High-throughput fabrication of hepatic cell clusteroids with enhanced growth and functionality for tissue engineering applications

Culturing of cells as three-dimensional (3D) clusters can enhance in vitro tests for basic biological research as well as for therapeutics development. Such 3D culture models, however, are often more complicated, cumbersome and expensive than two-dimensional (2D) cultures. Current methods for the preparation of tissue spheroids require complex materials, involve tedious facilities and are generally not scalable. We report a novel inexpensive and up-scalable method for the preparation of large quantities of viable cell clusters (clusteroids) of hepatocytes (Hep-G2). The method has a high throughput potential and is based on an aqueous two-phase system (ATPS) of stable water-in-water (w/w) Pickering emulsions, formed of dextran (DEX) drops and poly ethylene oxide (PEO) continuous phase stabilized with whey protein particles. This system enabled the rapid fabrication of cell clusteroids from Hep-G2 cells. Here, the interfacial tension of the aqueous phase in the emulsion droplets, where the cells partition preferentially, is used to wrap the cells in separate compartments, and then the droplets are shrank by changing the balance of ATPS, thus rapidly driving the cells from larger and loosely packed DEX drops to mostly spherical clusters. Cell-Cell adhesion was strongly promoted within the w/w Pickering emulsion droplets which helped the formation of the 3D clusteroids. These were collected after subsequent dilution of the emulsion with culture media. The collected hepatic clusteroids were incorporated into an alginate hydrogel in media to study their proliferation and the cell function compared with individual cells under the same conditions. Our results confirmed that urea and albumin production, which are both linked to hepatocyte cell function, was strongly reinforced in the clusteroid based tissues compared to the one formulated with individual cells. This methodology could potentially extend the w/w emulsion cell clustering platform in tissue generation and preparation of large quantities of organoids for drug tests and replacement of animal models

Advanced biomedical applications based on emerging 3D cell culturing platforms

It is of great value to develop reliable in vitro models for cell biology and toxicology. However, ethical issues and the decreasing number of donors restrict the further use of traditional animal models in various fields, including the emerging fields of tissue engineering and regenerative medicine. The huge gap created by the restrictions in animal models has pushed the development of the increasingly recognized three-dimensional (3D) cell culture, which enables cells to closely simulate authentic cellular behaviour such as close cell-to-cell interactions and can achieve higher functionality. Furthermore, 3D cell culturing is superior to the traditional 2D cell culture, which has obvious limitations and cannot closely mimic the structure and architecture of tissues. In this study, we review several methods used to form 3D multicellular spheroids. The extracellular microenvironment of 3D spheroids plays a role in many aspects of biological sciences, including cell signalling, cell growth, cancer cell generation, and anti-cancer drugs. More recently, they have been explored as basic construction units for tissue and organ engineering. We review this field with a focus on the previous research in different areas using spheroid models, emphasizing aqueous two-phase system (ATPS)-based techniques. Multi-cellular spheroids have great potential in the study of biological systems and can closely mimic the in vivo environment. New technologies to form and analyse spheroids such as the aqueous two-phase system and magnetic levitation are rapidly overcoming the technical limitations of spheroids and expanding their applications in tissue engineering and regenerative medicine

An Overview of the Dwarf Galaxy Survey

The Dwarf Galaxy Survey (DGS) program is studying low-metallicity galaxies using 230 hr of far-infrared (FIR) and submillimetre (submm) photometric and spectroscopic observations of the Herschel Space Observatory and draws from this a rich database of a wide range of wavelengths tracing the dust, gas and stars. This sample of 50 galaxies includes the largest metallicity range achievable in the local Universe including the lowest metallicity (Z) galaxies, 1/50 Z_⊙, and spans four orders of magnitude in star formation rates. The survey is designed to get a handle on the physics of the interstellar medium (ISM) of low metallicity dwarf galaxies, especially their dust and gas properties and the ISM heating and cooling processes. The DGS produces PACS and SPIRE maps of low-metallicity galaxies observed at 70, 100, 160, 250, 350, and 500 μm with the highest sensitivity achievable to date in the FIR and submm. The FIR fine-structure lines, [CII] 158 μm, [OI] 63 μm, [OI] 145 μm, [OIII] 88 μm, [NIII] 57 μm, and [NII] 122 and 205 μm have also been observed with the aim of studying the gas cooling in the neutral and ionized phases. The SPIRE FTS observations include many CO lines (J = 4–3 to J = 13–12), [NII] 205 μm, and [CI] lines at 370 and 609 μm. This paper describes the sample selection and global properties of the galaxies and the observing strategy as well as the vast ancillary database available to complement the Herschel observations. The scientific potential of the full DGS survey is described with some example results included

Physical conditions in the gas phases of the giant HII region LMC-N11 unveiled by Herschel - I. Diffuse [CII] and [OIII] emission in LMC-N11B

(Abridged) The Magellanic Clouds provide a nearby laboratory for metal-poor dwarf galaxies. The low dust abundance enhances the penetration of UV photons into the interstellar medium (ISM), resulting in a relatively larger filling factor of the ionized gas. Furthermore, there is likely a hidden molecular gas reservoir probed by the [CII]157um line. We present Herschel/PACS maps in several tracers, [CII], [OI]63um,145um, [NII]122um, [NIII]57um, and [OIII]88um in the HII region N11B in the Large Magellanic Cloud. Halpha and [OIII]5007A images were used as complementary data to investigate the effect of dust extinction. Observations were interpreted with photoionization models to infer the gas conditions and estimate the ionized gas contribution to the [CII] emission. Photodissociation regions (PDRs) are probed through polycyclic aromatic hydrocarbons (PAHs). We first study the distribution and properties of the ionized gas. We then constrain the origin of [CII]157um by comparing to tracers of the low-excitation ionized gas and of PDRs. [OIII] is dominated by extended emission from the high-excitation diffuse ionized gas; it is the brightest far-infrared line, ~4 times brighter than [CII]. The extent of the [OIII] emission suggests that the medium is rather fragmented, allowing far-UV photons to permeate into the ISM to scales of >30pc. Furthermore, by comparing [CII] with [NII], we find that 95% of [CII] arises in PDRs, except toward the stellar cluster for which as much as 15% could arise in the ionized gas. We find a remarkable correlation between [CII]+[OI] and PAH emission, with [CII] dominating the cooling in diffuse PDRs and [OI] dominating in the densest PDRs. The combination of [CII] and [OI] provides a proxy for the total gas cooling in PDRs. Our results suggest that PAH emission describes better the PDR gas heating as compared to the total infrared emission.Comment: Accepted for publication in Astronomy and Astrophysics. Fixed inverted line ratio in Sect. 5.

Gravitational Instantons, Confocal Quadrics and Separability of the Schr\"odinger and Hamilton-Jacobi equations

A hyperk\"ahler 4-metric with a triholomorphic SU(2) action gives rise to a family of confocal quadrics in Euclidean 3-space when cast in the canonical form of a hyperk\"ahler 4-metric metric with a triholomorphic circle action. Moreover, at least in the case of geodesics orthogonal to the U(1) fibres, both the covariant Schr\"odinger and the Hamilton-Jacobi equation is separable and the system integrable.Comment: 10 pages Late

Enhanced clearing of Candida biofilms on a 3D urothelial cell in vitro model using lysozyme-functionalized fluconazole-loaded shellac nanoparticles

Candida urinary tract biofilms are increasingly witnessed in nosocomial infections due to reduced immunity of patients and the hospital ecosystem. The indwelling devices utilized to support patients with urethral diseases that connect the unsterilized external environment with the internal environment of the patient are another significant source of urinary tract biofilm infections. Recently, nanoparticle (NP)-associated therapeutics have gained traction in a number of areas, including fighting antibiotic-resistant bacterial biofilm infection. However, most studies on nanotherapeutic delivery have only been carried out in laboratory settings rather than in clinical trials due to the lack of precise in vitro and in vivo models for testing their efficiency. Here we develop a novel biofilm-infected 3D human urothelial cell culture model to test the efficiency of nanoparticle (NP)-based antifungal therapeutics. The NPs were designed based on shellac cores, loaded with fluconazole and coated with the cationic enzyme lysozyme. Our formulation of 0.2 wt% lysozyme-coated 0.02 wt% fluconazole-loaded 0.2 wt% shellac NPs, sterically stabilised by 0.25 wt% poloxamer 407, showed an enhanced efficiency in removing Candida albicans biofilms formed on 3D layer of urothelial cell clusteroids. The NP formulation exhibited low toxicity to urothelial cells. This study provides a reliable in vitro model for Candida urinary tract biofilm infections, which could potentially replace animal models in the testing of such antifungal nanotechnologies. The reproducibility and availability of a well-defined biofilm-infected 3D urothelial cell culture model give valuable insights into the formation and clearing of fungal biofilms and could accelerate the clinical use of antifungal nanotherapeutics

The 158 micron (CII) mapping of galaxies: Probing the atomic medium

Using the MPE/UCB Far-infrared Imaging Fabry-Perot Interferometer (FIFI) on the Kuiper Airborne Observatory (KAO), we have made large scale maps of (CII) in the spiral galaxies NGC 6946, NGC 891, M83 and the peculiar elliptical Cen A, thus allowing for the first time, detailed studies of the spatial distribution of the FIR line emission in external galaxies. We find that the (CII) emission comes from a mixture of components of interstellar gas. The brightest emission is associated with the nuclear regions, a second component traces the spiral arms as seen in the nearly face on spiral galaxies NGC 6946 and M83 and the largest star forming/H2 regions contained within them, and another extended component of low brightness can be detected in all of the galaxies far from the nucleus, beyond the extent of CO emission

Milk production functions in relation to feed inputs, cow characteristics and environmental conditions

The main emphasis of this study is on milk production functions as they relate to various levels and proportions of grain and hay in a ration restricted to these two feeds. However, certain auxiliary variables representing cow characteristics and environmental conditions are also incorporated into the production function. The auxiliary variables include: stage of lactation, milk-producing ability, temperature, age, maturity, body weight and a coefficient of inbreeding. These auxiliary variables are fixed at different levels to allow expression of milk production functions when the characteristics of cows are set at different magnitudes. This study considers the economic optima in ration specification for dairy cows and represents a cooperative interdisciplinary effort. The estimates allow derivation of production surfaces, milk isoclines and isoquants, marginal rates of feed substitution and profit-maximizing rations for cows of different characteristics (maturity, ability, inbreeding, weight) producing under specified environmental conditions

In Silico Prediction of Organ Level Toxicity: Linking Chemistry to Adverse Effects

In silico methods to predict toxicity include the use of (Quantitative) Structure-Activity Relationships ((Q)SARs as well as grouping (category formation) allowing for read-across. A challenging area for in silico modelling is the prediction of chronic toxicity and the No Observed (Adverse) Effect Level (NO(A)EL) in particular. A proposed solution to the prediction of chronic toxicity is to consider organ level effects, as opposed to modelling the NO(A)EL itself. This study has focussed on the use of structural alerts to identify potential liver toxicants. In silico profilers, or groups of structural alerts, were developed based on mechanisms of action and informed by current knowledge of Adverse Outcome Pathways. These profilers are robust and can be coded computationally to allow for prediction. However, they do not cover all mechanisms or modes of liver toxicity and recommendations for the improvement of these approaches are given