scholarly journals Homogeneous and Reproducible Mixing of Highly Viscous Biomaterial Inks and Cell Suspensions to Create Bioinks

Gels ◽  
2021 ◽  
Vol 7 (4) ◽  
pp. 227
Author(s):  
Sophie Dani ◽  
Tilman Ahlfeld ◽  
Franziska Albrecht ◽  
Sarah Duin ◽  
Petra Kluger ◽  
...  
Keyword(s):  
Cell Suspension ◽  
Three Dimensional ◽  
Human Mesenchymal Stem Cells ◽  
High Viscosity ◽  
Model System ◽  
Human Blood Plasma ◽  
Viable Cells ◽  
Fresh Frozen ◽  
A Cell ◽  
Novel Method

Highly viscous bioinks offer great advantages for the three-dimensional fabrication of cell-laden constructs by microextrusion printing. However, no standardised method of mixing a high viscosity biomaterial ink and a cell suspension has been established so far, leading to non-reproducible printing results. A novel method for the homogeneous and reproducible mixing of the two components using a mixing unit connecting two syringes is developed and investigated. Several static mixing units, based on established mixing designs, were adapted and their functionality was determined by analysing specific features of the resulting bioink. As a model system, we selected a highly viscous ink consisting of fresh frozen human blood plasma, alginate, and methylcellulose, and a cell suspension containing immortalized human mesenchymal stem cells. This bioink is crosslinked after fabrication. A pre-crosslinked gellan gum-based bioink providing a different extrusion behaviour was introduced to validate the conclusions drawn from the model system. For characterisation, bioink from different zones within the mixing device was analysed by measurement of its viscosity, shape fidelity after printing and visual homogeneity. When taking all three parameters into account, a comprehensive and reliable comparison of the mixing quality was possible. In comparison to the established method of manual mixing inside a beaker using a spatula, a significantly higher proportion of viable cells was detected directly after mixing and plotting for both bioinks when the mixing unit was used. A screw-like mixing unit, termed “HighVisc”, was found to result in a homogenous bioink after a low number of mixing cycles while achieving high cell viability rates.

Micromachined Ultrasonic Print-Head for Deposition of High-Viscosity Materials

2010 ◽  
Vol 132 (3) ◽  
Author(s):  
J. Mark Meacham ◽  
Amanda O’Rourke ◽  
Yong Yang ◽  
Andrei G. Fedorov ◽  
F. Levent Degertekin ◽  
...  
Keyword(s):  
Additive Manufacturing ◽  
Three Dimensional ◽  
High Viscosity ◽  
The Novel ◽  
Glycerol Water ◽  
Print Head ◽  
Cavity Resonances ◽  
Allowable Range ◽  
Novel Method ◽  
Manufacturing Techniques

The recent application of inkjet printing to fabrication of three-dimensional, multilayer and multimaterial parts has tested the limits of conventional printing-based additive manufacturing techniques. The novel method presented here, termed as additive manufacturing via microarray deposition (AMMD), expands the allowable range of physical properties of printed fluids to include important, high-viscosity production materials (e.g., polyurethane resins). AMMD relies on a piezoelectrically driven ultrasonic print-head that generates continuous streams of droplets from 45 μm orifices while operating in the 0.5–3.0 MHz frequency range. The device is composed of a bulk ceramic piezoelectric transducer for ultrasound generation, a reservoir for the material to be printed, and a silicon micromachined array of liquid horn structures, which make up the ejection nozzles. Unique to this new printing technique are the high frequency of operation, use of fluid cavity resonances to assist ejection, and acoustic wave focusing to generate the pressure gradient required to form and eject droplets. We present the initial characterization of a micromachined print-head for deposition of fluids that cannot be used with conventional printing-based rapid prototyping techniques. Glycerol-water mixtures with a range of properties (surface tensions of ∼58–73 mN/m and viscosities of 0.7–380 mN s/m2) were used as representative printing fluids for most investigations. Sustained ejection was observed in all cases. In addition, successful ejection of a urethane-based photopolymer resin (surface tension of ∼25–30 mN/m and viscosity of 900–3000 mN s/m2) was achieved in short duration bursts. Peaks in the ejection quality were found to correspond to predicted device resonances. Based on these results, we have demonstrated the printing of fluids that fall well outside of the accepted range for the previously introduced printing indicator. The micromachined ultrasonic print-head achieves sustained printing of fluids up to 380 mN s/m2, far above the typical printable range.

Three-dimensional distribution of ribulose-1, 5-bisphosphate carboxylase/oxygemase during the early development of proplastids in dark-grown Euglena cells transferred to an inorganic medium

1990 ◽  
Vol 48 (3) ◽  
pp. 906-907
Author(s):  
Tomoko Ehara ◽  
Shuji Sumida ◽  
Tetsuaki Osafune ◽  
Eiji Hase
Keyword(s):  
Cell Suspension ◽  
Euglena Gracilis ◽  
Carbon Materials ◽  
Three Dimensional ◽  
Peripheral Region ◽  
Plastid Development ◽  
Bisphosphate Carboxylase ◽  
Inorganic Medium ◽  
Ribulose 1 ◽  
Dimensional Distribution

As shown previously, Euglena cells grown in Hutner’s medium in the dark without agitation accumulate wax as well as paramylum, and contain proplastids showing no internal structure except for a single prothylakoid existing close to the envelope. When the cells are transferred to an inorganic medium containing ammonium salt and the cell suspension is aerated in the dark, the wax was oxidatively metabolized, providing carbon materials and energy 23 for some dark processes of plastid development. Under these conditions, pyrenoid-like structures (called “pro-pyrenoids”) are formed at the sites adjacent to the prolamel larbodies (PLB) localized in the peripheral region of the proplastid. The single prothylakoid becomes paired with a newly formed prothylakoid, and a part of the paired prothylakoids is extended, with foldings, in to the “propyrenoid”. In this study, we observed a concentration of RuBisCO in the “propyrenoid” of Euglena gracilis strain Z using immunoelectron microscopy.

scholarly journals Homogeneous 2D and 3D alignment of cardiomyocyte in dilated cardiomyopathy revealed by intravital heart imaging

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kiyoshi Masuyama ◽  
Tomoaki Higo ◽  
Jong-Kook Lee ◽  
Ryohei Matsuura ◽  
Ian Jones ◽  
...  
Keyword(s):  
Heart Failure ◽  
Dilated Cardiomyopathy ◽  
Cardiac Dysfunction ◽  
Three Dimensional ◽  
Single Layer ◽  
Specific Pattern ◽  
Two Dimensional ◽  
Heart Imaging ◽  
Novel Method ◽  
2D And 3D

AbstractIn contrast to hypertrophic cardiomyopathy, there has been reported no specific pattern of cardiomyocyte array in dilated cardiomyopathy (DCM), partially because lack of alignment assessment in a three-dimensional (3D) manner. Here we have established a novel method to evaluate cardiomyocyte alignment in 3D using intravital heart imaging and demonstrated homogeneous alignment in DCM mice. Whilst cardiomyocytes of control mice changed their alignment by every layer in 3D and position twistedly even in a single layer, termed myocyte twist, cardiomyocytes of DCM mice aligned homogeneously both in two-dimensional (2D) and in 3D and lost myocyte twist. Manipulation of cultured cardiomyocyte toward homogeneously aligned increased their contractility, suggesting that homogeneous alignment in DCM mice is due to a sort of alignment remodelling as a way to compensate cardiac dysfunction. Our findings provide the first intravital evidence of cardiomyocyte alignment and will bring new insights into understanding the mechanism of heart failure.

scholarly journals 3D Printing of High Viscosity Reinforced Silicone Elastomers

Polymers ◽  
2021 ◽  
Vol 13 (14) ◽  
pp. 2239
Author(s):  
Nicholas Rodriguez ◽  
Samantha Ruelas ◽  
Jean-Baptiste Forien ◽  
Nikola Dudukovic ◽  
Josh DeOtte ◽  
...  
Keyword(s):  
Mechanical Properties ◽  
High Performance ◽  
New Technologies ◽  
Three Dimensional ◽  
High Viscosity ◽  
Layer By Layer ◽  
Silicone Elastomers ◽  
Uv Curable ◽  
Octet Truss ◽  
Tunable Mechanical Properties

Recent advances in additive manufacturing, specifically direct ink writing (DIW) and ink-jetting, have enabled the production of elastomeric silicone parts with deterministic control over the structure, shape, and mechanical properties. These new technologies offer rapid prototyping advantages and find applications in various fields, including biomedical devices, prosthetics, metamaterials, and soft robotics. Stereolithography (SLA) is a complementary approach with the ability to print with finer features and potentially higher throughput. However, all high-performance silicone elastomers are composites of polysiloxane networks reinforced with particulate filler, and consequently, silicone resins tend to have high viscosities (gel- or paste-like), which complicates or completely inhibits the layer-by-layer recoating process central to most SLA technologies. Herein, the design and build of a digital light projection SLA printer suitable for handling high-viscosity resins is demonstrated. Further, a series of UV-curable silicone resins with thiol-ene crosslinking and reinforced by a combination of fumed silica and MQ resins are also described. The resulting silicone elastomers are shown to have tunable mechanical properties, with 100–350% elongation and ultimate tensile strength from 1 to 2.5 MPa. Three-dimensional printed features of 0.4 mm were achieved, and complexity is demonstrated by octet-truss lattices that display negative stiffness.

scholarly journals The Use of Right Angle Fluorescence Spectroscopy to Distinguish the Botanical Origin of Greek Common Honey Varieties

2021 ◽  
Vol 11 (9) ◽  
pp. 4047
Author(s):  
Marinos Xagoraris ◽  
Panagiota-Kyriaki Revelou ◽  
Eleftherios Alissandrakis ◽  
Petros A. Tarantilis ◽  
Christos S. Pappas
Keyword(s):  
Fluorescence Spectroscopy ◽  
External Validation ◽  
Three Dimensional ◽  
Excitation Wavelength ◽  
Botanical Origin ◽  
Linear Discriminant ◽  
Classification Score ◽  
Hydroxybenzoic Acids ◽  
Unifloral Honey ◽  
Novel Method

The standardization of the botanical origin of honey reflects the commercial value and quality of honey. Nowadays, most consumers are looking for a unifloral honey. The aim of the present study was to develop a novel method for honey classification using chemometric models based on phenolic compounds analyzed with right angle fluorescence spectroscopy, coupled with stepwise linear discriminant analysis (LDA). The deconstructed spectrum from three-dimensional-emission excitation matrix (3D-EEM) spectra provided a correct classification score of 94.9% calibration and cross-validation at an excitation wavelength (λex) of 330 nm. Subsequently, a score of 81.4% and 79.7%, respectively, at an excitation wavelength (λex) of 360 nm was achieved. Each chemometric model confirmed its power through the external validation with a score of 82.1% for both. Differentiation could be correlated with hydroxycinnamic and hydroxybenzoic acids, which absorb in this region of the spectrum. Fluorescence spectroscopy constitutes a rapid and sensitive technique, which, when combined with the stepwise algorithm and LDA method, can be used as a reliable and predictive authentication tool for honey. This study indicates that the developed methodology is a promising technique for determination of the botanical origin of common Greek honey varieties. Our long-term ambition is to support producers and suppliers to remain in a competitive national and international market.

scholarly journals A novel method for processing adipose-derived stromal stem cells using a closed cell washing concentration device with a hollow fiber membrane module

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Shinji Hayashi ◽  
Rieko Yagi ◽  
Shuhei Taniguchi ◽  
Masami Uji ◽  
Hidaka Urano ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hollow Fiber ◽  
Hollow Fiber Membrane ◽  
Membrane Module ◽  
Fiber Membrane ◽  
Cell Processing ◽  
Closed Cell ◽  
A Cell ◽  
Novel Method ◽  
Stromal Stem Cells

AbstractCell-assisted lipotransfer (CAL) is an advanced lipoinjection method that uses autologous lipotransfer with addition of a stromal vascular fraction (SVF) containing adipose-derived stromal stem cells (ASCs). The CAL procedure of manual isolation of cells from fat requires cell processing to be performed in clean environment. To isolate cells from fat without the need for a cell processing center, such as in a procedure in an operation theater, we developed a novel method for processing SVF using a closed cell washing concentration device (CCD) with a hollow fiber membrane module. The CCD consists of a sterilized closed circuit, bags and hollow fiber, semi-automatic device and the device allows removal of >99.97% of collagenase from SVF while maintaining sterility. The number of nucleated cells, ASCs and viability in SVF processed by this method were equivalent to those in SVF processed using conventional manual isolation. Our results suggest that the CCD system is as reliable as manual isolation and may also be useful for CAL. This approach will help in the development of regenerative medicine at clinics without a cell processing center.

scholarly journals The Effect of Lateral Column Lengthening on Subtalar Motion: Are We Trading Deformity for Stiffness?

2020 ◽  
Vol 5 (4) ◽  
pp. 2473011420S0004
Author(s):  
Brittany Hedrick ◽  
Anthony Riccio ◽  
Danielle M. Thomas ◽  
Claire Shivers ◽  
Matthew Siebert ◽  
...  
Keyword(s):  
Motion Capture ◽  
Three Dimensional ◽  
Deformity Correction ◽  
Porous Titanium ◽  
Testing System ◽  
Materials Testing ◽  
Lateral Column ◽  
Fresh Frozen ◽  
Lateral Column Lengthening ◽  
Paired T Test

Category: Hindfoot; Other Introduction/Purpose: While lengthening of the lateral column through a calcaneal neck osteotomy is an integral component of flatfoot reconstruction in younger patients with flexible planovalgus deformities, concern exists as to the effect of this intra- articular osteotomy on subtalar motion. The purpose of this study is to quantify the alterations in subtalar motion following lateral column lengthening (LCL). Methods: The subtalar motion of 14 fresh frozen cadaveric feet was assessed using a three-dimensional motion capture system and materials testing system (MTS). Following potting of the tibia and calcaneus, optic markers were placed into the tibia, calcaneus and talus. The MTS was used to apply a rotational force across the subtalar joint to a torque of 5Nm. Abduction/adduction, supination/pronation, and plantarflexion/dorsiflexion about the talus was recorded. Specimens then underwent LCL via a calcaneal neck osteotomy which was maintained with a 12mm porous titanium wedge. Repeat subtalar motion analysis was performed and compared to pre-LCL motion using a paired t-test. Results:: No statistically significant differences in subtalar abduction/adduction (10.9O vs. 11.8O degrees, p=.48), supination/pronation (3.5O vs. 2.7O, p=.31), or plantarflexion/dorsiflexion (1.6O vs 1.0O, p=.10) were identified following LCL. Conclusion:: No significant changes in subtalar motion were observed following lateral column lengthening in this biomechanical cadaveric study. While these findings do not obviate concerns of clinical subtalar stiffness following planovalgus deformity correction, they suggest that diminished postoperative subtalar motion may be due to soft tissue scarring rather than alterations of joint anatomy.

A Three-Dimensional Viscous Transonic Inverse Design Method

2000 ◽  
Author(s):  
W. T. Tiow ◽  
M. Zangeneh
Keyword(s):  
Design Method ◽  
Flow Analysis ◽  
Three Dimensional ◽  
Viscous Effects ◽  
Flow Equations ◽  
Flow Solution ◽  
Turbomachinery Blades ◽  
A Cell ◽  
Inverse Design Method ◽  
Euler Solver

The development and application of a three-dimensional inverse methodology is presented for the design of turbomachinery blades. The method is based on the mass-averaged swirl, rV~θ distribution and computes the necessary blade changes directly from the discrepancies between the target and initial distributions. The flow solution and blade modification converge simultaneously giving the final blade geometry and the corresponding steady state flow solution. The flow analysis is performed using a cell-vertex finite volume time-marching algorithm employing the multistage Runge-Kutta integrator in conjunction with accelerating techniques (local time stepping and grid sequencing). To account for viscous effects, dissipative forces are included in the Euler solver using the log-law and mixing length models. The design method can be used with any existing solver solving the same flow equations without any modifications to the blade surface wall boundary condition. Validation of the method has been carried out using a transonic annular turbine nozzle and NASA rotor 67. Finally, the method is demonstrated on the re-design of the blades.

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