Liem Pham Hieu, Morimoto Naoki, Ito Ran, Kawai Katsuya, Suzuki Shigehiko
Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan,
J Artif Organs. 2013 Sep;16(3):332-42. doi: 10.1007/s10047-013-0708-2. Epub 2013 May 5.
Giant congenital melanocytic nevi (GCMN) are defined as nevi greater than 20 cm in diameter. It is difficult to completely remove GCMN because of the lack of available skin grafts for covering the resultant defects. This study examined whether it is possible to produce reconstructed skin by combining epidermal and acellular dermal matrix (ADM) tissue derived from excised GCMN. GCMN skin samples were obtained with the informed consent of volunteer patients. The abilities of hypertonic saline (1 N NaCl), 0.05% trypsin, 0.1% SDS (sodium dodecyl sulfate), and phosphate buffered saline (PBS) to decellularize GCMN tissue were compared. The specimens were incubated in one of the test solutions at 37 °C for 48 h, before being washed with PBS at 4 °C for 14 days. Residual nuclei, residual DNA, nevus tissue viability, and the structural integrity of the basement membrane and capillaries were evaluated before treatment, and after 48 h' treatment with or without 7 or 14 days' washing. We tried to produce reconstructed skin by combining the resultant ADM with enzymatically separated GCMN epidermal tissue. The histological structure of the reconstructed skin was examined after it had been cultured for 5 days. In the SDS group, most cells had been removed after 48 h, and the DNA content of the ADM was significantly lower than in the other groups. As for viability, no significant difference was detected among the groups. The basement membrane and capillaries remained intact in all groups. After 5 days' culturing, the epidermis had become attached to the ADM in all groups, except the SDS group. SDS displayed a superior decellularization ability compared with the other methods; however, it cannot be used to produce reconstructed skin because of its toxicity. In conclusion, we produced reconstructed skin that was devoid of nevus cells by combining GCMN epidermal tissue with GCMN-derived ADM produced with NaCl or trypsin. This is a promising treatment strategy for giant nevus.
巨大先天性黑素细胞痣(GCMN)被定义为直径大于20厘米的痣。由于缺乏可用的皮肤移植物来覆盖由此产生的缺损,完全切除GCMN很困难。本研究探讨了通过将切除的GCMN来源的表皮组织和脱细胞真皮基质(ADM)相结合来制备重建皮肤的可能性。在志愿者患者知情同意的情况下获取GCMN皮肤样本。比较了高渗盐水(1N NaCl)、0.05%胰蛋白酶、0.1%十二烷基硫酸钠(SDS)和磷酸盐缓冲盐水(PBS)使GCMN组织脱细胞的能力。将标本在其中一种测试溶液中于37℃孵育48小时,然后在4℃用PBS洗涤14天。在处理前以及在48小时处理后(无论是否经过7天或14天洗涤)评估残留细胞核、残留DNA、痣组织活力以及基底膜和毛细血管的结构完整性。我们试图通过将所得的ADM与酶解分离的GCMN表皮组织相结合来制备重建皮肤。在培养5天后检查重建皮肤的组织结构。在SDS组中,48小时后大多数细胞已被去除,ADM的DNA含量显著低于其他组。关于活力,各组之间未检测到显著差异。所有组的基底膜和毛细血管均保持完整。培养5天后,除SDS组外,所有组的表皮均已附着于ADM。与其他方法相比,SDS显示出更强的脱细胞能力;然而,由于其毒性,它不能用于制备重建皮肤。总之,我们通过将GCMN表皮组织与用NaCl或胰蛋白酶制备的GCMN来源的ADM相结合,制备出了不含痣细胞的重建皮肤。这是一种治疗巨大痣的有前景的策略。