Application of a rotating bioreactor consisting of low‐cost and ready‐to‐use medical disposables for in vitro evaluation of the endothelialization efficiency of small‐caliber vascular prostheses

Abstract

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.

Optimized conditions for successful transfection of human endothelial cells with in vitro synthesized and modified mRNA for induction of protein expression

“…BJ human foreskin fibroblasts (Stemgent, Cambridge, USA) were cultivated in DMEM with high glucose containing 10% FBS, 2 mM L-glutamine, 1% penicillin/streptomycin, and 30 mM HEPES (Life Technologies, Darmstadt, Germany). Human endothelial cells (ECs) were isolated as described previously [12] and cultivated in flasks precoated with 0.1% gelatin in Vasculife® EnGS EC culture medium (CellSystems, Troisdorf, Germany) containing VascuLife EnGS LifeFactors Kit, 50 mg/ml gentamicin, and 0.05 mg/ml amphotericin B (PAA Laboratories). Cells were kept at 37°C with 5% CO 2 and media was changed every 3 days.…”

In vitro Study of a Novel Stent Coating Using Modified CD39 Messenger RNA to Potentially Reduce Stent Angioplasty-Associated Complications

“…Isolation of primary human ECs from residual saphenous vein biopsies was performed as previously described and approved by the ethics committee of the University of Tuebingen, Tuebingen, Germany [ 17 ]. One day prior to the transfection, HEK293 cells and human ECs were seeded with a density of 150.000 cells per well onto a 12-well plate (Corning Costar, NY, USA).…”

Strategies and Processes to Decellularize and Recellularize Hearts to Generate Functional Organs and Reduce the Risk of Thrombosis

“…The smooth muscle cells were cultured on the exterior of a decellularized matrix, while the endothelial cells were cultured on the lumen. Avci-Adali et al 86 discussed a rotating bioreactor based on ready-touse medical disposables to reendothelialize a vascular conduit. Their results demonstrated that expanded polytetrafluoroethylene (ePTFE) grafts with albumin and heparin coatings produced the most extensive endothelial cell adhesion and spreading.…”