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2D profiling of tumor chemotactic and molecular phenotype at single cell resolution using a SERS-Microfluidic chip

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Nano Research 2022年第5期15卷 4357-4365页

作者： Yizhi Zhang Lei Wu Kuo Yang Shenfei Zong Zhuyuan Wang Yiping Cui Advanced Photonics Center School of Electronic Science and EngineeringSoutheast UniversityNanjing 210096China Department of Biomedical Engineering Nanjing University of Aeronautics and AstronauticsNanjing 211106China

Emerging single-cell technologies create new opportunities for unraveling tumor heterogeneity.However,the development of high-content phenotyping platform is still at its infancy.Here,we develop a Microfluidic chip for two-dimensional(2D)profiling of tumor chemotactic and molecular features at single cell resolution.Individual cells were captured by the triangular micropillar arrays in the cell-loading channel,facilitating downstream single-cell analysis.For 2D phenotyping,the chemotactic properties of tumor cells were visualized through cellular migratory behavior in microchannels,while their protein expression was profiled with multiplex surface enhanced Raman scattering(SERS)nanovectors,in which Raman reporter-embedded gold@silver core-shell nanoparticles(Au@Ag REPs)were modified with DNA aptamers targeting cellular surface proteins.As a proof of concept,breast cancer cells with diverse phenotypes were tested on the chip,demonstrating the capability of this platform for simultaneous chemotactic and molecular analysis.The chip is expected to provide a powerful tool for investigating tumor heterogeneity and promoting clinical precision medicine.

关键词： gold@silver nanoparticles surface enhanced Raman spectroscopy Microfluidic chip single cell analysis two-dimensional(2D)phenotype profiling

The protective effect of icariin on glucocorticoid-damaged BMECs explored by microfluidic organ chip

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Chinese Chemical Letters 2022年第6期33卷 3177-3182页

作者： Tengqi Li Qingyu Zhang Fuqiang Gao Yadi Liu Wei Sun Yiyang Dong Department of Orthopedics Peking University China-Japan Friendship School of Clinical MedicineBeijing 100029China Department of Orthopedics Peking University Shougang HospitalBeijing 100144China Department of Orthopedics Shandong Provincial Hospital Affiliated to Shandong First Medical UniversityJi’nan 250021China Department of Orthopedics China-Japan Friendship HospitalBeijing 100029China Office of Academic Affairs Beijing University of Chemical TechnologyBeijing 100029China Department of Pharmaceutical Analysis College of Life Science and TechnologyBeijing University of Chemical TechnologyBeijing 100029China

Osteonecrosis of the femoral head(ONFH)is a devastating musculoskeletal disease characterized by the impaired circulation of bone.The purpose of this study was to explore the underlying mechanisms of the protective effect of icariin on the glucocorticoid-induced injury of bone microvascular endothelial cells(BMECs).Normal BMECs were extracted from the femoral heads by enzymatic isolation and magneticactivated cell sorting methods.Dexamethasone and icariin were used to intervene BMECs in microfluidic organ chips,and phalloidin staining was conducted to observe the cell morphology and viability.Then next-generation transcriptome sequencing and real-time quantitative reverse transcription-polymerase chain reaction(q RT-PCR)were performed to identify the differentially expressed genes(DEGs)in different groups.Through the microfluidic organ chip,it can be observed that after dexamethasone intervention,the filamentous structure in cell fibers disappeared and the cell morphology changed from spindle to round until death.Icariin could relieve these changes and showed a protective effect on glucocorticoiddamaged BMECs.In addition,201 DEGs were detected between the icariin protection group and the dexamethasone group,which were significantly enriched in 17 signaling pathways.8 of the top ten selected hub genes(IL6,PTGS2,VEGFA,etc.)were confirmed by q RT-PCR.Transcription factors(TFs)-gene network showed 63 connections between 18 TFs and 12 DEGs.For instance,GATA2 could regulate 5 DEGs.The associations between 92 mi RNA and 12 DEGs were visualized in a mi RNA-gene network.The hub mi RNA,has-mir-335–5 p was predicted to interact with 8 DEGs(PTGS2,VEGFA,etc.).microfluidic organ chips could provide excellent morphological results for cell experiments,by which it could be observed that icariin showed a protective effect on the glucocorticoid-induced injury of BMECs.Beside,these DEGs,possible regulatory TF(GATA2,FOXC1,etc.)and mi RNA(has-mir-335–5 p)might be dysregulated in the initiation of ONFH and have prospective importance

关键词： Osteonecrosis of the femoral head Bone microvascular endothelial cells Glucocorticoid Microfluidic chip Bioinformatic analysis

A modular single-cell pipette Microfluidic chip coupling to ETAAS and ICP-MS for single cell analysis

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Chinese Chemical Letters 2022年第3期33卷 1373-1376页

作者： Xing Wei Meng Yang Ze Jiang Jinhui Liu Xuan Zhang Mingli Chen Jianhua Wang Research Center for Analytical Sciences Department of ChemistryCollege of SciencesNortheastern UniversityShenyang 110819China

Accurate single-cell capture is a crucial step for single cell biological and chemical analysis. Conventional single-cell capturing often confront operational complexity, limited efficiency, cell damage, large scale but low accuracy, incompetence in the acquirement of nano-upgraded single-cell liquid. Flow cytometry has been widely used in large-scale single-cell detection, while precise single-cell isolation relies on both a precision operating platform and a microscope, which is not only extremely inefficient, but also not conducive to couple with modern analytical instruments. Herein, we develop a modular single-cell pipette(m SCP) Microfluidic chip with high efficiency and strong applicability for accurate direct capture of single viable cell from cell suspensions into nanoliter droplets(30-1000 n L). The m SCP is used as a sampling platform for the detection of Cd Te quantum dots in single cells with electrothermal atomic absorption spectrometry(ETAAS) for the first time. It also ensures precise single-cell sampling and detection by inductively coupled plasma mass spectrometry(ICP-MS).

关键词： Single-cell pipette Microfluidic chip Single-cell capture ETAAS ICP-MS CdTe QDs

Microfluidic chip of concentration gradient and fluid shear stress on a single cell level

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Chinese Chemical Letters 2022年第6期33卷 3133-3138页

作者： Xuexia Lin Jianlong Su Shufeng Zhou Department of Chemical Engineering&Pharmaceutical Engineering College of Chemical EngineeringHuaqiao UniversityXiamen 361021China

Concentration gradient and fluid shear stress(FSS)for cell microenvironment were investigated through microfluidic technology.The Darcy–Weisbach equation combined with computational fluid dynamics modeling was exploited to design the Microfluidic chip,and the FSS distribution on the cell model with varying micro-channels(triangular,conical,and elliptical).The diffusion with the incompressible laminar flow model by solving the time-dependent diffusion–convection equation was applied to simulate the gradient profiles of concentration in the micro-channels.For the study of single cell in-depth,the FSS was investigated by the usage of polystyrene particles and the concentration diffusion distribution was studied by the usage of different colors of dyes.A successful agreement between model simulations and experimental data was obtained.Finally,based on the established method,the communication between individual cells was envisaged and modeled.The developed method provides valuable insights and allows to continuously improve the design of microfluidic devices for the study of single cell,the occurrence and development of tumors,and therapeutic applications.

关键词： Concentration gradient Fluid shear stress Single particle Computational fluid dynamics Microfluidic chip

In vitro study of emodin-induced nephrotoxicity in human renal glomerular endothelial cells on a Microfluidic chip

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BIOCELL 2023年第1期47卷 125-131页

作者： ZHUO YANG WEN QIN DI CHEN JUNSHENG HUO JINGBO WANG LIYUAN WANG QIN ZHUO JIYONG YIN National Institute for Nutrition and Health Chinese Center for Disease Control and PreventionBeijing100050China

Emodin is an effective component of rhubarb with positive pharmacological effects on human health.However,it is also toxic to different cells or tissues to varying degrees.The effects of emodin on glomerular endothelial cells(GECs)remain to be tested,and the documented works were always performed in vitro and hardly reflect the real physiological situation.To study the effects of emodin on GECs in a biomimetic environment,we utilized a Microfluidic chip to assess the physiological reaction of human renal glomerular endothelial cells to various concentrations of emodin in this work.The results showed that emodin caused cytotoxicity,impaired glomerular filtration barrier integrity to macromolecules,and increased barrier permeability in a dose-dependent manner.With the increase in emodin concentration,the concentration of the pro-inflammatory cytokine tumor necrosis factor-α,interleukin(IL)-6,transforming growth factor-β1,and monocyte chemoattractant protein(MCP-1)increased while the production of inflammatory cytokine IL-6 first increased and then decreased with the increase in emodin concentration.Our findings shed new light on emodin-induced nephrotoxicity and provide insights for the application of Microfluidic chip devices to reveal drug-cell interactions.

关键词： Emodin Nephrotoxicity HRGECs Microfluidic chip

High Precision Temperature Measurement for Microfluidic chip Applications

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Journal of Shanghai Jiaotong university(Science) 2022年第5期27卷 699-705页

作者：熊越夫吴校生曾照丰黄山陈天培 National Key Laboratory of Science and Technology on Micro/Nano Fabrication Department of Micro/Nano ElectronicsShanghai Jiao Tong UniversityShanghai200240China

Biochemical reaction in Microfluidic chip is sensitive to temperature.Temperature precise control in a small size device requires the temperature measurement with high measurement precision.Traditional temperature measurement method usually measures the voltage drop of the thermistor,which is excited by a constant current source.This method requires the constant current source with high precision and stability.The output of the constant current source is influenced by environmental factors,resulting in a larger measurement error.To solve this problem,a proportion method,a two-layer filtering algorithm,and a power management technique were applied to improve the temperature measurement precision.The proportion method can reduce the low frequency fluctuation error.The two-layer filtering algorithm can reduce the high frequency fluctuation error furtherly.The power management technique used can improve the system stability.Through testing the temperature measurement system built,the experimental results show that the fluctuation error can be significantly decreased from 0.5◦C to 0.2◦C.

关键词： Microfluidic chip temperature measurement proportion method two-layer filtering algorithm fluctuation error

Inhibition of the Hedgehog Signaling Pathway Depresses the Cigarette Smoke-Induced Malignant Transformation of 16HBE Cells on a Microfluidic chip

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Chinese Medical Journal 2018年第10期131卷 1191-1198页

作者： Yong-Xin Qin Zhi-Hui Yang Xiao-Hui DU Hui Zhao Yuan-Bin Liu Zhe Guo Qi Wang Department of Respiratory Medicine The Second Affiliated Hospital Dalian Medical University Dalian Liaoning 116027 China Department of Critical Care Medicine The First Affiliated Hospital Dalian Medical University Dalian Liaoning 116021 China Department of Scientific Research Center The Second Affiliated Hospital Dalian Medical University Dalian Liaoning 116027 China

Background：The hedgehog signaling system （HHS） plays an important role in the regulation of cell proliferation and differentiation during the embryonic phases.However,little is known about the involvement of HHS in the malignant transformation of cells.This study aimed to detect the role of HHS in the malignant transformation of human bronchial epithelial （16HBE） cells.Methods：In this study,two Microfluidic chips were designed to investigate cigarette smoke extract （CSE）-induced malignant transformation of cells.chip A contained a concentration gradient generator,while chip B had four cell chambers with a central channel.The 16HBE cells cultured in chip A were used to determine the optimal concentration of CSE for inducing malignant transformation.The 16HBE cells in chip B were cultured with 12.25% CSE （Group A）,12.25% CSE ± 5 μmol/L cyclopamine （Group B）,or normal complete medium as control for 8 months （Group C）,to establish the in vitro lung inflammatory-cancer transformation model.The transformed cells were inoculated into 20 nude mice as cells alone （Group 1） or cells with cyclopamine （Group 2） for tumorigenesis testing.Expression of HHS proteins was detected by Western blot.Data were expressed as mean ± standard deviation.The t-test was used for paired samples,and the difference among groups was analyzed using a one-way analysis of variance.Results：The optimal concentration of CSE was 12.25%.Expression of HHS proteins increased during the process of malignant transformation （Group B vs.Group A,F =7.65,P 〈 0.05）.After CSE exposure for 8 months,there were significant changes in cellular morphology,which allowed the transformed cells to grow into tumors in 40 days after being inoculated into nude mice.Cyclopamine could effectively depress the expression of HHS proteins （Group C vs.Group B,F =6.47,P 〈 0.05） and prevent tumor growth in nude mice （Group 2 vs.Group 1,t=31.59,P〈 0.01）.Conclusions：The activity of HHS is upregulated during the CSE-induced malignant transformation of 16HBE cells.Cyclopamine can effect

关键词： Hedgehog Signaling System Lung Cancer Malignant Transformation Microfluidic chip

Flow field analyses of a porous membrane-separated,double-layered Microfluidic chip for cell co-culture

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Acta Mechanica Sinica 2020年第3期36卷 754-767页

作者： Shenbao Chen Jian Xue Jinrong Hu Qihan Ding Luwen Zhou Shiliang Feng Yuhong Cui Shouqin Lu Mian Long Center of Biomechanics and Bioengineering Key Laboratory of Microgravity(National Microgravity Laboratory)Beijing Key Laboratory of Engineered Construction and Mechanobiologyand CAS Center for Excellence in Complex System MechanicsInstitute of MechanicsChinese Academy of SciencesBeijing100190China School of Engineering Science University of Chinese Academy of SciencesBeijing 100049China State Key Laboratory of Nonlinear Mechanics(LNM) Institute of MechanicsChinese Academy of SciencesBeijing 100190China Faculty of Mechanical Engineering and Mechanics Ningbo UniversityNingbo 315211ZhejiangChina Department of Mechanics Tianjin UniversityTianjin 300072China

Organs-on-chips composed of a porous membrane-separated,double-layered channels are used widely in elucidating the effects of cell co-culture and flow shear on biological functions.While the diversity of channel geometry and membrane permeability is applied,their quantitative correlation with flow features is still unclear.Immersed boundary methods(IBM)simulations and theoretical modelling were performed in this study.Numerical simulations showed that channel length,height and membrane permeability jointly regulated the flow features of flux,penetration velocity and wall shear stress(WSS).Increase of channel length,lower channel height or membrane permeability monotonically reduced the flow flux,velocity and WSS in upper channel before reaching a plateau.While the flow flux in lower channel monotonically increased with the increase of each factor,the WSS surprisingly exhibited a biphasic pattern with first increase and then decrease with increase of lower channel height.Moreover,the transition threshold of maximum WSS was sensitive to the channel length and membrane permeability.Theoretical modeling,integrating the transmembrane pressure difference and inlet flow flux with chip geometry and membrane permeability,was in good agreement with IBM simulations.These analyses provided theoretical bases for optimizing flow-specified chip design and evaluating flow microenvironments of in vivo tissue.

关键词： Immersed boundary method Porous membrane Wall shear stress Penetration velocity Microfluidic chip

Application of adaptive pressure-driven Microfluidic chip in thyroid function measurement

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Chinese Chemical Letters 2021年第5期32卷 1747-1750页

作者： Xingshang Xu Nongyue He State Key Laboratory of Bioelectronics School of Biological Science and Medical EngineeringSoutheast UniversityNanjing 210096China Hunan Key Laboratory of Biomedical Nanomaterials and Devices College of Life Sciences and ChemistryHunan University of TechnologyZhuzhou 412007China Nanjing Lanyu Biotechnology Co. Ltd.Nanjing 210000China

The improvement in accuracy of in vitro diagnosis has always been the focus of early screening of thyroid dysfunction.We constructed a Microfluidic chip based on a polystyrene polymer substrate.Total triiodothyronine(TT3),total thyroxine(TT4),free triiodothyronine(FT3),free thyroxine(FT4),and thyrotropin(TSH) in human whole blood samples were analysed by fluorescence immunoassay to evaluate thyroid function.The results indicate that the Microfluidic chip shows a good linear relationship in the detection of TT3,TT4,FT3,FT4,and TS H standards,and the correlation coefficient(r) is not less than0.9900.In addition,the chip also has strong anti-interference(RSD%≤5%) and good repeatability(CV≤8%),and its inter-batch differences are small(CV ≤15%).The results of practical application in clinical thyroid function mea surement indicated its high accuracy(r≥0.9900).It provides a new method for the determination of thyroid function and lays a foundation for subsequent clinical application.

关键词： Microfluidic chip Adaptive pressure drive Thyroid dysfunction In vitro diagnosis Whole blood injection