Biedermann Thomas, Klar Agnieszka S, Böttcher-Haberzeth Sophie, Michalczyk Teresa, Schiestl Clemens, Reichmann Ernst, Meuli Martin
Tissue Biology Research Unit, University Children's Hospital Zurich, Zurich, Switzerland.
Pediatr Surg Int. 2015 Jan;31(1):69-76. doi: 10.1007/s00383-014-3622-7. Epub 2014 Oct 18.
Transplantation of pigmented tissue-engineered human autologous skin substitutes represents a promising procedure to cover skin defects. We have already demonstrated that we can restore the patient's native light or dark skin color by adding melanocytes to our dermo-epidermal skin analogs. In this long-term study, we investigated if melanocytes in our skin substitutes continue to express markers as BCL2, SOX9, and MITF, known to be involved in survival, differentiation, and function of melanocytes.
Human epidermal melanocytes and keratinocytes, as well as dermal fibroblasts from light- and dark-pigmented skin biopsies were isolated and cultured. Bovine collagen hydrogels containing fibroblasts were prepared, and melanocytes and keratinocytes were seeded in a 1:5 ratio onto the gels. Pigmented dermo-epidermal skin substitutes were transplanted onto full-thickness wounds of immuno-incompetent rats and analyzed for the expression of melanocyte markers after 15 weeks.
Employing immunofluorescence staining techniques, we observed that our light and dark dermo-epidermal skin substitutes expressed the same typical melanocyte markers including BCL2, SOX9, and MITF 15 weeks after transplantation as normal human light and dark skin.
These data suggest that, even in the long run, our light and dark dermo-epidermal tissue-engineered skin substitutes contain melanocytes that display a characteristic expression pattern as seen in normal pigmented human skin. These findings have crucial clinical implications as such grafts transplanted onto patients should warrant physiological numbers, distribution, and function of melanocytes.
移植色素沉着的组织工程化人自体皮肤替代物是一种覆盖皮肤缺损的有前景的方法。我们已经证明，通过在我们的真皮-表皮皮肤类似物中添加黑素细胞，可以恢复患者的天然浅色或深色皮肤颜色。在这项长期研究中，我们调查了我们皮肤替代物中的黑素细胞是否继续表达已知参与黑素细胞存活、分化和功能的BCL2、SOX9和MITF等标志物。
从浅色和深色皮肤活检中分离并培养人表皮黑素细胞、角质形成细胞以及真皮成纤维细胞。制备含有成纤维细胞的牛胶原蛋白水凝胶，并将黑素细胞和角质形成细胞以1:5的比例接种到凝胶上。将色素沉着的真皮-表皮皮肤替代物移植到免疫缺陷大鼠的全层伤口上，并在15周后分析黑素细胞标志物的表达。
采用免疫荧光染色技术，我们观察到，在移植15周后，我们的浅色和深色真皮-表皮皮肤替代物表达与正常人浅色和深色皮肤相同的典型黑素细胞标志物，包括BCL2、SOX9和MITF。
这些数据表明，即使从长远来看，我们的浅色和深色真皮-表皮组织工程皮肤替代物中的黑素细胞也表现出如正常色素沉着的人类皮肤中所见的特征性表达模式。这些发现具有关键的临床意义，因为移植到患者身上的此类移植物应保证黑素细胞的生理数量、分布和功能。
