The treatment of large-scale skin wounds remains a therapeutic challenge. In most cases there is not enough autologous material available for full coverage. Cultured epithelial autografts are efficient in restoring the lost epidermal cover; however, they have some disadvantages, such as difficult application and protracted cell cultivation periods. Transplanting a sprayed keratinocyte suspension in fibrin sealant as biological carrier is an option to overcome those disadvantages. Here, we studied different seeding techniques regarding their applicability and advantages on cell survival, attachment, and outgrowth in vitro and thereby improve the cell transfer to the wound bed. Human primary keratinocytes were suspended in a fibrin sealant. WST-8 assay was used to evaluate the vitality for 7 days. Furthermore, the cells were labeled with CellTracker™ CM-Di-I and stained with a life/dead staining. Cell morphology, shape, and distribution were microscopically analyzed. There was a significant increase in vitality while cultivating the cells in fibrin. Sprayed cells were considerably more homogenously distributed. Sprayed cells reached the confluent state earlier than dripped cells. There was no difference in the vitality and morphology in both groups over the observation period. These findings indicate that the sprayed keratinocytes are superior to the application of the cells as droplets. The sprayed application may offer a promising therapeutic option in the treatment of large chronic wounds.
Keywords: air pressure; fibrin; keratinocytes; spray application; wound healing.