In a recent study published in food, Researchers are investigating environmentally friendly and sustainable uses for processed tart cherries (Prunus genus Prunus L.) Useless. Their research focused on cosmetic applications, including the extraction of bioactive metabolites from cherries and the optimization of their beneficial effects on human skin.
study: Tart cherry (Prunus cerasus L.) pit extract protects human skin cells from oxidative stress: enabling sustainable use of food industry by-products. Image credit: Serhii Ivashchuk / Shutterstock.com
Cherries and the burden of environmental waste
cherries, especially cherries belonging to the species bird primrose land P. Cerasus L., processed in large quantities. In fact, the United States Department of Agriculture (USDA) National Agricultural Statistics Service (NASS) reported a staggering 275,000 tons of agricultural statistics. bird primrose L. and 114,600 tons of P. Cerasus L. cherry was processed in 2022 alone.
More than 90% of cherries are processed by the food industry to produce juice, salted products, and wine. This produces by-products such as cherry pomace and cherry pits (CP), most of which are incinerated or disposed of.
In addition to economic losses due to the disposal of organic resources that could be reused or recycled, conventional waste causes environmental pollution of soil, water, and air through greenhouse gas emissions.
The skin is the individual’s first line of defense against biological pathogens, non-biological environmental pollutants, and ultraviolet (UV) radiation. Long-term exposure to UV light can cause skin inflammation and the production of harmful reactive oxygen species (ROS), which induce cell damage and lead to premature aging and skin diseases such as psoriasis and cancer. may be caused.
Polyphenols, one of the largest classes of plant-derived metabolites, have been shown to exhibit antioxidant activity. in vitro, may reduce skin inflammation. Cherries are rich in polyphenols and could be used as raw materials for new cosmetics. Although previous studies have investigated the benefits of by-products from cherry stems and seeds, the antioxidant activity of CP still needs to be evaluated.
In this study, researchers developed a new method to extract bioactive compounds from tart cherries (P. Cerasus L.) CP evaluated the cytotoxic and anti-inflammatory properties of these bioactive substances using human keratinocyte (HaCaT) cells. Tart cherry seeds obtained from Peterson Farms in Michigan were oven dried and ground into a seed powder. Extraction was achieved by conventional extraction using 100% water and a new extraction approach using water:ethanol in a 1:1 ratio.
Extraction was performed at 150 °C with a solid-liquid ratio of 30 g of pit powder in 300 mL of liquid to produce CP water (CPW) or CP ethanol (CPE). CPW and CPE were centrifuged, and the resulting supernatant was used for analysis.
A double-beam Lambda 365 UV-Vis spectrophotometer was used to quantify the polyphenol (TP), anti-radical powder (ARP), and flavonoid (TP) content of the supernatant. To improve the accuracy of TP quantification, the colorimetric Folin-Ciocalteu assay (TP) was adopted. At the same time, a second colorimetric assay was performed to in vitro Antioxidant activities of TP, TF, and ARP toward 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS).
Metabolomic analysis was performed using liquid chromatography-mass spectrometry (LC-MS/MS). Quantification of individual polyphenols was achieved using a UV-visible spectrophotometer with dimethyl sulfoxide (DMSO) as the reference solution.
HaCaT cells and mouse fibroblast (NIH3T3) cells were treated with various concentrations of CPW and CPE or extract and hydrogen peroxide combinations (H2○2) for cell viability assay and ROS activity assay, respectively.
Intracellular ROS activity was measured using fluorescence microscopy. RNA isolation and real-time polymerase chain reaction (RT-PCR) analysis were also performed to identify HaCaT and NIH3T3 genes that were up- or down-regulated after extract treatment.
The new water:ethanol extraction protocol significantly outperformed the traditional water-only protocol. Addition of ethanol to the mixture increased the TP yield from the same starting amount of CP by 2.1 times.
As observed through ARP assay, CPE was associated with approximately 2.5 times more significant antioxidant activity compared to CPW. The antioxidant superiority of CPE was confirmed by ABTS assay, which showed a 2.4-fold increase in activity compared to CPW. Both extraction protocols revealed nearly 10-fold higher measured TP content compared to previous studies on his TP sources from other natural plants.
LC-MS/MS results identified and quantified multiple caffeoylquinate ester isomers, hydroxycinnamic acids, and flavonoids. CPE extracts showed higher concentrations of compounds than CPW. Lyophilized CP extracts similarly showed higher concentrations of bioactive substances than liquid extracts, suggesting future industrial optimization in extraction protocols.
Comparing the UV-Vis spectra of CPW and CPE yielded similar results corresponding to the peaks of apigenin, quercetin, and chlorogenic acid. These results support the enrichment of flavonoids and coumaroyl derivatives in CP and CP extracts.
CP extract concentrations of 50–400 nM did not induce toxicity in HaCaT cells and NIH3T3 fibroblasts even after 48 h of incubation. We tested the effects of common pure phenols detected during the characterization of CP extracts and found that only apigenin and catechin negatively affected cell viability at concentrations above 100 μM. This was more than 50 times higher than that found in CP extracts.
Both CPW and CPE significantly decreased ROS levels in HaCaT cells treated with H.2○2.
CP extract was effective in protecting cells from LPS-induced nitric oxide production, which was verified by 2′-7′-dichlorodihydrofluorescein diacetate (DCF-DA) staining assay . By RT-PCR assay, H2○2 Treatment decreased the expression levels of catalase (CAT) and superoxide dismutase (SOD1) in HaCaT cells. CPE and CPW pretreatment rescued HaCaT cells from these effects.
In this study, the researchers aimed to find alternative uses for by-products of cherry processing, thereby reducing the environmental and economic burden of the millions of tons of by-product waste that pollute ecosystems every year. To achieve this objective, they devised a new method to extract natural bioactive substances from his CP. This method was found to be two to three times more efficient than traditional extraction protocols.
CP extract is rich in metabolites that can effectively reduce skin oxidative stress caused by pollution and long-term UV exposure, preventing skin diseases such as premature aging, psoriasis, and even cancer. To do.
- Dekot, H., Sudhakaran, M., Bozmier, E., other. (2022). Tart cherry (Prunus cerasus L.) pit extract protects human skin cells from oxidative stress: enabling sustainable use of food industry by-products. food 12(20), 3748.doi:10.3390/foods12203748