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Self-standing in vitro multi-layered blood vessel model reproducing immunological tissue deformation (本文)

板井, 駿 慶應義塾大学

2022.03.23

概要

The biochemical reaction of blood vessels plays an essential role in immune response and various diseases. Thus, the construction of in vitro blood vessel models mimicking in vivo tissues has a high demand for biomedical research and pharmacokinetic testing. However, previously reported models were difficult to mimic the biomedical deformation reaction of blood vessels faithfully because the tissues were fixed to rigid scaffolds. The insufficient flexibility of the scaffold disturbs the contraction of blood vessels and leads to the deviation of the behaviors with in vivo tissues.

Here, the author proposes a flexibly deformable self-standing multi-layered blood vessel model to reproduce an inflammatory mediator-induced deformation reaction. A self- standing collagen tube enables the tissue to deform flexibly in biochemical reactions and achieves contraction and relaxation at tissue and cell levels. Also, by constructing a circumferentially aligned smooth muscle layer outside the endothelial layer, the immunological deformation reaction on both endothelial and smooth muscle layers is reproduced.

Chapter 1 introduces the background, the purpose, and the concept of this research.

Chapter 2 describes the fabrication of a double-layered collagen hydrogel tube device.
First, a simple repeated molding process precisely and easily fabricates the double-layered collagen tube device. After forming an endothelial layer inside the tube, the tissue formation is confirmed by evaluating the barrier function of the endothelium. Also, the cellular response of the endothelial cells is confirmed by co-culturing with vascular pericytes.

Chapter 3 reproduces the immunological deformation reaction of the endothelium layer with the tube device forming an endothelial layer inside. Exposure to histamine (an inflammatory mediator of allergy) confirms contraction and recovery reactions at both tissue and cell levels. Also, the ability of the device for drug testing is evaluated by exposing olopatadine hydrochloride (an anti-histamine material) and histamine. In addition, the traction force of the endothelial cells under reaction is calculated by numerical analysis with the finite element method.

Chapter 4 proposes a blood vessel model with a circumferentially aligned smooth muscle layer. By rolling up an axially aligned smooth muscle fiber, a circumferential alignment is easily formed in a collagen tube (Rolling up maturation molding (RuM-M) method). The alignment is evaluated by live imaging of actin. An immunological deformation reaction of the co-cultured endothelium and smooth muscle is reproduced with co-cultured tissue under exposure to histamine.

Chapter 5 concludes this dissertation by discussing the value of the proposed self- standing in vitro multi-layered blood vessel model and the prospective.

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