scholarly journals Acoustic Cell Patterning in Hydrogel for Three-Dimensional Cell Network Formation

Micromachines ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 3
Author(s):  
Kyo-in Koo ◽  
Andreas Lenshof ◽  
Le Thi Huong ◽  
Thomas Laurell

In the field of engineered organ and drug development, three-dimensional network-structured tissue has been a long-sought goal. This paper presents a direct hydrogel extrusion process exposed to an ultrasound standing wave that aligns fibroblast cells to form a network structure. The frequency-shifted (2 MHz to 4 MHz) ultrasound actuation of a 400-micrometer square-shaped glass capillary that was continuously perfused by fibroblast cells suspended in sodium alginate generated a hydrogel string, with the fibroblasts aligned in single or quadruple streams. In the transition from the one-cell stream to the four-cell streams, the aligned fibroblast cells were continuously interconnected in the form of a branch and a junction. The ultrasound-exposed fibroblast cells displayed over 95% viability up to day 10 in culture medium without any significant difference from the unexposed fibroblast cells. This acoustofluidic method will be further applied to create a vascularized network by replacing fibroblast cells with human umbilical vein endothelial cells.

Polymers ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1217
Author(s):  
Jang Ho Ha ◽  
Jae Hyun Lim ◽  
Ji Woon Kim ◽  
Hyeon-Yeol Cho ◽  
Seok Geun Jo ◽  
...  

Blended hydrogels play an important role in enhancing the properties (e.g., mechanical properties and conductivity) of hydrogels. In this study, we generated a conductive blended hydrogel, which was achieved by mixing gelatin methacrylate (GelMA) with collagen, and silver nanowire (AgNW). The ratio of GelMA, collagen and AgNW was optimized and was subsequently gelated by ultraviolet light (UV) and heat. The scanning electron microscope (SEM) image of the conductive blended hydrogels showed that collagen and AgNW were present in the GelMA hydrogel. Additionally, rheological analysis indicated that the mechanical properties of the conductive GelMA–collagen–AgNW blended hydrogels improved. Biocompatibility analysis confirmed that the human umbilical vein endothelial cells (HUVECs) encapsulated within the three-dimensional (3D), conductive blended hydrogels were highly viable. Furthermore, we confirmed that the molecule in the conductive blended hydrogel was released by electrical stimuli-mediated structural deformation. Therefore, this conductive GelMA–collagen–AgNW blended hydrogel could be potentially used as a smart actuator for drug delivery applications.


2017 ◽  
Vol 33 (9) ◽  
pp. 592-599 ◽  
Author(s):  
Francesca Felice ◽  
Ester Belardinelli ◽  
Alessandro Frullini ◽  
Tatiana Santoni ◽  
Egidio Imbalzano ◽  
...  

Objectives Aminaphtone, a naphtohydrochinone used in the treatment of capillary disorders, may affect oedema in chronic venous insufficiency. Aim of study is to investigate the effect of aminaphtone on vascular endothelial permeability in vitro and its effects on three-dimensional capillary-like structures formed by human umbilical vein endothelial cells. Method Human umbilical vein endothelial cells were treated with 50 ng/ml VEGF for 2 h and aminaphtone for 6 h. Permeability assay, VE-cadherin expression and Matrigel assay were performed. Results VEGF-induced permeability was significantly decreased by aminaphtone in a range concentration of 1–20 µg/ml. Aminaphtone restored VE-cadherin expression. Finally, 6 h pre-treatment with aminaphtone significantly preserved capillary-like structures formed by human umbilical vein endothelial cells on Matrigel up to 48 h compared to untreated cells. Conclusions Aminaphtone significantly protects endothelium permeability and stabilises endothelial cells organised in capillary-like structures, modulating VE-cadherin expression. These data might explain the clinical benefit of aminaphtone on chronic venous insufficiency.


2002 ◽  
Vol 11 (4) ◽  
pp. 369-377 ◽  
Author(s):  
Makarand V. Risbud ◽  
Erdal Karamuk ◽  
René Moser ◽  
Joerg Mayer

Three-dimensional (3-D) scaffolds offer an exciting possibility to develop cocultures of various cell types. Here we report chitosan–collagen hydrogel-coated fabric scaffolds with defined mesh size and fiber diameter for 3-D culture of human umbilical vein endothelial cells (HUVECs). These scaffolds did not require pre-coating with fibronectin and they supported proper HUVEC attachment and growth. Scaffolds preserved endothelial cell-specific cobblestone morphology and cells were growing in compartments defined by the textile mesh. HUVECs on the scaffold maintained the property of contact inhibition and did not exhibit overgrowth until the end of in vitro culture (day 6). MTT assay showed that cells had preserved mitochondrial functionality. It was also noted that cell number on the chitosan-coated scaffold was lower than that of collagen-coated scaffolds. Calcein AM and ethidium homodimer (EtD-1) dual staining demonstrated presence of viable and metabolically active cells, indicating growth supportive properties of the scaffolds. Actin labeling revealed absence of actin stress fibers and uniform distribution of F-actin in the cells, indicating their proper attachment to the scaffold matrix. Confocal microscopic studies showed that HUVECs growing on the scaffold had preserved functionality as seen by expression of von Willebrand (vW) factor. Observations also revealed that functional HUVECs were growing at various depths in the hydrogel matrix, thus demonstrating the potential of these scaffolds to support 3-D growth of cells. We foresee the application of this scaffold system in the design of liver bioreactors wherein hepatocytes could be cocultured in parallel with endothelial cells to enhance and preserve liver-specific functions.


2013 ◽  
Vol 06 (01) ◽  
pp. 1250053 ◽  
Author(s):  
YITENG ZHANG ◽  
DEJUN LI ◽  
MENGLI ZHAO ◽  
MEIXIAN GUO ◽  
XIANGYUN DENG ◽  
...  

Influence of carboxylic functionalization on the cytocompatibility of multiwalled carbon nanotubes (MWCNTs) was investigated in this work. Water contact angle assay showed that the surface of MWCNTs-containing carboxyl (MWCNTs-COOH) became much more hydrophilic compared with pure MWCNTs. In cell-adhesion assays, two cell lines, mouse fibroblast cells (L929) and human umbilical vein endothelial cells (EAHY926) were used to assess the cytocompatibility of materials. The MWCNTs-COOH displayed the improved cell proliferation, viability and adhesion due to the enhanced wettability, indicating their superior cytocompatibility over MWCNTs. The existence of carboxyl groups should be benefit to the adhesion and growth of both cells, which implied that MWCNTs-COOH were helpful for seeding both cells and could be used as the functional surface for the adhesion and growth of cells.


2009 ◽  
Vol 3 (2) ◽  
Author(s):  
A. Mohamed ◽  
A. Erdman ◽  
G. Timm

Previous biomechanical models of the penis that have attempted to simulate penile erections have either been limited to two-dimensional geometry, simplified three-dimensional geometry or made inaccurate assumptions altogether. Most models designed the shaft of the penis as a one-compartment pressurized vessel fixed at one end, when in reality it is a two-compartments pressurized vessel, in which the compartments diverge as they enter the body and are fixed at two separate points. This study began by designing simplified two-dimensional and three-dimensional models of the erect penis using Finite Element Analysis (FEA) methods with varying anatomical considerations for analyzing structural stresses, axial buckling and lateral deformation. The study then validated the results by building physical models replicating the computer models. Finally a more complex and anatomically accurate model of the penis was designed and analyzed. There was a significant difference in the peak von-Mises stress distribution between the one-compartment pressurized vessel and the more anatomically correct two-compartments pressurized vessel. Furthermore, the two-compartments diverging pressurized vessel was found to have more structural integrity when subject to external lateral forces than the one-compartment pressurized vessel. This study suggests that Mother Nature has favored an anatomy of two corporal cavernosal bodies separated by a perforated septum as opposed to one corporal body, due to better structural integrity of the tunica albuginea when subject to external forces.


2015 ◽  
Vol 2015 ◽  
pp. 1-14 ◽  
Author(s):  
Rui Zhang ◽  
Nan Wang ◽  
Man Zhang ◽  
Li-Nan Zhang ◽  
Zhi-Xia Guo ◽  
...  

Mesenchymal stem cells (MSCs) are known to undergo endothelial differentiation in response to treatment with vascular endothelial growth factor (VEGF), but their angiogenic ability is poorly characterized. In the present study, we aimed to further investigate the role of Rho/MRTF-A in angiogenesis by MSCs and the effect of the Rho/MRTF-A pathway on the expression of integrinsα1β1 andα5β1, which are known to mediate physiological and pathological angiogenesis. Our results showed that increased expression ofα1,α5, andβ1 was observed during angiogenesis of differentiated MSCs, and the Rho/MRTF-A signaling pathway was demonstrated to be involved in regulating the expression of integrinsα1,α5, andβ1. Luciferase reporter assay and ChIP assay determined that MRTF-A could bind to and transactivate the integrinα1 andα5 promoters. Treatment with the Rho inhibitor C3 transferase, the Rho-associated protein kinase (ROCK) inhibitor Y27632 or with shMRTF-A inhibited both the upregulation ofα1,α5, andβ1 as well as angiogenesis. Furthermore, in human umbilical vein endothelial cells (HUVECs), MRTF-A deletion led to marked reductions in cell migration and vessel network formation compared with the control. These data demonstrate that Rho/MRTF-A signaling is an important mediator that controls integrin gene expression during MSC-mediated angiogenic processes.


2016 ◽  
Vol 113 (12) ◽  
pp. 3179-3184 ◽  
Author(s):  
David B. Kolesky ◽  
Kimberly A. Homan ◽  
Mark A. Skylar-Scott ◽  
Jennifer A. Lewis

The advancement of tissue and, ultimately, organ engineering requires the ability to pattern human tissues composed of cells, extracellular matrix, and vasculature with controlled microenvironments that can be sustained over prolonged time periods. To date, bioprinting methods have yielded thin tissues that only survive for short durations. To improve their physiological relevance, we report a method for bioprinting 3D cell-laden, vascularized tissues that exceed 1 cm in thickness and can be perfused on chip for long time periods (>6 wk). Specifically, we integrate parenchyma, stroma, and endothelium into a single thick tissue by coprinting multiple inks composed of human mesenchymal stem cells (hMSCs) and human neonatal dermal fibroblasts (hNDFs) within a customized extracellular matrix alongside embedded vasculature, which is subsequently lined with human umbilical vein endothelial cells (HUVECs). These thick vascularized tissues are actively perfused with growth factors to differentiate hMSCs toward an osteogenic lineage in situ. This longitudinal study of emergent biological phenomena in complex microenvironments represents a foundational step in human tissue generation.


1995 ◽  
Vol 7 (6) ◽  
pp. 1499 ◽  
Author(s):  
N Osses ◽  
L Sobrevia ◽  
C Cordova ◽  
SM Jarvis ◽  
DL Yudilevich

Pregnancy complicated by diabetes is a relatively frequent event and may result in fetal embriopathy. However, little is known regarding whether placental transport functions are altered. In this study, we have compared the activity of the nitrobenzylthioinosine (NBMPR)-sensitive adenosine transporter and adenosine metabolism in human placental brush-border- and basal-membrane vesicles from placentas of normal and diabetic pregnancies. Neither [3H]NBMPR binding, a marker of the facilitative-diffusion nucleoside transporter in the human placenta, nor adenosine metabolism exhibited a significant difference in either the brush-border- or the basal-membrane vesicles between the normal and diabetic group, except for an increased affinity in [3H]NBMPR binding at the basal side in diabetic placenta. This result contrasts with an earlier finding using the same group of patients that adenosine transport is downregulated in umbilical vein endothelial cells from diabetic pregnancies. It is concluded that adenosine transport is modulated selectively in different tissues in diabetic pregnancies.


Author(s):  
Yasuhiro Yoshida ◽  
Toshiyuki Tsunoda ◽  
Yasuo Takashima ◽  
Takahiro Fujimoto ◽  
Keiko Doi ◽  
...  

AbstractZFAT, originally identified as a susceptibility gene for autoimmune thyroid disease, encodes a transcriptional regulator with one AT-hook and 18 C2H2-type zinc-finger domains. It is highly conserved among species. Here, we demonstrate that ZFAT is clearly expressed in human umbilical vein endothelial cells (HUVECs). Furthermore, we show that endothelial cell assembly and the branch point formation of capillary-like structures in HUVECs is impaired by the reduction of ZFAT expression through the use of ZFAT-miRNAs, whereas differences in cell proliferation or apoptotic features were not observed after the reduction in ZFAT expression. These results suggest that ZFAT may have critical roles in the capillary-like network formation that is involved in vascular remodeling. Elucidating the ZFAT-mediated transcriptional network will lead to a better understanding of the molecular mechanisms of angiogenesis.


2020 ◽  
Vol 982 ◽  
pp. 51-58
Author(s):  
Fu Liang Jiang ◽  
Hong Bo Zhang ◽  
Wen Jun Zhang

Microfluidics has great control over the size and uniformity of microspheres, which has been widely used in fabrication of different types of microspheres such as core-shell microbubbles. Gelatin Methacrylate (GelMa) as a biodegradable material that is closely resemble to native extracellular matrix (ECM). Photocrosslinked GelMa microspheres have gained numerous concerns in biomedical applications especially in three-dimensional cell culture and tissue engineering. In this article, we presented a suitable core-shell cell carrier based on biocompatible GelMa microbubbles. Highly monodispersed microbubbles were fabricated using a non-planar flow focusing microfluidic device. Both intact and collapsed microbubbles morphology were characterized through scanning electron microscopy (SEM), where clear hollow structures were found resulting from the gas core collapsing during the manipulation process. Furthermore, human umbilical vein endothelial cells (HUVEC) were seeded in the existence of microbubbles. Cells adhesion, migration and proliferation were observed in one week. It was notable that cells maintained high level of cell viability throughout the experiment. GelMa microbubble surface was also covered with cells, which became a facile carrier for cell culturing and targeted cell delivery.


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