AIM To develop a human in vitro model of non-alcoholic fatty liver disease(NAFLD), utilising primary hepatocytes cultured in a three-dimensional(3D) perfused platform. METHODS Fat and lean culture media were developed...AIM To develop a human in vitro model of non-alcoholic fatty liver disease(NAFLD), utilising primary hepatocytes cultured in a three-dimensional(3D) perfused platform. METHODS Fat and lean culture media were developed to directly investigate the effects of fat loading on primary hepatocytes cultured in a 3D perfused culture system. Oil Red O staining was used to measure fat loading in the hepatocytes and the consumption of free fatty acids(FFA) from culture medium was monitored. Hepatic functions, gene expression profiles and adipokine release were compared for cells cultured in fat and lean conditions. To determine if fat loading in the system could be modulated hepatocytes were treated with known anti-steatotic compounds. RESULTS Hepatocytes cultured in fat medium were found to accumulate three times more fat than lean cells and fat uptake was continuous over a 14-d culture. Fat loading of hepatocytes did not cause any hepatotoxicity and significantly increased albumin production. Numerous adipokines were expressed by fatty cells and genes associated with NAFLD and liver disease were upregulated including: Insulin-like growth factorbinding protein 1, fatty acid-binding protein 3 and CYP7A1. The metabolic activity of hepatocytes cultured in fatty conditions was found to be impaired and the activities of CYP3A4 and CYP2C9 were significantlyreduced, similar to observations made in NAFLD patients. The utility of the model for drug screening was demonstrated by measuring the effects of known antisteatotic compounds. Hepatocytes, cultured under fatty conditions and treated with metformin, had a reduced cellular fat content compared to untreated controls and consumed less FFA from cell culture medium.CONCLUSION The 3D in vitro NAFLD model recapitulates many features of clinical NAFLD and is an ideal tool for analysing the efficacy of anti-steatotic compounds.展开更多
The plausibility of human exposure to particulate matter(PM)has witnessed an increase within the last several years.PM of different sizes has been discovered in the atmosphere given the role of dust transport in weath...The plausibility of human exposure to particulate matter(PM)has witnessed an increase within the last several years.PM of different sizes has been discovered in the atmosphere given the role of dust transport in weather and climate composition.As a regulator,the lung epithelium orchestrates the innate response to local damage.Herein,we developed a lung epithelium-ona-chip platform consisting of easily moldable polydimethylsiloxane layers along with a thin,flexible,and transparent ionic liquid-based poly(hydroxyethyl)methacrylate gel membrane.The epithelium was formed through the culture of human lung epithelial cells(Calu-3)on this membrane.The mechanical stress at the air–liquid interface during inhalation/exhalation was recapitulated using an Arduino-based servo motor system,which applied a uniaxial tensile strength from the two sides of the chip with 10%strain and a frequency of 0.2 Hz.Subsequently,the administration of silica nanoparticles(PM0.5)with an average size of 463 nm to the on-chip platform under static,dynamic,and dynamic+mechanical stress(DMS)conditions demonstrated the effect of environmental pollutants on lung epithelium.The viability and release of lactate dehydrogenase were determined along with proinflammatory response through the quantification of tumor necrosis factor-α,which indicated alterations in the epithelium.展开更多
The timely establishment of functional neo-vasculature is pivotal for successful tissue development and regen-eration,remaining a central challenge in tissue engineering.In this study,we present a novel(micro)vascular...The timely establishment of functional neo-vasculature is pivotal for successful tissue development and regen-eration,remaining a central challenge in tissue engineering.In this study,we present a novel(micro)vascular-ization strategy that explores the use of specialized“vascular units”(VUs)as building blocks to initiate blood vessel formation and create perfusable,stroma-embedded 3D microvascular networks from the bottom-up.We demonstrate that VUs composed of endothelial progenitor cells and organ-specific fibroblasts exhibit high angiogenic potential when embedded in fibrin hydrogels.This leads to the formation of VUs-derived capillaries,which fuse with adjacent capillaries to form stable microvascular beds within a supportive,extracellular matrix-rich fibroblastic microenvironment.Using a custom-designed biomimetic fibrin-based vessel-on-chip(VoC),we show that VUs-derived capillaries can inosculate with endothelialized microfluidic channels in the VoC and become perfused.Moreover,VUs can establish capillary bridges between channels,extending the microvascular network throughout the entire device.When VUs and intestinal organoids(IOs)are combined within the VoC,the VUs-derived capillaries and the intestinal fibroblasts progressively reach and envelop the IOs.This promotes the formation of a supportive vascularized stroma around multiple IOs in a single device.These findings un-derscore the remarkable potential of VUs as building blocks for engineering microvascular networks,with ver-satile applications spanning from regenerative medicine to advanced in vitro models.展开更多
pioids are commonly used for treating chronic pain.However,with continued use,they may induce tolerance and/or hyperalgesia,which limits therapeutic efficacy.The human mechanisms of opioid-induced tolerance and hypera...pioids are commonly used for treating chronic pain.However,with continued use,they may induce tolerance and/or hyperalgesia,which limits therapeutic efficacy.The human mechanisms of opioid-induced tolerance and hyperalgesia are significantly understudied,in part,because current models cannot fully recapitulate human pathology.Here,we engineered novel human spinal microphysiological systems(MPSs)integrated with plug-and-play neural activity sensing for modeling human nociception and opioid-induced tolerance.Each spinal MPS consists of a flattened human spinal cord organoid derived from human stem cells and a 3D printed organoid holder device for plug-and-play neural activity measurement.We found that the flattened organoid design of MPSs not only reduces hypoxia and necrosis in the organoids,but also promotes their neuron maturation,neural activity,and functional development.We further demonstrated that prolonged opioid exposure resulted in neurochemical correlates of opioid tolerance and hyperalgesia,as measured by altered neural activity,and downregulation ofμ-opioid receptor expression of human spinal MPSs.The MPSs are scalable,cost-effective,easy-to-use,and compatible with commonly-used well-plates,thus allowing plug-and-play measurements of neural activity.We believe the MPSs hold a promising translational potential for studying human pain etiology,screening new treatments,and validating novel therapeutics for human pain medicine.展开更多
Organs-on-a-chip is a microfluidic microphysiological system that uses microfluidic technology to analyze the structure and function of living human cells at the tissue and organ levels in vitro.Organs-on-a-chip techn...Organs-on-a-chip is a microfluidic microphysiological system that uses microfluidic technology to analyze the structure and function of living human cells at the tissue and organ levels in vitro.Organs-on-a-chip technology,as opposed to traditional two-dimensional cell culture and animal models,can more closely simulate pathologic and toxicologic interactions between different organs or tissues and reflect the collaborative response of multiple organs to drugs.Despite the fact that many organs-on-a-chip-related data have been published,none of the current databases have all of the following functions:searching,downloading,as well as analyzing data and results from the literature on organs-on-a-chip.Therefore,we created an organs-on-a-chip database(OOCDB)as a platform to integrate information about organs-on-a-chip from various sources,including literature,patents,raw data from microarray and transcriptome sequencing,several open-access datasets of organs-on-a-chip and organoids,and data generated in our laboratory.OOCDB contains dozens of sub-databases and analysis tools,and each sub-database contains various data associated with organs-on-a-chip,with the goal of providing researchers with a comprehensive,systematic,and convenient search engine.Furthermore,it offers a variety of other functions,such as mathematical modeling,three-dimensional modeling,and citation mapping,to meet the needs of organs-on-a-chip.展开更多
.Organ-on-Chip(OoC)has emerged as a revolutionary approach to emulate human organ function-ality in vitro,offering unparalleled insights into physiological processes and disease modeling.The integration of artificial i....Organ-on-Chip(OoC)has emerged as a revolutionary approach to emulate human organ function-ality in vitro,offering unparalleled insights into physiological processes and disease modeling.The integration of artificial intelligence(AI)with OoC platforms presents a transformative synergy,combining the precision of microscale organ replication with the analytical prowess of intelligent algorithms,is emerging as a transforma-tive force in harnessing the full potential of OoC.This perspective investigates the multifaceted implications of integrating AI with OoC,examining its impact on biomedical research,acknowledging the synergistic po-tential that arises from combining the precision of microscale organ replication with the analytical capabilities of intelligent algorithms,and fostering a future where the intricate workings of the technology and biology.展开更多
In the human body,almost all cells interact with extracellular matrices(ECMs),which have tissue and organ-specific compositions and architectures.These ECMs not only function as cellular scaffolds,providing structural...In the human body,almost all cells interact with extracellular matrices(ECMs),which have tissue and organ-specific compositions and architectures.These ECMs not only function as cellular scaffolds,providing structural support,but also play a crucial role in dynamically regulating various cellular functions.This comprehensive review delves into the examination of biofabrication strategies used to develop bioactive materials that accurately mimic one or more biophysical and biochemical properties of ECMs.We discuss the potential integration of these ECM-mimics into a range of physiological and pathological in vitro models,enhancing our understanding of cellular behavior and tissue organization.Lastly,we propose future research directions for ECM-mimics in the context of tissue engineering and organ-on-a-chip applications,offering potential advancements in therapeutic approaches and improved patient outcomes.展开更多
基金Supported by Innovate UK(Technology Strategy Board)Advancing the Development and Application of Non-Animal Technologies Project:3D cell culture model for studying NonAlcoholic Fatty Liver Disease(NAFLD)-Ref:131720
文摘AIM To develop a human in vitro model of non-alcoholic fatty liver disease(NAFLD), utilising primary hepatocytes cultured in a three-dimensional(3D) perfused platform. METHODS Fat and lean culture media were developed to directly investigate the effects of fat loading on primary hepatocytes cultured in a 3D perfused culture system. Oil Red O staining was used to measure fat loading in the hepatocytes and the consumption of free fatty acids(FFA) from culture medium was monitored. Hepatic functions, gene expression profiles and adipokine release were compared for cells cultured in fat and lean conditions. To determine if fat loading in the system could be modulated hepatocytes were treated with known anti-steatotic compounds. RESULTS Hepatocytes cultured in fat medium were found to accumulate three times more fat than lean cells and fat uptake was continuous over a 14-d culture. Fat loading of hepatocytes did not cause any hepatotoxicity and significantly increased albumin production. Numerous adipokines were expressed by fatty cells and genes associated with NAFLD and liver disease were upregulated including: Insulin-like growth factorbinding protein 1, fatty acid-binding protein 3 and CYP7A1. The metabolic activity of hepatocytes cultured in fatty conditions was found to be impaired and the activities of CYP3A4 and CYP2C9 were significantlyreduced, similar to observations made in NAFLD patients. The utility of the model for drug screening was demonstrated by measuring the effects of known antisteatotic compounds. Hepatocytes, cultured under fatty conditions and treated with metformin, had a reduced cellular fat content compared to untreated controls and consumed less FFA from cell culture medium.CONCLUSION The 3D in vitro NAFLD model recapitulates many features of clinical NAFLD and is an ideal tool for analysing the efficacy of anti-steatotic compounds.
基金BK acknowledges the TUBITAK 2210-C National Graduate Scholarship Program and access to the laboratory of Prof.Dr.Sinan Akgol at Biochemistry Department of Ege UniversityThis work was supported by the Presidency of the Republic of Türkiye Strategy Budget Department(2019K12-149080).
文摘The plausibility of human exposure to particulate matter(PM)has witnessed an increase within the last several years.PM of different sizes has been discovered in the atmosphere given the role of dust transport in weather and climate composition.As a regulator,the lung epithelium orchestrates the innate response to local damage.Herein,we developed a lung epithelium-ona-chip platform consisting of easily moldable polydimethylsiloxane layers along with a thin,flexible,and transparent ionic liquid-based poly(hydroxyethyl)methacrylate gel membrane.The epithelium was formed through the culture of human lung epithelial cells(Calu-3)on this membrane.The mechanical stress at the air–liquid interface during inhalation/exhalation was recapitulated using an Arduino-based servo motor system,which applied a uniaxial tensile strength from the two sides of the chip with 10%strain and a frequency of 0.2 Hz.Subsequently,the administration of silica nanoparticles(PM0.5)with an average size of 463 nm to the on-chip platform under static,dynamic,and dynamic+mechanical stress(DMS)conditions demonstrated the effect of environmental pollutants on lung epithelium.The viability and release of lactate dehydrogenase were determined along with proinflammatory response through the quantification of tumor necrosis factor-α,which indicated alterations in the epithelium.
基金developed under the scope of the EndoSWITCH project(PTDC/BTMORG/5154/2020)supported by the Portuguese Founda-tion for Science and Technology(FCT).The authors thanks FCT for Iasmim Orge’s PhD scholarship SFRH/BD/2020.07458+5 种基金Sílvia Bidarra’s research contract DL 57/2016/CP1360/CT0006 and Silvia Ferreira’s research contract CEECINST/00132/2021/CP1774/CT0001Iasmim Orge thanks the training provided under the scope of the REMODEL project(European Union’s Horizon 2020 research and innovation pro-gramme,grant agreement 7857491)The authors also acknowledge the support of i3S Scientific Platforms:“Bioimaging”member of the PPBI(Grant No:PPBI-POCI-01-0145-FEDER-022122)“Biointerfaces and Nanotechnology”(Grant No:UID/BIM/04293/2019)“BioSciences Screening”(member of the PT-OPENSCREEN(NORTE-01-0145-FEDER-085468)PPBI(PPBI-POCI-01-0145-FEDER-022122)).
文摘The timely establishment of functional neo-vasculature is pivotal for successful tissue development and regen-eration,remaining a central challenge in tissue engineering.In this study,we present a novel(micro)vascular-ization strategy that explores the use of specialized“vascular units”(VUs)as building blocks to initiate blood vessel formation and create perfusable,stroma-embedded 3D microvascular networks from the bottom-up.We demonstrate that VUs composed of endothelial progenitor cells and organ-specific fibroblasts exhibit high angiogenic potential when embedded in fibrin hydrogels.This leads to the formation of VUs-derived capillaries,which fuse with adjacent capillaries to form stable microvascular beds within a supportive,extracellular matrix-rich fibroblastic microenvironment.Using a custom-designed biomimetic fibrin-based vessel-on-chip(VoC),we show that VUs-derived capillaries can inosculate with endothelialized microfluidic channels in the VoC and become perfused.Moreover,VUs can establish capillary bridges between channels,extending the microvascular network throughout the entire device.When VUs and intestinal organoids(IOs)are combined within the VoC,the VUs-derived capillaries and the intestinal fibroblasts progressively reach and envelop the IOs.This promotes the formation of a supportive vascularized stroma around multiple IOs in a single device.These findings un-derscore the remarkable potential of VUs as building blocks for engineering microvascular networks,with ver-satile applications spanning from regenerative medicine to advanced in vitro models.
基金The project was supported by the departmental start-up funds of Indiana University Bloomington,and in part by NSF grants(CCF-1909509,and CMMI-2025434)NIH awards(DP2AI160242,DA056242,and DA047858).
文摘pioids are commonly used for treating chronic pain.However,with continued use,they may induce tolerance and/or hyperalgesia,which limits therapeutic efficacy.The human mechanisms of opioid-induced tolerance and hyperalgesia are significantly understudied,in part,because current models cannot fully recapitulate human pathology.Here,we engineered novel human spinal microphysiological systems(MPSs)integrated with plug-and-play neural activity sensing for modeling human nociception and opioid-induced tolerance.Each spinal MPS consists of a flattened human spinal cord organoid derived from human stem cells and a 3D printed organoid holder device for plug-and-play neural activity measurement.We found that the flattened organoid design of MPSs not only reduces hypoxia and necrosis in the organoids,but also promotes their neuron maturation,neural activity,and functional development.We further demonstrated that prolonged opioid exposure resulted in neurochemical correlates of opioid tolerance and hyperalgesia,as measured by altered neural activity,and downregulation ofμ-opioid receptor expression of human spinal MPSs.The MPSs are scalable,cost-effective,easy-to-use,and compatible with commonly-used well-plates,thus allowing plug-and-play measurements of neural activity.We believe the MPSs hold a promising translational potential for studying human pain etiology,screening new treatments,and validating novel therapeutics for human pain medicine.
基金supported by the National Natural Science Foundation of China(Grant No.31871322)the National Key R&D Program of China(Grant No.2017YFA0700500)the Fundamental Research Funds for the Central Universities,China(Grant Nos.2242020 k10001 and 2242019 k10016).
文摘Organs-on-a-chip is a microfluidic microphysiological system that uses microfluidic technology to analyze the structure and function of living human cells at the tissue and organ levels in vitro.Organs-on-a-chip technology,as opposed to traditional two-dimensional cell culture and animal models,can more closely simulate pathologic and toxicologic interactions between different organs or tissues and reflect the collaborative response of multiple organs to drugs.Despite the fact that many organs-on-a-chip-related data have been published,none of the current databases have all of the following functions:searching,downloading,as well as analyzing data and results from the literature on organs-on-a-chip.Therefore,we created an organs-on-a-chip database(OOCDB)as a platform to integrate information about organs-on-a-chip from various sources,including literature,patents,raw data from microarray and transcriptome sequencing,several open-access datasets of organs-on-a-chip and organoids,and data generated in our laboratory.OOCDB contains dozens of sub-databases and analysis tools,and each sub-database contains various data associated with organs-on-a-chip,with the goal of providing researchers with a comprehensive,systematic,and convenient search engine.Furthermore,it offers a variety of other functions,such as mathematical modeling,three-dimensional modeling,and citation mapping,to meet the needs of organs-on-a-chip.
文摘.Organ-on-Chip(OoC)has emerged as a revolutionary approach to emulate human organ function-ality in vitro,offering unparalleled insights into physiological processes and disease modeling.The integration of artificial intelligence(AI)with OoC platforms presents a transformative synergy,combining the precision of microscale organ replication with the analytical prowess of intelligent algorithms,is emerging as a transforma-tive force in harnessing the full potential of OoC.This perspective investigates the multifaceted implications of integrating AI with OoC,examining its impact on biomedical research,acknowledging the synergistic po-tential that arises from combining the precision of microscale organ replication with the analytical capabilities of intelligent algorithms,and fostering a future where the intricate workings of the technology and biology.
基金funding from National Key Research and Development Program of China(No.2018YFA0703000)The National Natural Science Foundation of China No.52275294 and supports from Zhejiang University Global Partnership Fundthe financial support from Chinese Scholar Councils(CSC)Scholarship fund.We would also like to thank Dr.Zhaoying Li and Dr.Elisabeth Lauren Gill for their essential contributions.
文摘In the human body,almost all cells interact with extracellular matrices(ECMs),which have tissue and organ-specific compositions and architectures.These ECMs not only function as cellular scaffolds,providing structural support,but also play a crucial role in dynamically regulating various cellular functions.This comprehensive review delves into the examination of biofabrication strategies used to develop bioactive materials that accurately mimic one or more biophysical and biochemical properties of ECMs.We discuss the potential integration of these ECM-mimics into a range of physiological and pathological in vitro models,enhancing our understanding of cellular behavior and tissue organization.Lastly,we propose future research directions for ECM-mimics in the context of tissue engineering and organ-on-a-chip applications,offering potential advancements in therapeutic approaches and improved patient outcomes.
