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Background:Several patients experience persistent otorrhea after a flawless surgical procedure because of insufficient epithelial healing.Several efforts,such as autologous tissue allograft and xenograft,have been made to halt otorrhea.However,a stable technology to induce temporal epithelial repair is yet to be established.Therefore,this study aims to investigate whether implantation of seeding adipose-derived mesenchymal stem cell (ADMSC) aggregates on extracellular matrix (ECM;herein,ADMSC aggregate-ECM) into damaged skin wound promotes skin regeneration.Methods:ADMSC aggregate-ECM was prepared using a previously described procedure that isolated ADMSCs from rabbits and applied to the auricle and auditory meatus wound beds of New Zealand white rabbits.Wound healing was assessed by general observation and hematoxylin and eosin (H&E) staining.Secretion of growth factor of the tissue was evaluated by west blotting.Two other groups,namely,ECM and control,were used.Comparisons of three groups were conducted by one-way analysis of variance analysis.Results:ADMSCs adhered tighdy to the ECM and quickly formed cell sheets.At 2 weeks,general observation and H&E staining indicated that the wound healing rates in the ADMSC aggregate-ECM (69.02±6.36％) and ECM (59.32±4.10％) groups were higher than that in the control group (43.74± 12.15％;P=0.005,P＜ 0.001,respectively) in ear auricle excisional wounds.At 7 weeks,The scar elevation index was evidently reduced in the ADMSC aggregate-ECM (2.08±0.87) and ECM (2.31±0.33) groups compared with the control group (4.06 ± 0.45;P ＜ 0.001,P ＜ 0.001,respectively).In addition,the scar elevation index of the ADMSC aggregate-ECM group reached the lowest rate 4 weeks in advance.In auditory meatus excisional wounds,the ADMSC aggregate-ECM group had the largest range of normal skin-like structure at 4 weeks.The ADMSC aggregate-ECM and ECM groups secreted increased amounts of growth factors that contributed to skin regeneration at weeks 1 and 2,respectively.Conclusions:ADMSC aggregate-ECM and ECM are effective repair materials for wound healing,especially ADMSC aggregateECM.This approach will provide a meaningful experimental basis for mastoid epithelium repair in subsequent clinical trials.