用人血管细胞对脱细胞牛颈动脉进行表面修饰可显著降低其血栓形成性。
Surface modification of decellularized bovine carotid arteries with human vascular cells significantly reduces their thrombogenicity.
作者信息
Keshi Eriselda, Tang Peter, Weinhart Marie, Everwien Hannah, Moosburner Simon, Seiffert Nicolai, Lommel Michael, Kertzscher Ulrich, Globke Brigitta, Reutzel-Selke Anja, Strücker Benjamin, Pratschke Johann, Sauer Igor Maximillian, Haep Nils, Hillebrandt Karl Herbert
机构信息
Department of Surgery, Campus Charité Mitte | Campus Virchow-Klinikum, Experimental Surgery, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, Augustenburger Platz 1, 13353, Berlin, Germany.
Cluster of Excellence Matters of Activity. Image Space Material funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany's Excellence Strategy - EXC 2025 - 390648296, Berlin, Germany.
出版信息
J Biol Eng. 2021 Nov 24;15(1):26. doi: 10.1186/s13036-021-00277-2.
BACKGROUND
Since autologous veins are unavailable when needed in more than 20% of cases in vascular surgery, the production of personalized biological vascular grafts for implantation has become crucial. Surface modification of decellularized xenogeneic grafts with vascular cells to achieve physiological luminal coverage and eventually thromboresistance is an important prerequisite for implantation. However, ex vivo thrombogenicity testing remains a neglected area in the field of tissue engineering of vascular grafts due to a multifold of reasons.
METHODS
After seeding decellularized bovine carotid arteries with human endothelial progenitor cells and umbilical cord-derived mesenchymal stem cells, luminal endothelial cell coverage (LECC) was correlated with glucose and lactate levels on the cell supernatant. Then a closed loop whole blood perfusion system was designed. Recellularized grafts with a LECC > 50% and decellularized vascular grafts were perfused with human whole blood for 2 h. Hemolysis and complete blood count evaluation was performed on an hourly basis, followed by histological and immunohistochemical analysis.
RESULTS
While whole blood perfusion of decellularized grafts significantly reduced platelet counts, platelet depletion from blood resulting from binding to re-endothelialized grafts was insignificant (p = 0.7284). Moreover, macroscopic evaluation revealed thrombus formation only in the lumen of unseeded grafts and histological characterization revealed lack of CD41 positive platelets in recellularized grafts, thus confirming their thromboresistance.
CONCLUSION
In the present study we were able to demonstrate the effect of surface modification of vascular grafts in their thromboresistance in an ex vivo whole blood perfusion system. To our knowledge, this is the first study to expose engineered vascular grafts to human whole blood, recirculating at high flow rates, immediately after seeding.
背景
由于在血管外科手术中，超过20%的病例在需要时无法获得自体静脉，因此生产用于植入的个性化生物血管移植物变得至关重要。用血管细胞对脱细胞异种移植物进行表面修饰，以实现生理性管腔覆盖并最终获得抗血栓性，是植入的重要前提。然而，由于多种原因，体外血栓形成性测试在血管移植物组织工程领域仍然是一个被忽视的领域。
方法
在脱细胞牛颈动脉接种人内皮祖细胞和脐带间充质干细胞后，管腔内内皮细胞覆盖率（LECC）与细胞上清液中的葡萄糖和乳酸水平相关。然后设计了一个闭环全血灌注系统。将LECC>50%的再细胞化移植物和脱细胞血管移植物用人全血灌注2小时。每小时进行溶血和全血细胞计数评估，随后进行组织学和免疫组织化学分析。
结果
虽然脱细胞移植物的全血灌注显著降低了血小板计数，但与再内皮化移植物结合导致的血液中血小板消耗不显著（p = 0.7284）。此外，宏观评估显示仅在未接种移植物的管腔内形成血栓，组织学特征显示再细胞化移植物中缺乏CD41阳性血小板，从而证实了它们的抗血栓性。
结论
在本研究中，我们能够在体外全血灌注系统中证明血管移植物表面修饰对其抗血栓性的影响。据我们所知，这是第一项在接种后立即将工程化血管移植物暴露于高流速循环的人全血中的研究。
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