scholarly journals In Situ LSPR Sensing of Secreted Insulin in Organ-on-Chip

Biosensors ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 138
Author(s):  
María A. Ortega ◽  
Júlia Rodríguez-Comas ◽  
Ozlem Yavas ◽  
Ferran Velasco-Mallorquí ◽  
Jordina Balaguer-Trias ◽  
...  
Keyword(s):  
Pancreatic Islets ◽  
Metabolic Diseases ◽  
Disease Modeling ◽  
Localized Surface Plasmon ◽  
Animal Testing ◽  
Novel Approach ◽  
On Chip ◽  
External Stimuli

Organ-on-a-chip (OOC) devices offer new approaches for metabolic disease modeling and drug discovery by providing biologically relevant models of tissues and organs in vitro with a high degree of control over experimental variables for high-content screening applications. Yet, to fully exploit the potential of these platforms, there is a need to interface them with integrated non-labeled sensing modules, capable of monitoring, in situ, their biochemical response to external stimuli, such as stress or drugs. In order to meet this need, we aim here to develop an integrated technology based on coupling a localized surface plasmon resonance (LSPR) sensing module to an OOC device to monitor the insulin in situ secretion in pancreatic islets, a key physiological event that is usually perturbed in metabolic diseases such as type 2 diabetes (T2D). As a proof of concept, we developed a biomimetic islet-on-a-chip (IOC) device composed of mouse pancreatic islets hosted in a cellulose-based scaffold as a novel approach. The IOC was interfaced with a state-of-the-art on-chip LSPR sensing platform to monitor the in situ insulin secretion. The developed platform offers a powerful tool to enable the in situ response study of microtissues to external stimuli for applications such as a drug-screening platform for human models, bypassing animal testing.

scholarly journals New Endeavors of (Micro)Tissue Engineering: Cells Tissues Organs on-Chip and Communication Thereof

2021 ◽  
pp. 1-15
Author(s):  
Haysam M.M.A.M. Ahmed ◽  
Liliana S. Moreira Teixeira
Keyword(s):  
Tissue Engineering ◽  
Food Additives ◽  
Market Potential ◽  
Model Systems ◽  
Human Model ◽  
Human Stem Cells ◽  
Animal Testing ◽  
Preclinical Research ◽  
On Chip

The development of new therapies is tremendously hampered by the insufficient availability of human model systems suitable for preclinical research on disease target identification, drug efficacy, and toxicity. Thus, drug failures in clinical trials are too common and too costly. Animal models or standard 2D in vitro tissue cultures, regardless of whether they are human based, are regularly not representative of specific human responses. Approaching near human tissues and organs test systems is the key goal of organs-on-chips (OoC) technology. This technology is currently showing its potential to reduce both drug development costs and time-to-market, while critically lessening animal testing. OoC are based on human (stem) cells, potentially derived from healthy or disease-affected patients, thereby amenable to personalized therapy development. It is noteworthy that the OoC market potential goes beyond pharma, with the possibility to test cosmetics, food additives, or environmental contaminants. This (micro)tissue engineering-based technology is highly multidisciplinary, combining fields such as (developmental) biology, (bio)materials, microfluidics, sensors, and imaging. The enormous potential of OoC is currently facing an exciting new challenge: emulating cross-communication between tissues and organs, to simulate more complex systemic responses, such as in cancer, or restricted to confined environments, as occurs in osteoarthritis. This review describes key examples of multiorgan/tissue-on-chip approaches, or linked organs/tissues-on-chip, focusing on challenges and promising new avenues of this advanced model system. Additionally, major emphasis is given to the translation of established tissue engineering approaches, bottom up and top down, towards the development of more complex, robust, and representative (multi)organ/tissue-on-chip approaches.

scholarly journals In Situ Visualizing the Recognition Between Proteins and Platinum-Damaged DNA in Single-Cells by Correlated Optical and Secondary Ion Mass Spectrometric Imaging

2020 ◽  
Author(s):  
Yu Lin ◽  
Kui Wu ◽  
Feifei Jia ◽  
Ling Chen ◽  
Zhaoying Wang ◽  
...  
Keyword(s):  
Single Cells ◽  
Mass Spectrometric ◽  
Dependent Manner ◽  
Mass Spectrometric Imaging ◽  
Novel Approach ◽  
Molecular Mechanism Of Action ◽  
Secondary Ion ◽  
Damaged Dna

<p><b><i>In situ</i> visualization of the recognition and interaction between proteins and drug damaged DNA at single cell level is highly important for understanding the molecular mechanism of action of DNA targeting drugs, yet a great challenge. We herein report a novel approach, termed as correlated optical and secondary ion mass spectrometric imaging (COSIMSi), for exploring the recognition between proteins and cisplatin-damaged DNA in single cells. Genetically encoded EYFP-fused HMGB1, an <i>in vitro</i> well-known specific binder of cisplatin-damaged DNA, and dye-stained DNA, and cisplatin were mapped by LSCM and ToF-SIMS imaging, respectively. The LSCM and SIMS images were aligned with aiding of an addressable silicon wafer to generate fused images, in which the co-localization of the fluorescence and MS signals indicated the formation of HMGB1-Pt-DNA ternary complexes in a dose- and time-dependent manner. In contrast, COSIMSi showed that little HMGB1(F37A)-Pt-DNA complex was produced under the same conditions. Moreover, we demonstrated for the first time that cisplatin lesion on DNA prevented a DNA-binding protein Smad3 from interacting with DNA. These results verify that the COSIMSi is an effective and straightforward tool for <i>in situ</i> visualization of recognition and interaction between proteins and specific damaged DNA in single cells. </b><br></p>

scholarly journals SARS-CoV-2 Viral Replication in a High Throughput Human Primary Epithelial Airway Organ Model

2021 ◽  
Author(s):  
Christine R Fisher ◽  
Felix Mba Medie ◽  
Rebeccah J Luu ◽  
Rob Gaibler ◽  
Caitlin R Miller ◽  
...  
Keyword(s):  
Viral Replication ◽  
High Throughput ◽  
Disease Modeling ◽  
Rapid Development ◽  
The United States ◽  
Modern Medicine ◽  
Airway Epithelial Cells ◽  
Airway Epithelial ◽  
On Chip

COVID-19 emerged as a worldwide pandemic early in 2020, and at this writing has caused over 170 million cases and 3.7 million deaths worldwide, and almost 600,000 deaths in the United States. The rapid development of several safe and highly efficacious vaccines stands as one of the most extraordinary achievements in modern medicine, but the identification and administration of efficacious therapeutics to treat patients suffering from COVID-19 has been far less successful. A major factor limiting progress in the development of effective treatments has been a lack of suitable preclinical models for the disease, currently reliant upon various animal models and in vitro culture of immortalized cell lines. Here we report the first successful demonstration of SARS-CoV-2 infection and viral replication in a human primary cell-based organ-on-chip, leveraging a recently developed tissue culture platform known as PREDICT96. This successful demonstration of SARS-CoV-2 infection in human primary airway epithelial cells derived from a living donor represents a powerful new pathway for disease modeling and an avenue for screening therapeutic candidates in a high throughput platform.

In Situ Visualizing the Recognition Between Proteins and Platinum-Damaged DNA in Single-Cells by Correlated Optical and Secondary Ion Mass Spectrometric Imaging

2020 ◽  
Author(s):  
Yu Lin ◽  
Kui Wu ◽  
Feifei Jia ◽  
Ling Chen ◽  
Zhaoying Wang ◽  
...  
Keyword(s):  
Single Cells ◽  
Mass Spectrometric ◽  
Dependent Manner ◽  
Mass Spectrometric Imaging ◽  
Novel Approach ◽  
Molecular Mechanism Of Action ◽  
Secondary Ion ◽  
Damaged Dna

<p><b><i>In situ</i></b><b> visualization of the recognition and interactions between proteins and drug damaged DNA at single cell level is highly important for understanding the molecular mechanism of action of DNA targeting drugs, yet a great challenge. We report herein a novel approach, termed as correlated optical and secondary ion mass spectrometric imaging (COSIMSi), to explore the recognition between proteins and cisplatin-damaged DNA in single cells. Genetically encoded EYFP-fused HMGB1, an <i>in vitro</i> well-known specific binder of cisplatin-damaged DNA, dye-stained DNA, and platinum were mapped by LSCM and ToF-SIMS imaging, respectively. The LSCM and SIMS images were aligned with aiding of an addressable silicon wafer to generate fused images, in which the co-localization of the fluorescent and MS signals indicated the formation of HMGB1-Pt-DNA ternary complexes in a dose- and time-dependent manner. In contrast, COSIMSi results showed that little HMGB1<a></a><a>(F37A</a>)-Pt-DNA complex was produced under the same conditions. Moreover, we demonstrated for the first time that cisplatin lesions on DNA prevented DNA-binding proteins Smad3 and Smad7 from interacting with DNA. These results verify that the COSIMSi is an effective and straightforward tool for <i>in situ</i> visualization of recognition and interactions between proteins and specific damaged DNA in single cells. </b></p>

scholarly journals High-throughput screening of biomolecules using cell-free gene expression systems

2018 ◽  
Vol 3 (1) ◽  
Author(s):  
Luis E Contreras-Llano ◽  
Cheemeng Tan
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Method ◽  
Screening Methods ◽  
Free Gene ◽  
On Chip ◽  
Translation Systems ◽  
Selection Of

Abstract The incorporation of cell-free transcription and translation systems into high-throughput screening applications enables the in situ and on-demand expression of peptides and proteins. Coupled with modern microfluidic technology, the cell-free methods allow the screening, directed evolution and selection of desired biomolecules in minimal volumes within a short timescale. Cell-free high-throughput screening applications are classified broadly into in vitro display and on-chip technologies. In this review, we outline the development of cell-free high-throughput screening methods. We further discuss operating principles and representative applications of each screening method. The cell-free high-throughput screening methods may be advanced by the future development of new cell-free systems, miniaturization approaches, and automation technologies.

scholarly journals Airway-On-A-Chip: Designs and Applications for Lung Repair and Disease

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1602
Author(s):  
Tanya J. Bennet ◽  
Avineet Randhawa ◽  
Jessica Hua ◽  
Karen C. Cheung
Keyword(s):  
In Vitro Models ◽  
Chronic Obstructive ◽  
Obstructive Pulmonary Disease ◽  
Characterization Methods ◽  
Composition And Structure ◽  
Challenges And Opportunities ◽  
On Chip ◽  
Respiratory Conditions

The lungs are affected by illnesses including asthma, chronic obstructive pulmonary disease, and infections such as influenza and SARS-CoV-2. Physiologically relevant models for respiratory conditions will be essential for new drug development. The composition and structure of the lung extracellular matrix (ECM) plays a major role in the function of the lung tissue and cells. Lung-on-chip models have been developed to address some of the limitations of current two-dimensional in vitro models. In this review, we describe various ECM substitutes utilized for modeling the respiratory system. We explore the application of lung-on-chip models to the study of cigarette smoke and electronic cigarette vapor. We discuss the challenges and opportunities related to model characterization with an emphasis on in situ characterization methods, both established and emerging. We discuss how further advancements in the field, through the incorporation of interstitial cells and ECM, have the potential to provide an effective tool for interrogating lung biology and disease, especially the mechanisms that involve the interstitial elements.

scholarly journals In Situ-Forming Cellulose/Albumin-Based Injectable Hydrogels for Localized Antitumor Therapy

Polymers ◽  
2021 ◽  
Vol 13 (23) ◽  
pp. 4221
Author(s):  
Ying Chen ◽  
Xiaomin Wang ◽  
Yudong Huang ◽  
Peipei Kuang ◽  
Yushu Wang ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Antitumor Therapy ◽  
Injectable Hydrogels ◽  
External Temperature ◽  
Minimal Invasiveness ◽  
In Situ Forming ◽  
External Stimuli

Injectable hydrogels, which are formed in situ by changing the external stimuli, have the unique characteristics of easy handling and minimal invasiveness, thus providing the advantage of bypass surgical operation and improving patient compliance. Using external temperature stimuli to realize the sol-to-gel transition when preparing injectable hydrogel is essential since the temperature is stable in vivo and controllable during ex vivo, although the hydrogels obtained possibly have low mechanical strength and stability. In this work, we designed an in situ fast-forming injectable cellulose/albumin-based hydrogel (HPC-g-AA/BSA hydrogels) that responded to body temperature and which was a well-stabilized hydrogen-bonding network, effectively solving the problem of poor mechanical properties. The application of localized delivery of chemotherapeutic drugs of HPC-g-AA/BSA hydrogels was evaluated. In vitro and in vivo results show that HPC-g-AA/BSA hydrogels exhibited higher antitumor efficacy of reducing tumor size and seem ideal for localized antitumor therapy.

scholarly journals In Situ Visualizing the Recognition Between Proteins and Platinum-Damaged DNA in Single-Cells by Correlated Optical and Secondary Ion Mass Spectrometric Imaging

2020 ◽  
Author(s):  
Yu Lin ◽  
Kui Wu ◽  
Feifei Jia ◽  
Ling Chen ◽  
Zhaoying Wang ◽  
...  
Keyword(s):  
Single Cells ◽  
Mass Spectrometric ◽  
Dependent Manner ◽  
Mass Spectrometric Imaging ◽  
Novel Approach ◽  
Molecular Mechanism Of Action ◽  
Secondary Ion ◽  
Damaged Dna

<p><b><i>In situ</i> visualization of the recognition and interaction between proteins and drug damaged DNA at single cell level is highly important for understanding the molecular mechanism of action of DNA targeting drugs, yet a great challenge. We herein report a novel approach, termed as correlated optical and secondary ion mass spectrometric imaging (COSIMSi), for exploring the recognition between proteins and cisplatin-damaged DNA in single cells. Genetically encoded EYFP-fused HMGB1, an <i>in vitro</i> well-known specific binder of cisplatin-damaged DNA, and dye-stained DNA, and cisplatin were mapped by LSCM and ToF-SIMS imaging, respectively. The LSCM and SIMS images were aligned with aiding of an addressable silicon wafer to generate fused images, in which the co-localization of the fluorescence and MS signals indicated the formation of HMGB1-Pt-DNA ternary complexes in a dose- and time-dependent manner. In contrast, COSIMSi showed that little HMGB1(F37A)-Pt-DNA complex was produced under the same conditions. Moreover, we demonstrated for the first time that cisplatin lesion on DNA prevented a DNA-binding protein Smad3 from interacting with DNA. These results verify that the COSIMSi is an effective and straightforward tool for <i>in situ</i> visualization of recognition and interaction between proteins and specific damaged DNA in single cells. </b><br></p>

scholarly journals Modeling MyD88 Deficiency In Vitro Provides New Insights in Its Function

2020 ◽  
Vol 11 ◽  
Author(s):  
Nils Craig-Mueller ◽  
Ruba Hammad ◽  
Roland Elling ◽  
Jamal Alzubi ◽  
Barbara Timm ◽  
...  
Keyword(s):  
Cellular Response ◽  
Disease Modeling ◽  
Genetic Defect ◽  
Sleeping Beauty ◽  
Myeloid Differentiation ◽  
Toll Like Receptor ◽  
Monocytes And Macrophages ◽  
Induced Pluripotent ◽  
External Stimuli

Inherited defects in MyD88 and IRAK4, two regulators in Toll-like receptor (TLR) signaling, are clinically highly relevant, but still incompletely understood. MyD88- and IRAK4-deficient patients are exceedingly susceptible to a narrow spectrum of pathogens, with ∼50% lethality in the first years of life. To better understand the underlying molecular and cellular characteristics that determine disease progression, we aimed at modeling the cellular response to pathogens in vitro. To this end, we determined the immunophenotype of monocytes and macrophages derived from MyD88- and IRAK4-deficient patients. We recognized that macrophages derived from both patients were particularly poorly activated by streptococci, indicating that both signaling intermediates are essential for the immune response to facultative pathogens. To characterize this defect in more detail, we generated induced pluripotent stem cells (iPSCs) of fibroblasts derived from an MyD88-deficient patient. The underlying genetic defect was corrected using Sleeping Beauty transposon vectors encoding either the long (L) or the short (S) MYD88 isoform, respectively. Macrophages derived from these iPSC lines (iMacs) expressed typical macrophage markers, stably produced either MyD88 isoform, and showed robust phagocytic activity. Notably, iMacs expressing MyD88-L, but not MyD88-S, exhibited similar responses to external stimuli, including cytokine release patterns, as compared to genetically normal iMacs. Thus, the two MyD88 isoforms assume distinct functions in signaling. In conclusion, iPSC technology, in combination with efficient myeloid differentiation protocols, provides a valuable and inexhaustible source of macrophages, which can be used for disease modeling. Moreover, iPSC-derived macrophages may eventually aid in stabilizing MyD88-deficient patients during pyogenic infections.

scholarly journals Compliant 3D frameworks instrumented with strain sensors for characterization of millimeter-scale engineered muscle tissues

2021 ◽  
Vol 118 (19) ◽  
pp. e2100077118
Author(s):  
Hangbo Zhao ◽  
Yongdeok Kim ◽  
Heling Wang ◽  
Xin Ning ◽  
Chenkai Xu ◽  
...  
Keyword(s):  
Disease Modeling ◽  
Three Dimensional ◽  
High Sensitivity ◽  
Strain Sensors ◽  
High Temporal Resolution ◽  
On Chip ◽  
Contractile Forces ◽  
Traditional Approaches ◽  
And Control

Tissue-on-chip systems represent promising platforms for monitoring and controlling tissue functions in vitro for various purposes in biomedical research. The two-dimensional (2D) layouts of these constructs constrain the types of interactions that can be studied and limit their relevance to three-dimensional (3D) tissues. The development of 3D electronic scaffolds and microphysiological devices with geometries and functions tailored to realistic 3D tissues has the potential to create important possibilities in advanced sensing and control. This study presents classes of compliant 3D frameworks that incorporate microscale strain sensors for high-sensitivity measurements of contractile forces of engineered optogenetic muscle tissue rings, supported by quantitative simulations. Compared with traditional approaches based on optical microscopy, these 3D mechanical frameworks and sensing systems can measure not only motions but also contractile forces with high accuracy and high temporal resolution. Results of active tension force measurements of engineered muscle rings under different stimulation conditions in long-term monitoring settings for over 5 wk and in response to various chemical and drug doses demonstrate the utility of such platforms in sensing and modulation of muscle and other tissues. Possibilities for applications range from drug screening and disease modeling to biohybrid robotic engineering.

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