关键词: 营养不良性大疱性表皮松解, 皮肤, 人工, 皮肤移植, 移植, 同种, 挛缩, 活性皮肤替代物, 自体与异体细胞

Abstract: Yang Lyujun, Zhang Mingjun, Xie Sitian, Ye Danyan Department of Burns and Plastic Surgery, The Second Affiliated Hospital of Shantou University Medical College, Shantou 510041, Guangdong, China （Yang LJ, Zhang MJ, Xie ST）; Research Center for Translational Medicine, The Second Affiliated Hospital of Shantou University Medical College, Shantou 510041, Guangdong, China （Yang LJ, Ye DY） Corresponding author: Yang Lyujun, Email: yanglujun726@hotmail.com 【Abstract】 Objective To evaluate the effect of living skin equivalents （LSE） constructed of mixed autologous and allogeneic skin cells and human amnion which served as a matrix on repairing scar contracture of the hand in a patient with recessive dystrophic epidermolysis bullosa（RDEB）. Methods Skin tissues were obtained from a patient with RDEB and her mother, and epidermal keratinocytes and dermal fibroblasts were isolated from these tissues and cultured in vitro separately. Human amnion was obtained from the placenta of an unrelated healthy parturient undergoing cesarean delivery, and served as a matrix of the LSE. The autologous and allogeneic fibroblasts were mixed and seeded on the stromal side of the amnion, and then the autologous and allogeneic keratinocytes were mixed and seeded on the epithelial side of the amnion, so as to construct the human amnion-LSE （AM-LSE）. Histological examination was performed to observe the structure of the skin tissues obtained from the patient and her mother, and immunofluorescence staining was conducted to detect the of type Ⅶ collagen in the skin tissues of the patient and her mother and in the AM-LSE. The AM-LSE was grafted on the skin defects of the patient after release of scar contracture of the hand. After grafting, the survival status of the AM-LSE graft and repairing effect on the wounds were evaluated. Results The constructed AM-LSE consisted of dermis, multilayered and fully differentiated epidermis and well-developed basement membrane. Immunofluo-rescence examination revealed that type Ⅶ collagen was linearly distributed along the basement membrane. Half a year after grafting, the AM-LSE survived well, and no obvious rejection reaction was observed. No blisters or ulcers occurred at the recipient sites, and the scar contracture was mild. The grafted area showed normal skin color with soft texture. The patient could grab and hold things, which had met self-care requirements of daily living. Conclusions The AM-LSE constructed of mixed autologous and allogeneic skin cells have good histological structures, and can be grafted on the wounds after resection of the scars in a RDEB patient. After grafting, no obvious rejection reaction was observed, and the skin was not liable to develop blisters, ulcers or scar contracture due to friction.

Key words: Epidermolysis bullosa dystrophica, Skin, artificial, Skin transplantation, Transplan-tation, homologous, Contracture, Living skin equivalent, Autologous-allogeneic cells