Biomimetic approach to the design of artificial small‑diameter blood vessels

Author:

Nemets E. A.1,Belov Yu. V.2,Kiryakov K. S.1,Grudinin N. V.1,Bogdanov V. K.1,Filippov K. S.1,Nikolskaya A. O.1,Tyunyaeva I. Yu.1,Vypryshko A. A.1,Zaxarevich V. M.1,Basok Yu. B.1,Sevastianov V. I.2

Affiliation:

1. Shumakov National Medical Research Center of Transplantology and Artificial Organs

2. Shumakov National Medical Research Center of Transplantology and Artificial Organs; Institute of Biomedical Research and Technology

Abstract

Objective: To create 2-mm diameter multilayer porous tubular scaffolds (PTS) with characteristics that resemble small-diameter native blood vessels in terms of characteristics.Materials and methods. PTS made of polycaprolactone (PCL, MM 80000) with a PCL-made sealing coat/layer with gelatin addition (PCL-gelatin) with a diameter of 2 mm were created by electrospinning (NANON-01A). Bioactive coating was applied to the PTS surface by sequential incubation in solutions of bovine serum albumin, heparin (Hp), and platelet lysate (PL). Cytotoxicity was investigated under conditions of direct contact of PTS with a monolayer of NIH/3T3 mouse fibroblasts. Viability of human umbilical vein endothelial cells (EA.hy926) was evaluated using Live/Dead® Viability/Cytotoxicity Kit. Permeability and blood flow parameters of the PTS implanted in the infrarenal section of the rat aorta were recorded using Doppler imaging.Results. A three-layer PTS construct with an inner diameter of 2 mm was developed. Its inner and outer layers were formed from 0.2 mL of PCL solution, and the middle sealing coat/layer was from 0.5 mL of PCL with addition of 30% (by weight of polymer) gelatin. Introduction of the sealing coat/layer reduced surgical porosity (SP) from 56.2 ± 8.7 mL/(cm2·min) for a single-layer PTS made of pure PCL to 8.9 ± 2.6 mL/(cm2·min) for a three-layer PTS. The resulting PTS demonstrated physicomechanical characteristics similar to those of native blood vessels; it also showed no cytotoxicity. Application of a bioactive coating of Hp and PL allowed for increased in vitro adhesion and proliferation of endothelial cells. The technique of implantation of 10 mm long fragments of three-layer PTS into the infrarenal section of a rat aorta was corrected, thus minimizing blood loss and narrowing the anastomosis site. In an acute experiment, it was proven that the prostheses were patent and that blood flow parameters (systolic and diastolic velocity, resistivity index) were close to the corresponding indicators of native rat aorta.Conclusion. The developed three-layer PTS constructs have low SP and physicomechanical properties close to those of native blood vessels. Bioactive coating improves the in vitro matrix properties of PTS relative to human endothelial cells. At short-term implantation into the aorta of experimental animals, PTS showed no early thrombosis, while blood flow parameters were close to those of native rat aorta. Thus, three-layer PTS with bioactive coating can be used as a scaffold for creation of in situ tissue-engineered construct of a small-diameter blood vessel.

Publisher

V.I. Shimakov Federal Research Center of Transplantology and Artificial Organs

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