Department of Plastic and Reconstructive Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Sapporo, Japan.
Department of Plastic and Reconstructive Surgery, Aichi Medical University, Nagakute, Japan.
Microsurgery. 2019 Mar;39(3):247-254. doi: 10.1002/micr.30394. Epub 2018 Dec 3.
Vascularized lymph node transfer is becoming more common in the treatment of lymphedema, but suitable small animal models for research are lacking. Here, we evaluated the feasibility of pedicled vascularized inguinal lymph node transfer in mice.
Twenty-five mice were used in the study. An inguinal lymph node-bearing flap with a vascular pedicle containing the superficial caudal epigastric vessels was transferred into the ipsilateral popliteal fossa after excision of the popliteal lymph node. Indocyanine green (ICG) angiography was used to confirm vascularity of the flap. ICG lymphography was performed to evaluate lymphatic flow at 3 and 4 weeks postoperatively. Patent blue dye was injected into the ipsilateral hind paw to observe staining of the transferred lymph node at 4 weeks postoperatively. All transferred lymph nodes were then harvested and histologically evaluated by hematoxylin and eosin staining.
In 16 of the 25 mice, ICG lymphography showed reconnection between the transferred lymph node and the afferent lymphatic vessels, as confirmed by patent blue staining. Histologically, these transferred lymph nodes with afferent lymphatic reconnection significantly regressed in size (0.37 ± 0.24 mm ) and showed clear follicle formation, whereas those without afferent lymphatic reconnection showed less size regression (1.31 ± 1.17 mm ); the cell population was too dense to allow identification of follicles.
We established a mouse model of vascularized lymph node transfer with predictable afferent lymphatic reconnection. Both the vascularization and reconnection might be necessary for functional regeneration of the transferred lymph node.
带血管淋巴结移植在治疗淋巴水肿中的应用越来越普遍,但缺乏合适的小动物模型进行研究。在此,我们评估了在小鼠中进行带蒂血管化腹股沟淋巴结移植的可行性。
本研究共使用了 25 只小鼠。在切除腘窝淋巴结后,将包含浅表尾侧腹壁血管的带血管蒂的腹股沟淋巴结皮瓣转移到同侧的腓肠窝。吲哚菁绿(ICG)血管造影用于确认皮瓣的血供。ICG 淋巴造影术用于评估术后 3 周和 4 周的淋巴流动情况。在术后 4 周,将专利蓝染料注入同侧后足,观察转移淋巴结的染色情况。所有转移的淋巴结均被采集并通过苏木精-伊红染色进行组织学评估。
在 25 只小鼠中,16 只的 ICG 淋巴造影显示转移淋巴结与输入淋巴管之间重新连接,这通过专利蓝染色得到了证实。组织学上,这些与输入淋巴管重新连接的转移淋巴结显著缩小(0.37±0.24mm),并显示出明显的滤泡形成,而那些没有输入淋巴管重新连接的淋巴结则缩小程度较小(1.31±0.17mm);细胞密度过高,无法识别滤泡。
我们建立了一种具有可预测性输入淋巴管再连接的血管化淋巴结移植小鼠模型。血管化和再连接可能是转移淋巴结功能再生所必需的。