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provide a feasible solution for this in cell culture. Methods: Morphological and biomechanical properties of cells undergoing chondrogenic differentiation from human adipose-derived stem cells (ADSCs) were measured at the nanometer scale using atomic force microscopy and laser confocal scanning microscopy. Gene expression was determined by real-time quantitative polymerase chain reaction. Results: The expression of COL II, SOX9, and Aggrecan mRNA began to increase gradually at the beginning of differentiation and reach a peak similar to that of normal chondrocytes on the 12th day, then dropped to the level of the 6th day at 18th day. Cell topography and mechanics trended resembled those of the genes' expression. Integrin β1 was expressed in ADSCs and rapidly upregulated during differentiation but downregulated after reaching maturity. Conclusions: The amount and distribution of integrin β1 may play a critical role in mediating both chondroid cell maturity and dedifferentiation. Integrin β1 is a possible new marker and target for phenotypic maintenance in chondroid cells. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to compare the difference between insulin-producing cells (IPCs) and normal human pancreatic beta cells both in physiological function and morphological features in cellular level. Methods: The levels of insulin secretion were measured by enzyme-linked immunosorbent assay. The insulin gene expression was determined by real-time quantitative polymerase chain reaction. The morphological features were detected by atomic force microscopy (AFM) and laser confocal scanning microscopy. Results: IPCs and normal human pancreatic beta cells were similar to each other under the observation in AFM with the porous structure features in the cytoplasm. Both number of membrane particle size and average roughness of normal human beta cells were higher than those of IPCs. Conclusions: Our results firstly revealed that the cellular ultrastructure of IPCs was closer to that of normal human pancreatic beta cells, but they still could not mimic the physiological regulation of insulin secretion performed by pancreatic beta cells. [ABSTRACT FROM AUTHOR]

T-cell immune responses modulated by T-cell immunoglobulin and mucin domain-containing molecule 3 (Tim-3) during Mycobacterium tuberculosis (Mtb) infection in humans remain poorly understood. Here, we found that active TB patients exhibited increases in numbers of Tim-3-expressing CD4+ and CD8+ T cells, which preferentially displayed polarized effector memory phenotypes. Consistent with effector phenotypes, Tim-3+ CD4+ and Tim-3+ CD8+ T-cell subsets showed greater effector functions for producing Th1/Th22 cytokines and CTL effector molecules than Tim-3- counterparts, and Tim-3- expressing T cells more apparently limited intracellular Mtb replication in macrophages. The increased effector functions for Tim-3-expressing T cells consisted with cellular activation signaling as Tim-3+ CD4+ and Tim-3+ CD8+ T-cell subsets expressed much higher levels of phosphorylated signaling molecules p38, stat3, stat5, and Erk1/2 than Tim-3- controls. Mechanistic experiments showed that siRNA silencing of Tim-3 or soluble Tim-3 treatment interfering with membrane Tim-3-ligand interaction reduced de novo production of IFN-γ and TNF-α by Tim-3-expressing T cells. Furthermore, stimulation of Tim-3 signaling pathways by antibody cross-linking of membrane Tim-3 augmented effector function of IFN-γ production by CD4+ and CD8+ T cells, suggesting that Tim-3 signaling helped to drive stronger effector functions in active TB patients. This study therefore uncovered a previously unknown mechanism for T-cell immune responses regulated by Tim-3, and findings may have implications for potential immune intervention in TB. [ABSTRACT FROM AUTHOR]

The photodynamic antimicrobial chemotherapy as a promising approach for efficiently killing pathogenic microbes is attracting increasing interest. In this study, the cytotoxic and phototoxic effects of hematoporphyrin monomethyl ether (HMME) on the Gram-positive and Gram-negative bacteria were investigated. The cell viability was assessed by colony-forming unit method, and the results indicated that there was no significant cytotoxicity but high phototoxicity in the examined concentrations. Notably, the Gram-positive bacteria were more sensitive to HMME in phototoxicity. Simultaneously, an atomic force microscope (AFM) was used to detect the changes in morphological and nanomechanical properties of bacteria before and after HMME treatment. AFM images indicate that upon photoinactivation, the bacterial surface changed from a smooth, homogeneous architecture to a heterogenous, crackled morphology. The force spectroscopy measurements reveal that the cell wall became less rigid and the Young’s modulus decreased about 50%, whereas the tip-cell-surface adhesion forces increased significantly compared to those of native cells. It was speculated that the photodynamic effects of HMME induced the changes in the chemical composition of the outer membrane and exposure of some proteins inside the envelope. AFM can be utilized as a powerful and sensitive method for studying the interaction between bacteria and drugs. [ABSTRACT FROM AUTHOR]

This article presents a new route for preparing collagen/hydroxyapatite (Coll/HA) composite. Urease-catalyzed decomposition of urea was used for the first time to prepare Coll/HA. Acting as an automatic pH controller, urea totally eliminated the traditional requirement of an external base additive and facilitated procedures for synthesis as a result. This Coll/HA complex was characterized by Fourier transformed infrared spectroscopy, X-ray diffraction, laser diffraction particle size analyzer, scanning electron microscope (SEM), and transmission electron microscopy. The results indicated that chemical and crystallographic properties of this complex were similar to those of the natural bone and other Coll/HA systems. Laser diffraction particle size measurement and SEM showed that this Coll/HA sample evidenced smaller particle size and narrower particle size distribution than other Coll/HA complex prepared by the traditional co-precipitation method. [ABSTRACT FROM AUTHOR]

The activation of T lymphocytes plays a very important role in T-cell-mediated immune response. Though there are many related literatures, the changes of membrane surface nanostructures and adhesion property of T lymphocytes at different activation stages have not been reported yet. However, these investigations will help us further understand the biophysical and immunologic function of T lymphocytes in the context of activation. In the present study, the membrane architectures of peripheral blood T lymphocytes were obtained by AFM, and adhesion force of the cell membrane were measured by acquiring force–distance curves. The results indicated that the cell volume increased with the increases of activation time, whereas membrane surface adhesion force decreased, even though the local stiffness for resting and activated cells is similar. The results provided complementary and important data to further understand the variation of biophysical properties of T lymphocytes in the context of in vitro activation. [ABSTRACT FROM AUTHOR]

Biomaterial-induced human platelet activation remains one of the most crucial factors to determine the procoagulant properties of the biomaterial. In this experiment, a new type of biomacromolecule complex film (hyaluronic acid-collagen (I)/chitosan, HCC) was prepared using the electrostatic self-assembly method. Then the procoagulant properties of this complex film were characterized. Based on the nano-resolution of atomic force microscopy, the platelet-derived microparticles (PMPs) that present the activation of platelets were clearly visualized on the membrane surface of platelets for the first time, and the measurement indicated that the size of PMPs is around 50–110 nm. Furthermore, the results of AFM measurement were confirmed by flow cytometry analysis. The expression of CD62P (P-selectin) dramatically increased after the platelet-rich plasma interacted with the biomaterial solution. From the results, we could draw the conclusion that this biomacromolecule complex film has promising procoagulant properties, and has the potential to be practically used as procoagulant material. [ABSTRACT FROM AUTHOR]

Biomacromolecule has been widely used as biomedical material. Because different biomacromolecules possess different properties, how to exhibit the respective advantages of different components on one type of biomaterial becomes the hot spot in the field of biomaterial studying. This work reported a type of complex film that consisted of hyaluronic acid (HA), type I collagen (Col-I), and chitosan (CS) (HA–Col-I/CS, HCC). Then, a series of experiments were performed, such as inverted microscopic observation, atomic force microscopic (AFM) imaging, flow cytometry (FCM) measurement, MTT assay, and MIC assay. In the present work, we observed the growing condition of 3T3 fibroblasts on the surface of the HCC complex film, visualized the morphological changes of platelets during the coagulation process, and discovered microparticles on the platelet membrane. Moreover, we confirmed the microparticles are the platelet-derived microparticles (PMPs) using the FCM. In addition, the minimal inhibitory concentration (MIC) of HCC against Escherichia coli ( E. coli) 8099 was 0.025 mg/ml, against Staphylococcus aureus ( S. aureus) ATCC 6538 was 0.1 mg/ml. The results together indicated that the HCC film possessed promising coagulation property, cell compatibility and anti-bacteria property, and the potential in future clinical application such as wound healing and bandage. [ABSTRACT FROM AUTHOR]

The article focuses on influencing factors and comparative evaluation of chromosome imaging by atomic force microscopy (AFM). It is said that AFM has been used for analysis of chromosomes structures and has enable researchers to observe nanometer-scale structures. The morphology of samples by measuring the force difference between the microscope tip and the sample can be obtained by AFM.

In recent years, many mammalian cells, especially erythrocytes because of simpleness of their membrane surfaces, were widely studied by atomic force microscopy. In our study, diseased erythrocytes were taken from patients of lung cancer, myelodisplastic syndrome (MDS), and so on. We obtained many clear topographical images of numerous erythrocytes, single erythrocyte, and ultramicrostructure of erythrocyte membrane surfaces from normal persons and patients. By studying the red cells of lung cancer patients, we found that many erythrocytes of lung cancer patient have changed into echinocytes. One erythrocyte has 10–20 short projections, most of which, with a mean width of 589.0 nm and a length of 646.7 nm, are on the edge of cell. The projections in the center of echinocytes are lodged and embedded, but in conventional model of echinocytes, the projections in the center stretch outside cell membrane, so a novel model of erythrocytes was designed in our paper. After observation of microstructure of MDS patient's erythrocyte membrane surface, we found that many apertures with different diameters of tens to hundreds nanometers appeared on the surface of cell membrane. It can be concluded that AFM may be widely applied in clinic pathological inspection. [ABSTRACT FROM AUTHOR]

Nanoparticle aggregates induced by synergistic effect of electrostatic interaction and hydrogen-bonding recognition between melamine and 3-mercaptopriopionic acid (MA), which conjugated on the surface of gold nanoparticles (AuNPs) are used for detection of melamine. MA molecules are conjugated on AuNP surfaces to form MA-modified AuNPs (MA-AuNPs), acting as nanoprobes in the detection of melamine. Since such nanoparticle aggregates-mediated signal amplification can be measured by absorption spectroscopy, the method enables sensitive and real-time detection of melamine at the detection limit as low as 0.4µg/ml, and the linear detection ranging from 0.6µg/ml to 42µg/ml. The color change can be readily seen by the naked eye at 30µg/ml melamine without the aid of any advanced instruments. This method has been successfully applied to detect melamine in infant formula with good reproducibility, and it has the potential of being used in food safety detection. [ABSTRACT FROM AUTHOR]

In recent years, LPS activated RAW264.7 cells are widely used as an in vitro inflammatory model for the screen of effective anti-inflammation drugs and the investigation of exact anti-inflammation mechanism of these drugs. But up to now, there are few data about the effect of LPS on the morphology, especially on the membrane ultrastructure and bio-mechanical properties of RAW264.7 macrophages. In this work, the topographical and biophysical changes of RAW264.7 macrophages upon LPS stimulation are detected by high resolution atomic force microscopy (AFM). The AFM results suggested that LPS activated RAW264.7 macrophages changed to be much bigger than control cells with some holes emerged on cell surface. The size of membrane protein clusters and the roughness of membrane significantly increased after LPS exposure. In addition, the AFM force measurement results demonstrated that LPS stimulation increased the adhesion force of RAW264.7 macrophages, and also increased the stiffness of RAW264.7 macrophages, which were attributed to the re-distribution of intracellular F-actin structures induced by LPS. These findings suggested that LPS stimulation could also induce the pathophysiological changes of RAW264.7 macrophages, which would benefit our understanding of the inflammatory processes in macrophages upon pathogen stimulation at nano-scale. [ABSTRACT FROM AUTHOR]