The global demand for in vitro respiratory airway models has surged due to the coronavirus disease 2019(COVID-19)pandemic.Current state-of-the-art models use polymer membranes to separate epithelial cells from other c...The global demand for in vitro respiratory airway models has surged due to the coronavirus disease 2019(COVID-19)pandemic.Current state-of-the-art models use polymer membranes to separate epithelial cells from other cell types,creating a nonphysiological barrier.In this study,we applied three-dimensional(3D)printing and bioprinting to develop an in vitro model where endothelial and epithelial cells were in direct contact,mimicking their natural arrangement.This proof-ofconcept model includes a culture chamber,with an endothelial bioink printed and perfused through an epithelial channel.In silico simulations of the air velocity within the channel revealed shear stress values ranging from 0.13 to 0.39 Pa,aligning with the desired in vivo shear stress observed in the bronchi regions(0.1–0.4 Pa).Biomechanical movements during resting breathing were mimicked by incorporating a textile mesh positioned away from the cell–cell interface.The epithelial channel demonstrated a capacity for compression and expansion of up to−14.7%and+6.4%,respectively.Microscopic images showed that the epithelial cells formed a uniform monolayer within the lumen of the channel close to the bioprinted endothelial cells.Our novel model offers a valuable tool for future research into respiratory diseases and potential treatments under conditions closely mimicking those in the lung.展开更多
Currently,artificial-membrane lungs consist of thousands of hollow fiber membranes where blood flows around the fibers and gas flows inside the fibers,achieving diffusive gas exchange.At both ends of the fibers,the in...Currently,artificial-membrane lungs consist of thousands of hollow fiber membranes where blood flows around the fibers and gas flows inside the fibers,achieving diffusive gas exchange.At both ends of the fibers,the interspaces between the hollow fiber membranes and the plastic housing are filled with glue to separate the gas from the blood phase.During a uniaxial centrifugation process,the glue forms the“potting.”The shape of the cured potting is then determined by the centrifugation process,limiting design possibilities and leading to unfavorable stagnation zones associated with blood clotting.In this study,a new multiaxial centrifugation process was developed,expanding the possible shapes of the potting and allowing for completely new module designs with potentially superior blood flow guidance within the potting margins.Two-phase simulations of the process in conceptual artificial lungs were performed to explore the possibilities of a biaxial centrifugation process and determine suitable parameter sets.A corresponding biaxial centrifugation setup was built to prove feasibility and experimentally validate four conceptual designs,resulting in good agreement with the simulations.In summary,this study shows the feasibility of a multiaxial centrifugation process allowing greater variety in potting shapes,eliminating inefficient stagnation zones and more favorable blood flow conditions in artificial lungs.展开更多
During lung cancer metastasis,tumor cells undergo epithelial-to-mesenchymal transition(EMT),enabling them to intravasate through the vascular barrier and enter the circulation before colonizing secondary sites.Here,a ...During lung cancer metastasis,tumor cells undergo epithelial-to-mesenchymal transition(EMT),enabling them to intravasate through the vascular barrier and enter the circulation before colonizing secondary sites.Here,a human in vitro microphysiological model of EMT-driven lung cancer intravasation-on-a-chip was developed and coupled with machine learning(ML)-assisted automatic identification and quantification of intravasation events.A robust EMT-inducing cocktail(EMT-IC)was formulated by augmenting macrophage-conditioned medium with transforming growth factor-β1.When introduced into microvascular networks(MVNs)in microfluidic devices,EMT-IC did not affect MVN stability and physiologically relevant barrier functions.To model lung cancer intravasation on-a-chip,EMT-IC was supplemented into co-cultures of lung tumor micromasses and MVNs.Wihin 24 h of exposure,EMT-IC facilitated the insertion of membrane protrusions of migratory A549 cells into microvascular structures,followed by successful intravasation.EMT-IC reduced key basement membrane and vascular junction proteins-laminin and VE-Cadherin-rendering vessel walls more permissive to intravasating cells.ML-assisted vessel segmentation combined with co-localization analysis to detect intravasation events confirmed that EMT induction significantly increased the number of intravasation events.Introducing metastatic(NCI-H1975)and non-metastatic(BEAS-2B)cell lines demonstrated that both,baseline intravasation potential and responsiveness to EMT-IC,are reflected in the metastatic predisposition of lung cancer cell lines,highlighting the model’s universal applicability and cell-specific sensitivity.The reproducible detection of intravasation events in the established model provides a physiologically relevant platform to study processes of cancer metastasis with high spatio-temporal resolution and short timeframe.This approach holds promise for improved drug development and informed personalized patient treatment plans.展开更多
基金supported by the Volkswagen Foundation(Grant No.Az 99078 to DDC,ALT,and MT)funded by the Deutsche Forschungsgemeinschaft(DFG,German Research Foundation)under Germany’s Excellence Strategy–2082/1–390761711(to DDC)part of the research training group GRK 2415–Mechanobiology in Epithelial 3D Tissue Constructs(project number 363055819,to ALT and SJ).
文摘The global demand for in vitro respiratory airway models has surged due to the coronavirus disease 2019(COVID-19)pandemic.Current state-of-the-art models use polymer membranes to separate epithelial cells from other cell types,creating a nonphysiological barrier.In this study,we applied three-dimensional(3D)printing and bioprinting to develop an in vitro model where endothelial and epithelial cells were in direct contact,mimicking their natural arrangement.This proof-ofconcept model includes a culture chamber,with an endothelial bioink printed and perfused through an epithelial channel.In silico simulations of the air velocity within the channel revealed shear stress values ranging from 0.13 to 0.39 Pa,aligning with the desired in vivo shear stress observed in the bronchi regions(0.1–0.4 Pa).Biomechanical movements during resting breathing were mimicked by incorporating a textile mesh positioned away from the cell–cell interface.The epithelial channel demonstrated a capacity for compression and expansion of up to−14.7%and+6.4%,respectively.Microscopic images showed that the epithelial cells formed a uniform monolayer within the lumen of the channel close to the bioprinted endothelial cells.Our novel model offers a valuable tool for future research into respiratory diseases and potential treatments under conditions closely mimicking those in the lung.
文摘Currently,artificial-membrane lungs consist of thousands of hollow fiber membranes where blood flows around the fibers and gas flows inside the fibers,achieving diffusive gas exchange.At both ends of the fibers,the interspaces between the hollow fiber membranes and the plastic housing are filled with glue to separate the gas from the blood phase.During a uniaxial centrifugation process,the glue forms the“potting.”The shape of the cured potting is then determined by the centrifugation process,limiting design possibilities and leading to unfavorable stagnation zones associated with blood clotting.In this study,a new multiaxial centrifugation process was developed,expanding the possible shapes of the potting and allowing for completely new module designs with potentially superior blood flow guidance within the potting margins.Two-phase simulations of the process in conceptual artificial lungs were performed to explore the possibilities of a biaxial centrifugation process and determine suitable parameter sets.A corresponding biaxial centrifugation setup was built to prove feasibility and experimentally validate four conceptual designs,resulting in good agreement with the simulations.In summary,this study shows the feasibility of a multiaxial centrifugation process allowing greater variety in potting shapes,eliminating inefficient stagnation zones and more favorable blood flow conditions in artificial lungs.
基金supported by the research fund to the Center for Neuromusculoskeletal Restorative Medicine from Health@InnoHK program launched by Innovation and Technology Commission,the Government of the Hong Kong Special Administrative Region of the People’s Republic of China(AB),by the Health and Medical Research Fund(08191066,AB)a direct grant(4054732,AB)from the Faculty of Medicine,CUHK+1 种基金supported by the Lee Quo Wei and Lee Yick Hoi Lun Professorship in Tissue Engineering and Regenerative Medicine.A.J.receives a Walter Benjamin postdoctoral fellowship from the Deutsche Forschungsgemeinschaft(DFG,German Research Foundation,Germany521343357).
文摘During lung cancer metastasis,tumor cells undergo epithelial-to-mesenchymal transition(EMT),enabling them to intravasate through the vascular barrier and enter the circulation before colonizing secondary sites.Here,a human in vitro microphysiological model of EMT-driven lung cancer intravasation-on-a-chip was developed and coupled with machine learning(ML)-assisted automatic identification and quantification of intravasation events.A robust EMT-inducing cocktail(EMT-IC)was formulated by augmenting macrophage-conditioned medium with transforming growth factor-β1.When introduced into microvascular networks(MVNs)in microfluidic devices,EMT-IC did not affect MVN stability and physiologically relevant barrier functions.To model lung cancer intravasation on-a-chip,EMT-IC was supplemented into co-cultures of lung tumor micromasses and MVNs.Wihin 24 h of exposure,EMT-IC facilitated the insertion of membrane protrusions of migratory A549 cells into microvascular structures,followed by successful intravasation.EMT-IC reduced key basement membrane and vascular junction proteins-laminin and VE-Cadherin-rendering vessel walls more permissive to intravasating cells.ML-assisted vessel segmentation combined with co-localization analysis to detect intravasation events confirmed that EMT induction significantly increased the number of intravasation events.Introducing metastatic(NCI-H1975)and non-metastatic(BEAS-2B)cell lines demonstrated that both,baseline intravasation potential and responsiveness to EMT-IC,are reflected in the metastatic predisposition of lung cancer cell lines,highlighting the model’s universal applicability and cell-specific sensitivity.The reproducible detection of intravasation events in the established model provides a physiologically relevant platform to study processes of cancer metastasis with high spatio-temporal resolution and short timeframe.This approach holds promise for improved drug development and informed personalized patient treatment plans.
