By combining optical trapping with fluorescence imaging, the adhesion and deformation characteristics of suspension cells were probed on single cell level. We found that, after 24 h of co-culturing, stable attachment between non-adherent K562 cells and polystyrene beads coated with fibronectin, collagen I, or G-actin can all be formed with an adhesion energy density in the range of 1–3×10−2 mJ/m2, which is about one order of magnitude lower than the reported values for several adherent cells. In addition, it was observed that the formation of a stronger adhesion is accompanied with the appearance of a denser actin cell cortex, especially in the region close to the cell-bead interface, resulting in a significant increase in the apparent modulus of the cell. Findings here could be important for our understanding of why the aggregation of circulating cells, like that in leukostasis, takes place in vivo as well as how such clusters of non-adherent cells behave. The method proposed can also be useful in investigating adhesion and related phenomena for other cell types in the future.
This study aimed to determine the composition and diversity of bacterial communities on the ocular surface before and after the intervention with sodium hyaluronate eye drops (with or without preservatives) using 16S rRNA gene amplicon sequencing.Sixteen healthy adults were randomly divided into two groups and treated with sodium hyaluronate eye drops with or without preservatives for 2 weeks. The individuals used the same artificial tears in both eyes. The microbial samples from the conjunctival sac of each participant were collected at baseline and 2 weeks after intervention. The diversity and taxonomic differences among different groups before and after intervention were compared by sequencing the V3-V4 region of the 16S rRNA gene.The similarity in the binocular microbial community was high in 1 of the 16 volunteers (Bray-Curtis dissimilarity score < 0.3). At the genus level, 11 bacteria were detected in all samples with an average relative abundance of more than 1%. The bacterial community changed significantly after the use of sodium hyaluronate eye drops (with or without preservatives), whether within individuals or between individuals in different groups (P < 0.05, PERMANOVA). Different dosage forms of sodium hyaluronate eye drops significantly decreased the relative abundance of Flavobacterium caeni and Deinococcus antarcticus, respectively (P < 0.05).Healthy people had a rich diversity of the bacterial microbiota on the ocular surface, but the bacterial communities between the eyes were not completely similar. Irrespective of containing benzalkonium chloride (BAC), sodium hyaluronate eye drops can change the bacterial community on the ocular surface.
During tumor development, cancer cells constantly confront different types of extracellular barriers. However, fundamental questions like whether tumor cells will continue to grow against confinement or away from it and what key factors govern this process remain poorly understood. To address these issues, here we examined the growth dynamics of human lung epithelial carcinoma A549 cells partially confined in micrometer-sized cylindrical pores with precisely controlled wall stiffness. It was found that, after reaching confluency, the cell monolayer enclosed by a compliant wall was able to keep growing and pushing the boundary, eventually leading to a markedly enlarged pore. In contrast, a much reduced in-plane growth and elevated strain level among cells were observed when the confining wall becomes stiff. Furthermore, under such circumstance, cells switched their growth from within the monolayer to along the out-of-plane direction, resulting in cell stacking. We showed that these observations can be well explained by a simple model taking into account the deformability of the wall and the threshold stress for inhibiting cell growth. Interestingly, cadherins were found to play an important role in the proliferation and stress buildup within the cell monolayer by aggregating at cell–cell junctions. The stiff confinement led to an elevated expression level of cadherins. Furthermore, inhibition of N-cadherin resulted in a significantly suppressed cell growth under the same confining conditions.
The mechanical and electrical properties of cells serve as critical indicators of their physiological and pathological state. Currently, distinct setups are required to measure the electrical and mechanical responses of cells. In addition, most existing methods such as optical trapping (OT) and atomic force microscopy (AFM) are labor-intensive, expensive, and low-throughput. Here, we developed a microdevice that integrates automated cell trapping, deformation, and electric impedance spectroscopy to overcome these limitations. Our device enables parallel aspiration of tens of trapped cells in a highly scalable manner by simply adjusting the applied pressures, allowing for rapid probing of the dynamic viscoelastic properties of cells. Furthermore, embedded microelectrodes enable concurrent investigations of the electrical impedance of the cells. Through testing on different cell types, our platform demonstrated superior capabilities in comprehensive cell characterization and phenotyping, highlighting its great potential as a versatile tool for single cell analysis, drug screening, and disease detection.
Abstract Cell collectives migrate and undergo morphological transitions during development and diseases. How cells collectively invade a tissue barrier and its immediate consequence remains elusive. An outstanding challenge is to achieve a comprehensive understanding of the invasion cascade by integrating and applying principles from material science, biophysics, and cell biology. Using a three-dimensional model system of cancer spheroids invading the mesothelium, we show that collective cell invasion emerges from an interplay between spheroid fluidization and fracturing of the multicellular barrier. We found that mesothelial cell- cell contacts undergo tensile fracturing due to spheroid invasion. At the subcellular level, force transmission via intercellular integrin complexes between spheroid and mesothelial cells induces mesothelial cell apical constriction that leads to fracturing. Concurrently, fluidization and directed spheroid cells motility expand the mesothelial fracture. In response, the deformed mesothelium retards spheroid spreading, leading to spheroid cells crowding with contact inhibition of locomotion and proliferation. Our results demonstrate that collective invasion is a morphogenic cascade, revealing that the multicellular barrier undergoing invasion-induced fracture can subsequently force the invaded cells into a dormant state of reduced proliferative potential. Thus, our conclusion may help explain why cancer cells that have invaded a foreign microenvironment tend to enter dormancy.
The influence for flow granulating of traditional Chinese medicine to be taken after being mixed with boiling water: spray air pressure greater than powder granularity greater than inlet, exit temperature greater than wind speed. Optimum condition: inlet temperature 110 degrees C, exit temperature 50 degrees C; spray air pressure 1.0 kgf/cm2; sugar powder granularity: 40-60 eye; wind speed: 1.5 gear. It is necessary to provide reference basis for production by pilot test.