scholarly journals Quantification of Cigarette Smoke Particle DepositionIn VitroUsing a Triplicate Quartz Crystal Microbalance Exposure Chamber

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Jason Adamson ◽  
David Thorne ◽  
John McAughey ◽  
Deborah Dillon ◽  
Clive Meredith
Keyword(s):  
Quartz Crystal Microbalance ◽  
Cell Cultures ◽  
Quartz Crystal ◽  
Biological Effects ◽  
Exposure Chamber ◽  
Regional Deposition ◽  
Respiratory Toxicology ◽  
A Cell ◽  
Air Liquid Interface

There are a variety of smoke exposure systems available to the tobacco industry and respiratory toxicology research groups, each with their own way of diluting/delivering smoke to cell cultures. Thus a simple technique to measure dosein vitroneeds to be utilised. Dosimetry—assessment of dose—is a key element in linking the biological effects of smoke generated by various exposure systems. Microbalance technology is presented as a dosimetry tool and a way of measuring whole smoke dose. Described here is a new tool to quantify diluted smoke particulate depositionin vitro. The triplicate quartz crystal microbalance (QCM) chamber measured real-time deposition of smoke at a range of dilutions 1 : 5–1 : 400 (smoke : air). Mass was read in triplicate by 3 identical QCMs installed into onein vitroexposure chamber, each in the location in which a cell culture would be exposed to smoke at the air-liquid interface. This resulted in quantification of deposited particulate matter in the range 0.21–28.00 μg/cm2. Results demonstrated that the QCM could discriminate mass between dilutions and was able to give information of regional deposition where cell cultures would usually be exposed within the chamber. Our aim is to use the QCM to support the preclinical (in vitro) evaluation of tobacco products.

Particle deposition efficiency at air–liquid interface of a cell exposure chamber

2015 ◽  
Vol 81 ◽  
pp. 90-99 ◽  
Author(s):  
Yuji Fujitani ◽  
Yutaka Sugaya ◽  
Masanori Hashiguchi ◽  
Akiko Furuyama ◽  
Seishiro Hirano ◽  
...  
Keyword(s):  
Particle Deposition ◽  
Deposition Efficiency ◽  
Liquid Interface ◽  
Exposure Chamber ◽  
A Cell ◽  
Air Liquid Interface

scholarly journals In vitro selection of N peptide-binding RNA on a quartz-crystal microbalance

2002 ◽  
Vol 2 (1) ◽  
pp. 207-208
Author(s):  
A. Murakawa ◽  
S. Fukusho ◽  
H. Furusawa ◽  
Y. Okahata
Keyword(s):  
Quartz Crystal Microbalance ◽  
Quartz Crystal ◽  
In Vitro Selection ◽  
Peptide Binding ◽  
Selection Of

MATTERS NEEDING ATTENTION FOR APPLYING THE QUARTZ CRYSTAL MICROBALANCE TECHNIQUE TO DETECT THE CELL MORPHOLOGY

2009 ◽  
Vol 21 (06) ◽  
pp. 415-420 ◽  
Author(s):  
Hung-Che Chou ◽  
Tsong-Rong Yan ◽  
Chao-Fa Lee
Keyword(s):  
Experimental Data ◽  
Cell Culture ◽  
System Design ◽  
Quartz Crystal Microbalance ◽  
Quartz Crystal ◽  
Important Application ◽  
The Other ◽  
Cell Detection ◽  
A Cell ◽  
Culture Environment

Using quartz crystal microbalance (QCM) to design a cell detection platform is an important application of QCM. In the process of system design and application, there still have some points, which are easily overlooked or are not understood by researchers. Most of the problems can be separated into three types. One is the cell culture environment conditions that affect the system signals, another is the system unification or lack of communication over regulations, and the other is the over-analysis of the experimental data.

scholarly journals Human Keratinocyte Growth and Differentiation on Acellular Porcine Dermal Matrix in relation to Wound Healing Potential

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Robert Zajicek ◽  
Vaclav Mandys ◽  
Ondrej Mestak ◽  
Jan Sevcik ◽  
Radana Königova ◽  
...  
Keyword(s):  
Wound Healing ◽  
Human Keratinocytes ◽  
Dermal Matrix ◽  
Keratinocyte Proliferation ◽  
Proliferation And Differentiation ◽  
A Cell ◽  
Cell Culture Assay ◽  
Air Liquid Interface

A number of implantable biomaterials derived from animal tissues are now used in modern surgery. Xe-Derma is a dry, sterile, acellular porcine dermis. It has a remarkable healing effect on burns and other wounds. Our hypothesis was that the natural biological structure of Xe-Derma plays an important role in keratinocyte proliferation and formation of epidermal architecturein vitroas well asin vivo. The bioactivity of Xe-Derma was studied by a cell culture assay. We analyzed growth and differentiation of human keratinocytes culturedin vitroon Xe-Derma, and we compared the results with formation of neoepidermis in the deep dermal wounds treated with Xe-Derma. Keratinocytes cultured on Xe-Derma submerged in the culture medium achieved confluence in 7–10 days. After lifting the cultures to the air-liquid interface, the keratinocytes were stratified and differentiated within one week, forming an epidermis with basal, spinous, granular, and stratum corneum layers. Immunohistochemical detection of high-molecular weight cytokeratins (HMW CKs), CD29, p63, and involucrin confirmed the similarity of organization and differentiation of the cultured epidermal cells to the normal epidermis. The results suggest that the firm natural structure of Xe-Derma stimulates proliferation and differentiation of human primary keratinocytes and by this way improves wound healing.

scholarly journals Quartz Crystal Microbalance as a Sensor to Characterize Macromolecular Assembly Dynamics

2009 ◽  
Vol 2009 ◽  
pp. 1-17 ◽  
Author(s):  
K. Kanazawa ◽  
Nam-Joon Cho
Keyword(s):  
Cell Membrane ◽  
Quartz Crystal Microbalance ◽  
Quartz Crystal ◽  
Analytical Techniques ◽  
Top Down ◽  
Macromolecular Assembly ◽  
Basic Measurement ◽  
Quartz Crystal Microbalance Sensor ◽  
A Cell ◽  
Peptide Interaction

The quartz crystal microbalance sensor has a resonant frequency and a quality factor which can be used to probe the properties of nanometer thick film loads. A recent review by Arnau (2008) has discussed many of the considerations necessary to accurately probe for these properties. To avoid needless duplication but to still provide an adequate background for the new user, we briefly outline the basic measurement methodologies and analytical techniques that were covered in the review. Details will be provided on some specific perspectives of the authors. For example, the special precautions necessary when dealing with soft films (polymeric and biological) under liquid are overviewed. To illustrate applications of the QCM technique, simple bilayer and vesicle behaviors are discussed, along with the structural transformation resulting from protein adsorption onto an intact vesicle adlayer. The amphipathic -helical (AH) peptide interaction is given as a particular example. Lastly, we summarize a top-down approach to functionalize a surface with a cell membrane and to study its interaction with proteins.

scholarly journals A Custom-Made Device for Reproducibly Depositing Pre-metered Doses of Nebulized Drugs on Pulmonary Cells in vitro

2021 ◽  
Vol 9 ◽  
Author(s):  
Justus C. Horstmann ◽  
Chelsea R. Thorn ◽  
Patrick Carius ◽  
Florian Graef ◽  
Xabier Murgia ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Pharmaceutical Formulations ◽  
Barrier Properties ◽  
Liquid Interface ◽  
Bronchial Epithelial ◽  
Pulmonary Epithelial Cells ◽  
Custom Made ◽  
Air Liquid Interface ◽  
Phosphate Buffered Saline

The deposition of pre-metered doses (i.e., defined before and not after exposition) at the air–liquid interface of viable pulmonary epithelial cells remains an important but challenging task for developing aerosol medicines. While some devices allow quantification of the deposited dose after or during the experiment, e.g., gravimetrically, there is still no generally accepted way to deposit small pre-metered doses of aerosolized drugs or pharmaceutical formulations, e.g., nanomedicines. Here, we describe a straightforward custom-made device, allowing connection to commercially available nebulizers with standard cell culture plates. Designed to tightly fit into the approximately 12-mm opening of either a 12-well Transwell® insert or a single 24-well plate, a defined dose of an aerosolized liquid can be directly deposited precisely and reproducibly (4.8% deviation) at the air–liquid interface (ALI) of pulmonary cell cultures. The deposited dose can be controlled by the volume of the nebulized solution, which may vary in a range from 20 to 200 μl. The entire nebulization-deposition maneuver is completed after 30 s and is spatially homogenous. After phosphate-buffered saline (PBS) deposition, the viability and barrier properties transepithelial electrical resistance (TEER) of human bronchial epithelial Calu-3 cells were not negatively affected. Straightforward in manufacture and use, the device enables reproducible deposition of metered doses of aerosolized drugs to study the interactions with pulmonary cell cultures grown at ALI conditions.

In Vitro High-Content Imaging-Based Phenotypic Analysis of Bronchial 3D Organotypic Air–Liquid Interface Cultures

2020 ◽  
Vol 25 (3) ◽  
pp. 247-252
Author(s):  
Diego Marescotti ◽  
David Bovard ◽  
Moran Morelli ◽  
Antonin Sandoz ◽  
Karsta Luettich ◽  
...  
Keyword(s):  
Biological Effects ◽  
Liquid Interface ◽  
Phenotypic Analysis ◽  
Cell Hyperplasia ◽  
High Content Imaging ◽  
Differentiation Status ◽  
Third Dimension ◽  
Air Liquid Interface ◽  
Ciliated Epithelial Cells

High-content imaging (HCI) is a powerful method for quantifying biological effects in vitro. Historically, HCI has been applied to adherent cells growing in monolayers. With the advent of confocal versions of HCI devices, researchers now have the option of performing analyses on 3D cell cultures. However, some obstacles remain in integrating the third dimension, such as limited light penetration and less sophisticated image analysis. Here, we report the development of an HCI technique for imaging human bronchial 3D organotypic air–liquid interface (ALI) cultures (hBR-ALI). In this method, we monitored differentiation status through HCI evaluation markers representative of ciliated epithelial cells and goblet cells (Muc5AC [mucin 5AC]). As a second use case for demonstrating the utility of this technique, we induced goblet cell hyperplasia in hBR-ALI by using interleukin (IL)-13. Our results demonstrate the utility of the HCI technique for imaging hBR-ALI grown on Transwell inserts. This technique may be expanded to other cell culture systems, such as skin epithelia and 3D intestinal systems.

Expression of the heparin-binding cytokines, midkine (MK) and HB-GAM (pleiotrophin) is associated with epithelial-mesenchymal interactions during fetal development and organogenesis

Development ◽  
1995 ◽  
Vol 121 (1) ◽  
pp. 37-51 ◽  
Author(s):  
T.A. Mitsiadis ◽  
M. Salmivirta ◽  
T. Muramatsu ◽  
H. Muramatsu ◽  
H. Rauvala ◽  
...  
Keyword(s):  
Solid Phase ◽  
Biological Effects ◽  
Mesenchymal Cell ◽  
Basement Membranes ◽  
Dependent Manner ◽  
Heparin Binding ◽  
Limb Buds ◽  
New Family ◽  
A Cell

Midkine (MK) and heparin binding-growth associated molecule (HB-GAM or pleiotrophin), constitute a new family of heparin-binding proteins implicated in the regulation of growth and differentiation (T. Muramatsu (1993) Int. J. Dev. Biol. 37, 183–188). We used affinity-purified antibodies against MK and HB-GAM to analyze their distribution during mouse embryonic development. From 9 to 14.5 day post-coitum (dpc), both proteins were detected in central and peripheral nervous systems, facial processes, limb buds, sense organs, respiratory, digestive, urogenital, and skeletal systems. MK and HB-GAM were often localized on the surface of differentiating cells and in basement membranes of organs undergoing epithelial-mesenchymal interactions. The level of MK protein decreased considerably in the 16.5 dpc embryo, whereas HB-GAM staining persisted in many tissues. Our in situ hybridization results revealed a widespread expression of MK transcripts that was not always consistent with the distribution of MK protein in developing tissues. In many epithelio-mesenchymal organs MK and HB-GAM were codistributed with syndecan-1, a cell surface proteoglycan. In limb buds and facial processes, MK, HB-GAM, and syndecan-1 were localized to the apical epithelium and the adjacent proliferating mesenchyme. Both MK and HB-GAM bound syndecan-1 in solid-phase assays in a heparan sulfate-dependent manner. The biological effects of MK and HB-GAM on limb and facial mesenchyme were studied in vitro by application of beads preloaded with the proteins. Neither MK nor HB-GAM stimulated mesenchymal cell proliferation or induced syndecan-1 expression. Taken together these results indicate that MK and HB-GAM may play regulatory roles in differentiation and morphogenesis of the vertebrate embryo, particularly in epithelio-mesenchymal organs, and suggest molecular interactions with syndecan-1.

scholarly journals Protein Adsorption to Titanium and Zirconia Using a Quartz Crystal Microbalance Method

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
You Kusakawa ◽  
Eiji Yoshida ◽  
Tohru Hayakawa
Keyword(s):  
Surface Roughness ◽  
Protein Adsorption ◽  
Quartz Crystal Microbalance ◽  
Quartz Crystal ◽  
Adsorption Rate ◽  
Force Microscopy ◽  
Atomic Force ◽  
A Cell ◽  
Addition Amount

Protein adsorption onto titanium (Ti) or zirconia (ZrO2) was evaluated using a 27 MHz quartz crystal microbalance (QCM). As proteins, fibronectin (Fn), a cell adhesive protein, and albumin (Alb), a cell adhesion-inhibiting protein, were evaluated. The Ti and ZrO2 sensors for QCM were characterized by atomic force microscopy and electron probe microanalysis observation, measurement of contact angle against water, and surface roughness. The amounts of Fn and Alb adsorbed onto the Ti and ZrO2 sensors and apparent reaction rate were obtained using QCM measurements. Ti sensor showed greater adsorption of Fn and Alb than the ZrO2 sensor. In addition, amount of Fn adsorbed onto the Ti or ZrO2 sensors was higher than that of Alb. The surface roughness and hydrophilicity of Ti or ZrO2 may influence the adsorption of Fn or Alb. With regard to the adsorption rate, Alb adsorbed more rapidly than Fn onto Ti. Comparing Ti and ZrO2, Alb adsorption rate to Ti was faster than that to ZrO2. Fn adsorption will be effective for cell activities, but Alb adsorption will not. QCM method could simulate in vivo Fn and Alb adsorption to Ti or ZrO2.

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