Computational fluid dynamic analysis of bioprinted self‐supporting perfused tissue models

Sego, T. J.; Prideaux, Matthew; Sterner, Jane; McCarthy, Brian P.; Li, Ping; Bonewald, Lynda F.; Ekser, Burçin; Tovar, Andrés; Smith, Lester J.

doi:10.1002/bit.27238

Cited by 15 publications

(12 citation statements)

References 61 publications

(97 reference statements)

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“…Comparison of static and dynamic culture of barrier tissues with the addition of shear stress is technically challenging. Reported fluidic systems are complicated to setup and require specialized equipment [ 27 , 40 , 41 ]. In addition to being expensive compared to the standard static Transwell-based models, the throughput of these systems remains typically very low [ 42 ].…”

Section: Discussionmentioning

confidence: 99%

Online Measurement System for Dynamic Flow Bioreactors to Study Barrier Integrity of hiPSC-Based Blood–Brain Barrier In Vitro Models

et al. 2022

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Electrochemical impedance spectroscopy (EIS) is a noninvasive, reliable, and efficient method to analyze the barrier integrity of in vitro tissue models. This well-established tool is used most widely to quantify the transendothelial/epithelial resistance (TEER) of Transwell-based models cultured under static conditions. However, dynamic culture in bioreactors can achieve advanced cell culture conditions that mimic a more tissue-specific environment and stimulation. This requires the development of culture systems that also allow for the assessment of barrier integrity under dynamic conditions. Here, we present a bioreactor system that is capable of the automated, continuous, and non-invasive online monitoring of cellular barrier integrity during dynamic culture. Polydimethylsiloxane (PDMS) casting and 3D printing were used for the fabrication of the bioreactors. Additionally, attachable electrodes based on titanium nitride (TiN)-coated steel tubes were developed to perform EIS measurements. In order to test the monitored bioreactor system, blood–brain barrier (BBB) in vitro models derived from human-induced pluripotent stem cells (hiPSC) were cultured for up to 7 days. We applied equivalent electrical circuit fitting to quantify the electrical parameters of the cell layer and observed that TEER gradually decreased over time from 2513 Ω·cm2 to 285 Ω·cm2, as also specified in the static control culture. Our versatile system offers the possibility to be used for various dynamic tissue cultures that require a non-invasive monitoring system for barrier integrity.

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Section: Discussionmentioning

confidence: 99%

Online Measurement System for Dynamic Flow Bioreactors to Study Barrier Integrity of hiPSC-Based Blood–Brain Barrier In Vitro Models

et al. 2022

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“…For example, the design of a scaffold-free and perfused bioreactor can be optimized using computational fluid dynamic (CFD) simulations, allowing for tissue-specific designs to be optimized in silico. When tissue constructs with built-in microchannels are cultured in CFD-optimized bioreactors, effective nutrient perfusion and tissue maturation could be achieved over weeks of culture ( Sego et al, 2020 ). This approach was utilized in the fabrication of autologous cartilage-bone grafts engineered for temporomandibular joint regeneration (TMJ).…”

Section: From Bioreactors To Microphysiological Systemsmentioning

confidence: 99%

Human Organ-on-a-Chip Microphysiological Systems to Model Musculoskeletal Pathologies and Accelerate Therapeutic Discovery

Ajalik

Alenchery

Cognetti

et al. 2022

Front. Bioeng. Biotechnol.

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Human Microphysiological Systems (hMPS), otherwise known as organ- and tissue-on-a-chip models, are an emerging technology with the potential to replace in vivo animal studies with in vitro models that emulate human physiology at basic levels. hMPS platforms are designed to overcome limitations of two-dimensional (2D) cell culture systems by mimicking 3D tissue organization and microenvironmental cues that are physiologically and clinically relevant. Unlike animal studies, hMPS models can be configured for high content or high throughput screening in preclinical drug development. Applications in modeling acute and chronic injuries in the musculoskeletal system are slowly developing. However, the complexity and load bearing nature of musculoskeletal tissues and joints present unique challenges related to our limited understanding of disease mechanisms and the lack of consensus biomarkers to guide biological therapy development. With emphasis on examples of modeling musculoskeletal tissues, joints on chips, and organoids, this review highlights current trends of microphysiological systems technology. The review surveys state-of-the-art design and fabrication considerations inspired by lessons from bioreactors and biological variables emphasizing the role of induced pluripotent stem cells and genetic engineering in creating isogenic, patient-specific multicellular hMPS. The major challenges in modeling musculoskeletal tissues using hMPS chips are identified, including incorporating biological barriers, simulating joint compartments and heterogenous tissue interfaces, simulating immune interactions and inflammatory factors, simulating effects of in vivo loading, recording nociceptors responses as surrogates for pain outcomes, modeling the dynamic injury and healing responses by monitoring secreted proteins in real time, and creating arrayed formats for robotic high throughput screens. Overcoming these barriers will revolutionize musculoskeletal research by enabling physiologically relevant, predictive models of human tissues and joint diseases to accelerate and de-risk therapeutic discovery and translation to the clinic.

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“…Future studies will also attempt to generate low oxygenation levels needed for some tissue types such as bone and investigate if oxygenation stability at these levels is coupled to perfusion flow rate. Additionally, we recently generated self-supporting perfused (SSuPer) tissues 27 , which possess a microchannel structure conducive to perfusion of oxygenated media, and performed computational analysis in order to determine the ability of oxygen to reach the tissue bulk between the channels since the maximum tissue thickness should be less than the maximum perfusion distance of the perfused metabolites, specifically oxygen 34 . Since oxygenation, structure, and perfusion parameters change with cell/tissue type, cell concentration, and cell density, the oxygenated FABRICA coupled with the SSuPer Tissue represent a developing solution for generating robust engineered tissues with in situ-like structure which can be cultured in vitro with in vivo-like conditions.…”

Section: The Oxygenated Fabrica Exhibits Perfusion Partially Decouplementioning

confidence: 99%

“…Many bioreactor platforms lack modularity which is needed to account for design updates or new modules intended to increase functionality. In our previous work, we demonstrated our ability to control and characterize perfusive flow applied to tissues of different types and designs cultured within our FABRICA bioreactor platform, a bioprinting method-independent tissue perfusion system 1,27 . However, the oxygenation status of the tissues cultured within the FABRICA was not characterized.…”

mentioning

confidence: 99%

“…Since the FABRICA is both bioprinting method and tissue design agnostic, and studying tissue oxygenation (i.e., oxygen distribution and tissue survival) is futile without first providing and characterizing media oxygenation, we decided to investigate oxygenation of the perfused media without including biofabricated tissues. The complexities associated with designing, fabricating, culturing, and analyzing complex tissues with perfusible structures (i.e., vascular-like channels) are vast enough that they deserve their own investigation in parallel with oxygenation, which we have begun 27 .…”

mentioning

confidence: 99%

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Oxygenation Profiles of Human Blood, Cell Culture Medium, and Water for Perfusion of 3D-Bioprinted Tissues using the FABRICA Bioreactor Platform

Chen

Lashmet

Isidan

et al. 2020

Sci Rep

Self Cite

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Persistent and saturated oxygen distribution from perfusion media (i.e., blood, or cell culture media) to cells within cell-dense, metabolically-active biofabricated tissues is required to keep them viable. Improper or poor oxygen supply to cells within the tissue bulk severely limits the tissue culturing potential of many bioreactors. We added an oxygenator module to our modular FABRICA bioreactor in order to provide stable oxygenation to biofabricated tissues during culture. In this proof of concept study of an oxygenated and perfused bioreactor, we characterized the oxygenation of water, cell culture medium, and human blood in the FABRICA as functions of augmenting vacuum (air inlet) pressure, perfusion (volumetric flow) rate, and tubing/oxygenator components. The mean oxygen levels for water and cell culture media were 27.7 ± 2.1% and 27.6 ± 4.1%, respectively. The mean oxygen level for human blood was 197.0 ± 90.0 mmHg, with near-physiologic levels achieved with low-permeability PharMed tubing alone (128.0 ± 14.0 mmHg). Hematologic values pre-and post-oxygenation, respectively were (median ± IQR): Red blood cell: 6.0 ± 0.5 (10 6 /μL) and 6.5 ± 0.4 (10 6 /μL); Hemoglobin: 17.5 ± 1.2 g/dL and 19.2 ± 3.0 g/dL; and Hematocrit: 56.7 ± 2.4% and 61.4 ± 7.5%. The relative stability of the hematologic parameters indicates that blood function and thus blood cell integrity were maintained throughout oxygenation. Already a versatile research tool, the now oxygenated FABRICA provides easy-to-implement, in vivo-like perfusion and stable oxygenation culture conditions in vitro semi-independently of one another, which means the bioreactor has the potential to serve as a platform for investigating the behavior of 3D tissue models (regardless of biofabrication method), performing drug toxicity-testing, and testing pharmaceutical efficacy/safety. Recent 3D bioprinting/biofabrication advances allow generation of scaffold-free engineered tissue constructs with customizable 3D design and high cell density 1-4. These engineered tissues are comprised of cells in contact with one another and with extracellular matrix (ECM) in dense 3D structures, thus making them similar to natural tissues in vivo with the same requirements of controlled, well-distributed oxygenation, nutrition, mechanical (shear, compressive, and tensile) forces, temperature, pH, etc. 5-15 so that all cells receive proper metabolic and stimulatory support. Compared to cells cultured in 2 dimensions, engineered tissues with 3D structures, biofabricated by means of bioprinting or other methods, can plausibly be tuned to meet specific genetic, cellular, structural, extracellular, and metabolic parameters, and thus have incredible potential to provide tissues for clinical implants, research models, and drug testing platforms. Importantly, given that media (cell culture media or blood), used to perfuse biofabricated tissue cultured in bioreactors, is one constituent providing multiple critical metabolic factors (convective heat, nutrients, shear stress, and oxygen), the...

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Computational fluid dynamic analysis of bioprinted self‐supporting perfused tissue models

Cited by 15 publications

References 61 publications

Online Measurement System for Dynamic Flow Bioreactors to Study Barrier Integrity of hiPSC-Based Blood–Brain Barrier In Vitro Models

Online Measurement System for Dynamic Flow Bioreactors to Study Barrier Integrity of hiPSC-Based Blood–Brain Barrier In Vitro Models

Human Organ-on-a-Chip Microphysiological Systems to Model Musculoskeletal Pathologies and Accelerate Therapeutic Discovery

Oxygenation Profiles of Human Blood, Cell Culture Medium, and Water for Perfusion of 3D-Bioprinted Tissues using the FABRICA Bioreactor Platform

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