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Effect of glucose concentration and culture substrate on HUVECs viability in in vitro cultures: A literature review and own results

Ciechanowska Anna, Gora Ilona, Sabalinska Stanislawa, Foltynski Piotr, Ladyzynski Piotr

Biocybernetics and Biomedical Engineering

2021

Vol. 41, no. 4

1390--1405

Some clinical studies reported that glucose variability increased the risk of developing diabetes-related late complications more than constant hyperglycemia, while others claimed that the evidence was not strong enough to support such a conclusion. A few in vitro studies investigated the effect of constantly high or variable glucose levels (VGLs) on endothelial cells (EC). The first aim of this work was to review these studies and demonstrate that most of them support the notion that viability and other metabolic parameters of EC deteriorate faster in cell cultures with VGLs than in cultures with stable normal or high glucose concentration. The second aim was to verify whether the effect of glucose concentration is the same regardless of other culture conditions such as the substrate on which the cells are grown. We cultured Human Umbilical Vein Endothelial Cells (HUVECs) for 7 or 14 days in constant (5 mM or 20 mM) or variable (switching between 5 mM and 20 mM once a day) glucose concentration in culture plates, which were either not-covered with any additional substrate or were covered with fibronectin or gelatin. We assessed the cell viability using a propidium iodide test. The ANOVA revealed that HUVECs viability was affected not only by glucose concentration and duration of the cell culturing but also by the type of substrate and interactions of these three factors. In conclusion, the effect of glucose level on EC viability should not be analyzed in isolation from other culture conditions that may amplify or attenuate this effect.

Endothelial cells on pet vascular prostheses impregnated with polyester-based copolymers and coated with cell-adhesive protein assemblies

Chlupac J., Filova E., Riedel T., Brynda E., Pamuła E., Lisa V., Bacakova L.

Engineering of Biomaterials

2008

Vol. 11, no. 81-84

108--111

Arterial bypass surgery with synthetic vascular prostheses achieves poor patency rates compared to autogenous natural materials, and this is a challenge for tissue engineering research concerning small caliber vascular grafts. Modifications of the prosthetic surface followed by endothelial cell seeding may reduce thrombogenicity and intimal hyperplasia. Planar polyethylene terephthalate (PET) vascular prosthetic samples were impregnated with the copolymer poly(glycolide-L-lactide) (PGL) or with the terpolymer poly(glycolide-L-lactide-(e)caprolactone) (PGLCap) in order to lower the permeability of the knitted fabrics and ensure a less adhesive background. Subsequent modification with adhesive protein assemblies composed of collagen type I (Co) in conjunction with laminin (LM), fibronectin (FN) or fibrin (Fb) gel was performed to enhance cell adhesion. Bovine pulmonary artery endothelial cells (EC) of the CPAE line were seeded on to the coatings and subjected to static tissue culture conditions for 7 days. Impregnation of the PET prostheses decreased the initial adhesion and proliferation of the EC. After coating with the protein assemblies, the impregnated PET provided better substrates for cell culture than the protein-coated PET, on which the EC population started decreasing after 4 days of culture. The cells proliferated better on the CoFN, CoFb and CoFbFN coatings than on the Co and CoLM coatings. Impregnation type and adhesive matrix protein deposition may play an important role in successful endothelialization, healing and clinical performance of vascular grafts.