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A Mixing System for Uniform, Reproducible Viscous Bioinks Preparation
Ryan Singer1,2, David A González-Martínez3, Aidee Verónica Arizpe Tafoya2,3
1Firestone Institute for Respiratory Health─Division of Respirology, Department of Medicine, McMaster University, Hamilton, Ontario L8N 4A6, Canada.
ACS Biomaterials Science & Engineering
|November 14, 2025
Summary
This study introduces a novel static mixer system for preparing cell-laden bioinks used in extrusion 3D bioprinting. The optimized SMX-style mixer ensures high cell viability and uniform distribution for consistent tissue engineering.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Additive Manufacturing
Background:
- Extrusion 3D bioprinting relies on bioinks, which are challenging to prepare due to high viscosity.
- Manual mixing of cells into viscous biomaterials can be inconsistent and detrimental to cell viability.
- Static mixers offer a potential solution for reproducible bioink preparation, but their efficacy for viscous applications needs thorough investigation.
Purpose of the Study:
- To evaluate and compare the performance of four different static mixers for preparing cell-laden bioinks.
- To assess mixing uniformity, shear stress, and cell viability across various static mixer designs.
- To demonstrate the utility of the optimized mixing system in extrusion 3D bioprinting for tissue construct fabrication.
Main Methods:
- Three novel static mixers (SMX-style, Ross ISG, serpentine) were 3D printed and compared with a commercial CELLINK mixer.
- Biomaterial inks (PEGDA, methacrylated gelatin) were characterized using rheology.
- Fluid dynamics simulations estimated shear stress, while fluorescent beads assessed mixing effectiveness.
- Cell viability assays were performed on A549, NIH-3T3, and primary human lung fibroblasts post-mixing.
Main Results:
- The SMX-style mixer demonstrated superior mixing uniformity and the lowest simulated shear stress compared to other designs.
- Cell viability remained above 96% for all tested cell types when using the SMX-style mixer.
- The SMX-style mixer achieved a more homogeneous cell distribution than the commercial comparator.
- Successful extrusion 3D bioprinting of viable tissue constructs with evenly distributed cells was achieved.
Conclusions:
- A simple, reproducible, and flexible static mixing system was developed for incorporating cells into viscous bioinks.
- This system enhances consistency in cell distribution and viability during bioink preparation.
- The optimized static mixer facilitates standardized fabrication of cell-laden constructs for extrusion 3D bioprinting applications.
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