scholarly journals Optimized Cell Mixing Facilitates the Reproducible Bioprinting of Constructs with High Cell Viability

2021 ◽  
Vol 12 (1) ◽  
pp. 326
Author(s):  
Axel Pössl ◽  
David Hartzke ◽  
Peggy Schlupp ◽  
Frank E. Runkel

Bioprinting with cell-laden hydrogels (bioink) requires the careful mixing of cells with the hydrogel carrier to ensure that the bioink is homogeneous and functional, and the printing results are reproducible. Bioink preparation is therefore a critical process step that must accommodate the specific rheological properties of different bioinks. Here, we developed a reproducible method for the optimized mixing of cells and hydrogel carriers that can be integrated into current bioprinting processes. First, we tested and optimized different mixing devices for their effect on bioink homogeneity and rheological properties, resulting in a low-shear process for the preparation of homogenous bioinks. Based on these findings, we evaluated the impact of different cell densities on the rheological profile of bioinks according to shear and temperature, and estimated the impact of shear stress intensity and duration on 1.1B4 cells. Finally, we integrated the optimized mixing method into a current printing process and monitored the printed construct for 14 days to confirm cell viability. We found that the cell viability in the printed cell-laden constructs remained in excess of 91% after 14 days.

2018 ◽  
Vol 18 (2) ◽  
pp. 255-262 ◽  
Author(s):  
Aikebaier Maimaiti ◽  
Amier Aili ◽  
Hureshitanmu Kuerban ◽  
Xuejun Li

Aims: Gallic acid (GA) is generally distributed in a variety of plants and foods, and possesses cell growth-inhibiting activities in cancer cell lines. In the present study, the impact of GA on cell viability, apoptosis induction and possible molecular mechanisms in cultured A549 lung carcinoma cells was investigated. Methods: In vitro experiments showed that treating A549 cells with various concentrations of GA inhibited cell viability and induced apoptosis in a dose-dependent manner. In order to understand the mechanism by which GA inhibits cell viability, comparative proteomic analysis was applied. The changed proteins were identified by Western blot and siRNA methods. Results: Two-dimensional electrophoresis revealed changes that occurred to the cells when treated with or without GA. Four up-regulated protein spots were clearly identified as malate dehydrogenase (MDH), voltagedependent, anion-selective channel protein 1(VDAC1), calreticulin (CRT) and brain acid soluble protein 1(BASP1). VDAC1 in A549 cells was reconfirmed by western blot. Transfection with VDAC1 siRNA significantly increased cell viability after the treatment of GA. Further investigation showed that GA down regulated PI3K/Akt signaling pathways. These data strongly suggest that up-regulation of VDAC1 by GA may play an important role in GA-induced, inhibitory effects on A549 cell viability.


Metals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1085
Author(s):  
Williams Leiva ◽  
Norman Toro ◽  
Pedro Robles ◽  
Edelmira Gálvez ◽  
Ricardo Ivan Jeldres

This research aims to analyze the impact of sodium tripolyphosphate (STPP) as a rheological modifier of concentrated kaolin slurries in seawater at pH 8, which is characteristic of copper sulfide processing operations. The dispersion phenomenon was analyzed through chord length measurements using the focused beam reflectance measurement (FBRM) technique, complementing size distributions in unweighted and square-weighted modes. The reduction of the rheological properties was significant, decreasing from 231 Pa in a reagent-free environment to 80 Pa after the application of STPP. A frequency sweep in a linear viscoelastic regime indicated that by applying a characteristic dosage of 0.53 kg/t of STPP, the pulp before yielding increases its phase angle, which increases its liquid-like character. Measurements of the chord length verified the dispersion of particles, which showed an apparent increase in the proportion of fine particles and a reduction of the coarser aggregates when STPP was applied. Measurements of the zeta potential suggested that the high anionic charge of the reagent (pentavalent) increases the electrostatic repulsions between particles, overcoming the effect of cations in seawater. The results are relevant for the mining industry, especially when the deposits have high contents of complex gangues, such as clays, that increase the rheological properties. This increases the energy costs and water consumption needed for pumping the tailings from thickeners to the tailing storages facilities. The strategies that allow for the improvement of the fluidity and deformation of the tailings generate slack in order to maximize water recovery in the thickening stages.


Tumor Biology ◽  
2021 ◽  
Vol 43 (1) ◽  
pp. 11-26
Author(s):  
Maike Busch ◽  
Natalia Miroschnikov ◽  
Jaroslaw Thomas Dankert ◽  
Marc Wiesehöfer ◽  
Klaus Metz ◽  
...  

BACKGROUND: Retinoblastoma (RB) is the most common childhood eye cancer. Chemotherapeutic drugs such as etoposide used in RB treatment often cause massive side effects and acquired drug resistances. Dysregulated genes and miRNAs have a large impact on cancer progression and development of chemotherapy resistances. OBJECTIVE: This study was designed to investigate the involvement of retinoic acid receptor alpha (RARα) in RB progression and chemoresistance as well as the impact of miR-138, a potential RARα regulating miRNA. METHODS: RARα and miR-138 expression in etoposide resistant RB cell lines and chemotherapy treated patient tumors compared to non-treated tumors was revealed by Real-Time PCR. Overexpression approaches were performed to analyze the effects of RARα on RB cell viability, apoptosis, proliferation and tumorigenesis. Besides, we addressed the effect of miR-138 overexpression on RB cell chemotherapy resistance. RESULTS: A binding between miR-138 and RARα was shown by dual luciferase reporter gene assay. The study presented revealed that RARα is downregulated in etoposide resistant RB cells, while miR-138 is endogenously upregulated. Opposing RARα and miR-138 expression levels were detectable in chemotherapy pre-treated compared to non-treated RB tumor specimen. Overexpression of RARα increases apoptosis levels and reduces tumor cell growth of aggressive etoposide resistant RB cells in vitro and in vivo. Overexpression of miR-138 in chemo-sensitive RB cell lines partly enhances cell viability after etoposide treatment. CONCLUSIONS: Our findings show that RARα acts as a tumor suppressor in retinoblastoma and is downregulated upon etoposide resistance in RB cells. Thus, RARα may contribute to the development and progression of RB chemo-resistance.


Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1088
Author(s):  
Katarzyna Lipska ◽  
Agata Filip ◽  
Anna Gumieniczek

Malignant cells in chronic lymphocytic leukemia (CLL) show resistance to apoptosis, as well as to chemotherapy, which are related to deletions or mutations of TP53, high expression of MCL1 and BCL2 genes and other abnormalities. Thus, the main goal of the present study was to assess the impact of chlorambucil (CLB) combined with valproic acid (VPA), a known antiepileptic drug and histone deacetylation inhibitor, on apoptosis of the cells isolated from 17 patients with CLL. After incubation with CLB (17.5 µM) and VPA (0.5 mM), percentage of apoptosis, as well as expression of two TP53 target genes (p21 and HDM2) and two genes from Bcl-2 family (BCL2 and MCL1), were tested. As a result, an increased percentage of apoptosis was observed for CLL cells treated with CLB and VPA, and with CLB alone. Under the treatment with the drug combination, the expression of p21 gene was visibly higher than under the treatment with CLB alone. At the same time, the cultures under CLB treatment showed visibly higher expression of BCL2 than the cultures with VPA alone. Thus, the present study strongly suggests further investigations on the CLB and VPA combination in CLL treatment.


BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Aleksandra Majchrzak-Celińska ◽  
Julia O. Misiorek ◽  
Nastassia Kruhlenia ◽  
Lukasz Przybyl ◽  
Robert Kleszcz ◽  
...  

Abstract Background Glioblastoma (GBM) is the deadliest and the most common primary brain tumor in adults. The invasiveness and proliferation of GBM cells can be decreased through the inhibition of Wnt/β-catenin pathway. In this regard, celecoxib is a promising agent, but other COXIBs and 2,5-dimethylcelecoxib (2,5-DMC) await elucidation. Thus, the aim of this study was to analyze the impact of celecoxib, 2,5-DMC, etori-, rofe-, and valdecoxib on GBM cell viability and the activity of Wnt/β-catenin pathway. In addition, the combination of the compounds with temozolomide (TMZ) was also evaluated. Cell cycle distribution and apoptosis, MGMT methylation level, COX-2 and PGE2 EP4 protein levels were also determined in order to better understand the molecular mechanisms exerted by these compounds and to find out which of them can serve best in GBM therapy. Methods Celecoxib, 2,5-DMC, etori-, rofe- and valdecoxib were evaluated using three commercially available and two patient-derived GBM cell lines. Cell viability was analyzed using MTT assay, whereas alterations in MGMT methylation level were determined using MS-HRM method. The impact of COXIBs, in the presence and absence of TMZ, on Wnt pathway was measured on the basis of the expression of β-catenin target genes. Cell cycle distribution and apoptosis analysis were performed using flow cytometry. COX-2 and PGE2 EP4 receptor expression were evaluated using Western blot analysis. Results Wnt/β-catenin pathway was attenuated by COXIBs and 2,5-DMC irrespective of the COX-2 expression profile of the treated cells, their MGMT methylation status, or radio/chemoresistance. Celecoxib and 2,5-DMC were the most cytotoxic. Cell cycle distribution was altered, and apoptosis was induced after the treatment with celecoxib, 2,5-DMC, etori- and valdecoxib in T98G cell line. COXIBs and 2,5-DMC did not influence MGMT methylation status, but inhibited COX-2/PGE2/EP4 pathway. Conclusions Not only celecoxib, but also 2,5-DMC, etori-, rofe- and valdecoxib should be further investigated as potential good anti-GBM therapeutics.


Author(s):  
Tetsuo Takehara ◽  
Naoki Mizutani ◽  
Hayato Hikita ◽  
Yoshinobu Saito ◽  
Yuta Myojin ◽  
...  

Grb2-associated binder 1 (Gab1) is an adaptor protein that is important for intracellular signal transduction by receptor tyrosine kinases that are receptors for various growth factors and plays an important role in rapid liver regeneration after partial hepatectomy and during acute hepatitis. On the other hand, mild liver regeneration is induced in livers of individuals with chronic hepatitis, where hepatocyte apoptosis is persistent; however, the impact of Gab1 on such livers remains unclear. We examined the role of Gab1 in chronic hepatitis. Gab1 knockdown enhanced the decrease in cell viability and apoptosis induced by ABT-737, a Bcl-2/-xL/-w inhibitor, in BNL.CL2 cells, while cell viability and caspase activity were unchanged in the absence of ABT-737. ABT-737 treatment induced Gab1 cleavage to form p35-Gab1. p35-Gab1 was also detected in the livers of mice with hepatocyte-specific Mcl-1 knockout (KO), which causes persistent hepatocyte apoptosis. Gab1 deficiency exacerbated hepatocyte apoptosis in Mcl-1 KO mice with posttranscriptional downregulation of Bcl-XL. In BNL.CL2 cells treated with ABT-737, Gab1 knockdown posttranscriptionally suppressed Bcl-xL expression, and p35-Gab1 overexpression enhanced Bcl-xL expression. Gab1 deficiency in Mcl-1 KO mice activated STAT3 signaling in hepatocytes, increased hepatocyte proliferation, and increased the incidence of liver cancer with the exacerbation of liver fibrosis. In conclusion, Gab1 is cleaved in the presence of apoptotic stimuli and forms p35-Gab1 in hepatocytes. In chronic liver injury, the role of Gab1 in suppressing apoptosis and reducing liver damage, fibrosis, and tumorigenesis is more important than its role in liver regeneration.


2018 ◽  
Vol 234 ◽  
pp. 04004 ◽  
Author(s):  
Alexey Lobiak ◽  
Andrii Plugin ◽  
Larisa Kravtsiv ◽  
Oksana Kovalova

The paper presents a computer modelling technique for modernization of bridgework operations by building-up a mounted reinforced concrete slab. It implements the technique of the evolutionary transformation of a model in one calculation cycle with redistribution of forces between the elements of the built-up section, and consideration of the impact of elastoplastic and rheological properties of the materials. Consideration of the concrete creep implies the application of the generalized kinetic curve of prolonged deformation and phenomenological deformation development equations based on the colloid-chemical concept of the prolonged concrete deformation mechanism. The creep control was implemented through new structural coefficients which determined the structure of matrix interlayers between the sand grains, and mortar between the crushed stone grains. The technique proposed was realized in the program complex “LIRA-SAPR” based on the building information modelling (BIM) and the finite element method (FEM). The multistage modelling technique was shown by an example of calculation of a motorway bridge slab span within the transport structures under modernization along the Lev Landau Avenue in Kharkiv (Ukraine).


2018 ◽  
Vol 21 (3) ◽  
pp. 407-422 ◽  
Author(s):  
Débora da Costa Simões ◽  
José Vicente Caixeta-Filho ◽  
Udatta S. Palekar

This study analyzes the impact of anticipated investments to alter Brazilian port infrastructure on fertilizer flows and fertilizer transportation logistics costs using a linear programming model designed for the task. The most notable among these investments are directed toward accelerating port development in Brazil’s “Northern-Arc”, thereby increasing fertilizer supply to new markets opening throughout the country’s expanding agricultural frontier, particularly in northern Mato Grosso state, while increasing supply to existing markets. Results from model runs show that the anticipated port infrastructure investments should ensure nationwide fertilizer logistics savings of US$ 845 million over the 2017 through 2025 period. Although these estimated benefits are outstanding, the study indicates that further expansion of Brazil’s port system, particularly in the Northern-Arc, presents additional opportunities. Model projections were that in 2025, after all planned infrastructure improvements are operational, port terminals will be near full capacity, which should make planning for future projects a current priority.


2021 ◽  
Vol 877 (1) ◽  
pp. 012047
Author(s):  
Hind M. Ewadh ◽  
Mustafa J. Al Imari ◽  
Sabrean F. Jawad ◽  
Hayfaa A. Mubarak

Abstract A modest quantity of fluoride can increase the mineralization of teeth and reduce their cavities. But the presomerence of fluoride in excess in water can lead to severe disease infertility. In the past few decades, scientists have thus been preoccupied with developing ways to reduce sewage fluoride concentrations and reduce their effects on human health. The present study is aimed at using the technology of electrocoagulation to remove fluoride from polluted water. Tests have been done to examine the elimination of fluoride with a rectangular electrocoagulation cell and examine the impact of the experimental aspects on fluoride extraction, specifically electrical current, electrode spacing, and pH. The authors found that 93% of the fluoride has been extracted using 5mm spaced electrodes with a current density of 2 mA/cm2 and a level of pH of 7 from the polluted water after 20 min of processing. Experimental factors considerably impact the efficacy of fluoride removal. In the acidic environment, greater effectiveness of fluoride removal is being attained. The elimination effectiveness depends directly on the electric current, whereas the distance between poles is adversely linked to fluoride elimination.


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