JoVE Bioengineering merges both physical and life sciences to understand and predict biological processes. Applying physical science tools to life science questions allow for the discovery of better technologies to measure, diagnose, and clinically treat disease.
1Department of Orthopaedic Surgery, University of California Los Angeles, 2Department of Bioengineering, University of California Los Angeles
Animal models are important tools for the evaluation of tissue-engineered grafts. This paper presents the protocol for preparing an electrospun biodegradable polymer graft for use in anterior cruciate ligament tissue engineering, as well as a surgical protocol for implantation in a rat model.
Published March 26, 2015. Keywords: Bioengineering, Anterior cruciate ligament, tissue engineering, animal model, biodegradable scaffold, rat, knee
1Biomedical Engineering Department, Saint Louis University
Here, a method that enables quick, efficient, and inexpensive preparation of polyacrylamide gels in a multiwell plate format is described. The method does not require any specialized equipment and could be easily adopted by any research laboratory. It would be particularly useful in research focused on understanding stiffness-dependent cell responses.
Published March 25, 2015. Keywords: Bioengineering, Multiwell, substrate stiffness, drug screening, polyacrylamide, Young’s modulus, high-throughput
1Cardiovascular Division, King's College London BHF Centre
Here, we present a protocol to generate tissue engineered vessel grafts that are functional for grafting into mice by double seeding partially induced pluripotent stem cell (PiPSC) - derived smooth muscle cells and PiPSC - derived endothelial cells on a decellularized vessel scaffold bioreactor.
Published March 18, 2015. Keywords: Bioengineering, stem cells, partially induced pluripotent stem cells, tissue engineering, bioreactor, vascular differentiation, vessel graft, mouse models
1Division of Chemistry and Chemical Engineering, California Institute of Technology, 2Department of Molecular Medicine, Beckman Research Institute of the City of Hope
Gallium(III) 5,10,15-(tris)pentafluorophenylcorrole and its freebase analogue exhibit low micromolar cell cytotoxicity. This manuscript describes an RNA transcription reaction, imaging RNA with an ethidium bromide-stained gel, and quantifying RNA with UV-Vis spectroscopy, in order to assess transcription inhibition by corroles and demonstrates a straightforward method of evaluating anticancer candidate properties.
Published March 18, 2015. Keywords: Bioengineering, Corrole, RNA, transcription, inhibition, anti-cancer, DNA, binding, Actinomycin D, triptolide
1Bredesen Center, University of Tennessee, Knoxville, 2Center for Nanophase Materials Sciences, Oak Ridge National Laboratory, 3Department of Materials Science and Engineering, University of Tennessee, Knoxville
A microfabricated device with sealable femtoliter-volume reaction chambers is described. This report includes a protocol for sealing cell-free protein synthesis reactants inside these chambers for the purpose of understanding the role of crowding and confinement in gene expression.
Published March 11, 2015. Keywords: Bioengineering, Cell-free, synthetic biology, microfluidics, noise biology, soft lithography, femtoliter volumes
1Department of Civil and Environmental Engineering, Northwestern University, 2Department of Chemical and Biological Engineeering, Northwestern University, 3Department of Applied Mathematics and Engineering Sciences, Northwestern University
Biofilms have complex interactions with their surrounding environment. To comprehensively investigate biofilm-environment interactions, we present here a series of methods to create heterogeneous chemical environment for biofilm development, to quantify local flow velocity, and to analyze mass transport in and around biofilm colonies.
Published March 11, 2015. Keywords: Bioengineering, microfluidic flow cell, chemical gradient, biofilm development, particle tracking, flow characterization, fluorescent tracer, solute transport
1Department of Chemistry and Applied Biosciences, ETH Zurich
We present a discrete droplet sample introduction system for inductively coupled plasma mass spectrometry (ICPMS). It is based on a cheap and disposable microfluidic chip that generates highly monodisperse droplets in a size range of 40−60 µm at frequencies from 90 to 7,000 Hz.
Published March 5, 2015. Keywords: Bioengineering, mass spectrometry, ICPMS, microfluidics, droplet microfluidics, monodisperse, sample introduction, chip, red blood cells, erythrocytes, single cell analysis
1Department for Tissue Engineering and Regenerative Medicine, University Hospital Würzburg, 2Translational Center Würzburg, Regenerative Therapies in Oncology and Musculoskelettal Disease, Würzburg Branch of the Fraunhofer-Institute Interfacial Engineering and Biotechnology, IGB
The goal of this protocol is to build up a three-dimensional full thickness skin equivalent, which resembles natural skin. With a specifically constructed automated wounding device, precise and reproducible wounds can be generated under maintenance of sterility.
Published February 26, 2015. Keywords: Bioengineering, Tissue engineering, 3D in vitro models, test system, alternative to animal testing, full thickness, skin equivalent, skin injury, wound model, automation, wounding device
1Centre for Vascular Research, University of New South Wales, 2School of Medical Sciences, University of New South Wales
Here, we present a protocol to continuously quantify cell adhesion and de-adhesion processes with high temporal resolution in a non-invasive manner by cell-substrate impedance and live cell imaging analyses. These approaches reveal the dynamics of cell adhesion/de-adhesion processes triggered by matrix modification and their temporal relationship to adhesion-dependent signaling events.
Published February 19, 2015. Keywords: Bioengineering, Cell adhesion, biosensor, live cell imaging, extracellular matrix, fibronectin, mechanobiology, cell signaling, redox signaling, oxidative stress, myeloperoxidase, endothelium
1School for Engineering of Matter, Transport and Energy, Arizona State University, 2Department of Neurology, Georgetown University Medical Center, 3Department of Pathology, Georgetown University Medical Center
Using atomic force microscopy in combination with biopanning technology we created a negative and positive biopanning system to acquire antibodies against disease-specific protein variants present in any biological material, even at low concentrations. We were successful in obtaining antibodies to TDP-43 protein variants involved in Amyotrophic Lateral Sclerosis.
Published February 12, 2015. Keywords: Bioengineering, Amyotrophic Lateral Sclerosis, TDP-43, Biopanning, Atomic Force Microscopy, scFv, Neurodegenerative diseases