A Net Mold-Based Method of Biomaterial-Free Three-Dimensional Cardiac Tissue Creation

Abstract

Ischemic cardiomyopathy poses a significant public health burden due to the irreversible loss of functional cardiac tissue. Alternative treatment strategies include creation of three-dimensional (3D) cardiac tissues to both replace and augment injured native tissue. In this study, we utilize a net mold-based method to create a biomaterial-free 3D cardiac tissue and compare it to current methods using biomaterials. Cardiomyocytes, fibroblasts, and endothelial cells were combined using a hanging drop method to create spheroids. For the net mold patch method, spheroids were seeded into a net mold-based system to create biomaterial-free 3D cardiac patches. For the gel patch, spheroids were embedded in a collagen gel. Immunohistochemistry revealed increased alignment, vascularization, collagen I expression, cell viability, and higher density of cells in the net mold patch compared with the gel patch. Furthermore, in vivo testing in a left anterior descending artery ligation rat model found increased ejection fraction and smaller scar area following implantation of the net mold patch. We present a novel and simple reproducible method to create biomaterial-free 3D net mold patches that may potentially improve the treatment of heart failure in the future.

Applications of 3D Bioprinted-Induced Pluripotent Stem Cells in Healthcare

“…Bioengineers and surgeons are looking for novel methods to synthesize artificial skin substitutes that is readily available and easily implantable in burn injury patients[ 85 , 86 ]. Scaffold-free cellular spheroids obtained from a coculture of human iPSC-derived cardiomyocytes, fibroblasts, and endothelial cells were 3D printed and these cardiac cellular patches were tested successfully in rat models of myocardial infarction[ 87 ]. Bioprinted organ substitutes such as pancreas, ovary, liver, kidney, and nervous tissues also will be in high demand in the near future.…”

Extracellular Vesicles in Cardiac Regeneration: Potential Applications for Tissues-on-a-Chip

“…Supplementation with neurohormonal factors, such as angiotensin II or catecholamines, has been used to simulate the overstimulation of cardiomyocytes contributing to the progression to heart failure [51,103] and transforming growth factor beta (TGF-β) supplementation has been used to activate fibroblasts to model the development of cardiac fibrosis [104]. Perhaps most importantly for this review, groups have introduced conditioned media to their platforms and discovered that EVs in these media impact the function of engineered tissues [66,[104][105][106][107][108].…”

Devices and techniques used to obtain and analyze three‐dimensional cell cultures

“…g 3D bio-printing is the latest modality to be developed for the purpose of 3D cell culture where entire tissues can be generated by customizing the "bio-inks" used in the process of assembly assays and the handling may result in damage to the spheroids that potentially affects and data that may be acquired. 17,18 Low amounts of starting cell numbers and medium are the advantages of this technique enabling one to control the size of the spheroid seed cultures, as evidenced by the work of Bai et al, wherein the size of the spheroids was optimized by varying cell suspension concentrations. The cultivated spheroids can then be seeded into a biomaterial free collagen net-mold patch for the purpose of cardiac tissue engineering.…”