Journal «Angiology and Vascular Surgery» • 

2015 • VOLUME 21 • №4

Effect of extracellular matrix components on adhesion of bone marrow multipotent mesenchymal stromal cells to polytetrafluoroethylene

Karpenko A.A.1, Rozanova I.A.1, Povechshenko O.V.1,2, Lykov A.P.1,2, Bondarenko N.A.1,2, Kim I.I.1,2, Nikonorova Yu.V.1,2, Podkhvatilina N.A.1, Sergeevichev D.S.1, Popova I.V.1, Konenkov V.I.1,2

1) Laboratory of Experimental Surgery and Morphology, Centre for Vascular and Hybrid Surgery, Novosibirsk Scientific Research Institute for Circulatory Pathology named after Academician E.N. Meshalkin under the RF Ministry of Public Health,
2) Laboratory of Cellular Technologies, Scientific Research Institute of Clinical and Experimental Lymphology under the Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia

Search for new bioengineering materials for creation of small-diameter vascular grafts is currently a priority task. One of the promising trends of creating tissue engineering constructions is coating the internal layer of implants made of polytetrafluoroethylene (PTFE) with autologous mesenchymal multipotent stromal cells.

In the study we assessed the ability of separate components of the extracellular matrix such as fibronectin, type I collagen and type IV collagen to influence adhesion, proliferation and morphology of mesenchymal multipotent stromal cells being cultured on PTFE. Bone marrow multipotent stromal cells taken from second-passage Wistar rats in the amount of 106 per 1 cm2 were applied onto PTFE. We used the following variants of preliminary treatment of the material prior to seeding: fibronectin with type I collagen, fibronectin with type IV collagen, fibronectin with a mixture of type I and IV collagens, as well as a control group without coating. After six weeks of cell growth on PTFE patches the samples were subjected to fixation in 10% formalin followed by haematoxylin-eosin stain and morphometric assessment of adhered cells by calculation using the software AxioVision (Carl Zeiss), assessing the number of cells, area of the cellular monolayer, dimensions and ratios of the area of separate cells and the area of cellular nuclei.

The maximal area of the monolayer from mesenchymal multipotent stromal cells on the PTFE surface was revealed while culturing with a mixture of fibronectin and type I and IV collagens. Cell colonization density while treatment of the synthetic material with mixtures of fibronectin with type I collagen, type IV collagen and type I and IV collagens demonstrated the results exceeding the parameters of the control specimen 5-, 2.5- and 7-fold, respectively. Hence, extracellular matrix components considerably increase enhance adhesion of cells to PTFE, as well as improve formation of a monolayer from mesenchymal multipotent stromal cells.

KEY WORDS:polytetrafluoroethylene, multipotent mesenchymal stromal cells, synthetic vascular graft, tissue engineering constructions, biocompatible transplants, collagen, fibronectin, adhesion .

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