Department of Thoracic, Cardiac, and Vascular Surgery, University Hospital Tuebingen, Calwerstr. 7/1, 72076 Tuebingen, Germany.
J Biomed Mater Res B Appl Biomater. 2013 Aug;101(6):1061-8. doi: 10.1002/jbm.b.32916. Epub 2013 May 7.
The incomplete endothelialization of especially small-caliber vascular prostheses after implantation in patients is a major disadvantage in cardiovascular interventions. The lack of an endothelium leads to the occurrence of thrombosis at the luminal surface of artificial vascular prostheses. Thus, the development of new graft materials and coatings for induction of complete endothelialization on the implant surfaces is a promising approach to improve hemocompatibility and maintain long-term graft patency. In this study, we designed a rotation model to evaluate the early endothelial cell (EC) seeding efficiency of different small-caliber vascular devices, such as stents and vascular grafts. The suitability of the designed rotation model for endothelialization studies was investigated by seeding and cultivation of prostheses with ECs followed by scanning electron microscopy. Furthermore, the viability of attached ECs was determined by calcein acetoxymethyl ester (AM) staining. The rotation model consisting of low-cost medical disposables enabled sterile incubation and cultivation of ECs with vascular devices. Simultaneously, the rotation of the bioreactor ensured a uniform distribution and adhesion of cells to the devices. Calcein AM staining of adherent cells on prostheses revealed excellent cell viability. Moreover, using the designed rotation model, an influence of different coatings and materials on the adhesion and spreading of ECs was demonstrated. The rotating bioreactor described and used in this study not only saves time and money but is also eminently useful for the accelerated preclinical evaluation of the endothelialization efficiency of different materials and surface coatings of small-caliber vascular devices.
在将尤其小口径血管移植物植入患者体内后,内皮化不完全是心血管介入治疗中的一个主要缺点。缺乏内皮会导致人工血管移植物内腔表面发生血栓形成。因此,开发新的移植物材料和涂层以诱导在植入物表面完全内皮化是一种有前途的方法,可以改善血液相容性并维持移植物的长期通畅性。在这项研究中,我们设计了一种旋转模型来评估不同小口径血管器械(如支架和血管移植物)的早期内皮细胞(EC)接种效率。通过用 EC 接种和培养移植物,然后进行扫描电子显微镜检查,研究了设计的旋转模型在内皮化研究中的适用性。此外,通过 calcein 乙酰氧甲酯(AM)染色来确定附着的 EC 的活力。由低成本医疗耗材组成的旋转模型能够对 EC 和血管器械进行无菌孵育和培养。同时,生物反应器的旋转确保了细胞均匀分布并附着在器械上。对器械上附着细胞进行 calcein AM 染色显示出极好的细胞活力。此外,使用设计的旋转模型,还证明了不同涂层和材料对 EC 粘附和铺展的影响。本研究中描述和使用的旋转生物反应器不仅节省了时间和金钱,而且对于加速不同材料和小口径血管器械表面涂层的内皮化效率的临床前评估也非常有用。