ABSTRACT
In the realm of large-area trauma flap transplantation, averting ischaemic necrosis emerges as a pivotal concern. Several key mechanisms, including the promotion of angiogenesis, the inhibition of oxidative stress, the suppression of cell death, and the mitigation of inflammation, are crucial for enhancing skin flap survival. Apoptotic bodies (ABs), arising from cell apoptosis, have recently emerged as significant contributors to these functions. This study engineered three-dimensional (3D)-ABs using tissue-like mouse adipose-derived stem cells (mADSCs) cultured in a 3D environment to compare their superior biological effects against 2D-ABs in bolstering skin flap survival. The findings reveal that 3D-ABs (85.74 ± 4.51) % outperform 2D-ABs (76.48 ± 5.04) % in enhancing the survival rate of ischaemic skin flaps (60.45 ± 8.95) % (all p < 0.05). Mechanistically, they stimulated angiogenesis, mitigated oxidative stress, suppressed apoptosis, and facilitated the transition of macrophages from M1 to M2 polarization (all p < 0.05). A comparative analysis of microRNA (miRNA) profiles in 3D- and 2D-ABs identified several specific miRNAs (miR-423-5p-up, miR30b-5p-down, etc.) with pertinent roles. In summary, ABs derived from mADSCs cultured in a 3D spheroid-like arrangement exhibit heightened biological activity compared to those from 2D-cultured mADSCs and are more effective in promoting ischaemic skin flap survival. These effects are attributed to their influence on specific miRNAs.
Subject(s)
Adipose Tissue , Apoptosis , Cell Culture Techniques , Ischemia , Stem Cells , Cells, Cultured , Humans , Animals , Mice , Stem Cells/cytology , Stem Cells/metabolism , Male , Mice, Inbred C57BL , Cell Culture Techniques/methods , Cell Separation/methods , Adipose Tissue/cytology , Adipose Tissue/metabolism , Ischemia/genetics , Ischemia/pathology , Cell Hypoxia , Cell Survival , MicroRNAs/genetics , Oxidative Stress , Neovascularization, Pathologic , Gene Expression ProfilingABSTRACT
Preventing and treating avascular necrosis at the distal end of the flaps are critical to surgery success, but current treatments are not ideal. A recent study shows that apoptotic bodies (ABs) generated near the site of apoptosis can be taken up and promote cell proliferation. The study reveals that ABs derived from fibroblast-like cells in the subcutaneous connective tissue (FSCT cells) of skin flaps promoted ischaemic flap survival. It is also found that ABs inhibited cell death and oxidative stress and promoted M1-to-M2 polarization in macrophages. Transcriptome sequencing and protein level testing demonstrated that ABs promoted ischaemic flap survival in endothelial cells and macrophages by inhibiting ferroptosis via the KEAP1-Nrf2 axis. Furthermore, microRNA (miR) sequencing data and in vitro and in vivo experiments demonstrated that ABs inhibited KEAP1 by delivering miR-339-5p to exert therapeutic effects. In conclusion, FSCT cell-derived ABs inhibited ferroptosis, promoted the macrophage M1-to-M2 transition via the miR-339-5p/KEAP1/Nrf2 axis and promoted ischaemic flap survival. These results provide a potential therapeutic strategy to promote ischaemic flap survival by administering ABs.
Subject(s)
Ferroptosis , Fibroblasts , Kelch-Like ECH-Associated Protein 1 , MicroRNAs , NF-E2-Related Factor 2 , Surgical Flaps , Animals , Mice , Kelch-Like ECH-Associated Protein 1/metabolism , Kelch-Like ECH-Associated Protein 1/genetics , NF-E2-Related Factor 2/metabolism , NF-E2-Related Factor 2/genetics , Ferroptosis/genetics , MicroRNAs/genetics , MicroRNAs/metabolism , Fibroblasts/metabolism , Disease Models, Animal , Ischemia/metabolism , Ischemia/genetics , Male , Apoptosis/genetics , Connective Tissue/metabolism , Signal Transduction/geneticsABSTRACT
PURPOSE: The excision of plantar malignant melanoma frequently leads to wide skin defects on the plantar surface. This study aimed to investigate the advantages and feasibility of dermal regenerative template reconstructing plantar blemishes caused by malignant melanoma. METHODS: 28 patients identified with plantar malignant melanoma were included in this retrospective article. Eighteen patients received immediate skin grafts after wide excision skin graft (SG) group), whereas the remaining 10 patients were treated with dermal regenerative template (DRT) (Lando ®, Shenzhen TsingCare Medical Co. Ltd) 14 days before skin grafts (DRT group) and the postoperative survival rate in the two groups was analyzed. During the 6-month follow-up, we compared the scar index, plantar pain, and recurrent skin graft ulcer incidence on the skin grafts area. RESULTS: Postoperative survival rate in the DRT group (91.75% ± 7.64%) was higher than in the SG group (80.51% ± 7.17%). The DRT group showed less scar formation on Vancouver scar scale (VSS index): 3.40 ± 1.07 than the SG group (VSS index: 6.33 ± 0.68). The dermal regenerative template alleviated plantar pain and decreased the incidence of ulcer on the skin grafts area. CONCLUSIONS: The dermal regenerative template not only improves the survival rate of skin grafts but also alleviates scar condition, plantar pain and recurrent skin graft ulcer. This study provides a new reconstructive strategy in plantar skin defects after the excision of malignant melanoma.
