scholarly journals Towards the Experimentally-Informed In Silico Nozzle Design Optimization for Extrusion-Based Bioprinting of Shear-Thinning Hydrogels

2021 ◽  
Vol 9 ◽  
Author(s):  
Esther Reina-Romo ◽  
Sourav Mandal ◽  
Paulo Amorim ◽  
Veerle Bloemen ◽  
Eleonora Ferraris ◽  
...  
Keyword(s):  
Shear Stress ◽  
Cell Viability ◽  
Cell Survival ◽  
Material Properties ◽  
In Silico ◽  
Shear Thinning ◽  
Nozzle Geometry ◽  
3D Bioprinting ◽  
Printing Process ◽  
Constant Flow Rate

Research in bioprinting is booming due to its potential in addressing several manufacturing challenges in regenerative medicine. However, there are still many hurdles to overcome to guarantee cell survival and good printability. For the 3D extrusion-based bioprinting, cell viability is amongst one of the lowest of all the bioprinting techniques and is strongly influenced by various factors including the shear stress in the print nozzle. The goal of this study is to quantify, by means of in silico modeling, the mechanical environment experienced by the bioink during the printing process. Two ubiquitous nozzle shapes, conical and blunted, were considered, as well as three common hydrogels with material properties spanning from almost Newtonian to highly shear-thinning materials following the power-law behavior: Alginate-Gelatin, Alginate and PF127. Comprehensive in silico testing of all combinations of nozzle geometry variations and hydrogels was achieved by combining a design of experiments approach (DoE) with a computational fluid dynamics (CFD) of the printing process, analyzed through a machine learning approach named Gaussian Process. Available experimental results were used to validate the CFD model and justify the use of shear stress as a surrogate for cell survival in this study. The lower and middle nozzle radius, lower nozzle length and the material properties, alone and combined, were identified as the major influencing factors affecting shear stress, and therefore cell viability, during printing. These results were successfully compared with those of reported experiments testing viability for different nozzle geometry parameters under constant flow rate or constant pressure. The in silico 3D bioprinting platform developed in this study offers the potential to assist and accelerate further development of 3D bioprinting.

scholarly journals EGFRvIII Promotes Cell Survival during Endoplasmic Reticulum Stress through a Reticulocalbin 1-Dependent Mechanism

Cancers ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 1198
Author(s):  
Juliana Gomez ◽  
Zammam Areeb ◽  
Sarah F. Stuart ◽  
Hong P. T. Nguyen ◽  
Lucia Paradiso ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Viability ◽  
Er Stress ◽  
Cell Survival ◽  
Glioblastoma Cells ◽  
Over Expression ◽  
Stress Markers ◽  
Apoptotic Protein ◽  
Novel Association ◽  
Reduced Activity

Reticulocalbin 1 (RCN1) is an endoplasmic reticulum (ER)-residing protein, involved in promoting cell survival during pathophysiological conditions that lead to ER stress. However, the key upstream receptor tyrosine kinase that regulates RCN1 expression and its potential role in cell survival in the glioblastoma setting have not been determined. Here, we demonstrate that RCN1 expression significantly correlates with poor glioblastoma patient survival. We also demonstrate that glioblastoma cells with expression of EGFRvIII receptor also have high RCN1 expression. Over-expression of wildtype EGFR also correlated with high RCN1 expression, suggesting that EGFR and EGFRvIII regulate RCN1 expression. Importantly, cells that expressed EGFRvIII and subsequently showed high RCN1 expression displayed greater cell viability under ER stress compared to EGFRvIII negative glioblastoma cells. Consistently, we also demonstrated that RCN1 knockdown reduced cell viability and exogenous introduction of RCN1 enhanced cell viability following induction of ER stress. Mechanistically, we demonstrate that the EGFRvIII-RCN1-driven increase in cell survival is due to the inactivation of the ER stress markers ATF4 and ATF6, maintained expression of the anti-apoptotic protein Bcl-2 and reduced activity of caspase 3/7. Our current findings identify that EGFRvIII regulates RCN1 expression and that this novel association promotes cell survival in glioblastoma cells during ER stress.

scholarly journals Rapid Fabrication of Microfluidic Devices for Biological Mimicking: A Survey of Materials and Biocompatibility

Micromachines ◽  
2021 ◽  
Vol 12 (3) ◽  
pp. 346
Author(s):  
Hui Ling Ma ◽  
Ana Carolina Urbaczek ◽  
Fayene Zeferino Ribeiro de Souza ◽  
Paulo Augusto Gomes Garrido Carneiro Leão ◽  
Janice Rodrigues Perussi ◽  
...  
Keyword(s):  
Shear Stress ◽  
Cell Viability ◽  
Cellular Response ◽  
Fluid Shear Stress ◽  
Umbilical Vein ◽  
Microfluidic Devices ◽  
In Vitro Assays ◽  
Stress Effect

Microfluidics is an essential technique used in the development of in vitro models for mimicking complex biological systems. The microchip with microfluidic flows offers the precise control of the microenvironment where the cells can grow and structure inside channels to resemble in vivo conditions allowing a proper cellular response investigation. Hence, this study aimed to develop low-cost, simple microchips to simulate the shear stress effect on the human umbilical vein endothelial cells (HUVEC). Differentially from other biological microfluidic devices described in the literature, we used readily available tools like heat-lamination, toner printer, laser cutter and biocompatible double-sided adhesive tapes to bind different layers of materials together, forming a designed composite with a microchannel. In addition, we screened alternative substrates, including polyester-toner, polyester-vinyl, glass, Permanox® and polystyrene to compose the microchips for optimizing cell adhesion, then enabling these microdevices when coupled to a syringe pump, the cells can withstand the fluid shear stress range from 1 to 4 dyne cm2. The cell viability was monitored by acridine orange/ethidium bromide (AO/EB) staining to detect live and dead cells. As a result, our fabrication processes were cost-effective and straightforward. The materials investigated in the assembling of the microchips exhibited good cell viability and biocompatibility, providing a dynamic microenvironment for cell proliferation. Therefore, we suggest that these microchips could be available everywhere, allowing in vitro assays for daily laboratory experiments and further developing the organ-on-a-chip concept.

Shear-Thinning and Thermo-Reversible Nanoengineered Inks for 3D Bioprinting

2017 ◽  
Vol 9 (50) ◽  
pp. 43449-43458 ◽  
Author(s):  
Scott A. Wilson ◽  
Lauren M. Cross ◽  
Charles W. Peak ◽  
Akhilesh K. Gaharwar
Keyword(s):  
Shear Thinning ◽  
3D Bioprinting

scholarly journals Sensitivity analysis of FDA´s benchmark nozzle regarding in vitro imperfections - Do we need asymmetric CFD benchmarks?

2020 ◽  
Vol 6 (3) ◽  
pp. 78-81
Author(s):  
Michael Stiehm ◽  
Christoph Brandt-Wunderlich ◽  
Stefan Siewert ◽  
Klaus-Peter Schmitz ◽  
Niels Grabow ◽  
...  
Keyword(s):  
Reynolds Number ◽  
Flow Field ◽  
In Silico ◽  
Nozzle Geometry ◽  
Geometrical Imperfection ◽  
Custom Made ◽  
Inlet Conditions ◽  
Silicone Model ◽  
Bench Testing

AbstractModern technologies and methods such as computer simulation, so-called in silico methods, foster the development of medical devices. For accelerating the uptake of computer simulations and to increase credibility and reliability the U.S. Food and Drug Administration organized an inter-laboratory round robin study of a generic nozzle geometry. In preparation of own bench testing experiment using Particle Image Velocimetry, a custom made silicone nozzle was manufactured. By using in silico computational fluid dynamics method the influence of in vitro imperfections, such as inflow variations and geometrical deviations, on the flow field were evaluated. Based on literature the throat Reynolds number was varied Rethroat = 500 ± 50. It could be shown that the flow field errors resulted from variations of inlet conditions can be largely eliminated by normalizing if the Reynolds number is known. Furthermore, a symmetric imperfection of the silicone model within manufacturing tolerance does not affect the flow as much as an asymmetric failure such as an unintended curvature of the nozzle. In brief, we can conclude that geometrical imperfection of the reference experiment should be considered accordingly to in silico modelling. The question arises, if an asymmetric benchmark for biofluid analysis needs to be established. An eccentric nozzle benchmark could be a suitable case and will be further investigated.

Investigation on the Rheological Behavior of Polyamide6/Montmorillonite Nanocomposites by Reactive Extrusion

2011 ◽  
Vol 233-235 ◽  
pp. 1998-2001 ◽  
Author(s):  
Ming Zhao ◽  
Xiao Zhong Lu ◽  
Kai Gu ◽  
Xiao Min Sun ◽  
Chang Qing Ji
Keyword(s):  
Activation Energy ◽  
Shear Stress ◽  
Shear Rate ◽  
Rheological Behavior ◽  
Reactive Extrusion ◽  
Shear Thinning ◽  
Experimental Results ◽  
Montmorillonite Nanocomposites

The rheological behavior of PA6/montmorillonite(MMT) by reactive extrusion was investigated using cone-and-plate rheometer. The experimental results indicated that PA6/MMT exhibited shear-thinning behavior. The shear stress of both neat PA6 and PA6/MMT increased with the increase in the shear rate. The reduction of the viscous activation energy with the increase of shear stress reflected PA6/MMT can be processed over a wider temperature.

A bioprinted composite hydrogel with controlled shear stress on cells

2020 ◽  
pp. 095441192097668
Author(s):  
Amirhossein Bakhtiiari ◽  
Rezvan Khorshidi ◽  
Fatemeh Yazdian ◽  
Hamid Rashedi ◽  
Meisam Omidi
Keyword(s):  
Shear Stress ◽  
Cell Viability ◽  
Room Temperature ◽  
Degradation Rate ◽  
Diffusion Rate ◽  
Sol Gel ◽  
Three Dimensional ◽  
Simulation Software ◽  
Sol Gel Transition ◽  
Gel Transition

In recent decades, three dimensional (3D) bio-printing technology has found widespread use in tissue engineering applications. The aim of this study is to scrutinize different parameters of the bioprinter – with the help of simulation software – to print a hydrogel so much so that avoid high amounts of shear stress which is detrimental for cell viability and cell proliferation. Rheology analysis was done on several hydrogels composed of different percentages of components: alginate, collagen, and gelatin. The results have led to the combination of percentages collagen:alginate:gelatin (1:4:8)% as the best condition which makes sol-gel transition at room temperature possible. The results have shown the highest diffusion rate and cell viability for the cross-linked sample with 1.5% CaCl2 for the duration of 1 h. Finally, we have succeeded in printing the hydrogel that is mechanically strong with suitable degradation rate and cell viability.

scholarly journals Application of feed forward and recurrent neural networks in simulation of left ventricular mechanics

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yaghoub Dabiri ◽  
Alex Van der Velden ◽  
Kevin L. Sack ◽  
Jenny S. Choy ◽  
Julius M. Guccione ◽  
...  
Keyword(s):  
Real Time ◽  
Material Properties ◽  
In Silico ◽  
Prediction Models ◽  
Heart Function ◽  
Active Material ◽  
Experimental Studies ◽  
Left Ventricular ◽  
Feed Forward ◽  
Ventricular Mechanics

AbstractAn understanding of left ventricle (LV) mechanics is fundamental for designing better preventive, diagnostic, and treatment strategies for improved heart function. Because of the costs of clinical and experimental studies to treat and understand heart function, respectively, in-silico models play an important role. Finite element (FE) models, which have been used to create in-silico LV models for different cardiac health and disease conditions, as well as cardiac device design, are time-consuming and require powerful computational resources, which limits their use when real-time results are needed. As an alternative, we sought to use deep learning (DL) for LV in-silico modeling. We used 80 four-chamber heart FE models for feed forward, as well as recurrent neural network (RNN) with long short-term memory (LSTM) models for LV pressure and volume. We used 120 LV-only FE models for training LV stress predictions. The active material properties of the myocardium and time were features for the LV pressure and volume training, and passive material properties and element centroid coordinates were features of the LV stress prediction models. For six test FE models, the DL error for LV volume was 1.599 ± 1.227 ml, and the error for pressure was 1.257 ± 0.488 mmHg; for 20 LV FE test examples, the mean absolute errors were, respectively, 0.179 ± 0.050 for myofiber, 0.049 ± 0.017 for cross-fiber, and 0.039 ± 0.011 kPa for shear stress. After training, the DL runtime was in the order of seconds whereas equivalent FE runtime was in the order of several hours (pressure and volume) or 20 min (stress). We conclude that using DL, LV in-silico simulations can be provided for applications requiring real-time results.

scholarly journals Harnessing shear stress preconditioning to improve cell viability in 3D post-printed biostructures using extrusion bioprinting

Bioprinting ◽  
2021 ◽  
pp. e00184
Author(s):  
Selwa Boularaoui ◽  
Aya Shanti ◽  
Kamran A. Khan ◽  
Saverio Iacoponi ◽  
Nicolas Christoforou ◽  
...  
Keyword(s):  
Shear Stress ◽  
Cell Viability ◽  
Improve Cell Viability

scholarly journals Comparison of Linear Poly Ethylene Imine (LPEI) and Poly L-Lysine (PLL) in Fabrication of CHOK1 Cell-Loaded Multilayer Alginate Microcapsules

2020 ◽  
Vol 10 (2) ◽  
pp. 290-296
Author(s):  
Fariba Hajifathaliha ◽  
Arash Mahboubi ◽  
Elham Mohit ◽  
Noushin Bolourchian ◽  
Vahid Khalaj ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cell Survival ◽  
Metabolic Activity ◽  
Encapsulation Efficiency ◽  
Cell Encapsulation ◽  
High Price ◽  
Ethylene Imine ◽  
Linear Poly ◽  
Cellular Metabolic Activity ◽  
Poly Ethylene

Purpose: Poly l-lysine (PLL) has been introduced as a strengthening covering layer for alginate microcapsules which are the most convenient way for cell encapsulation. Some disadvantages of PLL such as high price and low biocompatibility have prompted scientists to find better alternatives. Linear poly ethylene imine (LPEI), thanks to its highly similar structure to PLL, could be considered as a proper cost-effective alternative. In this study LPEI and PLL were compared as covering layers of cell-loaded alginate-LPEI-alginate (cALA) and alginate-PLL-alginate (cAPA) microcapsules. Methods: In addition to the physico-mechanical properties, the encapsulation efficiency, cell survival post encapsulation, cell viability, and cellular metabolic activity within the microcapsules were evaluated using trypan blue, live/dead cell staining, and MTT test, respectively. Results: Physico-mechanical evaluation of the microcapsules revealed that the cell microencapsulation process did not affect their shape, size, and mechanical stability. Although the encapsulation efficiency for cALA and cAPA was not different (P>0.05), cell survival post encapsulation was higher in cALA than in cAPA (P<0.05) which could be the reason for the higher cell viability and also cellular metabolic activity within these microcapsules in comparison to cAPA. Conclusion: Here, based on these results, ALA could be introduced as a preferable alternative to APA for cell encapsulation.

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