Kljenak Antun, Tominac Trcin Mirna, Bujić Marina, Dolenec Tamara, Jevak Martina, Mršić Gordan, Zmiš Gordana, Barčot Zoran, Muljačić Ante, Popović Maja
Zagreb Children's Hospital, Zagreb, Croatia
Tissue Bank, University Department of Traumatology, Sestre milosrdnice University Hospital Center, Zagreb, Croatia
Acta Clin Croat. 2016 Jun;55(2):279-89. doi: 10.20471/acc.2016.55.02.15.
The purpose of this study was to create a fibrin-based human skin substitute in vitro with epidermal and dermal component and to assess its healing potential in deep partial and full thickness burns. Fibrin scaffolds were prepared from commercial fibrin glue kits. Human fibroblasts were cultured in fibrin gel. Human keratinocytes were seeded on the top of the gel. Viability of cells was determined fluorimetrically. Scanning electron microscope and immunocytochemistry analysis of cultured cells were performed. After hydrosurgical preparation of deep burn necrotic tissue, wound bed was prepared for skin substitutes. Progress of healing was documented using visual estimation and photos. Scanning electron microscope images showed good cell attachment and colony spreading of keratinocytes and fibroblasts on fibrin scaff old. Immunofluorescent staining of cell cultures on fibrin scaffold showed expression of vimentin, a marker of fibroblast cells, cytokeratin 19, a marker of epithelial stem cells, as well as involucrin, a marker of differentiated keratinocytes. Clinical results clearly showed that appearance of the skin did not differ significantly from the areas of transplanted skin using split-thickness skin graft techniques. In conclusion, using these fibrin-cultured autografts on massive full-thickness burn resulted in good healing.
本研究的目的是在体外创建一种具有表皮和真皮成分的基于纤维蛋白的人工皮肤替代品,并评估其在深Ⅱ度和全层烧伤中的愈合潜力。纤维蛋白支架由商用纤维蛋白胶试剂盒制备。人成纤维细胞在纤维蛋白凝胶中培养。人角质形成细胞接种在凝胶顶部。通过荧光法测定细胞活力。对培养的细胞进行扫描电子显微镜和免疫细胞化学分析。在对深度烧伤坏死组织进行水刀清创后,为皮肤替代品准备创面床。使用视觉评估和照片记录愈合过程。扫描电子显微镜图像显示角质形成细胞和成纤维细胞在纤维蛋白支架上有良好的细胞附着和集落扩散。纤维蛋白支架上细胞培养物的免疫荧光染色显示波形蛋白(成纤维细胞的标志物)、细胞角蛋白19(上皮干细胞的标志物)以及兜甲蛋白(分化角质形成细胞的标志物)的表达。临床结果清楚地表明,使用该人工皮肤的部位外观与采用中厚皮片移植技术的移植皮肤区域相比无显著差异。总之,在大面积全层烧伤中使用这些纤维蛋白培养的自体移植物可实现良好愈合。