Ischemic stroke disrupts the endothelial glycocalyx through activation of proHPSE via acrolein exposure

Infiltration of peripheral immune cells after blood-brain barrier dysfunction causes severe inflammation after a stroke. Although the endothelial glycocalyx, a network of membrane-bound glycoproteins and proteoglycans that covers the lumen of endothelial cells, functions as a barrier to circulating cells, the relationship between stroke severity and glycocalyx dysfunction remains unclear. In this study, glycosaminoglycans, a component of the endothelial glycocalyx, were studied in the context of ischemic stroke using a photochemically induced thrombosis mouse model. Decreased levels of heparan sulfate and chondroitin sulfate and increased activity of hyaluronidase 1 and heparanase (HPSE) were observed in ischemic brain tissues. HPSE expression in cerebral vessels increased after stroke onset and infarct volume greatly decreased after co-administration of N-acetylcysteine + glycosaminoglycan oligosaccharides as compared with N-acetylcysteine administration alone. These results suggest that the endothelial glycocalyx was injured after the onset of stroke. Interestingly, scission activity of proHPSE produced by immortalized endothelial cells and HEK293 cells transfected with hHPSE1 cDNA were activated by acrolein (ACR) exposure. We identified the ACR-modified amino acid residues of proHPSE using nano LC–MS/MS, suggesting that ACR modification of Lys¹³⁹ (6-kDa linker), Lys¹⁰⁷, and Lys¹⁶¹, located in the immediate vicinity of the 6-kDa linker, at least in part is attributed to the activation of proHPSE. Because proHPSE, but not HPSE, localizes outside cells by binding with heparan sulfate proteoglycans, ACR-modified proHPSE represents a promising target to protect the endothelial glycocalyx.     

Effect of polymer differences on fungal penetration of hydrogel lenses

Summary Two chemically distinct types of hydrogel lenses, vifilcon A and bufilcon A, each with a water content of 55%, were challenged in a balanced salts solution withAspergillus fumigatus, Cladosporium cladosporioides, Curvularia lunata andFusarium solani. The lenses were cleaned, disinfected and stained after varying periods of incubation and examined with light microscopy and scanning electron microscopy. For three of the four fungi, the bufilcon A lens was more susceptible to fungal attack than the vifilcon A lens.Curv. lunata produced the greatest number of penetration pegs within 72 h for both lens types. Etching of lens surfaces was observed withC. cladosporioides. In general, the susceptibility of a hydrogel lens to penetration with a fungus appeared to vary with the species of fungus and the chemical composition of the lens.     

Biochemical analyses of the cement float of the goose barnacle Dosima fascicularis – a preliminary study

The goose barnacle Dosima fascicularis produces an excessive amount of adhesive (cement), which has a double function, being used for attachment to various substrata and also as a float (buoy). This paper focuses on the chemical composition of the cement, which has a water content of 92%. Scanning electron microscopy with EDX was used to measure the organic elements C, O and N in the foam-like cement. Vibrational spectroscopy (FTIR, Raman) provided further information about the overall secondary structure, which tended towards a β-sheet. Disulphide bonds could not be detected by Raman spectroscopy. The cystine, methionine, histidine and tryptophan contents were each below 1% in the cement. Analyses of the cement revealed a protein content of 84% and a total carbohydrate content of 1.5% in the dry cement. The amino acid composition, 1D/2D-PAGE and MS/MS sequence analysis revealed a de novo set of peptides/proteins with low homologies with other proteins such as the barnacle cement proteins, largely with an acidic pI between 3.5 and 6.0. The biochemical composition of the cement of D. fascicularis is similar to that of other barnacles, but it shows interesting variations.     

Serum‐free,chemically defined medium with TGF‐β3 enhances functional properties of nucleus pulposus cell‐laden carboxymethylcellulose hydrogel constructs

Degeneration of the nucleus pulposus (NP) has been implicated as a major cause of low back pain. Tissue engineering strategies may provide a viable NP replacement therapy; however, culture conditions must be optimized to promote functional tissue development. In this study, a standard serum‐containing medium formulation was compared to a chemically defined, serum‐free medium to determine the effect on matrix elaboration and functional properties of NP cell‐laden carboxymethylcellulose (CMC) hydrogels. Additionally, both media were further supplemented with transforming growth factor‐beta 3 (TGF‐β₃). Glycosaminoglycan (GAG) content increased in both TGF‐β₃‐treated groups and was highest for treated, serum‐free constructs (9.46 ± 1.51 µg GAG/mg wet weight), while there were no quantifiable GAGs in untreated serum‐containing samples. Histology revealed uniform, interterritorial staining for chondroitin sulfate proteoglycan throughout the treated, serum‐free constructs. Type II collagen content was greater in both serum‐free groups and highest in treated, serum‐free constructs. The equilibrium Young's modulus was highest in serum‐free samples supplemented with TGF‐β₃ (18.54 ± 1.92 kPa), and the equilibrium weight swelling ratio of these constructs approached that of the native NP tissue (22.19 ± 0.46 vs. 19.94 ± 3.09, respectively). Taken together, these results demonstrate enhanced functional matrix development by NP cells when cultured in CMC hydrogels maintained in serum‐free, TGF‐β₃ supplemented medium, indicating the importance of medium formulation in NP construct development. Biotechnol. Bioeng. 2010; 105: 384–395. © 2009 Wiley Periodicals, Inc.     

Poly(2-oxazoline) hydrogels as next generation three-dimensional cell supports

Synthetic hydrogels selectively decorated with cell adhesion motifs are rapidly emerging as promising substrates for 3D cell culture. When cells are grown in 3D they experience potentially more physiologically relevant cell–cell interactions and physical cues compared with traditional 2D cell culture on stiff surfaces. A newly developed polymer based on poly(2-oxazoline)s has been used for the first time to control attachment of fibroblast cells and is discussed here for its potential use in 3D cell culture with particular focus on cancer cells toward the ultimate aim of high-throughput screening of anticancer therapies. Advantages and limitations of using poly(2-oxazoline) hydrogels are discussed and compared with more established polymers, especially polyethylene glycol (PEG).     

Zwitterionic Sulfobetaine Hydrogel Electrolyte Building Separated Positive/Negative Ion Migration Channels for Aqueous Zn‐MnO2 Batteries with Superior Rate Capabilities

Hydrogel electrolytes have attracted increasing attention due to their potential uses in the fabrication of flexible solid‐state batteries. However, the development of hydrogel electrolytes is still in the initial stage and the number of available strategies is limited. Ideally, the hydrogel electrolyte should exhibit suitable ionic conductivity rate, mechanical strength, and biocompatibility for safety. In this study, a zwitterionic sulfobetaine/cellulose hydrogel electrolyte is fabricated using raw materials from natural plants, which exhibits a good biocompatibility with mammalian cells. The intrinsic zwitterionic groups on sulfobetaine chains can provide separated ion migration channels for positive and negative ions, which largely facilitates electrolyte ion transport. A solid‐state Zn‐MnO₂ battery with a fabricated zwitterionic gel electrolyte exhibits a very high rate performance. It exhibits a specific capacity of 275 mA h ${\text{g}}_{{\text{MnO}}_2}^{?1}$ at 1 C. Even up to 30 C, a high capacity of 74 mA h ${\text{g}}_{{\text{MnO}}_2}^{?1}$ is maintained during the charging–discharging for up to 10 000 cycles. For wearable applications, the flexible solid‐state batteries can be used as reliable and portable sources to power different wearable electronics such as a commercial smart watch, electroluminescent panel, and color electroluminescence line, which shows their large potentials for use in next‐generation flexible and wearable battery technologies.