The effect of gradually graded shear stress on the morphological integrity of a huvec-seeded compliant small-diameter vascular graft.
The premature endothelialization of tissue-engineered grafts had often induced cellular detachment at an early period of implantation in arterial circulation, resulting in occlusion at an early period of implantation. This study was aimed to determine whether gradually increased shear stress applied ex vivo improves cell retention and tissue morphological integrity including cell shape and alignment, actin fiber alignment and expression of vascular endothelial (VE) cadherin. Tissue-engineered grafts used for this study were human umbilical vein endothelial cell (HUVEC)-seeded compliant small-diameter grafts made of poly(L-lactide-co-epsilon-caprolactone) fiber meshes fabricated by electrospinning. The shear stresses applied to grafts, generated using a custom-designed mock circulatory apparatus, were 3.2, 8.7 and 19.6 dyn/cm(2). The grafts completely monolayered prior to shear stress exposure exhibited a polygonal cobblestone morphology with randomly distributed actin fibers and VE cadherin at the continuous peripheral region of adjacent cells. The 24-h-loading of high shear stresses (8.7 and 19.6 dyn/cm(2)) equivalent to those of the arterial circulatory system resulted in severe cellular damage resulting in the complete loss of cells. However, a gradually increased graded exposure from a low (3.2 dyn/cm(2)) to a high shear stress (19.6 dyn/cm(2)) resulted in a markedly reduced cell detachment, a highly elongated cell shape, and orientation or alignment of both cells and actin fibers, which were parallel to the direction of flow. Although VE-cadherin expression was not detected yet, a higher degree of tissue integrity was achieved, which may greatly improve the performance particularly at an early period of implantation.
Inoguchi
Hiroyuki
H
Division of Biomedical Engineering, Graduate School of Medicine, Kyushu University, 3-1-1 Maidashi, Fukuoka 812-8582, Japan.
Tanaka
Takashi
T
Maehara
Yoshihiko
Y
Matsuda
Takehisa
T
D001158:Arteries / Q000473:pathology; D001807:Blood Vessel Prosthesis; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry; D003599:Cytoskeleton / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology* / Q000473:pathology; D006801:Humans; D008422:Materials Testing; D008855:Microscopy, Electron, Scanning; D011091:Polyesters / Q000737:chemistry; D013314:Stress, Mechanical; D013997:Time Factors; D023822:Tissue Engineering / Q000379:methods*; D014471:Umbilical Veins / Q000166:cytology*
D011091:Polyesters; D003094:Collagen