We report about rationally designed ultrashort peptide bioinks, overcoming severe limitations in current bioprinting procedures. Bioprinting is increasingly relevant in tissue engineering, regenerative and personalized medicine due to its ability to fabricate complex tissue scaffolds through an automated deposition process. Printing stable large-scale constructs with high shape fidelity and enabling long-term cell survival are major challenges that most existing bioinks are unable to solve. Additionally, they require chemical or UV-cross-linking for the structure-solidifying process which compromises the encapsulated cells, resulting in restricted structure complexity and low cell viability. Using ultrashort peptide bioinks as ideal bodylike but synthetic material, we demonstrate an instant solidifying cellembedding printing process via a sophisticated extrusion procedure under true physiological conditions and at cost-effective low bioink concentrations. Our printed large-scale cell constructs and the chondrogenic differentiation of printed mesenchymal stem cells point to the strong potential of the peptide bioinks for automated complex tissue fabrication.
Tetrameric peptide-based bioinks allow the printing of 3D cell-laden scaffolds under true physiological conditions avoiding harsh UV or chemical treatment.
Injured skeletal muscles which lose more than 20% of their volume, known as volumetric muscle loss, can no longer regenerate cells through self-healing. The traditional solution for recovery is through regenerative therapy. As the technology of three-dimensional (3D) bioprinting continues to advance, a new approach for tissue transplantation is using biocompatible materials arranged in 3D scaffolds for muscle repair. Ultrashort self-assembling peptide hydrogels compete as a potential biomaterial for muscle tissue formation due to their biocompatibility. In this study, two sequences of ultrashort peptides were analyzed with muscle myoblast cells (C2C12) for cell viability, cell proliferation, and differentiation in 3D cell culture. The peptides were then extruded through a custom-designed robotic 3D bioprinter to create cell-laden 3D structures. These constructs were also analyzed for cell viability through live/dead assay. Results showed that 3D bioprinted structures of peptide hydrogels could be used as tissue platforms for myotube formation – a process necessary for muscle repair.
The field of three-dimensional (3D) bioprinting is rapidly emerging as an additive manufacturing method for tissue and organ fabrication. The demand for tissues and organ transplants is ever increasing, although donors are not as readily available. Consequently, tissue engineering is gaining much attention to alleviate this problem. The process of achieving well-structured 3D bioprinted constructs using hydrogel bioinks depends on symmetrical precision, regulated flow rates, and viability of cells. Even with the mentioned parameters optimized, the printed structures need additional refining by removing excessive liquids, as peptide hydrogel bioprints encapsulate water. However, it is challenging to eliminate the confined fluids without compromising the printing process. In this paper, we introduced a vacuum system to our 3D bioprinting robotic arm and thus optimized the printing quality for complex and refined 3D scaffolds. Moreover, the proposed vacuum system supports printing with cells. Our results show improved printing resolution which facilitates the printing of higher and more stable structures.
The technology of 3D bioprinting has gained significant interest in biomedical engineering, regenerative medicine, and the pharmaceutical industry. Providing a new scope in tissue and organ printing, 3D bioprinters are becoming commercialized for biological processes. However, the current technology is costly, ranging from USD$9,000-$30,000 and is limited to customized extrusion methods. Multiple microfluidic pump systems for bioink extrusion are commercially available at USD$30,000. Additionally, the use of Cartesian systems for 3D printing restricts the user to three axes of movement and makes multi-material modeling a challenge. Consequently, it was proposed to design a cost effective robotic 3D bioprinting system, compatible with peptide bioinks which were developed at KAUST Laboratory for Nanomedicine. The components of the system included a programmable robotic arm, an extruder for bioprinting, and multiple microfluidic pumps. The extruder was designed using a coaxial nozzle made of three inlets and one outlet. The programmable microfluidic pumps transported the peptide bioink, phosphate buffer saline (PBS) and human skin fibroblast cells (in cell culture media solution) through the nozzle to extrude a peptide nanogel thread. Model cell structures were printed and monitored for a period of two weeks and subsequently found to be alive and healthy. The system was kept well under a budget of USD$3,500. Future modifications of the current system will include adding a custom bioprinting arm to allow multi-material printing which can fully integrate and synchronize between the pumps and the robotic arm. This system will allow the production of a more advanced robotic arm-based 3D bioprinting system in the future.
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