Cystoseira is abundant in bioactive chemicals that have been emphasized for their pharmacological value as plentiful and typically safe alternatives, offering a viable technique for treating cancer and inflammatory diseases. However, evaluating the effect of seasonality and extraction methods on Cystoseira chemical composition and cytotoxicity activity are still not thoroughly investigated. This study investigates the effect of seasonal variations and extraction methods of the Cystoseira extracts on the chemical composition and cytotoxicity activity on human cancer cell lines. C. tamariscifolia, C. crinita and C. compressa, were collected from the UK and Libya and colourimetric techniques were used to measure the chemical components. The cell vitality was measured in vitro using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and Trypan Blue (TB) assays. The results showed that both chemical composition and anticancer activity were subject to the seasonality and extraction method. The study also confirmed that the three Cystoseira spices demonstrated potent anticancer activity against human leukaemia HL60 and THP-1, prostate PC3 and lymphoma REC-1 cancer cell lines. In an effort to investigate the anti-proinflammatory activities of Cystoseira extracts, human macrophage-like cells were exposed to the crude extracts and fractions and the release of TNF-α and IL-1β was measured using the enzyme-linked immunosorbent assay (ELISA). The results reveal that the methanol extract of C. tamariscifolia and C. crinita extracts significantly inhibited the release of TNF-α and IL-1β. C. crinita methanolic extract was partitioned using HPLC, and the fractions were tested for their anti-TNF-α release effect. The result showed that the sup-fraction C1c from C. crinita had a significant anti-proinflammatory effect and the LC-MS analysis of the fraction showed that it contains fatty acids and mannitol. These data suggest that sampling times and extraction methods should be considered to maximise the possibility of finding novel bioactive compounds in Cystoseira. The study also suggests that C. crinita is a valuable source of anti-inflammatory agents. This should provide helpful information for medicinal chemists in their attempts to develop anticancer and immunomodulatory agents. However, more research on this topic is needed to understand Cystoseira bioactive compounds mechanisms of action.

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