Abstract
Tissue engineered vascular grafts (TEVG) are potentially clear from ethical and epidemiological concerns sources for reconstructive surgery for small diameter blood vessels replacement. Here, we proposed a novel method to create three-layered TEVG on biocompatible glass fiber scaffolds starting from flat sheet state into tubular shape and to train the resulting tissue by our developed bioreactor system. Constructed tubular tissues were matured and trained under 3 types of individual flow programs, and their mechanical and biological properties were analyzed. Training in the bioreactor significantly increased the tissue burst pressure resistance (up to 18 kPa) comparing to untrained tissue. Fluorescent imaging and histological examination of trained vascular tissue revealed that each cell layer has its own individual response to training flow rates. Histological analysis suggested reverse relationship between tissue thickness and shear stress, and the thickness variation profiles were individual between all three types of cell layers. Concluding: a three-layered tissue structure similar to physiological can be assembled by seeding different cell types in succession; the following training of the formed tissue with increasing flow in a bioreactor is effective for promoting cell survival, improving pressure resistance, and cell layer formation of desired properties.
Highlights
Tissue engineered vascular grafts (TEVG) are potentially clear from ethical and epidemiological concerns sources for reconstructive surgery for small diameter blood vessels replacement
The clinical application of TEVG requires reconstruction of vascular function based on the three-layered tissue structure, as well as scalability in size and the strength to withstand blood flow pulses and surgery suturing, 1Graduate School of Systems and Information Engineering, University of Tsukuba, 1‐1‐1 Tennodai, Tsukuba, Ibaraki 305‐8573, Japan. 2Center for Cybernics Research, University of Tsukuba, Tsukuba, Japan. 3Faculty of Engineering, Information and Systems, University of Tsukuba, Tsukuba, Japan. *email: baba@
Layers of cells on glass fiber sheets were assembled in following succession: fibroblasts (NHDFc), red fluorescent protein (RFP) expressing Human Aortic Smooth Muscle Cells, and green fluorescent protein (GFP) expressing Human Umbilical Vein Endothelial Cells (HUVECs)
Summary
Tissue engineered vascular grafts (TEVG) are potentially clear from ethical and epidemiological concerns sources for reconstructive surgery for small diameter blood vessels replacement. Concluding: a three-layered tissue structure similar to physiological can be assembled by seeding different cell types in succession; the following training of the formed tissue with increasing flow in a bioreactor is effective for promoting cell survival, improving pressure resistance, and cell layer formation of desired properties. 3D bio-printing could make layer structure in principle, since gravity applied in the height direction, the stacked length is limited whether a bio-printer handles individual cells or spheroids as a single dispensing u nit[14]. We developed a novel bioreactor and housing devices which allowed to form the cell layers, to mature the layer structure, and to train the assembled vascular tissues with various perfusion flows. We examined mechanical and biological features of the constructed tubular tissue and the relationship between the properties of the composed layers and the training flow rate inside the bioreactor
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