Research for wound healing using an hybrid type artificial skin model composed with human immune cells

用培养的人皮肤细胞制成的人工皮肤培养作为实验模型和临床应用都引起了人们的关注。然而，人造皮肤几乎没有受到损害和观察到进展的化学物品。我们用人类免疫细胞创造了许多人造皮肤模型。这些人造皮肤模型被不同强度的二氧化碳激光照射。我们用病理学和免疫组织化学技术评估了这些模型的损伤程度和组织再生的时间进程，并以生物相容多肽水凝胶为支架，制备了不含动物源性材料或病原体的三维人造皮肤。本研究对人造真皮中成纤维细胞的制备方法、增殖、功能表达以及表皮中角质形成细胞的分化进行了研究。合成真皮是由成纤维细胞与多肽水凝胶混合制成的，合成真皮是由角质形成细胞在合成真皮上形成一层表皮层而制成的。一层富含成纤维细胞的泡沫层，由均一的多肽水凝胶组成，随后在合成真皮中形成，成纤维细胞在隔膜内聚集成簇。表皮由3~5层角质形成细胞组成。免疫组织化学染色显示人I型胶原蛋白，表明合成真皮中成纤维细胞周围有功能表达，表皮中角质形成细胞分化，基底膜蛋白表达。成纤维细胞数量在第2周有增加趋势，第4周维持，第5周迅速减少。在合成真皮培养液中，人I型胶原的浓度在第二周到第五周后增加。这些发现表明，多肽水凝胶作为一种合成的皮肤支架，为成纤维细胞和角质形成细胞的增殖和功能表达提供了平台。

Artificial skin cultures made using cultured human skin cells have attracted attention as experimental models, as well as for clinical applications. However artificial skin has few chemical articles that undergo damage and observed progress. We created a number of artificial skin models with human immune cells. These artificial skin models were irradiated with a CO2 laser at various strengths. We evaluated the degree of damage and the time course of tissue regeneration in these models using pathological and immunohistochemical techniques.Also using biocompatible peptide hydrogel as a scaffold, we prepared three -dimensional synthetic skin that does not contain animal-derived materials or pathogens. The present study investigated preparation methods, proliferation, and functional expression of fibroblasts in the synthetic dermis and differentiation of keratinocytes in the epidermis. Synthetic dermis was prepared by mixing fibroblasts with peptide hydrogel, and synthetic skin was prepared by forming an epidermal layer using keratinocytes on the synthetic dermis. A fibroblast-rich foamy layer consisting of homogeneous peptide hydrogel subsequently formed in the synthetic dermis, with fibroblasts aggregating in clusters within the septum. The epidermis consisted of 3-5 keratinocyte layers. Immunohistochemical staining showed human type I collagen, indicating functional expression around fibroblasts in the synthetic dermis, keratinocyte differentiation in the epidermis, and expression of basement membrane proteins. The number of fibroblasts tended to increase up to the second week and was maintained up to the fourth week, but rapidly decreased in the fifth week. In the synthetic dermis medium, the human type I collagen concentration increased after the second week to the fifth week. These findings suggest that peptide hydrogel acts as a synthetic skin scaffold that offers a platform for the proliferation and functional expression of fibroblasts and keratinocytes.