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

To eliminate the limitation of the conventional acoustic radiation torque theory, which is only applicable to a disklike scatterer in a plane sound field, a new theory is established to calculate the radiation torque on any irregularly shaped scatterer in any arbitrary sound field. First, with the aid of the conservation law of angular momentum, the acoustic radiation torque is expressed as the angular momentum flux through a spherical surface with the center at the scatterer's centroid. Second, the velocity potential of the scattered field is derived, taking into account the influences of the translational and rotational movements of the scatterer induced by the first order stress of the incident sound field. Finally, a general calculating formula of the acoustic radiation torque is achieved. For a disklike scatterer in a plane sound filed, results from the above formula are well identical with those conventional formulas. By studying the case of a semicircular cylinder scatterer in a standing-wave sound field, it is found that for an irregularly shaped scatterer its rotation velocity is normally nonzero and the radiation torque changes with the spatial attitude.

Acoustic radiation torque on an irregularly shaped scatterer in an arbitrary sound field.

To eliminate the limitations of the conventional sound field separation methods which are only applicable to regular surfaces, a sound field separation method based on combined integral equations is proposed to separate sound fields directly in the spatial domain. In virtue of the Helmholtz integral equations for the incident and scattering fields outside a sound scatterer, combined integral equations are derived for sound field separation, which build the quantitative relationship between the sound fields on two arbitrary separation surfaces enclosing the sound scatterer. Through boundary element discretization of the two surfaces, corresponding systems of linear equations are obtained for practical application. Numerical simulations are performed for sound field separation on different shaped surfaces. The influences induced by the aspect ratio of the separation surfaces and the signal noise in the measurement data are also investigated. The separated incident and scattering sound fields agree well with the original corresponding fields described by analytical expressions, which validates the effectiveness and accuracy of the combined integral equations based separation method. 

Sound field separating on arbitrary surfaces enclosing a sound scatterer based on combined integral equations.

Now, more and more patients are suffering cerebral aneurysm. However, long training time limits the rapid growth of cerebrovascular neurosurgeons. Here we developed a novel cerebral aneurysm simulator which can be better represented the dynamic bulging process of cerebral aneurysm The proposed simulator features the integration of a hollow elastic vascular model, a skull model and a brain model, which can be affordably fabricated at the clinic (Fab@Clinic), under $25.00 each with the help of a low-cost desktop 3D printer. Moreover, the clinical blood flow and pulsation pressure similar to the human can be well simulated, which can be used to train the neurosurgical residents how to clip aneurysms more effectively.

Fabrication of cerebral aneurysm simulator with a desktop 3D printer.

The precision spindle is a core component of high-precision machine tools, and the accurate measurement of its error motions is important for improving its rotation accuracy as well as the work performance of the machine. This paper presents a T-type capacitive sensor (T-type CS) with an integrated structure. The proposed sensor can measure the 5-degree-of-freedom (5-DOF) error motions of a spindle in-situ and simultaneously by integrating electrode groups in the cylindrical bore of the stator and the outer end face of its flange, respectively. Simulation analysis and experimental results show that the sensing electrode groups with differential measurement configuration have near-linear output for the different types of rotor displacements. What's more, the additional capacitance generated by fringe effects has been reduced about 90% with the sensing electrode groups fabricated based on flexible printed circuit board (FPCB) and related processing technologies. The improved signal processing circuit has also been increased one times in the measuring performance and makes the measured differential output capacitance up to 93% of the theoretical values.

A T-Type Capacitive Sensor Capable of Measuring5-DOF Error Motions of Precision Spindles.

As promising carbonaceous films, graphene films (GF) have exhibited many remarkable mechanical and electrical/thermal properties of great potential for wide functional applications. However, the electrical conductivity of GF still needs to be improved and the limitation lies in the low carrier density of pure graphene. Here, we presented a post-doping method for large-area potassium doped graphene films (GF-K) and promoted the electrical conductivity of GF approaching benchmark metals. The macroscopic-assembled GF-K shows a similar color to graphite intercalation compounds. The potassium doping increased the carrier density of the GF without undermining the electronic quality of the graphene unit. The doping concentration was optimized to prepare stage-2 GF-K (CK) with the highest electrical conductivity (1.49 × 10 S m), holding merits of low density (1.63 g cm), and high flexibility. Doped GF with better specific electrical conductivity than copper showed outstanding electromagnetic interference shielding performance. Shielding effectiveness (SE) increased from 70-85 dB for graphene film (GF) to over 130 dB for GF-K only at 31 μm thickness, which is among the best SEs in previous reports. The combination of high specific conductivity, mechanical flexibility, high EMI SE, light weight, and facile productivity enables GF-K to be promising in many high-end EMI applications such as aerospace and wearable devices.

Large-area potassium-doped highly conductive graphene films for electromagnetic interference shielding.

Graphene has shown great potential in microwave absorption (MA) owing to its high surface area, low density, tunable electrical conductivity and good chemical stability. To fully realize graphene's MA ability, the microstructure of graphene should be carefully addressed. Here we prepared graphene microflowers (Gmfs) with highly porous structure for high-performance MA filler material. The efficient absorption bandwidth (reflection loss ≤ -10 dB) reaches 5.59 GHz and the minimum reflection loss is up to -42.9 dB, showing significant increment compared with stacked graphene. Such performance is higher than most graphene-based materials in the literature. Besides, the low filling content (10 wt%) and low density (40-50 mg cm) are beneficial for the practical applications. Without compounding with magnetic materials or conductive polymers, Gmfs show outstanding MA performance with the aid of rational microstructure design. Furthermore, Gmfs exhibit advantages in facile processibility and large-scale production compared with other porous graphene materials including aerogels and foams.

Porous Graphene Microflowers for High-Performance Microwave Absorption.

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.

Photo-crosslinkable gelatin methacrylate (GelMA) has become an attractive ink in 3D printing due to its excellent biological performance. However, limited by low viscosity and long cross-linking time, it is still a challenge to directly print GelMA by extrusion-based 3D printing. Here, to balance the printability and biocompatibility, biomaterial ink composed of GelMA and nanoclay was specially designed. Using this ink, complex scaffolds with high shape fidelity can be easily printed based on the thixotropic property of nanoclay. In this study, we tried to answer some basic printing-required questions of this ink, including the printability window, general properties (porosity, mechanical strength, et al), and biocompatibility. We found that the GelMA/Nanoclay ink enabled printing complex 3D scaffolds, such as a bionic ear and a branched vessel. Furthermore, the addition of nanoclay improved the porosity, increased the mechanical strength, reduced the degradation ratio, and maintained a good biocompatibility of the printed scaffolds. Therefore, this method offers an easy way to print complex scaffolds with good shape fidelity and biological performance, and it might open up new potential applications for the customized therapy of tissue defects.

3D printing of complex GelMA-based scaffolds with nanoclay.

Hierarchical tissues composed of spheroid and fiber structures such as tumors, embryos and glomeruli widely exist in organisms. Methods have been developed to build spheroid and fiber structures, independently, as tissue models in vitro. However, it is still a challenge to print them simultaneously and integrated for effectively mimicking the complicated situations in vivo. Here, we propose a novel 3D cell co-culturing system, "microfiber-laden minispheroid", applying two fluidic phenomena, namely the "rope coiling effect" and "electrohydrodynamics", with a co-axial bioprinting nozzle and high voltage system. Gelatin methacryloyl (GelMA) hydrogels were extruded from the outer nozzle and separated by electrical attraction to form spheroids (∼1800 μm). GelMA mixed with sodium alginate was extruded from the inner nozzle to form fibers (∼180 μm) inside spheroids, whose morphology could be controlled by the ratio of inner and outer nozzle extruding flow rates. We analyzed the fabrication process and the material system in detail, verifying the fabrication feasibility and suitable microenvironment. The encapsulated cells possessed high viabilities. Importantly, the actin of human umbilical vein endothelial cells tended to elongate towards the co-cultured tumor cells, in contrast to the HUVECs cultured alone. We believe that "microfiber-laden minispheroids" could be a potential system for 3D cell co-culturing research in the future.

Zichen Chen

Zhejiang University

Protocols of 3D Bioprinting of Gelatin Methacryloyl Hydrogel Based Bioinks