Microwave- and Ultrasound-Assisted Extraction of _Cucurbita pepo_ Seeds: A Comparison Study of Antioxidant Activity, Phenolic Profile, and In-Vitro Cells Effects  
[PERSON]  
CEA CEA Saclay  
referred to as \"summer squash\" or \"zucchini\", originates from Central and South America, being currently cultivated worldwide in warm regions [18]. It is the most produced species of the Cucurbitaceae family, and one of the oldest known cultivated species, with Mexican archaeological evidence dating from 7000 BC [19; 20]. C. _pepo_ is an economically important crop in which immature fruits, leaves, peel, blossoms, and seeds are consumed due to their nutritional and medicinal benefits [21]. The seeds of the Cucurbitaceae family are globally used for the treatment of different diseases, particularly due to their antiviral, anti-inflammatory, anti-ulcerative, antidiabetic and antioxidant activities as well as analgesic for urinary disorders [17]. A few studies using _C. pepo_ seed extracts were published in the past years, revealing wound healing [22], hair growth promotion [23] and anthelmintic [24] properties, besides their ability to inhibit the cell growth of hyperplastic and cancer cells [25]. Moreover, the treatment with _C. pepo_ extract conducted to a substantial improvement of the lower urinary tract symptoms that are suggestive of benign prostatic hyperplasia [26]. According to different authors, _C. pepo_ seeds are rich in several bioactive compounds, such as polyphenols [27], which have anti-aging and anticarcinogenic effects and may protect against vascular inflammation and cardiovascular and neurodegenerative diseases [28; 29; 30; 31]. \(\alpha\)- and \(\gamma\)-tocopherols [32] are also present, defending cells from oxidative stress and inflammation [33], while carotenoids (e.g., \(\beta\)-carotene and \(\beta\)-cryptoxanthin) may protect against different chronic diseases, including cancer [34] and cardiovascular diseases [35]. Besides that, carotenoids improve the cognitive and visual functions [36]. In addition, _C. pepo_ seeds are rich in polyunsaturated fatty acids [37], zinc [38], and phytosterols such as \(\beta\)-sitosterol [32], which may reduce the blood cholesterol [39] and decrease the risk of certain types of cancer [40]. \(\beta\)-sitosterol is also the bioactive compound responsible for the successful use of _C. pepo_ seeds in the treatment of benign prostatic hyperplasia [41; 42; 43]. Berberine and palmatine are also present in considerable amounts [24], conferring to nematocidal [24], antimalarial [44], anti-tilieishmaniasis [45], anti-schistosomiasis [46] and _Toxoplasma gondii_ inhibitory properties [47]. Nevertheless, most of the authors employed organic solvents in the extractive step or, at least, used cold pressing methods aiming to obtain the lipidic fraction. Therefore, the use of water as an alternative solvent led to a greener extraction process and, simultaneously, avoided the extraction of a significant amount of lipids that are naturally present in _C. pepo_ seeds. Allied to the selection of less-polluting solvents, it is imperative to select eco-friendly extraction methods. Therefore, technologies such as microwave-assisted extraction (MAE) and ultrasound-assisted extraction (UAE) arise as an alternative to the traditional ones [48]. MAE is a conventional, automated green extraction technique that allows the extraction of active components from different matrices [49; 50]. By using microwave energy, MAE heats the solvents in contact with samples, disrupts the cell membrane, and releases the intracellular components into the solvents [50; 51]. This is opposite to the conventional methods, such as Soxhlet extraction, that often requires 12-24 h of extraction periods and hundreds of milliliters of organic solvents [50]. When compared to such methods, MAE requires shorter extraction times, decreasing the degradation of the extracted components as well as the costs and the volume of the solvents used, and therefore improving the purity of the final extracts [52; 53; 54; 55; 56].