Publications by authors named "Matthew Ishahak"

6 Publications

  • Page 1 of 1

Modular Microphysiological System for Modeling of Biologic Barrier Function.

Front Bioeng Biotechnol 2020 12;8:581163. Epub 2020 Nov 12.

Department of Biomedical Engineering, University of Miami, Coral Gables, FL, United States.

Microphysiological systems, also known as organs-on-chips, are microfluidic devices designed to model human physiology . Polydimethylsiloxane (PDMS) is the most widely used material for organs-on-chips due to established microfabrication methods, and properties that make it suitable for biological applications such as low cytotoxicity, optical transparency, gas permeability. However, absorption of small molecules and leaching of uncrosslinked oligomers might hinder the adoption of PDMS-based organs-on-chips for drug discovery assays. Here, we have engineered a modular, PDMS-free microphysiological system that is capable of recapitulating biologic barrier functions commonly demonstrated in PDMS-based devices. Our microphysiological system is comprised of a microfluidic chip to house cell cultures and pneumatic microfluidic pumps to drive flow with programmable pressure and shear stress. The modular architecture and programmable pumps enabled us to model multiple microenvironments. First, we demonstrate the ability to generate cyclic strain on the culture membrane and establish a model of the alveolar air-liquid interface. Next, we utilized three-dimensional finite element analysis modeling to characterize the fluid dynamics within the device and develop a model of the pressure-driven filtration that occurs at the glomerular filtration barrier. Finally, we demonstrate that our model can be used to recapitulate sphingolipid induced kidney injury. Together, our results demonstrate that a multifunctional and modular microphysiological system can be deployed without the use of PDMS. Further, the bio-inert plastic used in our microfluidic device is amenable to various established, high-throughput manufacturing techniques, such as injection molding. As a result, the development plastic organs-on-chips provides an avenue to meet the increasing demand for organ-on-chip technology.
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http://dx.doi.org/10.3389/fbioe.2020.581163DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7693638PMC
November 2020

Microelectrode Array based Functional Testing of Pancreatic Islet Cells.

Micromachines (Basel) 2020 May 17;11(5). Epub 2020 May 17.

Department of Biomedical Engineering, University of Miami, Coral Gables, FL 33146, USA.

Electrophysiological techniques to characterize the functionality of islets of Langerhans have been limited to short-term, one-time recordings such as a patch clamp recording. We describe the use of microelectrode arrays (MEAs) to better understand the electrophysiology of dissociated islet cells in response to glucose in a real-time, non-invasive method over prolonged culture periods. Human islets were dissociated into singular cells and seeded onto MEA, which were cultured for up to 7 days. Immunofluorescent imaging revealed that several cellular subtypes of islets; β, δ, and γ cells were present after dissociation. At days 1, 3, 5, and 7 of culture, MEA recordings captured higher electrical activities of islet cells under 16.7 mM glucose (high glucose) than 1.1 mM glucose (low glucose) conditions. The fraction of the plateau phase (FOPP), which is the fraction of time with spiking activity recorded using the MEA, consistently showed distinguishably greater percentages of spiking activity with high glucose compared to the low glucose for all culture days. In parallel, glucose stimulated insulin secretion was measured revealing a diminished insulin response after day 3 of culture. Additionally, MEA spiking profiles were similar to the time course of insulin response when glucose concentration is switched from 1.1 to 16.7 mM. Our analyses suggest that extracellular recordings of dissociated islet cells using MEA is an effective approach to rapidly assess islet functionality, and could supplement standard assays such as glucose stimulate insulin response.
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http://dx.doi.org/10.3390/mi11050507DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7281363PMC
May 2020

Integrated human pseudoislet system and microfluidic platform demonstrate differences in GPCR signaling in islet cells.

JCI Insight 2020 05 21;5(10). Epub 2020 May 21.

Division of Diabetes, Endocrinology and Metabolism, Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA.

Pancreatic islets secrete insulin from β cells and glucagon from α cells, and dysregulated secretion of these hormones is a central component of diabetes. Thus, an improved understanding of the pathways governing coordinated β and α cell hormone secretion will provide insight into islet dysfunction in diabetes. However, the 3D multicellular islet architecture, essential for coordinated islet function, presents experimental challenges for mechanistic studies of intracellular signaling pathways in primary islet cells. Here, we developed an integrated approach to study the function of primary human islet cells using genetically modified pseudoislets that resemble native islets across multiple parameters. Further, we developed a microperifusion system that allowed synchronous acquisition of GCaMP6f biosensor signal and hormone secretory profiles. We demonstrate the utility of this experimental approach by studying the effects of Gi and Gq GPCR pathways on insulin and glucagon secretion by expressing the designer receptors exclusively activated by designer drugs (DREADDs) hM4Di or hM3Dq. Activation of Gi signaling reduced insulin and glucagon secretion, while activation of Gq signaling stimulated glucagon secretion but had both stimulatory and inhibitory effects on insulin secretion, which occur through changes in intracellular Ca2+. The experimental approach of combining pseudoislets with a microfluidic system allowed the coregistration of intracellular signaling dynamics and hormone secretion and demonstrated differences in GPCR signaling pathways between human β and α cells.
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http://dx.doi.org/10.1172/jci.insight.137017DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7259531PMC
May 2020

Evaluating Vascularization of Heterotopic Islet Constructs for Type 1 Diabetes Using an In Vitro Platform.

Integr Biol (Camb) 2019 11;11(8):331-341

Department of Biomedical Engineering, University of Miami, Coral Gables, FL, USA.

Type 1 diabetes (T1D) results from the autoimmune destruction of β-cells within the pancreatic islets of Langerhans. Clinical islet transplantation from healthy donors is proposed to ameliorate symptoms, improve quality of life, and enhance the life span of afflicted T1D patients. However, post-transplant outcomes are dependent on the survival of the transplanted islets, which relies on the engraftment of the islets with the recipient's vasculature among other factors. Treatment strategies to improve engraftment include combining islets with supporting cells including endothelial cells (EC) and mesenchymal stem cells (MSC), dynamic cells capable of robust immunomodulatory and vasculogenic effects. In this study, we developed an in vitro model of transplantation to investigate the cellular mechanisms that enhance rapid vascularization of heterotopic islet constructs. Self-assembled vascular beds of fluorescently stained EC served as reproducible in vitro transplantation sites. Heterotopic islet constructs composed of islets, EC, and MSC were transferred to vascular beds for modeling transplantation. Time-lapsed imaging was performed for analysis of the vascular bed remodeling for parameters of neo-vascularization. Moreover, sampling of media following modeled transplantation showed secretory profiles that were correlated with imaging analyses as well as with islet function using glucose-stimulated insulin secretion. Together, evidence revealed that heterotopic constructs consisting of islets, EC, and MSC exhibited the most rapid recruitment and robust branching of cells from the vascular beds suggesting enhanced neo-vascularization compared to islets alone and control constructs. Together, this evidence supports a promising cell transplantation strategy for T1D and also demonstrates a valuable tool for rapidly investigating candidate cellular therapies for transplantation.
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http://dx.doi.org/10.1093/intbio/zyz027DOI Listing
November 2019

Comparison of temperature change and resulting ablation size induced by a 902-928 MHz and a 2450 MHz microwave ablation system in in-vivo porcine kidneys.

Int J Hyperthermia 2019 5;36(1):313-321. Epub 2019 Mar 5.

a Joint Bioengineering and Endourology Developmental Surgical Laboratory, Division of Endourology, Laparoscopy, and Minimally-Invasive Surgery, Department of Urology , University of Miami Miller School of Medicine , Miami , FL , USA.

Introduction: Microwave ablation (MWA) uses heat to ablate undesired tissue. Development of pre-planning algorithms for MWA of small renal masses requires understanding of microwave-tissue interactions at different operating parameters. The objective of this study was to compare the performance of two MWA systems in in-vivo porcine kidneys.

Methods: Five ablations were performed using a 902-928 MHz system (24 W, 5 min) and a 2450 MHz system (180 W, 2 min). Nonlinear regression analysis of temperature changes measured 5 mm from the antenna axis was completed for the initial 10 s of ablation using the power equation and after the inflection point using an exponential equation. Thermal damage was calculated using the Arrhenius equation. Long and short axis ablation diameters were measured.

Results: The average 'a' varied significantly between systems (902-928 MHz: 0.0299 ± 0.027, 2450 MHz: 0.1598 ± 0.158), indicating proportionality to the heat source, but 'b' did not (902-928 MHz: 1.910 ± 0.372, 2450 MHz: 2.039 ± 0.366), signifying tissue type dependence. Past the inflection point, average steady-state temperature increases were similar between systems but reached more quickly with the 2450 MHz system. Complete damage was reached at 5 mm for both systems. The 2450 MHz system produced significantly larger short axis ablations (902-928 MHz: 2.40 ± 0.54 cm, 2450 MHz: 3.32 ± 0.41cm).

Conclusion: The 2450 MHz system achieved similar steady state temperature increases compared to the 902-928 MHz system, but more quickly due to higher output power. Further investigations using various treatment parameters and precise thermal sensor placement are warranted to refine equation parameters for the development of an ablation model.
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http://dx.doi.org/10.1080/02656736.2019.1565788DOI Listing
January 2020

Engineered Microenvironments for Maturation of Stem Cell Derived Cardiac Myocytes.

Theranostics 2018 1;8(1):124-140. Epub 2018 Jan 1.

Department of Biomedical Engineering, University of Miami.

Through the use of stem cell-derived cardiac myocytes, tissue-engineered human myocardial constructs are poised for modeling normal and diseased physiology of the heart, as well as discovery of novel drugs and therapeutic targets in a human relevant manner. This review highlights the recent bioengineering efforts to recapitulate microenvironmental cues to further the maturation state of newly differentiated cardiac myocytes. These techniques include long-term culture, co-culture, exposure to mechanical stimuli, 3D culture, cell-matrix interactions, and electrical stimulation. Each of these methods has produced various degrees of maturation; however, a standardized measure for cardiomyocyte maturation is not yet widely accepted by the scientific community.
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http://dx.doi.org/10.7150/thno.19441DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5743464PMC
November 2018
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