scholarly journals Development and Characterization of Novel Porous 3D Alginate-Cockle Shell Powder Nanobiocomposite Bone Scaffold

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
B. Hemabarathy Bharatham ◽  
Md. Zuki Abu Bakar ◽  
Enoch Kumar Perimal ◽  
Loqman Mohamed Yusof ◽  
Muhajir Hamid
Keyword(s):  
Bone Growth ◽  
Three Dimensional ◽  
Future Application ◽  
Bone Scaffold ◽  
Composition Ratios ◽  
Evaluation Techniques ◽  
Shell Powder ◽  
Cockle Shell

A novel porous three-dimensional bone scaffold was developed using a natural polymer (alginate/Alg) in combination with a naturally obtained biomineral (nano cockle shell powder/nCP) through lyophilization techniques. The scaffold was developed in varying composition mixture of Alg-nCP and characterized using various evaluation techniques as well as preliminaryin vitrostudies on MG63 human osteoblast cells. Morphological observations using SEM revealed variations in structures with the use of different Alg-nCP composition ratios. All the developed scaffolds showed a porous structure with pore sizes ideal for facilitating new bone growth; however, not all combination mixtures showed subsequent favorable characteristics to be used for biological applications. Scaffolds produced using the combination mixture of 40% Alg and 60% nCP produced significantly promising results in terms of mechanical strength, degradation rate, and increased cell proliferation rates making it potentially the optimum composition mixture of Alg-nCP with future application prospects.

scholarly journals In Vitro Selection and Characterization of Hepatitis C Virus Serine Protease Variants Resistant to an Active-Site Peptide Inhibitor

2003 ◽  
Vol 77 (6) ◽  
pp. 3669-3679 ◽  
Author(s):  
Caterina Trozzi ◽  
Linda Bartholomew ◽  
Alessandra Ceccacci ◽  
Gabriella Biasiol ◽  
Laura Pacini ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Serine Protease ◽  
In Vitro Selection ◽  
Three Dimensional ◽  
Host Cells ◽  
Dimensional Structure ◽  
In Vitro Systems

ABSTRACT The hepatitis C virus (HCV) serine protease is necessary for viral replication and represents a valid target for developing new therapies for HCV infection. Potent and selective inhibitors of this enzyme have been identified and shown to inhibit HCV replication in tissue culture. The optimization of these inhibitors for clinical development would greatly benefit from in vitro systems for the identification and the study of resistant variants. We report the use HCV subgenomic replicons to isolate and characterize mutants resistant to a protease inhibitor. Taking advantage of the replicons' ability to transduce resistance to neomycin, we selected replicons with decreased sensitivity to the inhibitor by culturing the host cells in the presence of the inhibitor and neomycin. The selected replicons replicated to the same extent as those in parental cells. Sequence analysis followed by transfection of replicons containing isolated mutations revealed that resistance was mediated by amino acid substitutions in the protease. These results were confirmed by in vitro experiments with mutant enzymes and by modeling the inhibitor in the three-dimensional structure of the protease.

Preparation and Characterization of Porous Nanosized Hydroxyapatite/Collagen Composite as Bone Scaffold

2013 ◽  
Vol 647 ◽  
pp. 62-66 ◽  
Author(s):  
Ting Li Lu ◽  
Yu Hui Li ◽  
Yun Qing Wang ◽  
Yu Fan Ma
Keyword(s):  
Porous Scaffold ◽  
Good Mechanical Property ◽  
Bone Scaffold ◽  
Binding Agent ◽  
Average Pore ◽  
Bioactive Materials ◽  
Physical Chemical ◽  
Composite Nature

Inorganic-organic composites could mimic the composite nature of real bone and combine the toughness of a polymer with the strength of an inorganic one to generate bioactive materials with improved mechanical properties and degradation profiles. In this paper, HAp/Col porous scaffold was prepared based on inorganic nano-sized hydoroxyapatite (nHAp) and organic collagen (Col) by solvent casting/particulate leaching. Sodium chloride (NaCl) and ethyl cellulose (EC) were performed as the porogenic agent and binding agent, respectively. The physical, chemical and biodegradation property of this scaffold were investigated in vitro and its co-culture with cells was also studied. The results showed that the scaffold had good mechanical property with the average pore sizes about 150 μm and porosities as high as 75%. This nHAp/Col porous scaffold had no cytotoxicity to mouse pre-osteoblast MC3T3-E1 and the content of alkaline phosphatase (ALP) was ascending with the extension of culture time. The results of mineralization indicated that HAp/Col scaffold could promote the proliferation, differentiation and biological mineralization of MC3T3-E1.

scholarly journals Functional Characterization of Three-Dimensional Cortical Cultures for In Vitro Modeling of Brain Networks

iScience ◽  
2020 ◽  
Vol 23 (8) ◽  
pp. 101434
Author(s):  
Yu-Ting L. Dingle ◽  
Volha Liaudanskaya ◽  
Liam T. Finnegan ◽  
Kyler C. Berlind ◽  
Craig Mizzoni ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Brain Networks ◽  
Three Dimensional ◽  
In Vitro Modeling ◽  
Cortical Cultures

Design and Characterization of a Fluorescent Reporter Enabling Live-cell Monitoring of MCT8 Expression

2021 ◽  
Author(s):  
Adina Sophie Graffunder ◽  
Sarah Paisdzior ◽  
Robert Opitz ◽  
Kostja Renko ◽  
Peter Kühnen ◽  
...  
Keyword(s):  
Expression Patterns ◽  
Functional Characterization ◽  
Live Cell ◽  
Monocarboxylate Transporter ◽  
Start Codon ◽  
Future Application ◽  
Gene Assay ◽  
Cell Monitoring

AbstractThe monocarboxylate transporter 8 (MCT8) is a specific thyroid hormone transporter and plays an essential role in fetal development. Inactivating mutations in the MCT8 encoding gene SLC16A2 (solute carrier family 16, member 2) lead to the Allan-Herndon-Dudley syndrome, a condition presenting with severe endocrinological and neurological phenotypes. However, the cellular distribution pattern and dynamic expression profile are still not well known for early human neural development. Objective Development and characterization of fluorescent MCT8 reporters that would permit live-cell monitoring of MCT8 protein expression in vitro in human induced pluripotent stem cell (hiPSC)-derived cell culture models. Methods A tetracysteine (TC) motif was introduced into the human MCT8 sequence at four different positions as binding sites for fluorescent biarsenical dyes. Human Embryonic Kidney 293 cells were transfected and stained with fluorescein-arsenical hairpin-binder (FlAsH). Counterstaining with specific MCT8 antibody was performed. Triiodothyronine (T3) uptake was indirectly measured with a T3 responsive luciferase-based reporter gene assay in Madin-Darby Canine Kidney 1 cells for functional characterization. Results FlAsH staining and antibody counterstaining of all four constructs showed cell membrane expression of all MCT8 constructs. The construct with the tag after the first start codon demonstrated comparable T3 uptake to the MCT8 wildtype. Conclusion Our data indicate that introduction of a TC-tag directly after the first start codon generates a MCT8 reporter with suitable characteristics for live-cell monitoring of MCT8 expression. One promising future application will be generation of stable hiPSC MCT8 reporter lines to characterize MCT8 expression patterns during in vitro neuronal development.

scholarly journals Characterization of the proteasome from the extremely halophilic archaeonHaloarcula marismortui

Archaea ◽  
2002 ◽  
Vol 1 (1) ◽  
pp. 53-61 ◽  
Author(s):  
B. Franzetti ◽  
G. Schoehn ◽  
D. Garcia ◽  
R. W. H. Ruigrok ◽  
G. Zaccai
Keyword(s):  
Three Dimensional ◽  
Halophilic Archaea ◽  
20S Proteasome ◽  
Misfolded Protein ◽  
Haloarcula Marismortui ◽  
Low Salt ◽  
Dimensional Reconstruction ◽  
The Stability

A 20S proteasome, comprising two subunits α and β, was purified from the extreme halophilic archaeonHaloarcula marismortui, which grows only in saturated salt conditions. The three-dimensional reconstruction of theH. marismortuiproteasome (Hm proteasome), obtained from negatively stained electron micrographs, is virtually identical to the structure of a thermophilic proteasome filtered to the same resolution. The stability of the Hm proteasome was found to be less salt-dependent than that of other halophilic enzymes previously described. The proteolytic activity of the Hm proteasome was investigated using the malate dehydrogenase fromH. marismortui(HmMalDH) as a model substrate. The HmMalDH denatures when the salt concentration is decreased below 2 M. Under these conditions, the proteasome efficiently cleaves HmMalDH during its denaturation process, but the fully denatured HmMalDH is poorly degraded. These in vitro experiments show that, at low salt concentrations, the 20S proteasome from halophilic archaea eliminates a misfolded protein.

scholarly journals Novel Image Analysis Approach Quantifies Morphological Characteristics of 3D Breast Culture Acini with Varying Metastatic Potentials

2012 ◽  
Vol 2012 ◽  
pp. 1-16 ◽  
Author(s):  
Lindsey McKeen Polizzotti ◽  
Basak Oztan ◽  
Chris S. Bjornsson ◽  
Katherine R. Shubert ◽  
Bülent Yener ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Image Analysis ◽  
Three Dimensional ◽  
Morphological Characteristics ◽  
Tissue Samples ◽  
Metastatic Cascade ◽  
Automated Grading

Prognosis of breast cancer is primarily predicted by the histological grading of the tumor, where pathologists manually evaluate microscopic characteristics of the tissue. This labor intensive process suffers from intra- and inter-observer variations; thus, computer-aided systems that accomplish this assessment automatically are in high demand. We address this by developing an image analysis framework for the automated grading of breast cancer inin vitrothree-dimensional breast epithelial acini through the characterization of acinar structure morphology. A set of statistically significant features for the characterization of acini morphology are exploited for the automated grading of six (MCF10 series) cell line cultures mimicking three grades of breast cancer along the metastatic cascade. In addition to capturing both expected and visually differentiable changes, we quantify subtle differences that pose a challenge to assess through microscopic inspection. Our method achieves 89.0% accuracy in grading the acinar structures as nonmalignant, noninvasive carcinoma, and invasive carcinoma grades. We further demonstrate that the proposed methodology can be successfully applied for the grading ofin vivotissue samples albeit with additional constraints. These results indicate that the proposed features can be used to describe the relationship between the acini morphology and cellular function along the metastatic cascade.

scholarly journals Development and characterization of a human three-dimensional chondrosarcoma culture for in vitro drug testing

PLoS ONE ◽  
2017 ◽  
Vol 12 (7) ◽  
pp. e0181340 ◽  
Author(s):  
Aurélien Voissiere ◽  
Elodie Jouberton ◽  
Elise Maubert ◽  
Françoise Degoul ◽  
Caroline Peyrode ◽  
...  
Keyword(s):  
Drug Testing ◽  
Three Dimensional
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