protocols-3d-bioprinting-gelatin-methacryloyl-hydrogel-based

Protocols of 3D Bioprinting of Gelatin Methacryloyl Hydrogel Based Bioinks

Gelatin methacryloyl (GelMA) has become a popular biomaterial in the field of bioprinting. The derivation of this material is gelatin, which is hydrolyzed ...

Zhejiang University

Hydrogel microspheroids are widely used in tissue engineering, such as injection therapy and 3D cell culture, and among which, heterogeneous microspheroids are drawing much attention as a promising tool to carry multiple cell types in separated phases. However, it is still a big challenge to fabricate heterogeneous microspheroids that can reconstruct built-up tissues' microarchitecture with excellent resolution and spatial organization in limited sizes. Here, a novel airflow-assisted 3D bioprinting method is reported, which can print versatile spiral microarchitectures inside the microspheroids, permitting one-step bioprinting of fascinating hydrogel structures, such as the spherical helix, rose, and saddle. A microfluidic nozzle is developed to improve the capability of intricate cell encapsulation with heterotypic contact. Complex structures, such as a rose, Tai chi pattern, and single cell line can be easily printed in spheroids. The theoretical model during printing is established and process parameters are systematically investigated. As a demonstration, a human multicellular organoid of spirally vascularized ossification is reconstructed with this method, which shows that it is a powerful tool to build mini tissues on microspheroids.

Airflow-Assisted 3D Bioprinting of Human Heterogeneous Microspheroidal Organoids with Microfluidic Nozzle.

Low-concentration gelatin methacryloyl (GelMA) has excellent biocompatibility to cell-laden structures. However, it is still a big challenge to stably fabricate organoids (even microdroplets) using this material due to its extremely low viscosity. Here, a promising electro-assisted bioprinting method is developed, which can print low-concentration pure GelMA microdroplets with low cost, low cell damage, and high efficiency. With the help of electrostatic attraction, uniform GelMA microdroplets measuring about 100 μm are rapidly printed. Due to the application of lower external forces to separate the droplets, cell damage during printing is negligible, which often happens in piezoelectric or thermal inkjet bioprinting. Different printing states and effects of printing parameters (voltages, gas pressure, nozzle size, etc.) on microdroplet diameter are also investigated. The fundamental properties of low-concentration GelMA microspheres are subsequently studied. The results show that the printed microspheres with 5% w/v GelMA can provide a suitable microenvironment for laden bone marrow stem cells. Finally, it is demonstrated that the printed microdroplets can be used in building microspheroidal organoids, in drug controlled release, and in 3D bioprinting as biobricks. This method shows great potential use in cell therapy, drug delivery, and organoid building.

Electro-Assisted Bioprinting of Low-Concentration GelMA Microdroplets.

It is a dilemma that both strength and biocompatibility are requirements for an ideal scaffold in tissue engineering. The normal strategy is mixing or coating another material to improve the biocompatibility. Could we solve this dilemma by simply adjusting the scaffold structure? Here, a novel multi-scale scaffold was designed, in which thick fibers provide sufficient strength for mechanical support while the thin fibers provide a cell-favorable microenvironment to facilitate cell adhesion. Moreover, we developed a promising multi-scale direct writing system (MSDWS) for printing the multi-scale scaffolds. By switching the electrostatic field, scaffolds with fiber diameters from 3 μm to 600 μm were fabricated using one nozzle. Using this method, we proved that PCL scaffolds could also have excellent biocompatibility. BMSCs seeded on the scaffolds readily adhered to the thin fibers and maintained a high proliferation rate. Moreover, the cells bridged across the pores to form a cell sheet and gradually migrated to the thick fibers to cover the entire scaffold. We further combined the scaffolds with hydrogel for 3D cell culture and found that the fibers enhanced the strength and induced cell migration. We believe that the multi-scale scaffolds fabricated by an innovative 3D printing system have great potential for tissue engineering.

Article	Year
Airflow-Assisted 3D Bioprinting of Human Heterogeneous Microspheroidal Organoids with Microfluidic Nozzle. Small (Weinheim an der Bergstrasse, Germany)\| PubMed ID: 30133151	2018
Electro-Assisted Bioprinting of Low-Concentration GelMA Microdroplets. Small (Weinheim an der Bergstrasse, Germany)\| PubMed ID: 30569632	2019
3D printed multi-scale scaffolds with ultrafine fibers for providing excellent biocompatibility. Materials science & engineering. C, Materials for biological applications\| PubMed ID: 31761213	2020

Mingjun Xie

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