Effective stacking and transplantation of stem cell sheets using exogenous ROS-producing film for accelerated wound healing.

Extensive skin loss caused by burns or diabetic ulcers may lead to major disability or even death. Therefore, cell-based therapies that enhance skin regeneration are clinically needed. Previous approaches have been applied the injections of cell suspensions and the implantation of biodegradable three-dimensional scaffolds seeded cells. However, these treatments have limits due to poor localization of the injected cells and insufficient delivery of oxygen and nutrients to cells. Recently, cell sheet-based tissue engineering has been developed to transplant cell sheets, which are cell-dense tissues without scaffolds. Because cell density is one of the important factors for improving the therapeutic effect of cell transplantation, transplanting layered cell sheet constructs can promote the recovery of tissue function and tissue regeneration compared with a single cell sheet. Thus, this study designed ROS-induced cell sheet stacking method with newly fabricated hematoporphyrin-incorporated polyketone film (Hp-PK film) to enhance cell sheet delivery efficiency and application in wound healing. We have demonstrated the therapeutic effect of a multi-layered mesenchymal stem cell sheets onto a full-thickness wound defect in nude mice. Consequentially, three-layered cell sheets transplanted and stacked by ROS-induced method promoted angiogenesis and skin regeneration at the wound site. Thus, our strategy based on Hp-PK film, which allows for easy stacking and transplantation of cell sheets, could be applied to enhance tissue regeneration.