Dataset Information

Synthesis of microgel sensors for spatial and temporal monitoring of protease activity.

ABSTRACT: Proteases are involved in almost every important cellular activity, from embryonic morphogenesis to apoptosis. To study protease activity in situ, hydrogels provide a synthetic mimic of the extracellular matrix (ECM) and have utility as a platform to study activity, such as those related to cell migration, in three-dimensions. While 3-dimensional visualization of protease activity could prove quite useful to elucidate the proteolytic interaction at the interface between cells and their surrounding environment, there has been no versatile tool to visualize local proteolytic activity in real time. Here, micron-sized gels were synthesized by inverse suspension polymerization using thiolene photo-click chemistry. The size distribution was selected to avoid cellular uptake and to lower cytotoxicity, while simultaneously allowing the integration of peptide-based FRET sensors of local cell activity. Proteolytic activity of collagenase was detected within an hour via changes in fluorescence of embedded microgels; incubation of microgel sensors with A375 melanoma cells showed upregulated MMP activity in the presence of soluble fibronectins in media. The microgel sensors were readily incorporated into both gelatin and poly(ethylene glycol) (PEG) hydrogels and used to successfully detect spatiotemporal proteolytic activity of A375 melanoma cells. Finally, a tumor model was constructed from a hydrogel microwell array that was used to aggregate A375 melanoma cells, and local variations in proteolytic activity were monitored as a function of distance from the cell aggregate center.

SUBMITTER: Shin DS

PROVIDER: S-EPMC5842818 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Synthesis of microgel sensors for spatial and temporal monitoring of protease activity.

Shin Della S DS Tokuda Emi Y EY Leight Jennifer L JL Miksch Connor E CE Brown Tobin E TE Anseth Kristi S KS

ACS biomaterials science & engineering 20170313 2

Proteases are involved in almost every important cellular activity, from embryonic morphogenesis to apoptosis. To study protease activity <i>in situ</i>, hydrogels provide a synthetic mimic of the extracellular matrix (ECM) and have utility as a platform to study activity, such as those related to cell migration, in three-dimensions. While 3-dimensional visualization of protease activity could prove quite useful to elucidate the proteolytic interaction at the interface between cells and their su ...[more]

PMID: 29527570

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Project description:Littoral zones of freshwater lakes are exposed to environmental impacts from both terrestrial and aquatic sides, while substantial anthropogenic pressure also affects the high spatial, and temporal variability of the ecotone. In this study, the possibility of monitoring seasonal and spatial changes in reed (Phragmites australis) stands using an unmanned aerial vehicle (UAV) based remote sensing technique was examined. Stands in eutrophic and mesotrophic parts of Lake Balaton including not deteriorating (stable) and deteriorating (die-back) patches, were tracked throughout the growing season using a UAV equipped with a Normalized Difference Vegetation Index (NDVI) camera. Photophysiological parameters of P. australis were also measured with amplitude modulated fluorescence. Parameters characterizing the dynamics of seasonal changes in NDVI data were used for phenological comparison of eutrophic and mesotrophic, stable and die-back, terrestrial and aquatic, mowed and not-mowed patches of reed. It was shown that stable Phragmites plants from the eutrophic part of the lake reached specific phenological stages up to 3.5 days earlier than plants from the mesotrophic part of the lake. The phenological changes correlated with trophic (total and nitrate-nitrite nitrogen) and physical (organic C and clay content) properties of the sediment, while only minor relationships with air and water temperature were found. Phenological differences between the stable and die-back stands were even more pronounced, with ~34% higher rates of NDVI increase in stable than die-back patches, while the period of NDVI increase was 16 days longer. Aquatic and terrestrial parts of reed stands showed no phenological differences, although intermediate areas (shallow water parts of stands) were found to be less vigorous. Winter mowing of dried Phragmites sped up sprouting and growth of reed in the spring. This study showed that remote sensing-derived photophysiological and phenological variability within and between reed stands may provide valuable early indicators of environmental stress. The flexibility of the method makes it usable for mapping fine-scale temporal variability and spatial zonation within a stand, revealing ecophysiological hotspots that might require particular attention, and obtaining information vital for conservation and management of plants in the littoral zones.

Dataset Information

Synthesis of microgel sensors for spatial and temporal monitoring of protease activity.

Publications

Synthesis of microgel sensors for spatial and temporal monitoring of protease activity.

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