毛乳头细胞和表皮细胞构建带附属器的组织工程皮肤

Reconstruction of Tissue Engineered Skin Substitute Containing Appendages Using Dermal Papillae Cells And Keratinocytes

目的根据毛囊形成原理,以人毛乳头细胞和表皮细胞为种子细胞,构建带附属器的组织工程皮肤替代物。方法分实验组和对照组,实验组皮肤替代物真皮层接种人毛乳头细胞和表皮细胞;对照组真皮层不接种任何细胞。在裸鼠背部作一圆形全层皮肤缺损,将两组皮肤替代物气-液界面培养5 d后移植到裸鼠背部。观察创面愈合情况,并在移植后第4、6、8周取材,进行组织学观察。结果皮肤替代物移植后2周,实验组创面已完全愈合,移植后4周对照组创面才完全愈合,并且对照组创面收缩比实验组明显。移植后6周,实验组真皮层内见到毛囊样结构和皮脂腺结构,对照组未见毛囊样结构和皮脂腺样结构。结论将人的毛乳头细胞和表皮细胞共同种植在皮肤替代物的真皮层,可以构建出带附属器的组织工程皮肤替代物,这种替代物对创面的修复能力更强。

Objective To reconstruct tissue-engineered skin substitutes containing appendages using human dermal papillae cells and keratinocytes together according to the developmental mechanism of the hair follicles. Methods Dermis substitutes of the experimental group were reconstructed by mixing both keratinocytes and dermal papillae cells into collagen gel. The collagen gel was seeded onto the surface of dermal substitutes. No cells were seeded onto the dermis in control group. The composite skin substitutes were cultured at the air-liquid interface for 5 days. A rounded full-thickness skin defect in diameter of lcm was made on the back of the nude mice and then covered with the skin substitute. The healing and contraction of wound was observed. At 4, 6, 8 weeks after transplantion, samples were obtained. Results The wound healing time was 2 and 4 weeks in experimental and control group respectively. The wound contraction was more greater in control group than in experimental group. Hair follicles and sebaceous glands formed in the dermis of the experimental group at 6 weeks after transplantation. Conclusion In corporation of dermal papillae cells and keratinocytes into dermal substitutes is a promising method to reconstruct tissue-engineered skin substitutes containing appendage.