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Materials Today Bio · 2023 · IF 8.2

Immunosuppressive dead cell as lung-targeting vehicle and cytokine absorption material for cytokine storm attenuation of pneumonia

Tianyuan Ci,Yaoxuan Xiong,Jinniu Zhang,Jing Zang,Nianping Feng

Effectively controlling cytokine storm is important to reduce the mortality of severe pneumonia. In this work a bio-functional dead cell was engineered by one-time quick shock of live immune cells in liquid nitrogen, and the obtained immunosuppressive dead cell could server as both lung-targeting vehicle and cytokine absorption material. After loading the anti-inflammatory drugs of dexamethasone (DEX) and baicalin (BAI), the drug-loaded dead cell (DEX&BAI/Dead cell) could first passively target to the lung after intravenous administration and quickly release the drugs under high shearing stress of pulmonary capillaries, realizing drug enrichment in the lung. Then, the immunosuppressive dead cell acted as the camouflage of normal immune cells with various cytokine receptors exposing on their surface, to “capture” the cytokines and further reduce the state of inflammation. With above formulation design, a synergic anti-inflammatory effect between drugs and carrier could be achieved. In a lipopolysaccharide-induced pneumonia mice model, this system could calm down the cytokine storm with high efficacy and elongate the survival of mice.

研究领域: 药理学 · 免疫学 · 生物技术 · 治疗学
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ACS Biomaterials Science & Engineering · 2023 · IF 5.8

On-Chip Label-Free Sorting of Living and Dead Cells

Guowei Wang,Chengpan Li,Chunguang Miao,Shibo Li,Bensheng Qiu,Weiping Ding

With the emergence of various cutting-edge micromachining technologies, lab on a chip is growing rapidly, but it is always a challenge to realize the on-chip separation of living cells from cell samples without affecting cell activity and function. Herein, we report a novel on-chip label-free method for sorting living and dead cells by integrating the hypertonic stimulus and tilted-angle standing surface acoustic wave (T-SSAW) technologies. On a self-designed microfluidic chip, the hypertonic stimulus is used to distinguish cells by producing volume differences between living and dead cells, while T-SSAW is used to separate living and dead cells according to the cell volume difference. Under the optimized operation conditions, for the sample containing 50% of living human umbilical vein endothelial cells (HUVECs) and 50% of dead HUVECs treated with paraformaldehyde, the purity of living cells after the first separation can reach approximately 80%, while after the second separation, it can be as high as 93%; furthermore, the purity of living cells after separation increases with the initial proportion of living cells. In addition, the chip we designed is safe for cells and can robustly handle cell samples with different cell types or different causes of cell death. This work provides a new design of a microfluidic chip for label-free sorting of living and dead cells, greatly promoting the multi-functionality of lab on a chip.

CAS / MDL: 9048-46-8
研究领域: 微流控技术 · 生物医学工程 · 细胞生物学 · 芯片实验室技术
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PLoS One · 2024 ·Vol. 19 ·期 8 ·e0308132 · IF 2.9

Oligodendroglia-to-pericyte conversion after lipopolysaccharide exposure is gender-dependent

Qingting Yu,Linyuan Zhang,Ting Xu,Jiapeng Shao,Falei Yuan,Zuisu Yang,Yuncheng Wu,Haiyan Lyu

To investigate the sex-dependent differentiation of Sox10 cells and their response to pathological conditions such as lipopolysaccharide (LPS) exposure or ischemia, we utilized Sox10 Cre-ER T2 , tdTomato mice. Tamoxifen administration induced the expression of red fluorescent protein (RFP) in these cells, facilitating their subsequent tracking and analysis after LPS injection and ischemia via immunofluorescence staining. Propidium iodide (PI) was injected to label necrotic cells following LPS administration. We found that the conversion of Sox10 cells to pericytes in female mice was significantly higher than in male mice, especially in those exposed to LPS. After LPS injection, the number of PI + necrotic cells were significantly greater in females than in males. Moreover, RFP + cells did not co-localize with glial fibrillary acidic protein (GFAP) or cluster of differentiation 11b (CD11b). Similarly, after brain ischemia, RFP + cells did not express cluster of differentiation 13 (CD13), neuronal nuclei (NeuN), GFAP, or ionised calcium binding adaptor molecule 1 (Iba-1). These findings indicate that the conversion of Sox10 cells to pericytes following LPS exposure is sex-dependent, with neither male nor female groups showing differentiation into other cell types after LPS exposure or under ischemic conditions. The differences in LPS-induced necrosis of pericytes between sexes may explain the variations in the conversion of Sox10 cells to pericytes in both sexes.

研究领域: 神经科学 · 免疫学 · 细胞生物学
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SENSORS AND ACTUATORS B-CHEMICAL · 2020 · IF 7.1

High-throughput dielectrophoretic cell sorting assisted by cell sliding on scalable electrode tracks made of conducting-PDMS

Xiaofeng Nie,Yuan Luo,Penghui Shen,Chengwu Han,Duli Yu,Xiaoxing Xing

Dielectrophoresis (DEP) as a label-free cell separation approach in microdevices has been extensively investigated for a variety of applications. 3D microelectrodes made of conducting-PDMS inherit the merit of volumetric electrodes for generating influential DEP force throughout the entire channel depth and meanwhile, exploit low-cost fabrication process by soft lithography. However, the configuration of conducting-PDMS electrodes is limited to being embedded in sidewall of flow chamber, which leads to rather low flow rate and difficulties in extension of the flow rate. We previously reported a more effective configuration with 3D interdigitated electrodes made of silicon that assist cell sliding along solid tracks, yet such device requires expensive silicon dry etching and, moreover, the track appears to be patterned with non-straight and wavy outline, which not only hinders the flow rate but also allows cell sliding to occur only along its downstream side. Here we demonstrate low-cost silver-PDMS electrode-track featuring ideally straight outline that induces rather uniform drag to drive smooth cell sliding. Such design achieves live and dead cell separation at flow rate twice as that of silicon tracks with cell loading concentration 10 times higher. It also fully utilizes the track to enable cell sliding on both of the up- and down-stream sides. Notably, we also demonstrate that this track is expandable to be V-shape for more advanced bidirectional cell sliding, which is showcased here by tumor cells separation from lymphocytes at 1.2 mL/h. Such results greatly enhance the throughput as compared to the state-of-art conducting-PDMS based cell separator.

CAS / MDL: 148504-34-1
研究领域: 生物医学工程 · 微流控技术 · 细胞分离 · 生物技术
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CHEMOSPHERE · 2015 · IF 3.34

Necrotic cell death induced by the protein-mediated intercellular uptake of CdTe quantum dots

Lu Lai,Jian-Cheng Jin,Zi-Qiang Xu,Ping Mei,Feng-Lei Jiang,Yi Liu

The toxicity of CdTe QDs with nearly identical maximum emission wavelength but modified with four different ligands (MPA, NAC, GSH and dBSA) to HEK293 and HeLa cells were investigated using flow cytometry, spectroscopic and microscopic methods. The results showed that the cytotoxicity of QDs increased in a dose- and time-dependent manner. No appreciable fraction of cells with sub-G1 DNA content, the loss of membrane integrity, and the swelling of nuclei clearly indicated that CdTe QDs could lead to necrotic cell death in HEK293 cells. JC-1 staining and TEM images confirmed that QDs induced MPT, which resulted in mitochondrial swelling, collapse of the membrane potential. MPT is an important step in QDs-induced necrosis. Moreover, QDs induced MPT through the elevation of ROS. The fluorimetric assay and theoretical analysis demonstrated ROS production has been associated with the internalization of QDs with cells. Due to large surface/volume ratios of QDs, when QDs added in the culture medium, serum proteins in the culture medium will be adsorbed on the surface of QDs. This adsorption of serum protein will change the surface properties and size, and then mediate the cellular uptake of QDs via the clathrin-mediated endocytic pathway. After entering into cells, the translocation of QDs in cells is usually via endosomal or lysosomal vesicles. The rapid degradation of QDs in lysosome and the lysosomal destabilization induce cell necrosis. This study provides a basis for understanding the cytotoxicity mechanism of CdTe QDs, and valuable information for safe use of QDs in the future.

CAS / MDL: 298-93-1
研究领域: 毒理学 · 细胞生物学 · 材料科学 · 生物医学工程
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ANALYTICAL CHEMISTRY · 2024 · IF 6.7

User-Friendly Multifunctional Red-Emissive Carbon Dots for Rapid Cell Nucleus Staining via Targeting Nuclear Proteins

Jingbo Sun,Hongchen Li,Min Ouyang,Jing Cheng,Dong Xu,Xiaofeng Tan,Qinlu Lin

Cytoarchitectural staining is of great importance in disease diagnosis and cell biology research. This study developed user-friendly multifunctional red-emissive carbon dots (R-CDs) for rapid cell nucleus staining via targeting nuclear proteins. R-CDs, simply prepared by electrochemical treatment of 1,2,4-benzenetriamine, exhibit strong emission at 635 nm when excited at 507 nm. The R-CDs can rapidly stain the nucleus of human SH-SY5Y, HepG2, and HUH-7 cells with a high signal-to-noise ratio owing to fluorescence enhancement after entering the nucleus. Compared to conventional cytosolic dyes such as Hoechst and DAPI, R-CDs are cheaper, more highly dispersed in water, and more stable (requiring no stringent storage conditions). The R-CDs show stable optical properties with insignificant photobleaching over 7 days and salt resistance up to 2 M of NaCl. More importantly, R-CDs, possessing a positive charge, allow rapid staining of live cells (3 min) and dead cells (10 s) in saline. According to kinetic variation, R-CDs can distinguish live cells from dead cells. Staining exhibits high efficiency in onion epidermal cells, Aspergillus niger, Caenorhabditis elegans, and human spermatozoa. The mechanism for efficient staining is based on their fast accumulation in the nucleus due to their small size and positive charge and strong interaction with nuclear proteins at amino acid residues of histidine and arginine, resulting in fluorescence enhancement by dozens of times. The developed R-CDs do not bind to DNA and would not cause genetic damage and will find various safe applications in biological and medical fields.

研究领域: 细胞生物学 · 疾病诊断 · 材料科学 · 光学性质 · 生物医学应用
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