Bioactive films produced from self-assembling peptide amphiphiles as versatile substrates for tuning cell adhesion and tissue architecture in serum-free conditions

The development of versatile bioactive surfaces able to emulate in vivo conditions is of enormous importance to the future of cell and tissue therapy. Tuning cell behaviour on two-dimensional surfaces so that the cells perform as if they were in a natural three-dimensional tissue represents a significant challenge, but one that must be met if the early promise of cell and tissue therapy is to be fully realised. Due to the inherent complexities involved in the manufacture of biomimetic three-dimensional substrates, the scaling up of engineered tissue-based therapies may be simpler if based upon proven two-dimensional culture systems. In this work, we developed new coating materials composed of the self-assembling peptide amphiphiles (PAs) C16G3RGD (RGD) and C16G3RGDS (RGDS) shown to control cell adhesion and tissue architecture while avoiding the use of serum. When mixed with the C16ETTES diluent PA at 13 : 87 (mol mol-1) ratio at 1.25 times 10-3 M, the bioactive {PAs} were shown to support optimal adhesion, maximal proliferation, and prolonged viability of human corneal stromal fibroblasts ({hCSFs)}, while improving the cell phenotype. These {PAs} also provided stable adhesive coatings on highly-hydrophobic surfaces composed of striated polytetrafluoroethylene ({PTFE)}, significantly enhancing proliferation of aligned cells and increasing the complexity of the produced tissue. The thickness and structure of this highly-organised tissue were similar to those observed in vivo, comprising aligned newly-deposited extracellular matrix. As such, the developed coatings can constitute a versatile biomaterial for applications in cell biology, tissue engineering, and regenerative medicine requiring serum-free conditions.

Oxidized alginate hydrogels as niche environments for corneal epithelial cells

Chemical and biochemical modification of hydrogels is one strategy to create physiological constructs that maintain cell function. The aim of this study was to apply oxidised alginate hydrogels as a basis for development of a biomimetic niche for limbal epithelial stem cells that may be applied to treating corneal dysfunction. The stem phenotype of bovine limbal epithelial cells (LEC) and the viability of corneal epithelial cells (CEC) were examined in oxidised alginate gels containing collagen IV over a 3-day culture period. Oxidation increased cell viability (P corneal extracellular matrix proteins can influence corneal epithelial cell function in a manner that may impact beneficially on corneal wound healing therapy.

The secretome of alginate-encapsulated limbal epithelial stem cells modulates corneal epithelial cell proliferation

Limbal epithelial stem cells may ameliorate limbal stem cell deficiency through secretion of therapeutic proteins, delivered to the cornea in a controlled manner using hydrogels. In the present study the secretome of alginate-encapsulated limbal epithelial stem cells is investigated. Conditioned medium was generated from limbal epithelial stem cells encapsulated in 1.2% (w/v) calcium alginate gels. Conditioned medium proteins separated by 1-D gel electrophoresis were visualized by silver staining. Proteins of interest including secreted protein acidic and rich in cysteine, profilin-1, and galectin-1 were identified by immunoblotting. The effect of conditioned medium (from alginate-encapsulated limbal epithelial stem cells) on corneal epithelial cell proliferation was quantified and shown to significantly inhibit (Pcorneal epithelial cell proliferation. We conclude that limbal epithelial stem cells encapsulated in alginate gels may regulate corneal epithelialisation through secretion of inhibitory proteins.

The effects of retinoic acid on human corneal stromal keratocytes cultured in vitro under serum-free conditions

Purpose: Retinoic acid (RA) is a metabolite of vitamin A that plays a fundamental role in the development and function of the human eye. The purpose of this study was to investigate the effects of RA on the phenotype of corneal stromal keratocytes maintained in vitro for extended periods under serum-free conditions. Methods: Keratocytes isolated from human corneas were cultured up to 21 days in serum-free media supplemented with RA or DMSO vehicle. The effects of RA and of its removal after treatment on cell proliferation and morphology were evaluated. In addition, the expression of keratocyte markers was quantified at the transcriptional and protein levels by quantitative PCR and immunoblotting or ELISA, respectively. Furthermore, the effects of RA on keratocyte migration were tested using scratch assays. Results: Keratocytes cultured with RA up to 10×10-6 M showed enhanced proliferation and stratification, and reduced mobility. RA also promoted the expression of keratocyte-characteristic proteoglycans such as keratocan, lumican, and decorin, and increased the amounts of collagen type-I in culture while significantly reducing the expression of matrix metalloproteases 1, 3, and 9. RA effects were reversible, and cell phenotype reverted to that of control after removal of RA from media. Conclusions: RA was shown to control the phenotype of human corneal keratocytes cultured in vitro by regulating cell behaviour and extracellular matrix composition. These findings contribute to our understanding of corneal stromal biology in health and disease, and may prove useful in optimizing keratocyte cultures for applications in tissue engineering, cell biology, and medicine.

Optimization of a sea ice model using basinwide observations of Arctic sea ice thickness, extent, and velocity

A stand-alone sea ice model is tuned and validated using satellite-derived, basinwide observations of sea ice thickness, extent, and velocity from the years 1993 to 2001. This is the first time that basin-scale measurements of sea ice thickness have been used for this purpose. The model is based on the CICE sea ice model code developed at the Los Alamos National Laboratory, with some minor modifications, and forcing consists of 40-yr ECMWF Re-Analysis (ERA-40) and Polar Exchange at the Sea Surface (POLES) data. Three parameters are varied in the tuning process: Ca, the air–ice drag coefficient; P*, the ice strength parameter; and α, the broadband albedo of cold bare ice, with the aim being to determine the subset of this three-dimensional parameter space that gives the best simultaneous agreement with observations with this forcing set. It is found that observations of sea ice extent and velocity alone are not sufficient to unambiguously tune the model, and that sea ice thickness measurements are necessary to locate a unique subset of parameter space in which simultaneous agreement is achieved with all three observational datasets.

Improving the spatial distribution of modeled Arctic sea ice thickness

The spatial distribution of ice thickness/draft in the Arctic Ocean is examined using a sea ice model. A comparison of model predictions with submarine observations of sea ice draft made during cruises between 1987 and 1997 reveals that the model has the same deficiencies found in previous studies, namely ice that is too thick in the Beaufort Sea and too thin near the North Pole. We find that increasing the large scale shear strength of the sea ice leads to substantial improvements in the model's spatial distribution of sea ice thickness, and simultaneously improves the agreement between modeled and ERS-derived 1993–2001 mean winter ice thickness.

Dependence of sea ice yield-curve shape on ice thickness

In this note, the authors discuss the contribution that frictional sliding of ice floes (or floe aggregates) past each other and pressure ridging make to the plastic yield curve of sea ice. Using results from a previous study that explicitly modeled the amount of sliding and ridging that occurs for a given global strain rate, it is noted that the relative contribution of sliding and ridging to ice stress depends upon ice thickness. The implication is that the shape and size of the plastic yield curve is dependent upon ice thickness. The yield-curve shape dependence is in addition to plastic hardening/weakening that relates the size of the yield curve to ice thickness. In most sea ice dynamics models the yield-curve shape is taken to be independent of ice thickness. The authors show that the change of the yield curve due to a change in the ice thickness can be taken into account by a weighted sum of two thickness-independent rheologies describing ridging and sliding effects separately. It would be straightforward to implement the thickness-dependent yield-curve shape described here into sea ice models used for global or regional ice prediction.

Enhancement in corneal permeability of riboflavin using calcium sequestering compounds

Ethylenediaminetetraacetic acid, ethylenediamine-N,N′-disuccinic acid and ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid are polyaminocarboxylic acids that are able to sequester metal ions. Calcium is implicated in maintenance of intercellular matrix, zonula occludens (tight junctions) and zonula adherens of epithelium and endothelium cells. Corneal epithelium is impervious to many aqueous formulations due to it being lipophilic, whereby transcellular drug transit is resisted, whilst tight junctions restrict access via the paracellular route. Research has shown that integrity of tight junctions breaks down through loss of Ca2+ for endothelial and epithelial cells. This study investigates different Ca2+ sequestering compounds and their effect on corneal permeability of riboflavin at physiological pH. Riboflavin is a topically administered ocular drug applied during UV-induced corneal cross-linking for the treatment of keratoconus.

Will Arctic sea ice thickness initialization improve seasonal forecast skill?

Arctic sea ice thickness is thought to be an important predictor of Arctic sea ice extent. However, coupled seasonal forecast systems do not generally use sea ice thickness observations in their initialization and are therefore missing a potentially important source of additional skill. To investigate how large this source is, a set of ensemble potential predictability experiments with a global climate model, initialized with and without knowledge of the sea ice thickness initial state, have been run. These experiments show that accurate knowledge of the sea ice thickness field is crucially important for sea ice concentration and extent forecasts up to 8 months ahead, especially in summer. Perturbing sea ice thickness also has a significant impact on the forecast error in Arctic 2 m temperature a few months ahead. These results suggest that advancing capabilities to observe and assimilate sea ice thickness into coupled forecast systems could significantly increase skill.

Rapid changes in LLBL thickness

We employ data from two spacecraft, at the dawn flank of the magnetopause, to investigate fluctuations in the thickness of the low-latitude boundary layer (LLBL). We show the LLBL is considerably thinner shortly after the detection of a flux transfer event than it was during the event. These data are shown to be consistent with the theory of transient increases in the open LLBL thickness caused by a pulse of enhanced reconnection at the magnetopause.

Improved Arctic sea-ice thickness projections using bias corrected CMIP5 simulations

Projections of Arctic sea ice thickness (SIT) have the potential to inform stakeholders about accessibility to the region, but are currently rather uncertain. The latest suite of CMIP5 Global Climate Models (GCMs) produce a wide range of simulated SIT in the historical period (1979–2014) and exhibit various biases when compared with the Pan-Arctic Ice Ocean Modelling and Assimilation System (PIOMAS) sea ice reanalysis. We present a new method to constrain such GCM simulations of SIT via a statistical bias correction technique. The bias correction successfully constrains the spatial SIT distribution and temporal variability in the CMIP5 projections whilst retaining the climatic fluctuations from individual ensemble members. The bias correction acts to reduce the spread in projections of SIT and reveals the significant contributions of climate internal variability in the first half of the century and of scenario uncertainty from mid-century onwards. The projected date of ice-free conditions in the Arctic under the RCP8.5 high emission scenario occurs in the 2050s, which is a decade earlier than without the bias correction, with potentially significant implications for stakeholders in the Arctic such as the shipping industry. The bias correction methodology developed could be similarly applied to other variables to reduce spread in climate projections more generally.

Daily thin-ice thickness maps from modis thermal infrared imagery

Accurate knowledge of ice-production rates within the marginal ice zones of the Arctic Ocean requires monitoring of the thin-ice distribution within polynyas. The thickness of the ice layer controls the heat loss and hence the new-ice formation. An established thinice algorithm using high-resolution MODIS data allows deriving the ice-thickness distribution within polynyas. The average uncertainty is ±4.7 cm for ice thicknesses below 0.2 m. In this study, the ice-thickness distributions within the Laptev Sea polynya for the two winter seasons 2007/08 and 2008/09 are calculated. Then, a new method is applied to determine a daily MODIS thin-ice product.

Improvement and sensitivity analysis of thermal thin-ice thickness retrievals

Considering the sea ice decline in the Arctic during the last decades, polynyas are of high research interest since these features are core areas of new ice formation. The determination of ice formation requires accurate retrieval of polynya area and thin-ice thickness (TIT) distribution within the polynya.We use an established energy balance model to derive TITs with MODIS ice surface temperatures (Ts) and NCEP/DOE Reanalysis II in the Laptev Sea for two winter seasons. Improvements of the algorithm mainly concern the implementation of an iterative approach to calculate the atmospheric flux components taking the atmospheric stratification into account. Furthermore, a sensitivity study is performed to analyze the errors of the ice thickness. The results are the following: 1) 2-m air temperatures (Ta) and Ts have the highest impact on the retrieved ice thickness; 2) an overestimation of Ta yields smaller ice thickness errors as an underestimation of Ta; 3) NCEP Ta shows often a warm bias; and 4) the mean absolute error for ice thicknesses up to 20 cm is ±4.7 cm. Based on these results, we conclude that, despite the shortcomings of the NCEP data (coarse spatial resolution and no polynyas), this data set is appropriate in combination with MODIS Ts for the retrieval of TITs up to 20 cm in the Laptev Sea region. The TIT algorithm can be applied to other polynya regions and to past and future time periods. Our TIT product is a valuable data set for verification of other model and remote sensing ice thickness data.