Neutral, water-soluble poly(ester amide) hydrogels for cell encapsulation

© 2020 Elsevier Ltd Hydrogels are of significant interest for cell encapsulation and delivery in regenerative medicine. Poly(ester amide)s (PEAs) are a class of biodegradable polymers that exhibit promise for biomedical applications due to the degradability of the ester and amide linkages in their backbones, their preparation from biomolecules such as amino acids, and the ability to readily tune their properties through a modular synthesis approach. Water-soluble PEAs containing cationic arginine moieties have previously been developed, but to the best of our knowledge, neutral water-soluble PEAs based on non-charged amino acids have not been reported. Using a poly(ethylene glycol) (PEG)-based macromonomer, we describe here the syntheses of water-soluble amino acid-containing PEAs containing crosslinkable alkenes in their backbones. These PEAs were converted into hydrogels through photoinitiated crosslinking and their properties were compared, including gel content, water content, swelling, and Young\u27s moduli. Subsequent cell culture studies on a subset of hydrogels confirmed that human adipose-derived stromal cells (ASCs) showed \u3e 75% viability at 24 h post-encapsulation. To explore the potential of the hydrogels as cell delivery systems for applications in soft tissue regeneration, adipogenic differentiation of the encapsulated ASCs was probed in vitro at 7 days. Analysis of glycerol-3-phosphate dehydrogenase (GPDH) enzyme activity and intracellular lipid accumulation indicated that the hydrogels provided a supportive environment for ASC adipogenesis. Overall, these PEAs provide a new platform that warrants further development for regenerative medicine applications

Transesterification of Poly(ethyl glyoxylate): A Route to Structurally Diverse Polyglyoxylates

Polyglyoxylates are a class of self-immolative polymers that depolymerize in solution and the solid state. The glyoxylic acid degradation product is a metabolite in the glyoxylate cycle and can also be processed in the liver in humans, making polyglyoxylates attractive for applications in the environment and in medicine. Although expanding the scope of available polyglyoxylates would enable new properties and applications, highly pure glyoxylate monomers are required for polymerization, and this level of purity is difficult to achieve for many potential monomers. To address this challenge, we report here the 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD)-catalyzed post-polymerization transesterification of poly(ethyl glyoxylate) (PEtG) as a general method for the synthesis of directly inaccessible polyglyoxylates. Using a new end-capping strategy, PEtG compatible with the transesterification reaction was developed. n-Propanol, i-propanol, n-butanol, t-butanol, n-pentanol, n-hexanol, n-octanol, and benzyl alcohol were employed and the reactivities of these different alcohols were investigated. The resulting polyglyoxylates were characterized chemically and their thermal properties were compared. In all cases, the transesterified polyglyoxylates retained the stimuli-responsive depolymerization properties of the parent PEtG. In addition, functional polyglyoxylates based on allyl, propargyl, and furfuryl esters, which are suitable for subsequent click reactions, were prepared. The propargyl-functionalized polyglyoxylate was used to conjugate pyrene, and the resulting molecules underwent a change in fluorescence properties upon depolymerization

Investigating the Effects of Tissue-Specific Extracellular Matrix on the Adipogenic and Osteogenic Differentiation of Human Adipose-Derived Stromal Cells Within Composite Hydrogel Scaffolds

© Copyright © 2019 Shridhar, Amsden, Gillies and Flynn. While it has been postulated that tissue-specific bioscaffolds derived from the extracellular matrix (ECM) can direct stem cell differentiation, systematic comparisons of multiple ECM sources are needed to more fully assess the benefits of incorporating tissue-specific ECM in stem cell culture and delivery platforms. To probe the effects of ECM sourced from decellularized adipose tissue (DAT) or decellularized trabecular bone (DTB) on the adipogenic and osteogenic differentiation of human adipose-derived stem/stromal cells (ASCs), a novel detergent-free decellularization protocol was developed for bovine trabecular bone that complemented our established detergent-free decellularization protocol for human adipose tissue and did not require specialized equipment or prolonged incubation times. Immunohistochemical and biochemical characterization revealed enhanced sulphated glycosaminoglycan content in the DTB, while the DAT contained higher levels of collagen IV, collagen VI and laminin. To generate platforms with similar structural and biomechanical properties to enable assessment of the compositional effects of the ECM on ASC differentiation, micronized DAT and DTB were encapsulated with human ASCs within methacrylated chondroitin sulfate (MCS) hydrogels through UV-initiated crosslinking. High ASC viability (\u3e90%) was observed over 14 days in culture. Adipogenic differentiation was enhanced in the MCS+DAT composites relative to the MCS+DTB composites and MCS controls after 14 days of culture in adipogenic medium. Osteogenic differentiation studies revealed a peak in alkaline phosphatase (ALP) enzyme activity at 7 days in the MCS+DTB group cultured in osteogenic medium, suggesting that the DTB had bioactive effects on osteogenic protein expression. Overall, the current study suggests that tissue-specific ECM sourced from DAT or DTB can act synergistically with soluble differentiation factors to enhance the lineage-specific differentiation of human ASCs within 3-D hydrogel systems

Endogenous Skin Fluorescence Includes Bands that may Serve as Quantitative Markers of Aging and Photoaging

Aging and photoaging cause distinct changes in skin cells and extracellular matrix. Changes in hairless mouse skin as a function of age and chronic UVB exposure were investigated by fluorescence excitation spectroscopy. Fluorescence excitation spectra were measured in vivo, on heat-separated epidermis and dermis, and on extracts of mouse skin to characterize the absorption spectra of the emitting chromophores. Fluorescence excitation spectra obtained in vivo on 6 wk old mouse skin had maxima at 295, 340, and 360 nm; the 295 nm band was the dominant band. Using heat separated tissue, the 295 nm band predominantly originated in the epidermis and the bands at 340 and 360 nm originated in the dermis. The 295 nm band was assigned to tryptophan fluorescence, the 340 nm band to pepsin digestable collagen cross-links fluorescence and the 360 nm band to collagenase digestable collagen cross-links fluorescence. Fluorescence excitation maxima remained unchanged in chronologically aged mice (34–38 wk old), whereas the 295 nm band decreased in intensity with age and the 340 nm band increased in intensity with age. In contrast, fluorescence excitation spectra of chronically UVB exposed mice showed a large increase in the 295 nm band compared with age-matched controls and the bands at 340 and 350 nm were no longer distinct. Two new bands appeared in the chronically exposed mice at 270 nm and at 305 nm. These reproducible changes in skin autofluorescence suggest that aging causes predictable alterations in both epidermal and dermal fluorescence, whereas chronic UV exposure induces the appearance of new fluorphores

Connecting Subseasonal Movements of the Winter Mean Ridge in Western North America to Inversion Climatology in Cache Valley, Utah

A 10-yr record of PM2.5 (particulate matter of aerodynamic diameter ≤ 2.5 μm), collected in Cache Valley near downtown Logan, Utah, reveals a strong peak in the PM2.5 concentration climatology that is tightly localized in mid-January. The cause of this subseasonal variation in the PM2.5 climatology is investigated through dynamical downscaling and large-scale diagnostics. Climatological analysis of the U.S. winter mean ridge reveals a mid-January subseasonal shift in the zonal direction, likely in response to variations in the Rossby wave source over the central North Pacific Ocean. This displacement of the winter mean ridge, in turn, has an impact on regional-scale atmospheric conditions—specifically, subsidence with local leeside enhancements and midlevel warming over Cache Valley. The analyses of this study indicate that the subseasonal peak of long-term mean PM2.5 concentrations in Cache Valley is linked to the large-scale circulations’ subseasonal evolution, which involves remote forcing in the circumpolar circulations as well as possible tropical–midlatitude interactions. This subseasonal evolution of the winter mean circulation also affects precipitation along the West Coast

Tuning the hydrophobic cores of self-immolative polyglyoxylate assemblies

Polyglyoxylates are a recently-introduced class of self-immolative polymers, that depolymerize to small molecules upon the cleavage of a stimuli-responsive end-cap from the polymer terminus. The incorporation of different pendant ester groups or other aldehyde monomers offers the potential to tune the polymer properties, but this remains largely unexplored. With the goal of tuning the self-assembly and drug-loading properties of polyglyoxylate block copolymers, we explored the polymerization and copolymerization of n-butyl glyoxylate, L-menthyl glyoxylate, and chloral with ethyl glyoxylate to form UV light-responsive polyglyoxylates. The resulting polymers were coupled to poly(ethylene glycol) to afford amphiphilic block copolymers. Self-assembly of the different copolymers was studied and although each system formed solid particles, the cores of the assemblies differed in their stability, hydrophobicity, and their ability to load the hydrophobic drug celecoxib. All systems depolymerized and released the drug in response to UV light. The toxicity profiles for the assemblies were also evaluated using MDA-MB-231 cells. Overall, this work demonstrates that the properties of polyglyoxylates and their assemblies can be readily tuned through the incorporation of new monomers, thereby providing a promising platform for drug delivery and other applications

Polyesters based on aspartic acid and poly(ethylene glycol): Functional polymers for hydrogel preparation

Hydrogels are commonly used as scaffolds for the preparation of three-dimensional tissue constructs and for the encapsulation and delivery of cells in regenerative medicine. Polyesters are an attractive class of polymers for hydrogel preparation. However, most polyesters have hydrophobic backbones and lack pendent groups that can be chemically functionalized. We describe here the development of water-soluble polyesters based on aspartic acid and poly(ethylene glycol) (PEG) (600 or 1500 g/mol), having pendent reactive amines. The reactivity of these amines with methacrylic anhydride, maleic anhydride, and itaconic anhydride was explored for the introduction of crosslinkable groups. The resulting methacrylamide-functionalized polymers were successfully crosslinked to form hydrogels using a redox-initiated free radical polymerization. The use of 10% (weight/volume) of polymer, and 10 mM of potassium persulfate and tetramethylethylenediamine led to high (\u3e97%) gel content, and compressive moduli of 13–21 kPa. Human adipose-derived stromal cells were encapsulated during the crosslinking process and exhibited greater than 80% viability in the hydrogels prepared from the polyester containing 600 g/mol PEG, with lower viability observed for the polymer containing 1500 g/mol PEG. These results support the potential for aspartic acid-based copolymers with short PEG chains in the backbone to serve as a platform for cell encapsulation, with additional opportunities for further functionalization available in the future

Estimating the furrow infiltration characteristic from a single advance point

Management and control of surface irrigation, in particular furrow irrigation, is limited by spatio-temporal soil infiltration variability as well as the high cost and time associated with collecting intensive field data for estimation of the infiltration characteristics. Recent work has proposed scaling the commonly used infiltration function by using a model infiltration curve and a single advance point for every other furrow in an irrigation event. Scaling factors were calculated for a series of furrows at two sites and at four points down the length of the field (0.25 L, 0.5 L, 0.75 L and L). Differences in the value of the scaling factor with distance were found to be a function of the shape of the advance curves. It is concluded that use of points early in the advance results in a substantial loss of accuracy and should be avoided. The scaling factor was also strongly correlated with the furrow-wetted perimeter suggesting that the scaling is an appropriate way of both predicting and accommodating the effect of the hydraulic variability

The Campbells: lordship, literature and liminality

The Campbells have the potential to offer much to the theme of literature and borders, given that the kindred’s astonishing political success in the late medieval and early modern period depended heavily upon the ability to negotiate multiple frontiers: between Highlands and Lowlands; between Gaelic Scotland and Ireland, and, especially after the Reformation, with England and the matter of Britain. This paper will explore the literary dimension to Campbell expansionism, from the Book of the Dean of Lismore in the earlier sixteenth century, to poetry addressed to dukes of Argyll in the earlier eighteenth century. Particular attention will be paid to the literary proclivities of the household of the Campbells of Glenorchy on either side of what appears to be a major watershed in 1550; and to the agenda of the Campbell protégé John Carswell, first post-Reformation bishop of the Isles, and author of the first printed book in Gaelic in either Scotland or Ireland, Foirm na n-Urrnuidheadh (‘The Form of Prayers’), published at Edinburgh in 1567