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目的应用改良小室法在祼鼠体内构建出有正常组织结构的皮肤和毛发。方法应用16.5 d小鼠胚胎的表皮细胞、成纤维细胞和Ⅰ型鼠尾胶原蛋白构建毛囊重建模型。首先在裸鼠背部做直径1.0 cm的圆形创面,将小室固定于创面上;然后,在小室内加入成纤维细胞和Ⅰ型鼠尾胶原蛋白混合悬液形成真皮层,等待胶原凝固后加入表皮细胞形成表皮层。对照组采用传统小室法,16.5 d小鼠胚胎的表皮细胞和成纤维细胞混匀后加入裸鼠体内构建好的小室中。结果实验组和对照组在第10天拆除小室后创面皮肤均得到修复,两组无明显差异。拆除小室1周后,实验组在创面上形成均匀分布的毛发,对照组毛发呈现不均匀分布。拆除小室1个月后可观察到实验组和对照组创面均能够形成大量毛发。通过HE染色及免疫荧光检测发现改良小室法所构建的皮肤拥有正常的组织结构及完整的毛囊结构。结论改良小室法能够构建出有正常组织结构的皮肤组织,是一个高效稳定的毛囊重建模型。
Objective To construct the skin and hair with normal tissue structure by using modified chamber method. Methods Hair follicle reconstruction models were constructed using epidermal cells, fibroblasts and type Ⅰ murine collagen of 16.5 d mouse embryos. Firstly, a circular wound with a diameter of 1.0 cm was made on the back of the nude mouse, and the cell was fixed on the wound. Then, a mixed suspension of fibroblasts and collagen type Ⅰ was formed in the small chamber to form a dermis. After the collagen was coagulated, Cells form the epidermis. The control group using the traditional chamber method, 16.5 d mouse embryonic epidermal cells and fibroblasts after mixing into nude mice built in the chamber. Results In the experimental group and the control group, the skin of the wound was repaired on the 10th day after removal of the small cell. There was no significant difference between the two groups. One week after the chamber was removed, the experimental group formed a uniform distribution of hair on the wound, while the control group showed uneven distribution of hair. One month after removal of the cell, it was observed that both the experimental group and the control group wounds were able to form a large amount of hair. HE staining and immunofluorescence showed that the skin constructed by the modified chamber method had normal tissue structure and complete hair follicle structure. Conclusion Modified chamber method can construct skin tissue with normal tissue structure and is an efficient and stable hair follicle reconstruction model.