Department of Plastic and Reconstructive Surgery, Kansai Medical University, Hirakata, Osaka, Japan.
PLoS One. 2019 Feb 21;14(2):e0208658. doi: 10.1371/journal.pone.0208658. eCollection 2019.
We developed a novel skin regeneration therapy combining nevus tissue inactivated by high hydrostatic pressure (HHP) in the reconstruction of the dermis with a cultured epidermal autograft (CEA). The issue with this treatment is the unstable survival of CEA on the inactivated dermis. In this study, we applied collagen/gelatin sponge (CGS), which can sustain the release of basic fibroblast growth factor (bFGF), to the inactivated skin in order to accelerate angiogenesis. Murine skin grafts from C57BL6J/Jcl mice (8 mm in diameter) were prepared, inactivated by HHP and cryopreserved. One month later, the grafts were transplanted subcutaneously onto the back of other mice and covered by CGS impregnated with saline or bFGF. Grafts were harvested after one, two and eight weeks, at which point the engraftment was evaluated through the histology and angiogenesis-related gene expressions were determined by real-time polymerase chain reaction. Histological sections showed that the dermal cellular density and newly formed capillaries in the bFGF group were significantly higher than in the control group. The relative expression of FGF-2, PDGF-A and VEGF-A genes in the bFGF group was significantly higher than in the control group at Week 1. This study suggested that the angiogenesis into grafts was accelerated, which might improve the engraftment of inactivated dermis in combination with the sustained release of bFGF by CGSs.
我们开发了一种新的皮肤再生疗法,将经高静压(HHP)灭活的痣组织与培养的表皮移植物(CEA)结合用于真皮重建。这种治疗方法的问题是 CEA 在灭活真皮上的不稳定存活。在这项研究中,我们应用了胶原/明胶海绵(CGS),它可以持续释放碱性成纤维细胞生长因子(bFGF),以加速血管生成。从小鼠(C57BL6J/Jcl)背部制备 8mm 直径的皮肤移植物,经 HHP 灭活并冷冻保存。一个月后,将移植物皮下移植到其他小鼠的背部,并覆盖用生理盐水或 bFGF 浸渍的 CGS。在一周、两周和八周后收获移植物,并通过组织学评估移植物的植入情况,并通过实时聚合酶链反应确定与血管生成相关的基因表达。组织学切片显示,bFGF 组的真皮细胞密度和新形成的毛细血管明显高于对照组。在 bFGF 组中,FGF-2、PDGF-A 和 VEGF-A 基因的相对表达在第 1 周明显高于对照组。本研究表明,血管生成加速,这可能通过 CGS 持续释放 bFGF 来改善灭活真皮的植入。