3D Printable Modular Soft Elastomers from Physically Cross-linked Homogeneous Associative Polymers.

Three-dimensional (3D) printing of elastomers enables the fabrication of many technologically important structures and devices. However, there remains a critical need for the development of reprocessable, solvent-free, soft elastomers that can be printed without the need for post-treatment. Herein, we report modular soft elastomers suitable for direct ink writing (DIW) printing by physically cross-linking associative polymers with a high fraction of reversible bonds. We designed and synthesized linear-associative-linear (LAL) triblock copolymers; the middle block is an associative polymer carrying amide groups that form double hydrogen bonding, and the end blocks aggregate to hard glassy domains that effectively act as physical cross-links. The amide groups do not aggregate to nanoscale clusters and only slow down polymer dynamics without changing the shape of the linear viscoelastic spectra; this enables molecular control over energy dissipation by varying the fraction of the associative groups. Increasing the volume fraction of the end linear blocks increases the network stiffness by more than 100 times without significantly compromising the extensibility. We created elastomers with Young's moduli ranging from 8 kPa to 8 MPa while maintaining the tensile breaking strain around 150%. Using a high-temperature DIW printing platform, we transformed our elastomers to complex, highly deformable 3D structures without involving any solvent or post-print processing. Our elastomers represent the softest melt reprocessable materials for DIW printing. The developed LAL polymers synergize emerging homogeneous associative polymers with a high fraction of reversible bonds and classical block copolymer self-assembly to form a dual-cross-linked network, providing a versatile platform for the modular design and development of soft melt reprocessable elastomeric materials for practical applications.

Bottlebrush polyethylene glycol nanocarriers translocate across human airway epithelium via molecular architecture enhanced endocytosis.

Pulmonary drug delivery is critical to the treatment of respiratory diseases. However, the human airway surface presents multiscale barriers to efficient drug delivery. Here we report a bottlebrush polyethylene glycol (PEG-BB) nanocarrier that can translocate across all barriers within the human airway surface. Guided by the molecular theory, we design a PEG-BB molecule consisting of a linear backbone densely grafted by many (∼1,000) low molecular weight (∼1000 g/mol) PEG chains; this results in a highly anisotropic, wormlike nanocarrier featuring a contour length of ∼250 nm, a cross-section of ∼20 nm, and a hydrodynamic diameter of ∼40 nm. Using the classic air-liquid-interface culture system to recapitulate essential biological features of the human airway surface, we show that PEG-BB rapidly penetrates through endogenous airway mucus and periciliary brush layer (mesh size of 20-40 nm) to be internalized by cells across the whole epithelium. By quantifying the cellular uptake of polymeric carriers of various molecular architectures and manipulating cell proliferation and endocytosis pathways, we show that the translocation of PEG-BB across the epithelium is driven by bottlebrush architecture enhanced endocytosis. Our results demonstrate that large, wormlike bottlebrush PEG polymers, if properly designed, can be used as a novel carrier for pulmonary and mucosal drug delivery.

Dietary exposure assessment of paralytic shellfish toxins through shellfish consumption in Shenzhen population, China.

Paralytic shellfish toxins (PSTs) produced by certain marine dinoflagellates accumulate in filter-feeding marine bivalves. We used LC-MS/MS to detect and quantify 13 PSTs in 188 shellfish samples of 14 species collected from Shenzhen city's Buji seafood wholesale market from March 2019 to February 2020. Twenty-six of 188 shellfish samples (13.8%) were PSTs detectable. Within 14 species, 10 out of 34 noble clam Chlamys nobilis samples contain detectable PSTs with the highest detection rate 29.4%. Seven out of 17 samples from Nan'ao island contained detectable PSTs with the highest detection rate 41.2% among 11 origins. Samples containing PSTs were concentrated in spring and winter, with the highest levels in March>December>January. Among PSTs detected, C1 was dominant. Acute dietary exposure assessment for Shenzhen residents were based on mean adult body weight, 99th percentile daily shellfish consumption of Shenzhen food consumption survey 2008 and maximum PSTs concentration for each shellfish species. The outcome for Chlamys nobilis was 2.4~3.7-fold higher than recommended ARfDs. Mean PSTs concentration, P99, and mean shellfish consumption were used to assess chronic dietary exposure. The results were lower than recommended ARfDs. In conclusion, residents in Shenzhen are at risk for acute PSTs poisoning, while relatively safe from chronic PSTs exposure.

Shellfish contamination with lipophilic toxins and dietary exposure assessments from consumption of shellfish products in Shenzhen, China.

Lipophilic shellfish toxins (LSTs) can cause human illness and therefore represent a serious threat to public health. Shellfish are the main dietary source of LSTs, but very few studies have appraised dietary exposure to LSTs through shellfish consumption in China. We measured levels of multiple LSTs in shellfish samples sold in the principal wholesale seafood market in the southern coastal city of Shenzhen, and we estimated the potential for acute and chronic LST exposure of the Shenzhen population via ingestion of shellfish. LST contamination data were obtained from a total of 14 species of 188 commercial samples. Eleven individual LSTs, namely okadaic acid (OA), dinophysis toxin-1 and -2 (DTX1 and DTX2), pectenotoxin-2 (PTX2), yessotoxin and homo yessotoxin (YTX and hYTX), azaspiracid-1, -2 and -3 (AZA1, AZA2, AZA3), spirolides (SPXs), and gymnodimine (GYM), were determined using liquid chromatography electrospray-ionization tandem mass spectrometry (LC-ESI-MS/MS). More than two thirds of samples showed undetectable LSTs, while the detection rates (the proportion of samples with detectable LSTs) of individual LSTs ranged from 0% to 45.7%. Most shellfish samples had lower levels of LST contamination than the corresponding limits of detection (LODs), while some samples had levels of hYTX and GYM that exceeded the limits of quantification (LOQs). Overall, levels of LSTs in the 188 samples were below the regulatory limits set by most countries. Acute and chronic exposures of LST were estimated by a point-estimate modeling method that combined sample contamination data with consumption data from dietary survey of Shenzhen residents and consumption figures proposed by EFSA, the European Food Safety Authority. Seasonal variations in LST concentrations were noted in some instances. Overall, the estimated acute exposure to LSTs based on consumption of large-size shellfish portions and the maximum LSTs contamination level were below the provisional acute reference doses (ARfDs) proposed by the EFSA. Chronic exposure estimates based on mean and 99th percentile consumption of shellfish by Shenzhen residents and mean LSTs contamination levels in the collected samples were from 2452 to 74 times lower than those associated with estimated acute exposure levels.

Hemolytic and cytotoxic activity from cultures of Aureococcus anophagefferens-a causative species of brown tides in the north-western Bohai Sea, China.

Brown tides were first observed in 2009 in the north-western Bohai Sea (Qinhuangdao sea area), China, and blooms have occurred at different scales in late spring every year since then. Although the detrimental effects on marine organisms of the causative phytoplankton species Aureococcus anophagefferens have been extensively studied, the mechanism remains poorly understood. We used erythrocytes and adrenal gland chromaffin tumor cells (PC12) to explore the hemolytic activity and cytotoxicity, respectively, of chloroform and methanol extracts of cultured A. anophagefferens isolated from the north-western Bohai Sea area. The methanol extracts showed no hemolytic or cytotoxic activity. Chloroform extracts had a potent hemolytic effect on rabbit erythrocytes; thin layer chromatography (TLC) indicated that the hemolysin was a kind of glycolipid compound. Erythrocyte lysis assay showed that erythrocytes of sea bream were sensitive to the hemolysin, whereas those of human and chicken erythrocytes were insensitive. The hemolytic effects were elevated as temperatures rose from 4 °C to 37 °C. Hemolytic blocking experiments showed that sphingomyelin and d-xylose can inhibit hemolysis significantly, while osmotic protectants with different hydrated molecular diameters had no inhibition, and the hemolysins had no obvious phospholipase activity. The chloroform extracts of A. anophagefferens had significant inhibitory effects on the viability of PC12 cells, and can induce efflux of lactic dehydrogenase (LDH) of PC12 cells and lead to their necrosis.

Reactive oxygen species (ROS)-responsive biomaterials mediate tissue microenvironments and tissue regeneration.

Reactive oxygen species (ROS) have been considered the pivotal signaling molecules in many physiological processes, and are usually overproduced in various inflammatory tissues. Overproduction of ROS may disrupt cellular homeostasis, cause non-specific damage to critical components, and lead to a series of diseases. ROS are acknowledged as a type of emerging triggered event similar to acidic pH, overproduced enzymes, temperature and other specific stimuli found in pathological microenvironments. Recently, ROS-responsive biomaterials have been identified as a type of promising therapeutic substance to alleviate oxidative stress in tissue microenvironments, and for use as a vehicle triggered by inflammatory diseases to realize drug release under physiological oxidative microenvironments. In this review, we discuss mainly the mechanisms of ROS-responsive biomaterials with solubility switch and chemical degradation, and those ROS-responsive groups used in ROS-responsive biomaterials. The mechanism of ROS overproduction in pathophysiological conditions is introduced. The various applications of ROS-responsive biomaterials in tissue regeneration and disease therapy, such as cardiovascular diseases, osteoarthritis, chronic diabetic wounds, inflammatory bowel disease and other inflammatory diseases, are summarized.