Essential Oil and Supercritical Carbon Dioxide Extract of Grapefruit Peels Formulated for Candida albicans Infections: Evaluation by an in Vitro Model to Study Fungal-Host Interactions.

Resistance to currently available antifungal agents raises the need to develop alternative remedies. Candida albicans is the most common opportunistic pathogenic fungus of humans, colonizing in the genital and intestinal mucosa, skin, and oral-nasal cavity and reducing quality of life. Herein, essential oil from grapefruit (Citrus paradise) peels was obtained by hydrodistillation, and the remaining plant material was sequentially subjected to supercritical carbon dioxide (SC-CO2) extraction to determine the conditions for maximizing phenolic compounds. A statistical design was used to evaluate the effect of temperature (30, 50, 70 °C), pressure (80, 150, 220 bar), and ethanol as a cosolvent (0%, 10%, and 20% v/v). Essential oil and SC-CO2 extracts were mixed at various ratios to develop an effective antifungal formulation. Subsequently, fungal infection was modeled by coculturing C. albicans with human skin keratinocytes (HaCaT) to mimic dermal mycoses, endothelial cells (HUVEC) to evaluate vascular fate, and cervical adenocarcinoma (HeLa) cells to represent additional genital mycoses. Treatment with essential oil and extract (25:75%) formulation for 8 h exhibited slight cytotoxicity toward HeLa cells, no toxicity toward HaCaT and HUVECs, whereas inhibition of C. albicans. Considering the clinical significance, such in vitro models are essential to screen potential compounds for the treatment of opportunistic fungal infections.

Bilayered laponite/alginate-poly(acrylamide) composite hydrogel for osteochondral injuries enhances macrophage polarization: An in vivo study.

Addressing osteochondral defects, the objective of current study was to synthesize bilayered hydrogel, where the cartilage layer was formed by alginate (Alg)-polyacrylamide (PAAm) with and without the addition of TGF-ß3 and bone layer by laponite XLS/Alg-PAAm and characterize by in vitro and in vivo experiments. Exceeding the mechanical strength of Alg-PAAm (32.95 ± 1.23 kPa) and XLS based (317.5 ± 21.72 kPa) hydrogels, XLS/Alg-PAAm hydrogel (469.7 ± 6.1 kPa) activated macrophages towards M2 phenotype and stimulated the expression of anti-inflammatory factors. The addition of TGF-ß3 accelerated transition of macrophage polarization, especially between day 4 and 7. The expression levels of M1-related genes such as CD80, iNOS and TNF-α decreased gradually after day 4, reaching lowest values at day 13, whereas the expression levels of M2-related genes, CD206, Arg1 and STAT6 significantly increased promoting M2 macrophage polarization, which might be associated with accelerated bone repair. Moreover, bilayer structure exhibited a better cell viability as well as repairment thorough the XLS contents. In vivo histological examinations verified the significant surface regularity and hyaline like tissue formation employment, along with synchronized degradation profile of the hydrogel with tissue healing at the end of 12 weeks. A mechanically durable, biocompatible and immunocompatible hydrogel was formulated to be utilized in bone-cartilage engineering applications.