University Hospital Tuebingen, Department of Thoracic and Cardiovascular Surgery, Calwerstraße 7/1, 72076 Tuebingen, Germany.
University Hospital Tuebingen, Department of Thoracic and Cardiovascular Surgery, Calwerstraße 7/1, 72076 Tuebingen, Germany.
Biomater Adv. 2022 Jun;137:212824. doi: 10.1016/j.bioadv.2022.212824. Epub 2022 Apr 25.
Artificial lungs, also known as oxygenators, allow adequate oxygenation of the blood in patients with severe respiratory failure and enable patient survival. However, the insufficient hemocompatibility of the current of artificial lungs hampers their long-term use. Therefore, in this study, a novel strategy was developed to efficiently endothelialize blood-contacting surfaces to improve their hemocompatibility. Hollow fiber membranes (HFMs) were functionalized with dibenzylcyclooctyne (DBCO), and endothelial cells were glycoengineered for covalent conjugation to DBCO by a copper-free click reaction. Metabolic glycoengineering using azidoacetylmannosamine-tetraacylated (AcManNAz) resulted in highly efficient functionalization of endothelial cells with azide (N) molecules on the cell surface without negative impact on cell viability. After 48 h, significantly improved endothelialization was detected on the HFM surfaces functionalized with DBCO compared to unmodified HFMs. Endothelial cells were responsive to inflammatory stimulus and expressed adhesion-promoting molecules (E-selectin, VCAM-1, and ICAM-1). Furthermore, the hemocompatibility of HFMs was analyzed by dynamic incubation with fresh human blood. DBCO-coated and uncoated HFMs showed a comparable hemocompatibility, but the endothelialization of HFMs significantly reduced the activation of blood coagulation and platelets. Interestingly, the incubation of endothelialized HFMs with human blood further reduced the expression of E-selectin and VCAM-1 in endothelial cells. In this study, a highly efficient, cell-compatible method for endothelialization of artificial lungs was established. This click chemistry-based method can be also applied for the endothelialization of other artificial surfaces for tissue engineering and regenerative medicine applications.
人工肺，也称为氧合器，可在严重呼吸衰竭的患者中实现血液的充分氧合，并使患者存活下来。然而，目前人工肺的血液相容性不足限制了它们的长期使用。因此，在这项研究中，开发了一种新策略，通过高效内皮化血液接触表面来改善其血液相容性。将二苄基环辛炔（DBCO）功能化空心纤维膜（HFMs），并通过无铜点击反应使内皮细胞糖基工程化以共价连接到 DBCO。使用叠氮乙酰甘露糖胺四酰基（AcManNAz）进行代谢糖基工程化，可在不影响细胞活力的情况下，在细胞表面上高效地将叠氮化物（N）分子功能化到内皮细胞上。48 小时后，与未改性的 HFMs 相比，DBCO 功能化的 HFMs 表面的内皮化得到了显著改善。内皮细胞对炎症刺激有反应，并表达促进黏附的分子（E-选择素、VCAM-1 和 ICAM-1）。此外，通过与新鲜人血的动态孵育分析 HFMs 的血液相容性。DBCO 涂层和未涂层的 HFMs 显示出相当的血液相容性，但 HFMs 的内皮化显著降低了血液凝固和血小板的激活。有趣的是，内皮化的 HFMs 与人血孵育进一步降低了内皮细胞中 E-选择素和 VCAM-1 的表达。在这项研究中，建立了一种高效、细胞兼容的人工肺内皮化方法。这种基于点击化学的方法也可应用于其他人工表面的内皮化，用于组织工程和再生医学应用。
