Qingdao University; The Affiliated Hospital of Qingdao University.
Kardiologiia. 2022 Jul 31;62(7):38-43. doi: 10.18087/cardio.2022.7.n2100.
Aim Endothelial progenitor cells (EPCs) play important roles in heart valve replacement surgery. Up-regulation of Grb2‑associated binder 1 (Gab1) promotes hepatocyte growth factor (HGF) - induced endothelial progenitor cell proliferation and migration. This study aimed to investigate the effects of up-regulation of Gab1 in hepatocyte growth factor-induced EPCs in tissue-engineered heart valves (TEHV).Material and methods Fresh porcine aortic valves were placed in 1 % Triton X-100 and trypsin buffer for decellularization. EPCs in the control group were cultured normally, whereas those in the experimental group were both HGF stimulated and transfected with adenovirus containing the Gab1 gene. Cells in the two groups were seeded onto the decellularized valve scaffolds and cultured for 3 or 7 days. TEHV were analyzed by HE and AB-PAS staining.Results By day 3, the experimental group had formed confluent endothelial monolayers on top of the decellularized valves, on the basis of by HE staining and AB-PAS staining. One week later, the control group showed a imperfect endothelial layer.Conclusion HGF-induced EPCs overexpressing Gab1 can endothelialize the decellularized matrix and create functional TEHV, which may then be preconditioned in a bioreactor before clinical implantation.
目的 内皮祖细胞（EPCs）在心脏瓣膜置换术中发挥重要作用。Grb2 相关结合蛋白 1（Gab1）的上调促进了肝细胞生长因子（HGF）诱导的内皮祖细胞增殖和迁移。本研究旨在探讨上调 Gab1 对组织工程心脏瓣膜（TEHV）中 HGF 诱导的 EPC 的影响。
材料和方法 新鲜猪主动脉瓣置于 1%Triton X-100 和胰蛋白酶缓冲液中进行脱细胞处理。对照组的 EPC 正常培养，实验组既受 HGF 刺激又转染含有 Gab1 基因的腺病毒。两组细胞均接种于脱细胞瓣膜支架上，培养 3 或 7 天。通过 HE 和 AB-PAS 染色分析 TEHV。
结果 第 3 天，实验组通过 HE 染色和 AB-PAS 染色显示在脱细胞瓣膜顶部形成了连续的内皮单层。一周后，对照组显示出不完整的内皮层。
结论 HGF 诱导的过表达 Gab1 的 EPC 可使脱细胞基质内皮化，并构建具有功能的 TEHV，然后可在生物反应器中进行临床植入前预处理。
