnion grafts for treating difficult-to-heal wounds. The processing approach (comprising a unique combination of nonionic surfactants and trehalose lyopreservation) applied to develop these bioscaffolds maximized the retention of sulfated glycosaminoglycans while enhancing both tensile property and hydrophilicity. Postprocessing, the tensile properties were found to be similar to human skin (5.33 ± 2.45 MPa). Additionally, the surface hydrophilicity of the lyopreserved grafts was increased. It also exhibited optimum moisture transmissibility (evaluated as per BS EN 13726-2 standards), similar to moist wound dressing (1625 ± 375 g/m