Cellular Nutrition in Complex Three-Dimensional Scaffolds: A Comparison between Experiments and Computer Simulations

Studies on bone cell ingrowth into synthetic, porous three-dimensional (3D) implants showed difficulties arising from impaired cellular proliferation and differentiation in the core region of these scaffolds with increasing scaffold volumein vitro. Therefore, we developed anin vitroperfusion cell culture module, which allows the analysis of cells in the interior of scaffolds under different medium flow rates. For each flow rate the cell viability was measured and compared with results from computer simulations that predict the local oxygen supply and shear stress inside the scaffold based on the finite element method. We found that the local cell viability correlates with the local oxygen concentration and the local shear stress. On the one hand the oxygen supply of the cells in the core becomes optimal with a higher perfusion flow. On the other hand shear stress caused by high flow rates impedes cell vitality, especially at the surface of the scaffold. Our results demonstrate that both parameters must be considered to derive an optimal nutrient flow rate.

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

Micromachines ◽

10.3390/mi12030346 ◽

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.

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Pharmacokinetics and dialytic clearance of apixaban during in vitro continuous renal replacement therapy

BMC Nephrology ◽

10.1186/s12882-021-02248-7 ◽

2021 ◽

Vol 22 (1) ◽

Author(s):

Lauren Andrews ◽

Scott Benken ◽

Xing Tan ◽

Eric Wenzler

Keyword(s):

Renal Replacement Therapy ◽

Linear Regression ◽

Protein Binding ◽

Continuous Renal Replacement Therapy ◽

Replacement Therapy ◽

Flow Rate ◽

Systemic Exposure ◽

Flow Rates ◽

Filter Type

Abstract Background To evaluate the transmembrane clearance (CLTM) of apixaban during modeled in vitro continuous renal replacement therapy (CRRT), assess protein binding and circuit adsorption, and provide initial dosing recommendations. Methods Apixaban was added to the CRRT circuit and serial pre-filter bovine blood samples were collected along with post-filter blood and effluent samples. All experiments were performed in duplicate using continuous veno-venous hemofiltration (CVVH) and hemodialysis (CVVHD) modes, with varying filter types, flow rates, and point of CVVH replacement fluid dilution. Concentrations of apixaban and urea were quantified via liquid chromatography-tandem mass spectrometry. Plasma pharmacokinetic parameters for apixaban were estimated via noncompartmental analysis. CLTM was calculated via the estimated area under the curve (AUC) and by the product of the sieving/saturation coefficient (SC/SA) and flow rate. Two and three-way analysis of variance (ANOVA) models were built to assess the effects of mode, filter type, flow rate, and point of dilution on CLTM by each method. Optimal doses were suggested by matching the AUC observed in vitro to the systemic exposure demonstrated in Phase 2/3 studies of apixaban. Linear regression was utilized to provide dosing estimations for flow rates from 0.5–5 L/h. Results Mean adsorption to the HF1400 and M150 filters differed significantly at 38 and 13%, respectively, while mean (± standard deviation, SD) percent protein binding was 70.81 ± 0.01%. Effect of CVVH point of dilution did not differ across filter types, although CLTM was consistently significantly higher during CRRT with the HF1400 filter compared to the M150. The three-way ANOVA demonstrated improved fit when CLTM values calculated by AUC were used (adjusted R2 0.87 vs. 0.52), and therefore, these values were used to generate optimal dosing recommendations. Linear regression revealed significant effects of filter type and flow rate on CLTM by AUC, suggesting doses of 2.5–7.5 mg twice daily (BID) may be needed for flow rates ranging from 0.5–5 L/h, respectively. Conclusion For CRRT flow rates most commonly employed in clinical practice, the standard labeled 5 mg BID dose of apixaban is predicted to achieve target systemic exposure thresholds. The safety and efficacy of these proposed dosing regimens warrants further investigation in clinical studies.

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A bioprinted composite hydrogel with controlled shear stress on cells

Proceedings of the Institution of Mechanical Engineers Part H Journal of Engineering in Medicine ◽

10.1177/0954411920976682 ◽

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.

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In vivo / in vitro Correlation of Pharmacokinetics of Gentamicin, Vancomycin, Teicoplanin and Doripenem in a Bovine Blood Hemodialysis Model

Frontiers in Pharmacology ◽

10.3389/fphar.2021.702455 ◽

2021 ◽

Vol 12 ◽

Author(s):

M G Vossen ◽

S Pferschy ◽

C Milacek ◽

M Haidinger ◽

Mario Karolyi ◽

...

Keyword(s):

Blood Flow ◽

Renal Replacement Therapy ◽

Protein Binding ◽

Replacement Therapy ◽

Flow Rate ◽

Blood Flow Rate ◽

Bovine Blood ◽

Flow Rates ◽

Dialysate Flow Rate

Background: Elimination of a drug during renal replacement therapy is not only dependent on flow rates, molecular size and protein binding, but is often influenced by difficult to predict drug membrane interactions. In vitro models allow for extensive profiling of drug clearance using a wide array of hemofilters and flow rates. We present a bovine blood based in vitro pharmacokinetic model for intermittent renal replacement therapy.Methods: Four different drugs were analyzed: gentamicin, doripenem, vancomicin and teicoplanin. The investigated drug was added to a bovine blood reservoir connected to a hemodialysis circuit. In total seven hemofilter models were analyzed using commonly employed flow rates. Pre-filter, post-filter and dialysate samples were drawn, plasmaseparated and analyzed using turbidimetric assays or HPLC. Protein binding of doripenem and vancomycin was measured in bovine plasma and compared to previously published values for human plasma.Results: Clearance values were heavily impacted by choice of membrane material and surface as well as by dialysis parameters such as blood flow rate. Gentamicin clearance ranged from a minimum of 90.12 ml/min in a Baxter CAHP-170 diacetate hemofilter up to a maximum of 187.90 ml/min in a Fresenius medical company Fx80 polysulfone model (blood flow rate 400 ml/min, dialysate flow rate 800 ml/min). Clearance of Gentamicin vs Vancomicin over the F80s hemofilter model using the same flow rates was 137.62 mL vs 103.25 ml/min. Doripenem clearance with the Fx80 was 141.25 ml/min.Conclusion: Clearance values corresponded very well to previously published data from clinical pharmacokinetic trials. In conjunction with in silico pharmacometric models. This model will allow precise dosing recommendations without the need of large scale clinical trials.

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The Influence of Flow Rate on the Wake in a Centrifugal Impeller

Volume 1: Turbomachinery ◽

10.1115/82-gt-45 ◽

1982 ◽

Author(s):

M. W. Johnson ◽

J. Moore

Keyword(s):

Flow Rate ◽

Incidence Angle ◽

Three Dimensional ◽

Stagnation Pressure ◽

Centrifugal Impeller ◽

Suction Surface ◽

Cross Sectional ◽

Flow Rates ◽

Pressure Losses ◽

Compressor Impeller

Three-dimensional flows and their influence on the stagnation pressure losses in a centrifugal compressor impeller have been studied. All 3 mutally perpendicular components of relative velocity and stagnation pressure on 5 cross-sectional planes, between the inlet and outlet of a 1 m dia shrouded impeller running at 500 rpm were measured. Comparisons were made between results for a flow rate corresponding to nearly zero incidence angle and two other flows, with increased and reduced flow rates. These detailed measurements show how the position of separation of the shroud boundary layer moved downstream and the wake’s size decreased, as the flow rate was increased. The wake’s location, at the outlet of the impeller, was also observed to move from the suction surface at the lowest flow rate, to the shroud at higher flow rates.

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Facile fabrication and characterization on alginate microfibres with grooved structure via microfluidic spinning

Royal Society Open Science ◽

10.1098/rsos.181928 ◽

2019 ◽

Vol 6 (5) ◽

pp. 181928 ◽

Cited By ~ 3

Author(s):

Xiaolin Zhang ◽

Lin Weng ◽

Qingsheng Liu ◽

Dawei Li ◽

Bingyao Deng

Keyword(s):

Flow Rate ◽

Loss Modulus ◽

Fibre Diameter ◽

Morphological Structure ◽

Flow Rates ◽

Alginate Solution ◽

The Core ◽

Facile Fabrication ◽

Fabrication And Characterization ◽

Alginate Fibres

Alginate microfibres were fabricated by a simple microfluidic spinning device consisting of a coaxial flow. The inner profile and spinnability of polymer were analysed by rheology study, including the analysis of viscosity, storage modulus and loss modulus. The effect of spinning parameters on the morphological structure of fibres was studied by SEM, while the crystal structure and chemical group were characterized by FTIR and XRD, respectively. Furthermore, the width and depth of grooves on the fibres was investigated by AFM image analysis and the formation mechanism of grooves was finally analysed. It was illustrated that the fibre diameter increased with an increase in the core flow rate, whereas on the contrary of sheath flow rate. Fibre diameter exhibited an increasing tendency as the concentration of alginate solution increased, and the minimum spinning concentration of alginate solution was 1% with the finest diameter being around 25 µm. Importantly, the grooved structure was obtained by adjusting the concentration of solutions and flow rates, the depth of groove increased from 278.37 ± 2.23 µm to 727.52 ± 3.52 µm as the concentration varied from 1 to 2%. Alginate fibres, with topological structure, are candidates for wound dressing or the engineering tissue scaffolds.

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Effect of shear stress on microvessel network formation of endothelial cells with in vitro three-dimensional model

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00400.2003 ◽

2004 ◽

Vol 287 (3) ◽

pp. H994-H1002 ◽

Cited By ~ 47

Author(s):

Akinori Ueda ◽

Masaki Koga ◽

Mariko Ikeda ◽

Susumu Kudo ◽

Kazuo Tanishita

Keyword(s):

Endothelial Cells ◽

Shear Stress ◽

Network Formation ◽

Three Dimensional ◽

Collagen Gel ◽

Flow Chamber ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Collagen Gels

Shear stress stimulus is expected to enhance angiogenesis, the formation of microvessels. We determined the effect of shear stress stimulus on three-dimensional microvessel formation in vitro. Bovine pulmonary microvascular endothelial cells were seeded onto collagen gels with basic fibroblast growth factor to make a microvessel formation model. We observed this model in detail using phase-contrast microscopy, confocal laser scanning microscopy, and electron microscopy. The results show that cells invaded the collagen gel and reconstructed the tubular structures, containing a clearly defined lumen consisting of multiple cells. The model was placed in a parallel-plate flow chamber. A laminar shear stress of 0.3 Pa was applied to the surfaces of the cells for 48 h. Promotion of microvessel network formation was detectable after ∼10 h in the flow chamber. After 48 h, the length of networks exposed to shear stress was 6.17 (±0.59) times longer than at the initial state, whereas the length of networks not exposed to shear stress was only 3.30 (±0.41) times longer. The number of bifurcations and endpoints increased for networks exposed to shear stress, whereas the number of bifurcations alone increased for networks not exposed to shear stress. These results demonstrate that shear stress applied to the surfaces of endothelial cells on collagen gel promotes the growth of microvessel network formation in the gel and expands the network because of repeated bifurcation and elongation.

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Identification of Wake Convection and Flow Outline in the Interface Region of Blade Rows With Axial Gap in a Counter Rotating Turbine

Volume 1: Compressors, Fans, and Pumps; Turbines; Heat Transfer; Structures and Dynamics ◽

10.1115/gtindia2019-2634 ◽

2019 ◽

Author(s):

Rayapati Subbarao ◽

M. Govardhan

Keyword(s):

Flow Pattern ◽

Mass Flow Rate ◽

Mass Flow ◽

Flow Rate ◽

Three Dimensional ◽

Interface Region ◽

Turbine Stage ◽

Deviation Angle ◽

Flow Rates ◽

Improved Performance

Abstract In a Counter Rotating Turbine (CRT), the stationary nozzle is trailed by two rotors that rotate in the opposite direction to each other. Flow in a CRT stage is multifaceted and more three dimensional, especially, in the gap between nozzle and rotor 1 as well as rotor 1 and rotor 2. By varying this gap between the blade rows, the flow and wake pattern can be changed favorably and may lead to improved performance. Present work analyzes the aspect of change in flow field through the interface, especially the wake pattern and deviation in flow with change in spacing. The components of turbine stage are modeled for different gaps between the components using ANSYS® ICEM CFD 14.0. Normalized flow rates ranging from 0.091 to 0.137 are used. The 15, 30, 50 and 70% of the average axial chords are taken as axial gaps in the present analysis. CFX 14.0 is used for simulation. At nozzle inlet, stagnation pressure boundary condition is used. At the turbine stage or rotor 2 outlet, mass flow rate is specified. Pressure distribution contours at the outlets of the blade rows describe the flow pattern clearly in the interface region. Wake strength at nozzle outlet is more for the lowest gap. At rotor 1 outlet, it is less for x/a = 0.3 and increases with gap. Incidence angles at the inlets of rotors are less for the smaller gaps. Deviation angle at the outlet of rotor 1 is also considered, as rotor 1-rotor 2 interaction is more significant in CRT. Deviation angle at rotor 1 outlet is minimum for this gap. Also, for the intermediate mass flow rate of 0.108, x/a = 0.3 is giving more stage performance. This suggests that at certain axial gap, there is better wake convection and flow outline, when compared to other gap cases. Further, it is identified that for the axial gap of x/a = 0.3 and the mean mass flow rate of 0.108, the performance of CRT is maximum. It is clear that the flow pattern at the interface is changing the incidence and deviation with change in axial gap and flow rate. This study is useful for the gas turbine community to identify the flow rates and gaps at which any CRT stage would perform better.

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Inflammatory Response of Endothelial Cells to Wall Shear Stress in Three Dimensional Stenotic Models

ASME 2009 Summer Bioengineering Conference, Parts A and B ◽

10.1115/sbc2009-206669 ◽

2009 ◽

Author(s):

Leonie Rouleau ◽

Joanna Rossi ◽

Jean-Claude Tardif ◽

Rosaire Mongrain ◽

Richard L. Leask

Keyword(s):

Endothelial Cells ◽

Shear Stress ◽

Wall Shear Stress ◽

Three Dimensional ◽

Realistic Model ◽

In Vitro Models ◽

Steady Flows ◽

Wall Shear

Endothelial cells (ECs) are believed to respond differentially to hemodynamic forces in the vascular tree. Once atherosclerotic plaque has formed in a vessel, the obstruction creates complex spatial gradients in wall shear stress (WSS). In vitro models have used mostly unrealistic and simplified geometries, which cannot reproduce accurately physiological conditions. The objective of this study was to expose ECs to the complex WSS pattern created by an asymmetric stenosis. Endothelial cells were grown and exposed for different times to physiological steady flows in straight dynamic controls and in idealized asymmetric stenosis models. Cell morphology was noticeably different in the regions with spatial WSS gradients, being more randomly oriented and of cobblestone shape. Inflammatory molecule expression was also altered by exposure to shear and endothelial nitric oxide synthase (eNOS) was upregulated by its presence. A regional response in terms of inflammation was observed through confocal microscopy. This work provides a more realistic model to study endothelial cell response to spatial and temporal WSS gradients that are present in vivo and is an important advancement towards a better understanding of the mechanisms involved in coronary artery disease.

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