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Background The aim of this research was to determine the efficacy of combination therapy using an alginate dressing and mouse epidermal growth factor (mEGF) on proliferation and differentiation of epidermal stem cells (ESCs) in patients with refractory wounds.Methods Eighteen patients (12 males and 6 females,aged from 18 to 61 years (mean 36.4 years)) with various skin wounds,were treated by dressing changing for one month.The wounds were located in the foot (11),calf (3),thigh (2)and forearm (2).The patients were randomly divided into 3 groups:alginate dressing and mEGF (group A; n=6),mEGF (group B; n=6) and control (group C; n=6).Wound closure indexes were measured at 7,14,21 and 28 days.Samples were harvested for pathologic examination,at 7 and 14 days following treatment.Cytokeratin 10 (CK10) and cytokeratin 15 (CK15) positive cells were evaluated using the super-sensitivity (SP) immunohistochemical staining technique.Results Wound healing was promoted in groups A and B.In group A,the wound closure index was increased significantly (P ＜0.05),and in one case the maximum cure area reached 102 cm2.Pathological examination identified a thicker epidermis,active angiogenesis and enhanced granulation in group A compared with groups B and C.Using the SP immunohistochemical staining technique,we showed that ESCs in group A were bigger in size and larger in number than in groups B and C.Overall,there was a significant difference in ESCs proliferation and differentiation between group A and group B (or C).Conclusions Combination therapy using an alginate dressing and mEGF shows increased proliferation and differentiation of ESCs in patients with refractory wounds compared with those treated with mEGF alone.