Abstract
Severe skin wounds are often associated with large areas of damaged tissue, resulting in substantial loss of fluids containing electrolytes and proteins. The net result is a vulnerability clinically to skin infections. Therapies aiming to close these large openings are effective in reducing the complications of severe skin wounds. Recently, cell transplantation therapy showed the potential for rapid re-epithelialization of severe skin wounds. Here, we show the improved effects of cell transplantation therapy using a robust protocol of efficient expansion and delivery of epidermal cells for treatment of severe skin wounds. Human skin tissues were used to generate human epidermal organoids maintained under newly established culture conditions. The human epidermal organoids showed an improved capacity of passaging for at least 10 rounds, enabling organoids to expand to cell numbers required for clinical applications. A newly designed auto micro-atomization device (AMAD) was developed for delivery of human epidermal organoids onto the sites of severe skin wounds enhancing uniform and concentrated delivery of organoids, facilitating their engraftment and differentiation for skin reconstitution. With the optimal design and using pneumatic AMAD, both survival and functions of organoids were effectively protected during the spraying process. Cells in the sprayed human epidermal organoids participated in the regeneration of the epidermis at wound sites in a mouse model and accelerated wound healing significantly. The novel AMAD and out new protocol with enhanced effects with respect to both organoid expansion and efficient transplantation will be used for clincal treatments of complex, uneven, or large-area severe skin wounds.
Highlights
Severe skin wounds seriously influence the quality of life of patients and even threaten their lives, especially those with extensive skin injuries and difficult-to-heal burns
In order to ensure the survival of cells or organoids after spraying, the maximum pressure of air is controlled to be within the limitation of standard atmospheric pressure (∼101 kPa), which is designed with the rate of 4 L/min of air flow for spraying cells or organoids contained in medium
We present here a novel and facile set-up, the atomization device (AMAD), for spray delivery of skin cells in pre-expanded organoids and that can be used on skin wounds
Summary
Severe skin wounds seriously influence the quality of life of patients and even threaten their lives, especially those with extensive skin injuries and difficult-to-heal burns. During the last three decades, skin substitutes with inplanted cells inside three dimensional (3D) scaffolds have gradually demonstrated advantages at enhancing therapeutic effects for skin wound healing. Such types of skin substitutes were developed to replace epidermis to restore barrier functions of skin, leading to tissue engineered skin (TE-skin) grafts adapted and introduced into clinical applications (Sun et al, 2014; Singh et al, 2017). The TE-skins are usually fragile with poor flexibility and stiffness, which results in difficulties for TE-skin grafts to cover the wound bed completely and evenly, leading to poor graft adhesion and integration. This can result in bullae formation, increasing the risks of wound contracture and wound colonization by microbes, resulting in infections and with difficulties for storage and transportation (Sood et al, 2010; McHeik et al, 2014)
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