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CARF (CDKN2AIP) regulates cellular fate in response to various stresses. However, its role in metabolic stress is unknown. We found that fatty livers from mice exhibit low CARF expression. Similarly, overloaded palmitate inhibited CARF expression in HepG2 cells, suggesting that excess fat-induced stress downregulates hepatic CARF. In agreement with this, silencing and overexpressing CARF resulted in higher and lower fat accumulation in HepG2 cells, respectively. Furthermore, CARF overexpression lowered the ectopic palmitate accumulation in HepG2 cells. We were interested in understanding the role of hepatic CARF and underlying mechanisms in the development of NAFLD. Mechanistically, transcriptome analysis revealed that endoplasmic reticulum (ER) stress and oxidative stress pathway genes significantly altered in the absence of CARF. IRE1α, GRP78, and CHOP, markers of ER stress, were increased, and the treatment with TUDCA, an ER stress inhibitor, attenuated fat accumulation in CARF-deficient cells. Moreover, silencing CARF caused a reduction of GPX3 and TRXND3, leading to oxidative stress and apoptotic cell death. Intriguingly, CARF overexpression in HFD-fed mice significantly decreased hepatic steatosis. Furthermore, overexpression of CARF ameliorated the aberrant ER function and oxidative stress caused by fat accumulation. Our results further demonstrated that overexpression of CARF alleviates HFD-induced insulin resistance assessed with ITT and GTT assay. Altogether, we conclude that excess fat-induced reduction of CARF dysregulates ER functions and lipid metabolism leading to hepatic steatosis.

Cellulose nanofiber (CNF) hydrogels with low lignin (8%) (LL-CNF) and high lignin (18%) (HL-CNF) content were produced at nominal powers of 240, 720 and 1200 W using high-intensity ultrasound technology (HIUS). Freeze-dried CNF hydrogels were evaluated as scaffolds for gingival fibroblast cells proliferation aiming biomedical applications. HIUS processing improved the dispersibility of the CNF and increased the water retention value by more than 5 times. The LL-CNF had a maximum fibrillation yield of 46 wt.%, whereas the HL-CNF had a maximum fibrillation yield of 40 wt.% at nominal power of ≥720 W. Regardless of the lignin content, the CNF hydrogels exhibited a typical elastic gel-like behavior with the highest elasticity of 263 Pa. After freeze-drying, the CNF aerogels had porosity ≥ 96.8%, and swelling capacity up to 42.1 g PBS/g aerogel. Moreover, the cell proliferation assay showed no differences in proliferation among the LL-CNF and HL-CNF scaffolds up to 11 days. Therefore, CNF scaffolds prepared with lignin content up to 18% present promising application in the biomedical field., (Copyright © 2019. Published by Elsevier B.V.)

High-intensity ultrasound (HIUS) was used to produce emulsion-filled cellulose nanofiber (CNF) hydrogel using clove essential oil (0.1, 0.5 and 1.0 wt%) as dispersed phase towards tissue engineering applications. The novel encapsulating systems obtained using HIUS specific energy at the levels of 0.10, 0.17, and 0.24 kJ/g were characterized by oil entrapment efficiency, microstructure, water retention value, color parameters, and viscoelastic properties. Freeze-dried emulsion-filled CNF hydrogels were characterized by porosity and swelling capacity. In addition, human gingival fibroblast cell cytocompatibility tests were performed to evaluate their potential applications as tissue engineering scaffold. The clove essential oil content strongly affected the oil entrapment efficiency, water retention value, color difference and whiteness of the prepared emulsion-filled CNF hydrogel. And, the HIUS energy only affected the yellowness of the emulsion-filled CNF hydrogel. Via HIUS processing, the CNF hydrogel successfully acted as a continuous phase in the emulsion-filled gel system with maximum oil entrapment efficiency of 34% when 0.5 wt% clove essential oil was added to the system. The encapsulating systems had predominantly gel-like property with maximum elastic modulus of 411 Pa. Furthermore, the emulsion-filled CNF hydrogels with the addition of clove essential oil up to 0.5 wt% indicated good cell viability rates (74-101%) to human gingival fibroblast cells. The newly developed clove essential oil emulsion-filled CNF hydrogel shows desirable cytocompatibility characteristics and can be considered as an alternative scaffold for tissue engineering applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2019 Elsevier B.V. All rights reserved.)