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Objective: To determine the anticancer potential of the methanolic extract from Ephedra alata against breast cancer both in vitro and in vivo. Methods: The effects of the methanolic extract of Ephedra alata on the viability, migration as well as apoptosis of breast cancer 4T1 cells were measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Transwell assay, and annexin V-FITC staining assay, respectively. Histological examination was also carried out. Moreover, a murine breast cancer model was established to evaluate the inhibitory effect of the extract. Biochemical parameters including hepatic and non-hepatic enzymes, malondialdehyde, and glutathione were investigated. Results: The methanolic extract of Ephedra alata showed a strong anti-proliferative and anti-migratory activity against 4T1 cells in a dose-dependent manner. It also induced apoptosis in 4T1 cells. In an in vivo mouse model, the extract markedly inhibited tumor growth, reduced malondialdehyde, and hepatic and non-hepatic enzymes as well as increased glutathione level. Conclusions: The methanolic extract of Ephedra alata inhibits breast cancer in vitro and in vivo, which may be a promising anticancer agent.

Cancer is a class of diseases characterized by uncontrolled cell growth. One of the main aims of developing new therapies is to use natural resources to induce apoptosis. LC-ms/ms analysis of a methanolic extract of Ephedra alata (E.A.) allowed the identification of 20 secondary metabolites, including flavonoids, phenolic acids, and proanthocyanidins. Antiproliferative effect was assessed by crystal violet assay. Antimigration effect was tested by wound healing assay and apoptosis induction was determined by annexin binding assays, Hoechst staining, ROS production, and activation of apoptotic proteins. The results indicated that exposure of breast cancer cells to E.A. extract significantly reduced cell viability in a dose and time-dependent manner and inhibited the migration of 4T1 cells at a low dose. Moreover, treatment of cells with E.A. extract induced apoptosis, as it was detected by Annexin V/7 AAD, Hoechst staining, ROS production, and the activation of caspases. Abbreviation: BSA bovine serum albumin DMSO dimethyl sulfoxide EDTA ethylenediaminetetraacetic acid LC-ms/ms liquid chromatography-mass spectrometry NAC N-acetyl-l-cysteine PARP poly(ADP-ribose) polymerase PMSF phenylmethylsulfonyl fluoride RIPA radioimmunoprecipitation assay buffer ROS reactive oxygen species RPMI Roswell park memorial institute SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis. [ABSTRACT FROM AUTHOR]

Objective: To evaluate the effects of phenolic acids (caffeic, ferulic, and coumaric acids) and flavones (luteolin and apigenin) on the proliferation and melanogenesis in murine melanoma B16-F10 cells. Methods: Cell proliferation was determined after 24 and 48 hours of incubation using MTT assay. The effects of these tested compounds on cell cycle progression were analyzed by flow cytometry. Moreover, the melanin content and tyrosinase activity were measured spectrophotometrically at 475 nm. Results: Luteolin and apigenin exhibited significant anti-proliferative activity against B16-F10 cells, while caffeic, ferulic, and coumaric acids induced slight inhibition after 24 and 48 hours of incubation. The tested compounds disturbed cell cycle progression of B16-F10, by a subsequent decrease in G1 and arrested cycle progression in either G1/S or G2/M phase. Furthermore, apigenin provoked an increase in melanin content of B16-F10 cells. In contrast, luteolin, caffeic, ferulic and coumaric acids induced a decrease in melanin content of B16-F10 cells by inhibiting tyrosinase activity. Conclusions: These active polyphenols may be used as skin whitening agents or natural tanning agents to treat skin pigmentation disorders. [ABSTRACT FROM AUTHOR]

Tissue engineering involves the use of smart biomimetic hybrid matrices to reinforce the cellular interaction with the matrix and restore native properties after regeneration. In this study, we highlight the potential of 3D collagen sponges soaked with bioactive extract, to enhance the wound healing process in vivo . Acid-soluble collagen from two sources, marine and bovine, were extracted and characterized physiochemically using Fourier transform infrared spectroscopy (FTIR) and SDS-PAGE. Our results confirmed that the extracted collagens were mainly composed of collagen type I with slight molecular structure differences. Both collagens present two different α chains ( α 1 and α 2) and one β chain. Highly interconnected 3D scaffolds from collagen from the skin are designed and added by the widely known healing plants Pistacia lentiscus and Calendula officinalis . The resulting 3D collagen matrices possess fine biocompatibility with skin cells, Hacat (keratinocytes), and 3T3-L1 (fibroblasts) cells. To evaluate the potential wound healing effect, a collagen sponge soaked with the bioactive extract was tested on BALB/ c mice. Our findings confirmed that sponges significantly improve animal re-epithelialization by increasing wound closure. Consequently, spongy collagen scaffolds loaded with Pistacia lentiscus and Calendula officinalis could be used as potential wound dressing material., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2022 Aida Lahmar et al.)