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

使用培养的人皮肤细胞制成的人工皮肤培养物作为实验模型以及临床应用已经引起了关注。然而，人造皮肤几乎没有发生损伤和观察到进展的化学制品。我们用人类免疫细胞创造了许多人造皮肤模型。这些人工皮肤模型用不同强度的CO2激光照射。我们用病理学和免疫组化技术评估了这些模型的损伤程度和组织再生的时间过程。我们还使用生物相容性肽水凝胶作为支架，制备了不含动物源性材料或病原体的三维合成皮肤。本研究探讨了合成真皮中成纤维细胞的制备方法、增殖和功能表达以及表皮中角质形成细胞的分化。通过将成纤维细胞与肽水凝胶混合来制备合成真皮，并且通过在合成真皮上使用角质形成细胞形成表皮层来制备合成皮肤。随后在合成真皮中形成由均质肽水凝胶组成的富含成纤维细胞的泡沫层，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，表明在合成真皮中成纤维细胞周围的功能表达、表皮中的角质形成细胞分化和基底膜蛋白的表达。成纤维细胞的数量趋于增加，直到第二周，并保持到第四周，但迅速下降，在第五周。在合成真皮培养基中，人I型胶原蛋白浓度在第二周至第五周后增加。这些发现表明，肽水凝胶作为合成的皮肤支架，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.