Department of Plastic and Reconstructive Surgery, Otsu Red Cross Hospital, Shiga 520-8511, Japan.
Department of Plastic and Reconstructive Surgery, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
Biomed Res Int. 2021 Feb 24;2021:3485189. doi: 10.1155/2021/3485189. eCollection 2021.
Giant congenital melanocytic nevi (GCMN) treatment remains controversial. While surgical resection is the best option for complete removal, skin shortage to reconstruct the skin defect remains an issue. We report a novel treatment using a high hydrostatic pressurization (HHP) technique and a cryopreservation procedure. However, cryopreservation may inhibit revascularization of implanted nevus tissue and cultured epidermal autograft (CEA) take. We aimed to investigate the influence of the cryopreservation procedure on the HHP-treated dermis specimen and CEA take on cryopreserved tissue.
Nevus tissue harvested from a patient with GCMN was inactivated with HHP of 200 MPa and then cryopreserved at -30°C for 28 days. The cryopreserved specimen was compared with fresh (HHP-treated without cryopreservation) tissue and with untreated (without HHP treatment) tissue to evaluate the extracellular matrix, basal membranes, and capillaries. Cultured epidermis (CE) take on the cryopreserved tissue was evaluated following implantation of the cryopreserved nevus tissue with CE into the subcutis of nude mice.
No difference was observed between cryopreserved and fresh tissue in terms of collagen or elastic fibers, dermal capillaries, or basement membranes at the epidermal-dermal junction. In 4 of 6 samples (67%), applied CE took on the nevus tissues and regenerated the epidermis in the cryopreserved group compared with 5 of 6 samples (83%) in the fresh group.
Cryopreservation at -30°C for 28 days did not result in significant damage to inactivated nevus tissue, and applied CE on the cryopreserved nevus tissues took and regenerated the epidermis. Inactivated nevus tissue with HHP can be used as a dermal substitute after 28-day cryopreservation.
巨大先天性黑色素细胞痣（GCMN）的治疗仍存在争议。虽然手术切除是完全去除的最佳选择，但皮肤短缺仍然是重建皮肤缺损的一个问题。我们报告了一种使用高静水压力（HHP）技术和冷冻保存程序的新治疗方法。然而，冷冻保存可能会抑制植入性痣组织和培养表皮移植物（CEA）的再血管化。我们旨在研究冷冻保存程序对 HHP 处理的真皮标本和冷冻保存组织上 CEA 取的影响。
从患有 GCMN 的患者中采集痣组织，用 200 MPa 的 HHP 灭活，然后在-30°C 下冷冻保存 28 天。将冷冻保存的标本与新鲜（未冷冻保存的 HHP 处理）组织和未处理（未进行 HHP 处理）组织进行比较，以评估细胞外基质、基底膜和毛细血管。将冷冻保存的痣组织与 CEA 一起植入裸鼠皮下，评估冷冻保存组织上培养的表皮（CE）的取。
在胶原或弹性纤维、真皮毛细血管或表皮-真皮交界处的基底膜方面，冷冻保存组织与新鲜组织之间没有差异。在冷冻保存组的 6 个样本中有 4 个（67%），应用的 CEA 取了冷冻保存的痣组织，并在再生表皮，而在新鲜组的 6 个样本中有 5 个（83%）。
在-30°C 下冷冻保存 28 天不会对失活的痣组织造成显著损伤，并且应用于冷冻保存的痣组织上的 CEA 可以取并再生表皮。经过 28 天冷冻保存后，用 HHP 灭活的痣组织可作为真皮替代物使用。
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