Abstract
BackgroundAutologous epidermal basal cell suspension therapy has been proven to be one of the most effective treatments for full-thickness wounds. However, we found there remain obvious defects that significantly confined the utilization and function of the epidermal basal cells (EBCs), especially the epidermal stem cells (ESCs) in it. This study investigated whether precoating fibronectin (FN) on the wound bed before spraying EBCs could overcome these defects and further explored its possible mechanisms.MethodsIn the in vitro study, EBCs were isolated from the donor skin of patients who needed skin grafting. Different concentrations of FN were used to precoat culture dishes before cell culture; the adherent efficiency, proliferation and migration ability of ESCs were analyzed and compared with traditional collagen IV precoating.In the in vivo study, Sprague–Dawley (SD) rats with full-thickness skin wounds were selected as full-thickness wounds’ model. For the experiment groups, 20 μg/ml FN was precoated on the wound bed 10 min before EBC spray. The quality of wound healing was estimated by the residual wound area rate, wound healing time, and hematoxylin and eosin (H&E) staining. Expression of ESC markers, neovascular markers, inflammation markers, and collagen formation and degradation markers was elucidated by immunohistochemistry (IHC), immunofluorescence (IF), western blot (WB), and RT-qPCR analysis.ResultsThe in vitro study showed that the dishes precoated with 20 μg/ml FN had a similar adherent efficiency and colony formation rate with collagen IV, but it could improve the proliferation and migration of ESCs significantly. Similarly, in the in vivo study, precoating FN on wound bed before EBC spray also significantly promote wound healing by improving ESCs’ utilization efficiency, promoting angiogenesis, decreasing inflammations, and regulating collagen formation and degradation.ConclusionFN precoating wound bed before EBC spray could significantly promote full-thickness wound healing by improving the utilization and function of the ESCs and further by promoting angiogenesis, decreasing inflammations, and regulating collagen formation and degradation.Graphical abstract
Highlights
Autologous epidermal basal cell suspension therapy has been proven to be one of the most effective treatments for full-thickness wounds
The results of our study discovered an optimal concentration for FN precoating and its excellent effects on full-thickness wounds and demonstrated an exciting new way that may help us to explore more innovative approaches for full-thickness wound healing
When the wound bed was precoated by FN before epidermal basal cells (EBCs) spraying, the re-epithelialization, epidermal ridges, and epidermal thickness were significantly improved (P < 0.05). These results suggested that FN precoating wound bed had an active function in promoting re-epithelialization and improving wound healing quality, which might mainly be by increasing the epidermal stem cells (ESCs) utilization of EBCs suspension
Summary
Autologous epidermal basal cell suspension therapy has been proven to be one of the most effective treatments for full-thickness wounds. We found there remain obvious defects that significantly confined the utilization and function of the epidermal basal cells (EBCs), especially the epidermal stem cells (ESCs) in it. Effective skin wound healing becomes a major concern for global healthcare and requires further exploration. The residual bacteria in the wound bed might contaminate cells and lead them to die. Since these three defects significantly confined the utilization and function of the autologous EBCs, we hypothesized that overcoming these critical problems could further improve the therapeutic effect of full-thickness wounds
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