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 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
1Biological Systems Research, Philip Morris International R&D, Philip Morris Products S.A.
The aim of this protocol is to expose human organotypic 3D bronchial and nasal tissue models to mainstream cigarette smoke (CS) at the air-liquid interface. The impact of CS on the tissues is then investigated using a cytochrome P450 activity assay, a cilia beating measurement, and a systems biology approach.
Published February 12, 2015. Keywords: Bioengineering, human organotypic bronchial epithelial, 3D culture, in vitro exposure system, cigarette smoke, cilia beating, xenobiotic metabolism, network models, systems toxicology
1Department of Radiology, Memorial Sloan Kettering Cancer Center
The bioorthogonal inverse electron demand Diels-Alder cycloaddition has been harnessed to create an effective and modular pretargeted PET imaging strategy for cancer. In this protocol, the steps of this methodology are described in the context of a model system employing the colorectal cancer targeted antibody huA33 and a 64Cu-labeled radioligand.
Published February 3, 2015. Keywords: Bioengineering, Positron Emission Tomography, Click Chemistry, Pretargeting, Tetrazine, Trans-cyclooctene, Inverse Electron Demand Diels-Alder Cycloaddition
1School of Dentistry, University of California, Los Angeles, 2School of Medicine, Clinical Nutrition, University of California, Los Angeles
Exosomes are microvesicular structures found within biofluids that potentially carry important disease discriminatory biomarkers. Here, a novel method is used to specifically extract exosomes and rapidly test the exosomal cargo for both RNA/protein targets following the disruption of exosomes using non-uniform electric cyclic square waves.
Published January 23, 2015. Keywords: Bioengineering, Exosome, Electrochemical sensors, Tumor biomarkers, Lung cancer, Salivary diagnostics
1Department of Nanomedicine, Houston Methodist Research Institute, 2MOE Key Laboratory of Bioinorganic and Synthetic Chemistry, School of Chemistry and Chemical Engineering, Sun Yat-sen University, 3Pediatrics Department of Union Hospital, Huazhong University of Science and Technology, 4CAS Key Laboratory for Biomedical Effects of Nanomaterials & Nanosafety, National Center for Nanoscience & Technology of China, 5Department of Medicine, Weill Cornell Medical College, 6Department of Cell and Developmental Biology, Weill Cornell Medical College
Delivery remains the main challenge for the therapeutic implementation of small interfering RNA (siRNA). This protocol involves the use of a multifunctional and biocompatible siRNA delivery platform, consisting of arginine and polyethylenimine grafted porous silicon microparticles.
Published January 15, 2015. Keywords: Bioengineering, Porous silicon, siRNA, Nanodelivery system, Cancer therapy, Transfection, Polycation functionalization
1Experimental Radiology, Institute of Diagnostic and Interventional Radiology I, Jena University Hospital, 2Department of Pharmaceutical Technology, Friedrich-Schiller-University Jena, 3Center for Electron Microscopy, Jena University Hospital
The use of fluorophores for in vivo imaging can be greatly limited by opsonization, rapid clearance, low detection sensitivity and cytotoxic effects on the host. Encapsulation of fluorophores in liposomes by film hydration and extrusion leads to fluorescence quenching and protection which enables in vivo imaging with high detection sensitivity.
Published January 5, 2015. Keywords: Bioengineering, Drug-delivery, Liposomes, Fluorochromes, Fluorescence-quenching, Optical imaging, Inflammation
1Angiogenesis Laboratory, Department of Ophthalmology, Harvard Medical School, Massachusetts Eye & Ear Infirmary, 2St. Elizabeth's Medical Center
Here, we present a protocol that allows the investigator to assess leukocyte recruitment dynamics ex vivo by connecting a chamber coated with endothelial-derived adhesion molecules to the circulatory system of a mouse. This method offers significant advantages since it allows for leukocyte assessment under relative biological conditions.
Published December 30, 2014. Keywords: Bioengineering, Leukocyte recruitment, endothelium, flow chamber, adhesion, velocity, rolling, integrins, ex vivo
1Department of Chemistry and Biochemistry, University of Notre Dame, 2Harper Cancer Research Institute, University of Notre Dame
Immortalized cancer cell lines can be grown as 3D cell cultures, a valuable model for biological research. This protocol describes mass spectrometry imaging of 3D cell cultures, including improvements in the sample preparation platform. The goal of this protocol is to instruct users to prepare 3D cell cultures for mass spectrometry imaging analysis.
Published December 5, 2014. Keywords: Bioengineering, 3D cell culture, mass spectrometry, imaging, cell culture, sample preparation, spheroids