Biodegradable TEVGs (4 mm diameter, ≈400 μm thickness and 4 cm length) were fabricated using emulsion electrospinning (Nanon-01A, MECC, Tokyo, Japan) from PHBV (403105, Sigma-Aldrich, Saint Louis, MO, USA): PCL (440744, Sigma-Aldrich, Saint Louis, MO, USA) (5:10%)/chloroform (366927, Sigma-Aldrich, Saint Louis, MO, USA) solution using the following parameters: 23 kV voltage, 0.5 mL/h feed rate, 2 mm rotating drum diameter, 22G needle, and 150 mm tip-to-collector distance. Abovementioned polymer ratio was determined in our previous studies [25 (link),31 (link),32 (link)]. In all these investigations, PHBV/PCL vascular grafts did not show any signs of dissolution as long as 1 year after implantation into rat abdominal aorta. Either VEGF (V7259, Sigma-Aldrich, St. Louis, MO, USA), bFGF (SRP4037, Sigma-Aldrich, St. Louis, MO, USA), or SDF-1α (SRP3276, Sigma-Aldrich, St. Louis, MO, USA) were dissolved in phosphate buffered saline (10010023, Thermo Fisher Scientific, Waltman, MA, USA) to 10 µg/mL concentration and then added to PHBV/PCL/chloroform solution (1:20), with the final concentration of 500 ng/mL. Grafts with the combination of VEGF, bFGF, and SDF-1α were two-layered, with the inner layer fabricated using 27G needle and containing VEGF (500 ng/mL) and the outer layer prepared utilizing 22G needle and containing bFGF and SDF-1α (500 ng/mL each).
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