Responsive polymer-assisted 3D cryogel supports Huh7.5 as in vitro hepatitis C virus model and ectopic human hepatic tissue in athymic mice.

The three-dimensional (3D) cell culture models serve as the interface between conventional two-dimensional (2D) monolayer culture and animal models. 3D culture offers the best possible model system to understand the pathophysiology of human pathogens such as hepatitis C virus (HCV), which lacks a small animal model, due to narrow host tropism and non-permissiveness of murine hepatocytes. In this study, functionally robust spheroids of HCV permissive Huh7.5 cells were generated, assisted by the temperature or pH-responsive polymers PNIPAAm and Eudragit respectively, followed by the long-term growth of the multilayered 3D aggregates in poly(ethylene glycol) (PEG)-alginate-gelatin (PAG) cryogel. The human serum albumin (HSA), marker of hepatic viability was detected up to 600 ng/ml on 24th day of culture. The 3D spheroid culture exhibited a distinct morphology and transcript levels with the upregulation of hepato-specific transcripts, nuclear factor 4α (HNF4α), transthyretin (TTr), albumin (Alb), phase I and phase II drug-metabolizing genes. The two most important phase I enzymes CYP3A4 and CYP2D6, together responsible for 90% metabolism of drugs exhibited up to 9- and 12-fold increment, respectively in transcripts. The 3D culture was highly permissive to HCV infection and supported higher multiplicity of infection compared to monolayer Huh7.5 culture. Quantitation of high levels of HSA (500-200 ng/ml) in circulation in mice for 32 days asserted integration with host vasculature and in vivo establishment of 3D culture implants as an ectopic human hepatic tissue in mice. The study demonstrates the 3D spheroid Huh7.5 culture as a model for HCV studies and screening potential for anti-HCV drug candidates.

Sulfonated PAM/PPy Cryogels with Lowered Evaporation Enthalpy for Highly Efficient Photothermal Water Evaporation.

Because of the increasing scarcity of water resources, the desalination of seawater by photothermal evaporation with harvested solar energy has gradually become a popular research topic. The interconnected macroporous cryogel prepared from polymerization and crosslinking below the freezing temperature of the reactant solution has an excellent performance in photothermal water evaporation after loading photothermal materials. In this study, polyacrylamide (PAM) cryogels were prepared by cryo-polymerization and sulfonated in an alkaline solution containing formaldehyde and Na2SO3. Importantly, the evaporation enthalpy of water in sulfonated PAM cryogel was reduced to 1187 J·g-1 due to the introduction of sulfonate groups into PAM, which was beneficial to increase the photothermal evaporation rate and efficiency. The sulfonated PAM cryogels loaded with polypyrrole and the umbrella-shaped melamine foam substrate were combined to form a photothermal evaporation device, and the evaporation rate was as high as 2.50 kg·m-2·h-1 under one-sun radiation. Meanwhile, the evaporation rate reached 2.09 kg·m-2·h-1 in the 14 wt% high-concentration saline solution, and no salt crystals appeared on the surface of the cryogel after 5 h of photothermal evaporation. Therefore, it was evidenced that the presence of sulfonate groups not only reduced the evaporation enthalpy of water but also prevented salting-out from blocking the water delivery channel during photothermal evaporation, with a sufficiently high evaporation rate, providing a reliable idea of matrix modification for the design of high-efficiency photothermal evaporation materials.

Super-Macroporous Pulluan Cryogels as Controlled Active Delivery Systems with Controlled Degradability.

Here, super-macroporous cryogel from a natural polysaccharide, pullulan was synthesized using a cryo-crosslinking technique with divinyl sulfone (DVS) as a crosslinker. The hydrolytic degradation of the pullulan cryogel in various simulated body fluids (pH 1.0, 7.4, and 9.0 buffer solutions) was evaluated. It was observed that the pullulan cryogel degradation was much faster in the pH 9 buffer solution than the pH 1.0 and 7.4 buffer solutions in the same time period. The weight loss of the pullulan cryogel at pH 9.0 within 28 days was determined as 31% ± 2%. To demonstrate the controllable drug delivery potential of pullulan cryogels via degradation, an antibiotic, ciprofloxacin, was loaded into pullulan cryogels (pullulan-cipro), and the loading amount of drug was calculated as 105.40 ± 2.6 µg/mg. The release of ciprofloxacin from the pullulan-cipro cryogel was investigated in vitro at 37.5 °C in physiological conditions (pH 7.4). The amount of drug released within 24 h was determined as 39.26 ± 3.78 µg/mg, which is equal to 41.38% ± 3.58% of the loaded drug. Only 0.1 mg of pullulan-cipro cryogel was found to inhibit half of the growing Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) colonies for 10 min and totally eradicated within 2 h by the release of the loaded antibiotic. No significant toxicity was determined on L929 fibroblast cells for 0.1 mg drug-loaded pullulan cryogel. In contrast, even 1 mg of drug-loaded pullulan cryogel revealed slight toxicity (e.g., 66% ± 9% cell viability) because of the high concentration of released drug.