Hussaini Bader E, Lu Xiu-Gui, Wolfe J Alan, Thatte Hemant S
Cardiothoracic Surgery Division, Veterans Affairs Boston Healthcare System, Boston, MA, USA.
J Cardiothorac Surg. 2011 Jun 10;6:82. doi: 10.1186/1749-8090-6-82.
Endothelial injury during harvest influences graft patency post CABG. We have previously shown that endoscopic harvest causes structural and functional damage to the saphenous vein (SV) endothelium. However, causes of such injury may depend on the extraction technique. In order to assess this supposition, we evaluated the effect of VirtuoSaph endoscopic SV harvesting technique (VsEVH) on structural and functional viability of SV endothelium using multiphoton imaging, biochemical and immunofluorescence assays.
Nineteen patients scheduled for CABG were prospectively identified. Each underwent VsEVH for one portion and "No-touch" open SV harvesting (OSVH) for another portion of the SV. A two cm segment from each portion was immersed in GALA conduit preservation solution and transported overnight to our lab for processing. The segments were labeled with fluorescent markers to quantify cell viability, calcium mobilization and generation of nitric oxide. Morphology, expression, localization and stability of endothelial caveolin, eNOS, von Willebrand factor and cadherin were evaluated using immunofluorescence, Western blot and multiphoton microscopy (MPM).
Morphological, biochemical and immunofluorescence parameters of viability, structure and function were well preserved in VsEVH group as in OSVH group. However, tonic eNOS activity, agonist-dependent calcium mobilization and nitric oxide production were partially attenuated in the VsEVH group.
This study indicates that VirtuoSaph endoscopic SV harvesting technique preserves the structural and functional viability of SV endothelium, but may differentially attenuate the vasomotor function of the saphenous vein graft.
取材过程中的内皮损伤会影响冠状动脉旁路移植术(CABG)后移植物的通畅性。我们之前已经表明,内镜取材会对大隐静脉(SV)内皮造成结构和功能损伤。然而,这种损伤的原因可能取决于取材技术。为了评估这一假设,我们使用多光子成像、生化和免疫荧光分析,评估了VirtuoSaph内镜大隐静脉取材技术(VsEVH)对大隐静脉内皮结构和功能活力的影响。
前瞻性纳入19例计划行CABG的患者。每位患者的一条大隐静脉一部分采用VsEVH取材,另一部分采用“非接触式”开放大隐静脉取材(OSVH)。从每部分取材2厘米的节段,浸入GALA导管保存液中,并连夜运至我们实验室进行处理。这些节段用荧光标记物标记,以量化细胞活力、钙动员和一氧化氮的生成。使用免疫荧光、蛋白质印迹和多光子显微镜(MPM)评估内皮小窝蛋白、内皮型一氧化氮合酶(eNOS)、血管性血友病因子和钙黏蛋白的形态、表达、定位和稳定性。
VsEVH组的活力、结构和功能的形态学、生化和免疫荧光参数与OSVH组一样得到了良好的保留。然而,VsEVH组的静息eNOS活性、激动剂依赖性钙动员和一氧化氮生成部分减弱。
本研究表明,VirtuoSaph内镜大隐静脉取材技术保留了大隐静脉内皮的结构和功能活力,但可能会不同程度地减弱大隐静脉移植物的血管舒缩功能。