Chemotaxis Assay of Bone Marrow-Derived Macrophages.

Immune responses rely on efficient and coordinated migration of immune cells to the site of infection or injury. To reach the site of immunological threat often requires long-range navigation of immune cells through complex tissue and vascular networks. Chemotaxis, cell migration steered by gradients of cell-attractive chemicals that bind sensory receptors, is central to this response. Chemoattractant receptors mostly belong to the G-protein-coupled receptor (GPCR) family, but the way attractant-receptor signaling directs cell migration is not fully understood. Direct-viewing chemotaxis chambers combined with time-lapse microscopy give a powerful tool to study the dynamic details of cells' responses to different attractant landscapes. Here, we describe the application of one such chamber (the Dunn chamber) to study bone marrow-derived macrophage chemotaxis to gradients of complement C5a.

Droplet-Bijel-Droplet Transition in Aqueous Two-Phase Systems Stabilized by Oppositely Charged Nanoparticles: A Simple Pathway to Fabricate Bijels.

We demonstrate a novel yet straightforward methodology of stabilizing aqueous two-phase systems (ATPS) using oppositely charged nanoparticles (OCNPs). We employ commercial-grade, Ludox, OCNPs to induce self-assembly. This self-assembly route promotes the stronger adsorption of nanoparticles at the water-water interface by triggering the formation of 2D and 3D aggregates of varying sizes and shapes. The interplay of this size and shape promotes stability due to increased Gibbs detachment energy and modulates the resulting cluster adsorption at the interface, thereby the structural state of emulsions. We demonstrate the influence of polymers' and particles' composition on the structural transformation from droplet-bijel-droplet using a phase diagram. For the first time, such a structural transition and the single pathway are reported within the domain of ATPS to produce stable bijels or colloidal capsules. It is asserted that the essential condition of three-phase contact angle (θ) = 90° to favor the formation of bijels can be established by selecting a suitable experimental condition using a phase diagram without employing any complicated surface modification procedures reported in the literature. Further, the mechanistic route favoring the formation of bijels and emulsion droplets at different experimental regimes is presented based on the empirical study using turbidity and zeta potential measurements. These studies reveal that the formation of bijels will be most favored when the parameter M (ratio of weight fraction of positively charged nanoparticles to negatively charged nanoparticles) is chosen between 0.7 and 4. It is intriguing to note the fact that, while the droplets stabilized by OCNPs have shown good resilience under high centrifugal action, the bijels produced in this way continued to remain stable for a long time, offering a facile route to prepare the bijels with a hierarchical bicontinuous network structure.

Distinct Modes of Tissue Expansion in Free Versus Earlier-Confined Boundaries for More Physiological Modeling of Wound Healing, Cancer Metastasis, and Tissue Formation.

Collective cell migration is often seen in many biological processes like embryogenesis, cancer metastasis, and wound healing. Despite extensive experimental and theoretical research, the unified mechanism responsible for collective cell migration is not well known. Most of the studies have investigated artificial model wound to study the collective cell migration in an epithelial monolayer. These artificial model wounds possess a high cell number density compared to the physiological scenarios like wound healing (cell damage due to applied cut) and cancer metastasis (smaller cell clusters). Therefore, both systems may not completely relate to each other, and further investigation is needed to understand the collective cell migration in physiological scenarios. In an effort to fill this existing knowledge gap, we investigated the freely expanding monolayer that closely represented the physiological scenarios and compared it with the artificially created model wound. In the present work, we report the effect of initial boundary conditions (free and confined) on the collective cell migration of the epithelial cell monolayer. The expansion and migration aspects of the freely expanding and earlier-confined monolayer were investigated at the tissue and cellular levels. The freely expanding monolayer showed significantly higher expansion and lower migration in comparison to the earlier-confined monolayer. The expansion and migration rate of the monolayer exhibited a strong negative correlation. The study highlights the importance of initial boundary conditions in the collective cell migration of the expanding tissue and provides useful insights that might be helpful in the future to tune the collective cell migration in wound healing, cancer metastasis, and tissue formation.

Improved nanomechanical and in-vitro biocompatibility of graphene oxide-carbon nanotube hydroxyapatite hybrid composites by synergistic effect.

Hydroxyapatite (HAP) is an attractive bio-material for new bone growth process, hard tissue repair, bioactivity, osteoblast adhesion and proliferation due to its physicochemical resembles natural apatite. The intrinsic brittleness and poor mechanical properties of HAP restrict it for potential clinical applications. This problem is undertaken by exploiting the unique properties of carbon nanofillers (carbon nanotube (CNTs), graphene oxide (GO), graphene oxide-carbon nanotube (GCNTs) hybrid) which are used as reinforcement for preparing the carbon nanofillers based HAP composites. The nanomechanical and in-vitro biocompatibility of carbon nanofiller reinforced HAP composites have been studied. Carbon nanofiller reinforced HAP composites led to an improvement in nanomechanical and biocompatibility properties. The nanoindentation hardness and elastic modulus of GCNTs-HAP composites are significantly higher than other carbon nanofiller reinforced composites and pristine HAP powder. The in-vitro cytotoxicity of the prepared carbon nanofillers reinforced HAP composites is examined using MTT-assay on the MDCK cell line. The prepared GCNTs-HAP composites containing 2% of GCNTs nanofiller show higher cell viability, improved compatibility, and superior one cell proliferation induction than the other carbon nanofillers and HAP. These findings will provide the new prospects for utilizing the GO and its hybrid in HAP composites in bone repair, regeneration, augmentation and implantation.