Facilitation of adhesion and spreading of endothelial cells on silicone oxide-coated dacron material by microwave-excited low-pressure plasma
Authors: Tilkorn Daniel J. , Sorg Heiko , Sanders Agnes , Köller Manfred , Awakowicz Peter , Hauser Jörg 
Published: January 2022

Abstract

Objectives
Autologous transplants are still the means of choice for bypass surgery. In addition to good tolerability, there is a reduced thrombogenicity and fewer neointima hyperplasia compared to artificial materials. However, since viable transplants are limited, attempts are being made to improve existing artificial vascular prosthesis material. Next to the reduction of thrombogenicity, a rapid endothelialization of the vascular graft should reduce intimal hyperplasia and thus prevent stenoses. The effect of newly developed silicon oxide coatings on the growth of endothelial cells was therefore the goal of this work in a cell culture study.

Methods
A woven, uncoated polyethylene terephthalate (PET) vessel prosthesis was used. The coating process was carried out in a low-pressure plasma reactor in a multi-step process. After preparation of the vacuum chamber hexamethyldisiloxane (HDMSO) with oxygen was evaporated using argon plasma. By this an approx. 1 nm thin adhesion promoter layer was separated from plasma and HMDSO. The silicone oxide barrier layer was applied to the PET vessel samples. The carbon content of the layer could be selectively altered by changing the HMDSO oxygen flow ratio, resulting in coatings of 100 nm, 500 nm, and 1,000 nm. In addition, two different oxygen-to-HMDSO ratios were used. To achieve a carbon coating as low as possible, the ratio was set to 200:1. A carbon-rich layer was obtained with the 1:1 setting. The various coatings were then examined for their surface texture by scanning electron microscopy (SEM) as well as by cell culture experiments for cell viability and growth using EA.hy 926 cells.

Results
SEM showed no changes in the surface morphology; however a layer thickness of 1,000 nm showed peeled off coating areas. Alamar blue assays showed a significantly higher metabolic activity (p=0.026) for the coating 500 nm, ratio 200:1 compared to untreated control samples and a significantly lower metabolic activity (p=0.037) of the coating 500 nm, ratio 1:1 compared to the coating 500 nm, ratio 200:1. This underlines the apparent tendency of the 1:1 coating to inhibit the metabolic activity of the cells, while the 200:1 coating increases the activity. Fluorescence microscopy after calcein acetoxymethyl ester (AM) staining showed no significant difference between the different coatings and the uncoated PET material. However, a tendency of the increased surface growth on the coating 500 nm, ratio 200:1, is shown. The coatings with the ratio 1:1 tend to be less densely covered.

Conclusions
The results of this work indicate a great potential in the silicon coating of vascular prosthesis material. The plasma coating can be carried out easy and gently. Cell culture experiments demonstrated a tendency towards better growth of the cells on the 200:1 ratio coating and a poorer growth on the carbon-rich coating 1:1 compared to the uncoated material. The coating with silicon oxide with a thickness of 500 nm and an oxygen-HMDSO ratio of 200:1, a particularly low-carbon layer, appears to be a coating, which should therefore be further investigated for its effects on thrombogenicity and intimal hyperplasia.