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Background: There is not an ideal biomaterial for tissue‐engineered skin substitutes (TESSs), and most of the studies or existing therapies use xenogeneic origin natural biomaterials or biosynthetic scaffolds. Objective: To analyse clinical, histological integration and homeostasis parameters of a human TESS manufactured with fibrin–hyaluronic acid biomaterial (HA‐Skin), grafted in immunodeficient mice for 8 weeks, and compared with the gold standard treatment (Autograft), a human TESS manufactured with fibrin–agarose biomaterial (AG‐Skin) and secondary wound healing dressings. Methods: Human TESSs and autografts were implanted into BALB/c mice after surgical excision. Secondary wound healing approach was achieved with biosynthetic collagen wound dressing (Biobrane®) and fibrin–hyaluronic acid or fibrin–agarose biomaterial without cells (Total N = 44). Clinical integration and homeostasis parameters were evaluated every two weeks for two months. Histological and immunohistochemical analyses were performed four and eight weeks after grafting. Results: HA‐Skin, AG‐Skin and Autograft groups showed a proper clinical integration and epithelization eight weeks later. Scar evaluation revealed better results for Autograft and HA‐Skin. Homeostasis analysis indicated similar values of transepidermal water loss and elasticity between HA‐Skin (6.42 ± 0.75 g/h/m2, 0.42 ± 0.08 AU), Autograft (6.91 ± 1.28 g/h/m2, 0.40 ± 0.08 AU) and healthy mouse skin (6.40 ± 0.43 g/h/m2, 0.35 ± 0.03 AU). Histological results showed that human TESSs and autografts presented better skin structuration and higher expression of cytokeratins. Conclusions: This study suggests that human TESS based on fibrin–hyaluronic acid biomaterial could be suitable for clinical application in the treatment of several dermatological pathologies (wound healing). [ABSTRACT FROM AUTHOR]