Keratin, a natural biomaterial derived from wool or human hair, has the intrinsic ability to interact with different types of cells and the potential to serve as a controllable extracellular matrix that can be used a scaffold for tissue engineering. In this study, we demonstrated a simple and fast technique to construct 3D keratin scaffolds for accelerated wound healing using a lyophilization method based on extraction of keratin from human hair. The physical properties of the keratin scaffolds such as water uptake, pore size, and porosity can be adjusted by changing the protein concentrations during the fabrication process. The keratin scaffolds supported human adipose stem cells (hASCs) adhesion, proliferation, and differentiation. In vivo study performed on ICR mice showed that keratin scaffolds with hASCs shortened skin wound healing time, accelerated epithelialization, and promoted wound remodeling. Therefore, keratin scaffolds alone or together with hASCs may serve as therapeutic agents for repairing wounded tissue.

Wound dressing, which prevents dehydration and provides a physical barrier against infection to wound beds, can improve wound healing. The interactions between extracellular matrix (ECM) and growth factors is critical to the healing process. Electrospun nanofibers are promising templates for wound dressings due to the structure similarity to ECM of skin. Otherwise, the ECM secreted by human adipose-derived stem cells (hASCs) is rich in growth factors known to enhance wound healing. Accordingly, we propose that the PLGA nanofibrous template incorporated with hASCs-secreted ECM may enhance wound healing. In this study, PLGA nanofibrous matrixes with an aligned or a random structure were prepared by electrospinning. Human ASCs cultured on the aligned matrix had a better viability and produced a larger amount of ECM relative to that of random one. After 7 days’ cultivation, the hASCs on aligned PLGA substrates underwent decellularization to fabricate cECM/PLGA dressings. By using immunohistochemical staining against F-actin and cell nucleus, the removal of cellular components was verified. However, the type I collagen and laminin were well preserved on the cECM/PLGA nanofibrous matrixes. In addition, this substrate was hydrophilic, with appropriate mechanical strength to act as a wound dressing. The L929 fibroblasts had good activity, survival and proliferation on the cECM/PLGA meshes. In addition, the cECM/PLGA nanofibrous dressings improved the wound healing of surgically created full-thickness skin excision in a mouse model. This hASCs-secreted ECM incorporated into electrospun PLGA nanofibrous could be a promising dressing for enhancing wound healing.