Treatment of Cutaneous Ulcers with Multilayered Mixed Sheets of Autologous Fibroblasts and Peripheral Blood Mononuclear Cells

Background/Aims: We have developed a mixed-cell sheet consisting of autologous fibroblasts and peripheral blood mononuclear cells with a high potency for angiogenesis and wound healing against refractory cutaneous ulcers in mouse and rabbit models. To increase the effectiveness of the mixed sheet, we developed a multilayered mixed sheet. Methods: We assessed the therapeutic effects of multilayered sheets on cutaneous ulcers in mice. Growth factors and chemokines were assessed by enzyme-linked immunosorbent assay. Angiogenesis and fibroblast migration were measured by using tube formation and migration assays. Wound healing rate of cutaneous ulcers was evaluated in mice with diabetes mellitus. Results: The concentration of secreted vascular endothelial growth factor, hepatocyte growth factor, transforming growth factor, C-X-C motif chemokine ligand (CXCL)-1, and CXCL-2 in multilayered sheets was much higher than that in single-layered mixed-cell sheets (single-layered sheets) and multilayered sheets of fibroblasts alone (fibroblast sheets). The supernatant in multilayered sheets enhanced angiogenic potency and fibroblast migration compared with single-layered and fibroblast sheets in an in vitro experiment. The wound healing rate in the multilayered sheet-treated group was higher compared with the no-treatment group (control) at the early stage of healing. Moreover, both vessel lumen area and microvessel density in tissues treated with multilayered sheets were significantly increased compared with tissues in the control group. Conclusion: Multilayered sheets promoted wound healing and microvascular angiogenesis in the skin by supplying growth factors and cytokines. Accordingly, our data suggest that multilayered sheets may be a promising therapeutic material for refractory cutaneous ulcers.

New approaches for full-thickness grafting of the face

In addition to the physical injury, deep burns of the face also can cause psychological injury. The purpose of this study was to develop new technical methods for improving engraftment of full-thickness autografts. Since 2017, autologous fibroblasts have been used for skin facioplasty of one full thickness autograft in Scientific Research Institue, Reginal Clinical Hospital No 1. Isolation of fibroblasts was performed by using an enzymatic method and a skin sample with an area of 10 cm2. Cultivation of autologous fibroblasts was performed for 22 days according to a standard procedure. The prepared culture of autologous fibroblasts was used together with a full-layer autologous transplant to close the wound on the face. We found that autologous fibroblasts provided faster adaptation of the autograft. There were no signs of fibrous tissue over the long term. We concluded that fibroblasts promoted rapid engraftment and adaptation of the full-thickness autograft. Improvement of facial plastic surgery methods can help achieve maximum cosmetic treatment results.