Biofabrication of muscle fibers enhanced with plant viral nanoparticles using surface chaotic flows

Abstract

<jats:title>Abstract</jats:title><jats:p>Multiple human tissues exhibit fibrous nature. Therefore, the fabrication of hydrogel filaments for tissue engineering is a trending topic. Current tissue models are made of materials that often require further enhancement for appropriate cell attachment, proliferation and differentiation. Here we present a simple strategy, based on the use surface chaotic flows amenable of mathematical modeling, to fabricate continuous, long and thin filaments of gelatin methacryloyl (GelMA).</jats:p><jats:p>The fabrication of these filaments is achieved by chaotic advection in a finely controlled and miniaturized version of the journal bearing (JB) system. A drop of GelMA pregel was injected on a higher-density viscous fluid (glycerin) and a chaotic flow is applied through an iterative process. The hydrogel drop is exponentially deformed and elongated to generate a fiber, which was then polymerized under UV-light exposure. Computational fluid dynamic (CFD) simulations are conducted to determine the characteristics of the flow and design the experimental conditions for fabrication of the fibers. GelMA fibers were effectively used as scaffolds for C2C12 myoblast cells. Experimental results demonstrate an accurate accordance with CFD simulations for the predicted length of the fibers.</jats:p><jats:p>Plant-based viral nanoparticles (i.e., <jats:italic>Turnip mosaic virus</jats:italic>; TuMV) were then integrated to the hydrogel fibers as a secondary nano-scaffold for cells for enhanced muscle tissue engineering. The addition of TuMV significantly increased the metabolic activity of the cell-seeded fibers (p*&lt;0.05), strengthened cell attachment throughout the first 28 days, improved cell alignment, and promoted the generation of structures that resemble natural mammal muscle tissues.</jats:p><jats:p>Chaotic 2D-printing is proven to be a viable method for the fabrication of hydrogel fibers. The combined use of thin and long GelMA hydrogel fibers enhanced with flexuous virions offers a promising alternative for scaffolding of muscle cells and show potential to be used as cost-effective models for muscle tissue engineering purposes.</jats:p>

Bioinks for 3D Bioprinting: A Scientometric Analysis of Two Decades of Progress

“…However, the technological basis to develop skin capable of producing hair de novo is at hand[ 64 ]. Furthermore, recent reports have shown the feasibility of mimicking this multi-scale structure and alignment of muscle-like fibers using emerging bioprinting techniques[ 12 , 65 - 67 ].…”

“…New ingredients have been recently added to the repertoire of bioink additives to provide relevant functionalities for hydrogel-based inks. Examples are the use of a flexuous filamentous plant virus[ 65 ] to enhance cell attachment and proliferation in the context of fabrication of muscle fibers; the incorporation of protease-degradable cross-linkers to enable cell remodeling[ 49 ] and oxygen-releasing agents to improve and prolong tissue viability[ 246 ]…”

A skeleton muscle model using GelMA-based cell-aligned bioink processed with an electric-field assisted 3D/4D bioprinting

“…Others have attempted to fabricate single fibers of GelMA with aligned surface structure for muscle tissue regeneration. Frías-Sánchez et al developed a chaotic flow system to generate fibers by elongation of GelMA drop 10 . Fibers from sizes ranging from 20 to 400 µm in diameter could be obtained.…”

Engineering bioactive synthetic polymers for biomedical applications: a review with emphasis on tissue engineering and controlled release

“…In recent years, researchers have extensively utilized 3D printing for the biofabrication of tissue engineering scaffolds, as 3D printing can fabricate complex scaffolds at the micro-and macro-scale in an easy, low-cost process. 84,[151][152][153][154][155][156][157][158] To date, 3D printing represents a major scaffold production technology, and an enormous number of papers now describe constructs achieved with different SyPs and their composites. In silico designed shapes, sizes, spatial dependence, and pore topologies that are impossible to produce in alternative ways can be extremely easily produced by AM, virtually without any limitation.…”

“…That is the case of microfluidics 310 or 3D bioprinting which are just gaining momentum. New 3D printing strategies such as chaotic printing process, [151][152][153][154]157,311 the use of chaotic flows instead of layer by layer deposition to produced multilayered microstructures, may be a powerful enabler in the near future for the engineering of SyP-based scaffolds for tissue engineering.…”