Construction and histological analysis of a 3D human arterial wall model containing vasa vasorum using a layer‐by‐layer technique

Abstract

There is considerable global demand for three-dimensional (3D) functional tissues which mimic our native organs and tissues for use as in vitro drug screening systems and in regenerative medicine. In particular, there has been an increasing number of patients who suffer from arterial diseases such as arteriosclerosis. As such, in vitro 3D arterial wall models that can evaluate the effects of novel medicines and a novel artificial graft for the treatment are required. In our previous study, we reported the rapid construction of 3D tissues by employing a layer-by-layer (LbL) technique and revealed their potential applications in the pharmaceutical fields and tissue engineering. In this study, we successfully constructed a 3D arterial wall model containing vasa vasorum by employing a LbL technique for the first time. The cells were coated with extracellular matrix nanofilms and seeded into a culture insert using a cell accumulation method. This model had a three-layered hierarchical structure: a fibroblast layer, a smooth muscle layer, and an endothelial layer, which resembled the native arterial wall. Our method could introduce vasa vasorum into a fibroblast layer in vitro and the 3D arterial wall model showed barrier function which was evaluated by immunostaining and transendothelial electrical resistance measurement. Furthermore, electron microscopy observations revealed that the vasa vasorum was composed of single-layered endothelial cells, and the endothelial tubes were surrounded by the basal lamina, which are known to promote maturation and stabilization in native blood capillaries. These models should be useful for tissue engineering, regenerative medicine, and pharmaceutical applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 814-823, 2017.

Engraftment and morphological development of vascularized human iPS cell-derived 3D-cardiomyocyte tissue after xenotransplantation

“…Previous reports were consulted for ultrastructural analyses using transmission electron microscopy (TEM) 41 . Briefly, iPSC-CM 3D-tissues in in vitro and resected rat hearts were fixed with 2.5% glutaraldehyde and 1% PFA in 0.1 M phosphate buffer (PB) containing 0.01% CaCl 2 and MgCl 2 , and cut into small tissue pieces.…”

Advanced Bottom‐Up Engineering of Living Architectures

“…Nevertheless, physical stress can be avoided by resorting to filtration‐assisted LbL technology which assures high viability and efficiency to the FN/G coated cells. The filtration‐assisted LbL methodology has been employed to engineer 3D human tissue constructs, including vascularized cardiac microtissues, liver, and blood vessels . Using a different approach, researchers also demonstrated that the LbL assembly technology can be combined with automatic inkjet printing of single cells and ECM proteins to precisely develop 3D human microtissue chips in a rapid and automatic mode .…”

Hylozoic by Design: Converging Material and Biological Complexities for Cell‐Driven Living Materials with 4D Behaviors

“…[ 135 ] In this fashion, living material constructs are created in which biomaterials initiate composite assembling, after which, cells and cellular assemblies can partake in biological complexities. In order to guide such organization, biomaterials have been employed in the form of microgels, [ 136 ] microparticles, [ 19b,71b ] nanoparticles, [ 71c,137 ] and multilayer film architectures. [ 71a,138 ] A relevant challenge among these systems is the relative difficulty found in securely regulating the assembly process so as to achieve specific scales of assembly or assembly shapes.…”

“…[ 18b,66a,b,70 ] In addition to directly assembling composite constructs, the added material may itself play a role in driving composite assembly. For example, cell surfaces may be functionalized with thin material layers or objects which promote aggregation, [ 71 ] and cells themselves may be directly attached to larger material building blocks that confer cues of assembly or aggregation. [ 72 ] Composites created using these techniques have been implemented in areas such as biosensing, [ 73 ] water‐treatment, [ 74 ] bioremediation, [ 2b,75 ] drug delivery, [ 76 ] energy, [ 77 ] bioproduction, [ 78 ] “smart packaging,” [ 79 ] tissue engineering, [ 80 ] immune engineering, [ 81 ] and cell encapsulation.…”

“…In order to guide such organization, biomaterials have been employed in the form of microgels, [ 136 ] microparticles, [ 19b,71b ] nanoparticles, [ 71c,137 ] and multilayer film architectures. [ 71a,138 ] A relevant challenge among these systems is the relative difficulty found in securely regulating the assembly process so as to achieve specific scales of assembly or assembly shapes. To strengthen their dynamic capacities and controllability, stimulus‐responsive mechanisms may also be incorporated, allowing for notable spatiotemporal control.…”