Mui Hoon Nai

Epithelial bridges maintain tissue integrity during collective cell migration.

The ability of skin to act as a barrier is primarily determined by the efficiency of skin cells to maintain and restore its continuity and integrity. In fact, during wound healing keratinocytes migrate collectively to maintain their cohesion despite heterogeneities in the extracellular matrix. Here, we show that monolayers of human keratinocytes migrating along functionalized micropatterned surfaces comprising alternating strips of extracellular matrix (fibronectin) and non-adherent polymer form suspended multicellular bridges over the non-adherent areas. The bridges are held together by intercellular adhesion and are subjected to considerable tension, as indicated by the presence of prominent actin bundles. We also show that a model based on force propagation through an elastic material reproduces the main features of bridge maintenance and tension distribution. Our findings suggest that multicellular bridges maintain tissue integrity during wound healing when cell-substrate interactions are weak and may prove helpful in the design of artificial scaffolds for skin regeneration.

Rapid, High-Throughput Tracking of Bacterial Motility in 3D via Phase-Contrast Holographic Video Microscopy

Tracking fast-swimming bacteria in three dimensions can be extremely challenging with current optical techniques and a microscopic approach that can rapidly acquire volumetric information is required. Here, we introduce phase-contrast holographic video microscopy as a solution for the simultaneous tracking of multiple fast moving cells in three dimensions. This technique uses interference patterns formed between the scattered and the incident field to infer the three-dimensional (3D) position and size of bacteria. Using this optical approach, motility dynamics of multiple bacteria in three dimensions, such as speed and turn angles, can be obtained within minutes. We demonstrated the feasibility of this method by effectively tracking multiple bacteria species, including Escherichia coli, Agrobacterium tumefaciens, and Pseudomonas aeruginosa. In addition, we combined our fast 3D imaging technique with a microfluidic device to present an example of a drug/chemical assay to study effects on bacterial motility.

Role of Cytoskeletal Tension in the Induction of Cardiomyogenic Differentiation in Micropatterned Human Mesenchymal Stem Cell

The role of biophysical induction methods such as cell micropatterning in stem cell differentiation has been well documented previously. However, the underlying mechanistic linkage of the engineered cell shape to directed lineage commitment remains poorly understood. Here, it is reported that micropatterning plays an important role in regulating the optimal cytoskeletal tension development in human mesenchymal stem cell (hMSC) via cell mechanotransduction pathways to induce cardiomyogenic differentiation. Cells are grown on fibronectin strip patterns to control cell polarization and morphology. These patterned cells eventually show directed commitment toward the myocardial lineage. The cells mechanical properties (cell stiffness and cell traction forces) are observed to be very different for cells that have committed to the myocardial lineage when compared with that of control. These committed cells have mechanical properties that are significantly lower indicating a correlation between the micropatterning-induced differentiation and actomyosin-generated cytoskeletal tension within patterned cells. To study this correlation, patterned cells are treated with RhoA pathway inhibitor. Severely down-regulated cardiomyogenic marker expression is observed in those treated patterned cells, thus emphasizing the direct dependence of hMSCs differentiation fate on the cytoskeletal tension.

Polysaccharide nanofibers with variable compliance for directing cell fate

Cells perceive their microenvironment through physical and mechanical cues, such as extracellular matrix topography or stiffness. In this study, we developed a polysaccharide scaffold that can provide combined substrate topography and matrix compliance signals to direct cell fate. Pullulan/dextran (P/D) nanofibers were fabricated with variable stiffness by in situ crosslinking during electrospinning. By varying the chemical crosslinking content between 10, 12, 14, and 16%, (denoted as STMP10, STMP12, STMP14, and STMP16 respectively), scaffold mechanical stiffness was altered. We characterized substrate stiffness by various methods. Under hydrated conditions, atomic force microscopy and tensile tests of bulk scaffolds were conducted. Under dry conditions, tensile tests of scaffolds and single nanofibers were examined. In addition, we evaluated the efficacy of the scaffolds in directing stem cell differentiation. Using human first trimester mesenchymal stem cells (fMSCs) cultured on STMP14 P/D scaffolds (Youngs modulus: 7.84 kPa) in serum-free neuronal differentiation medium exhibited greatest extent of differentiation. Cells showed morphological changes and significantly higher expression of motor neuron markers. Further analyses by western blotting also revealed the enhanced expression of choline acetyltransferase on STMP14 (7.84 kPa) and STMP16 (11.08 kPa) samples as compared to STMP12 (7.19 kPa). Taken together, this study demonstrates that the stiffness of P/D nanofibers can be altered by differential in situ crosslinking during electrospinning and suggests the feasibility of using such polysaccharide nanofibers in supporting fMSC neuronal commitment.

Nanoindentation Study of Polymer Based Nanocomposites

Nanoindentation is a useful technique to measure hardness as well as elastic and timedependent plastic properties of materials with nanometer resolution. The measurement of elastic modulus of polymeric materials remains challenging due to their viscoelastic behavior. Clay reinforced nylon6 nanocomposites are found to have great improvement in the elastic modulus and tensile strength due to exfoliated hybrid structure. However, its mechanical properties have not been well investigated. In the present study, hardness and elastic modulus of nylon6-5wt%clay nanocomposites were investigated using nanoindentation. Creep effects of the nanocomposites on the unloading stiffness, which directly relates to the elastic modulus, were studied under various unloading rates and holding periods. It was found that the elastic modulus and hardness of nylon6-5wt%clay nanocomposites increased by 58% and 80%, respectively, as compared to pure nylon6. Experimental results for both polycarbonate and nylon6-5wt%clay nanocomposites showed that loading rate had no significant effects on the unloading stiffness. However, stiffness decreased to more consistent values after longer holding periods (more than 30 sec) and at faster unloading rates. The results indicated that creep behavior of the polymers affects the measurement of the unloading stiffness and may possibly overestimate the elastic modulus. Errors in the stiffness measurements from nanoindentation could be minimized with appropriate loading, unloading and holding conditions.

Probing the Physical Origin of Anisotropic Thermal Transport in Black Phosphorus Nanoribbons

Black phosphorus (BP) has emerged as a promising candidate for next-generation electronics and optoelectronics among the 2D family materials due to its extraordinary electrical/optical/optoelectronic properties. Interestingly, BP shows strong anisotropic transport behavior because of its puckered honeycomb structure. Previous studies have demonstrated the thermal transport anisotropy of BP and theoretically attribute this to the anisotropy in both the phonon dispersion relation and the phonon relaxation time. However, the exact origin of such strong anisotropy lacks clarity and has yet to be proven experimentally. Here, the thermal transport anisotropy of BP nanoribbons is probed by an electron beam technique. Direct evidence is provided that the origin of this anisotropy is dominated by the anisotropic phonon group velocity, verified by Youngs modulus measurements along different directions. It turns out that the ratio of the thermal conductivity between zigzag (ZZ) and armchair (AC) ribbons is almost same as that of the corresponding Young modulus values. The results from first-principles calculation are consistent with this experimental observation, where the anisotropic phonon group velocity between ZZ and AC is shown. These results provide fundamental insight into the anisotropic thermal transport in low-symmetry crystals.