Antioxidant profile of strawberry tree honey and its marker homogentisic acid in several models of oxidative stress.

The antioxidant activity of several honeys was evaluated considering the different contribution of entire samples. The strawberry tree honey emerged as the richest in total phenols and the most active honey in the DPPH and FRAP tests, and could protect cholesterol against oxidative degradation (140°C). Homogentisic acid (2,5-dihydroxyphenylacetic acid, HGA), the main phenolic compound from strawberry tree honey, showed interesting antioxidant and antiradical activities, and protective effect against thermal-cholesterol degradation, comparable to those of well known antioxidants. Moreover, the pre-treatment with HGA significantly preserved liposomes and LDL from Cu(2+)-induced oxidative damage at 37°C for 2h, inhibiting the reduction of polyunsaturated fatty acids and cholesterol and the increase of their oxidative products. This phenol had no toxic effect in human intestinal epithelial Caco-2 cells within the concentration range tested (5-1000µM). HGA was able to pass through the Caco-2 monolayers, the apparent permeability coefficients (Papp) in the apical-to-basolateral and basolateral-to-apical direction were 3.48±1.22×10(-6) and 2.18±0.34×10(-6)cm/s, respectively, suggesting a passive diffusion pathway as the dominating process. The results of the work qualify HGA as natural antioxidant, able to exert a significant in vitro protective effect and to contribute to the strawberry tree honey antioxidant activity.

First characterization of Pompia intrea candied fruit: The headspace chemical profile, polar extract composition and its biological activities.

Extracts obtained from Pompia intrea (PI), a traditional candied fruit whose main ingredient is the pompia fruit (Citrus monstruosa L.) were evaluated for the first time. Volatile compounds were evaluated by headspace solid-phase microextraction (HS-SPME) followed by GC-FID/MS analyses. Polar compounds were identified by high-performance liquid chromatography coupled with electrospray mass spectrometry LC-ESI-Orbitrap-MS/MS and quantified by liquid chromatography coupled with ultraviolet/visible detection (LC-DAD). The antioxidant activity of these extracts was tested using the FRAP, CUPRAC, ABTS•+ and DPPH● assays. Moreover, their ability to protect intestinal cells against lipid peroxidation was studied. The HS-SPME GC-FID/MS confirmed the presence of typical molecules originating from the fruit (mainly terpenes, but particularly limonene). The LC-DAD and LC-ESI-(HR)MSn profiles showed high levels of neohesperidin (45.7 ±â€¯11.1 mg/L) and 5-(hydroxymethyl)furfural (40.8 ±â€¯23.5 mg/L). The results showed that the PI extracts contained high levels of total phenols and exhibited considerable antioxidant activity, which was significantly correlated to the presence of specific compounds such as neoeriocitrin and neohesperidin. Furthermore, pretreatments with different concentrations of PI extracts preserved enterocytes from oxidative damage by scavenging reactive oxygen species, thus counteracting lipid peroxidation. This study suggests that consumption of PI could provide intake of compounds with ascertained biological activity.

Protective effect of the oligomeric acylphloroglucinols from Myrtus communis on cholesterol and human low density lipoprotein oxidation.

Myrtle (Myrtus communis L.), a culinary spice and flavouring agent for alcoholic beverages widespread in the Mediterranean area and especially in Sardinia, contains the structurally unique oligomeric non-prenylated acylphloroglucinols, semimyrtucommulone and myrtucommulone A, whose antioxidant activity was investigated during the oxidative modification of lipid molecules implicated in the onset of cardiovascular diseases. Both acylphloroglucinols showed powerful antioxidant properties during the thermal (140 degrees C), solvent-free degradation of cholesterol. Moreover, the pre-treatment with semimyrtucommulone and myrtucommulone A significantly preserved LDL from oxidative damage induced by Cu(2+) ions at 2h of oxidation, and showed remarkable protective effect on the reduction of polyunsaturated fatty acids and cholesterol, inhibiting the increase of their oxidative products (conjugated dienes fatty acids hydroperoxides, 7beta-hydroxycholesterol, and 7-ketocholesterol). Taking into account the widespread culinary use of myrtle leaves, the results of the present work qualify the natural compounds semimyrtucommulone and myrtucommulone A as interesting dietary antioxidants with potential antiatherogenicity.

Protective effect of vanilloids against tert-butyl hydroperoxide-induced oxidative stress in vero cells culture.

This study investigated the effect of synthetic capsiate, a simplified analogue of capsiate, and vanillyl alcohol on the oxidative stress induced by tert-butyl hydroperoxide (TBH) in a line of fibroblasts derived from monkey kidney (Vero cells). In response to the TBH-mediated oxidative stress, a reduction of the levels of total unsaturated fatty acids and cholesterol was observed, and a rise in the concentrations of conjugated dienes fatty acids hydroperoxides and 7-ketocholesterol. Pretreatment with both synthetic capsiate and vanillyl alcohol preserved Vero cells from oxidative damage and showed a remarkable protective effect on the reduction of the levels of total unsaturated fatty acids and cholesterol, inhibiting the increase of MDA, conjugated dienes fatty acids hydroperoxides, and 7-ketocholesterol. Both compounds were effective against peroxidation of cell membrane lipids induced by TBH, with synthetic capsiate essentially acting as a pro-drug of vanillyl alcohol, its hydrophilic hydrolytic derivative.

Protective effect of hydroxytyrosol and its metabolite homovanillic alcohol on H(2)O(2) induced lipid peroxidation in renal tubular epithelial cells.

We investigated the capacity of hydroxytyrosol (HT), 3,4-dihydroxyphenylethanol, and homovanillic alcohol (HVA), 4-hydroxy-3-methoxy-phenylethanol, to inhibit H(2)O(2) induced oxidative damage in LLC-PK1, a porcine kidney epithelial cell line, studying the effect of H(2)O(2) on specific cell membrane lipid targets, unsaturated fatty acids and cholesterol. Exposure to H(2)O(2) induced a significant increase of the level of MDA together with a disruption of the membrane structure, with the loss of unsaturated fatty acids, cholesterol and alpha-tocopherol, and the formation of fatty acids hydroperoxides and 7-ketocholesterol. Pretreatment with HT protected renal cells from oxidative damage: the level of membrane lipids was preserved and there was no significant detection of oxidation products. HVA exerted a comparable activity, thus both HT and HVA were able to prevent in renal cells the lipid peroxidation process that plays a central role in tubular cell injury.

Evaluation of the antioxidant and cytotoxic activity of arzanol, a prenylated alpha-pyrone-phloroglucinol etherodimer from Helichrysum italicum subsp.microphyllum.

Various phenolics and (mero)terpenoids from Helichrysum italicum subsp. microphyllum, a plant endemic to Sardinia, were investigated for their capacity to inhibit non-enzymatic lipid peroxidation. These compounds were studied in simple in vitro systems, under conditions of autoxidation and of iron (EDTA)-mediated oxidation of linoleic acid at 37 degrees C. Arzanol, a pyrone-phloroglucinol etherodimer, and helipyrone, a dimeric pyrone, showed antioxidant activity, and could protect linoleic acid against free radical attack in assays of autoxidation and EDTA-mediated oxidation. Methylarzanol, as well as the sesquiterpene alcohol rosifoliol, showed a decreased, but still significant, protective effect against linoleic acid oxidation. Arzanol and helipyrone were also tested in an assay of thermal (140 degrees C) autoxidation of cholesterol, where arzanol showed significant antioxidant activity. The cytotoxicity of arzanol was further evaluated in VERO cells, a line of fibroblasts derived from monkey kidney. Arzanol, at non-cytotoxic concentrations, showed a strong inhibition of TBH-induced oxidative stress in VERO cells. The results of the present work suggest that the natural compound arzanol exerts useful antioxidant properties in different in vitro systems of lipid peroxidation.

Protective effect of olive oil minor polar components against oxidative damage in rats treated with ferric-nitrilotriacetate.

The phenolic fraction of virgin olive oil exerts preventive effects against reactive oxygen species mediated degenerative diseases. To investigate its action as inhibitor of lipid peroxidation in vivo, we treated Wistar rats with olive oil minor polar components (MPC) (25-50 mg/kg bw) prior to the administration of a sublethal dose (15 mg Fe/kg bw) of ferric-nitrilotriacetate (Fe-NTA). Intraperitoneal injection (i.p.) of Fe-NTA lead to increased oxidative stress associated with extensive peroxidation of membrane lipids in plasma, kidney, and liver of treated rats. Fe-NTA treatment induced a significant decrease of the major oxidizable membrane lipids, alpha-tocopherol, fatty acids and cholesterol, together with an increase of fatty acids hydroperoxides (HP) and 7-ketocholesterol (7-keto). I.p. administration of MPC significantly inhibited fatty acids and cholesterol oxidation, and reduced the levels of HP and 7-keto. In summary, MPC administration protects organs against lipid peroxidation and conserves the endogenous alpha-tocopherol.

New insights into the antioxidant activity and cytotoxicity of arzanol and effect of methylation on its biological properties.

The heterodimeric phloroglucinyl pyrone arzanol (Arz) has raised considerable interest because of its antiviral, anti-inflammatory, and antioxidant activity. We have investigated the effect of methylation of the pyrone moiety on the antioxidant activity and cytotoxicity of Arz. This manoeuvre, that left the polyphenolic moiety unscathed, was nevertheless detrimental for antioxidant activity in both the cholesterol thermal degradation- and the Cu2+-induced liposome oxidation assays, providing evidence of structure-activity relationships that go beyond the preservation of the polyphenolic pharmacophore. The antioxidant activity of Arz was retained also in the Fe-NTA model of in vivo oxidative stress, with protective effect on the oxidative degradation of plasmatic lipids, unsaturated fatty acids and cholesterol. Both Arz and methylarzanol (Me-Arz) were devoid of toxic effect on colonic differentiated Caco-2 cells up to 100µM, but significantly reduced cancer Caco-2 cell viability at lower dosages. Arz could also selectively reduce viability of other cancer cell lines, [murine melanoma cells (B16F10 cells), human cervical carcinoma cells (HeLa cells)], suggesting that it can act as a selective modulator of cell processes typical of cancer cells. Taken together, our results qualify Arz as a lead structure for further in vivo investigation of its pharmacological potential.

Antioxidant Effect of Natural Table Olives Phenolic Extract Against Oxidative Stress and Membrane Damage in Enterocyte-Like Cells.

The phenolic fraction of a naturally fermented cultivar of table olives, "Tonda di Cagliari," was investigated for the ability to protect Caco-2 cells against oxidative stress and membrane damage induced by tert-butyl hydroperoxyde (TBH). TBH exposure resulted in an alteration of cellular redox status, with an increase in reactive oxygen species (ROS) and a decrease in reduced glutathione (GSH) level. A loss of the epithelial integrity, as indicated by the decrease of the transepithelial electrical resistance value, was also observed over time, together with an intense lipid peroxidation process. The olives phenolic extract significantly counteracted ROS generation and subsequent alteration of monolayer integrity and membrane oxidative damage. The protective action of the extract is likely due to the scavenging ability of its main components, as hydroxytyrosol, oleuropein, and verbascoside among the secoiridoids and derivatives. Since olives phenolic compounds concentrate in the intestinal lumen, they may be a useful tool in the prevention of intestinal disorders related to oxidative damage.

Chemical composition of Lycium europaeum fruit oil obtained by supercritical CO2 extraction and evaluation of its antioxidant activity, cytotoxicity and cell absorption.

We studied the total phenols and flavonoids, liposoluble antioxidants, fatty acid and triacylglycerol profiles, and oxidative status of oil obtained from Lycium europaeum fruits following supercritical CO2 extraction (at 30MPa and 40°C). Linoleic (52%), palmitic (18%), oleic (13%), and α-linolenic (6%) were the main oil fatty acids, while trilinolein and palmitodilinolein/oleodilinolein represented the main triacylglycerols. The oil was characterized by high levels of all-trans-zeaxanthin and all-trans-ß-carotene (755 and 332µg/g of oil, respectively), α-tocopherol (308µg/g of oil), total phenols (13.6mg gallic acid equivalents/g of oil), and total flavonoids (6.8mg quercetin equivalents/g of oil). The oil showed radical scavenging activities (ABTS and DPPH assays) and inhibited Caco-2 cell growth. Moreover, the incubation of differentiated Caco-2 cells with a non-toxic oil concentration (100µg/mL) induced a significant intracellular accumulation of essential fatty acids. The results qualify L. europaeum oil as a potential source for food/pharmaceutical applications.

Derangement of intestinal epithelial cell monolayer by dietary cholesterol oxidation products.

The emerging role of the diet in the incidence of intestinal inflammatory diseases has stimulated research on the influence of eating habits with pro-inflammatory properties in inducing epithelial barrier disturbance. Cholesterol oxidation products, namely oxysterols, have been shown to promote and sustain oxidative/inflammatory reactions in human digestive tract. This work investigated in an in vitro model the potential ability of a combination of dietary oxysterols representative of a hyper-cholesterol diet to induce the loss of intestinal epithelial layer integrity. The components of the experimental mixture were the main oxysterols stemming from heat-induced cholesterol auto-oxidation, namely 7-ketocholesterol, 5α,6α-and 5ß,6ß-epoxycholesterol, 7α- and 7ß-hydroxycholesterol. These compounds added to monolayers of differentiated CaCo-2 cells in combination or singularly, caused a time-dependent induction of matrix metalloproteinases (MMP)-2 and -9, also known as gelatinases. The hyperactivation of MMP-2 and -9 was found to be associated with decreased levels of the tight junctions zonula occludens-1 (ZO-1), occludin and Junction Adhesion Molecule-A (JAM-A). Together with such a protein loss, particularly evident for ZO-1, a net perturbation of spatial localization of the three tight junctions was observed. Cell monolayer pre-treatment with the selective inhibitor of MMPs ARP100 or polyphenol (-)-epicathechin, previously shown to inhibit NADPH oxidase in the same model system, demonstrated that the decrease of the three tight junction proteins was mainly a consequence of MMPs induction, which was in turn dependent on the pro-oxidant property of the oxysterols investigated. Although further investigation on oxysterols intestinal layer damage mechanism is to be carried on, the consequent - but incomplete - prevention of oxysterols-dependent TJs alteration due to MMPs inhibition, avoided the loss of scaffold protein ZO-1, with possible significant recovery of intestinal monolayer integrity.

Hydroxytyrosol and tyrosol sulfate metabolites protect against the oxidized cholesterol pro-oxidant effect in Caco-2 human enterocyte-like cells.

The aim of this study was to investigate the ability of the sulfate metabolites of hydroxytyrosol (HT) and tyrosol (TYR) to act as antioxidants counteracting the pro-oxidant effect of oxidized cholesterol in intestinal cells. For this purpose, we synthesized sulfate metabolites of HT and TYR using a chemical methodology and examined their antioxidant activity in Caco-2 monolayers in comparison with the parent compounds. Exposure to oxidized cholesterol led to ROS production, oxidative damage, as indicated by the MDA increase, a decrease of reduced glutathione concentration and an enhancement of glutathione peroxidase activity. All the tested compounds were able to counteract the oxidizing action of oxidized cholesterol; HT and TYR sulfate metabolites showed an efficiency in protecting intestinal cells comparable to that of the parent compounds, strengthening the assumption that the potential beneficial effect of the parent compounds is retained, although extensive metabolisation occurs, the resulting metabolites being able to exert a biological action themselves.

Extra virgin olive oil phenolic extracts counteract the pro-oxidant effect of dietary oxidized lipids in human intestinal cells.

The phenolic fraction of extra virgin olive oil (EVOO) concentrates before absorption in the intestinal lumen, where it may contribute to the modulation of enterocytes response to oxidative and inflammatory stimuli. We evaluated the ability of two monovarietal EVOOs phenolic extracts, Bosana and Nera di Gonnos/Tonda di Cagliari, typical and widespread varieties in Sardinia (Italy), to counteract in enterocytes like Caco-2 cells the pro-oxidant action of oxidized lipids, tert-butyl hydroperoxide (TBH) or a mixture of oxysterols of dietary origin. We confirmed that TBH treatment causes a significant increase of ROS production, GSH depletion, increase of MDA, fatty acids hydroperoxides and 7-ketocholesterol, and showed first evidence of oxidative imbalance and cell damage due to oxysterols exposure. Preincubation of cells with the phenolic extracts significantly attenuated oxidative modifications. Bosana extract showed the highest concentration of total phenols, mainly hydroxytyrosol and tyrosol, and was the most active in presence of TBH, where the free radical scavenging activity of these simple phenols seems to be a determining factor. The two extracts were equally effective, in spite of the different composition, in presence of oxysterols, where ROS production probably occurs according to different and more complex mechanisms.

Protective effect of capsinoid on lipid peroxidation in rat tissues induced by Fe-NTA.

The activity of a single IP administration (15 or 30 mg/Kg body weight) of vanillyl nonanoate, a simplified analog of capsiate, on ferric nitrilotriacetate (Fe-NTA)-mediated oxidative damage was investigated. A sub-lethal dose of Fe-NTA (15 mg Fe/Kg body weight) was administered IP to rats; animals were sacrificed, and kidney and plasma were collected 1 h after injection. In response to the Fe-NTA administration, a reduction of the levels of total lipids, total unsaturated fatty acids and cholesterol was observed, accompanied by a rise in the concentrations of malondialdehyde (MDA), conjugated dienes fatty acids hydroperoxides and 7-ketocholesterol in plasma and kidney 1 h after administration. A pre-treatment with synthetic capsiate (SCPT) showed remarkable protective effect on the reduction of the levels of total lipids, total unsaturated fatty acids and cholesterol, and the cellular antioxidant vitamin E, inhibiting the increase of MDA, conjugated dienes fatty acids hydroperoxides and 7-ketocholesterol in the plasma and kidney. The protective effect of SCPT and two analogues (vanillyl alcohol and vanillin) during the linoleic acid and cholesterol oxidation was investigated in in vitro systems, providing evidence of definite structure-activity relationships.

Cholesterol as target of Fe-NTA-induced lipid peroxidation in rat tissues.

Intraperitoneal injection of the iron-chelate, ferric-nitrilotriacetate (Fe-NTA), induces renal proximal tubular damage associated with oxidative damage in vivo. A sub-lethal dose of Fe-NTA (15 mg Fe/kg body weight) was administered IP to rats; animals were sacrificed and liver, kidney and plasma were collected 1-4 h after injection. In response to the Fe-NTA administration, there were significant time-dependent reductions of the levels of total lipids, cholesterol and total unsaturated fatty acids, and a rise in the concentrations of conjugated dienes, 7-ketocholesterol and fatty acids hydroperoxides, showing a pattern inversely correlated in plasma, kidney and liver. Cholesterol level decreased significantly from 1 h after injection in the kidney and 3-4 h in the plasma and liver of treated rats. This is the first report on cholesterol reduction and accumulated 7-ketocholesterol in the tissues of rats treated with Fe-NTA as a consequence of lipid peroxidation.

Maltese mushroom (Cynomorium coccineum L.) as source of oil with potential anticancer activity.

The present study aimed to examine the potential anticancer properties of fixed oil obtained from Maltese mushroom (Cynomorium coccineum L.), an edible, non-photosynthetic plant, used in traditional medicine of Mediterranean countries to treat various ailments and as an emergency food during the famine. We investigated the effect of the oil, obtained from dried stems by supercritical fractioned extraction with CO2, on B16F10 melanoma and colon cancer Caco-2 cell viability and lipid profile. The oil, rich in essential fatty acids (18:3n-3 and 18:2n-6), showed a significant growth inhibitory effect on melanoma and colon cancer cells. The incubation (24 h) with non-toxic oil concentrations (25 and 50 µg/mL) induced in both cancer cell lines a significant accumulation of the fatty acids 18:3n-3 and 18:2n-6 and an increase of the cellular levels of eicosapentaenoic acid (20:5n-3) with anticancer activity. Moreover, the oil exhibited the ability to potentiate the growth inhibitory effect of the antitumor drug 5-fluorouracil in Caco-2 cells and to influence the melanin content in B16F10 cells. The results qualify C. coccineum as a resource of oil, with potential benefits in cancer prevention, for nutraceutical and pharmaceutical applications.

Potential anti-tumor effects of Mugil cephalus processed roe extracts on colon cancer cells.

The salted-semidried mullet ovary product, bottarga, is a Mediterranean food rich in n-3 PUFA EPA and DHA. We studied and compared the effects on cell viability, sensitivity to the anti-tumor drug 5-fluorouracil, and lipid composition, in colon cancer Caco-2 cells after 24 h incubation with oils and hydrophilic extracts obtained from two bottarga samples stored at different conditions. The cellular absorption of bottarga lipids was assessed in cancer cells by the evaluation of lipid accumulation in cytoplasmic lipid droplets by fluorescence microscopy. Bottarga oil showed a significant in vitro inhibitory effect on the growth of cancer Caco-2 cells and the ability to potentiate, at non-toxic concentration, the growth inhibitory effect of 5-fluorouracil. Moreover, bottarga oil induced in cancer Caco-2 cells marked changes in fatty acid composition, with a significant accumulation of the n-3 PUFA EPA and DHA, and cytoplasmic lipid droplet formation. Also bottarga hydrophilic extract, characterized by means of ¹H NMR spectroscopy, exhibited a reduction in cancer cell viability, without affecting cell lipid profile. Cell cholesterol levels were unmodified by all treatments. The results showed interesting anti-tumor properties of bottarga lipids, and qualify this fish product as a food with nutraceutical properties and potential benefits in colon cancer prevention.

Effect of storage conditions on lipid components and color of Mugil cephalus processed roes.

UNLABELLED: The salted and semidried mullet (Mugil cephalus) ovary product (bottarga) is proposed as an important source of the long-chain n-3 polyunsaturated fatty acids (PUFA), eicosapentaenoic acid, and docosahexaenoic acid. In this work, we investigated the extent of lipid oxidation and browning of grated bottarga samples during 7 mo of storage at -20 °C, 2 to 3 °C in the absence of light, and at room temperature in the presence or absence of light. Modifications of the levels of total choline (as index of phospholipid breakdown), total sugars, and free amino acids such as lysine, methionine, and tryptophan (involved in nonenzymatic browning) were also studied at different storage conditions. Storage of bottarga did not significantly affect the n-3 PUFA and cholesterol levels with respect to the control; nevertheless, a significant hydroperoxide increase was observed during 7 mo in bottarga samples at all the storage conditions, while low malondialdeyde levels were measured. Samples placed at room temperature in the absence and in the presence of light showed over time a marked browning process, lipid breakdown, a sensible decrease in the levels of total sugars, tryptophan, and methionine with respect to control and samples stored at -20 °C and 2 to 3 °C. The resistance against the oxidation of the isolated bottarga lipids was also assessed in dry state at several temperatures (37, 75, and 140 °C). PRACTICAL APPLICATION: We evaluated the change in lipid compounds and color of dried and salted mullet roes under different storage conditions. The obtained results suggest the importance of the low temperatures to preserve the nutritional properties of this fish product during long storage.

Extraction and separation of volatile and fixed oils from seeds of Myristica fragrans by supercritical CO2: chemical composition and cytotoxic activity on Caco-2 cancer cells.

UNLABELLED: Isolation of volatile and fixed oils from nutmeg have been obtained by supercritical fractioned extraction with carbon dioxide. Extraction experiments were carried out at pressures of 90 and 250 bar and temperature of 40 °C. The extraction step performed at 90 bar produced a volatile fraction mainly formed by myristicin (32.8%), sabinene (16.1%), α-pinene (9.8%), ß-pinene (9.4%), ß-phellandrene (4.9%), safrole (4.1%) and terpinen-4-ol (3.6%). The oil yield relative to this step of the process was 1.4% by weight of the charge. The last extraction step at 250 bar produced a butter-like material (nutmeg butter). The yield of this step was 14.4% by weight. The most represented fatty acids of fixed oil from nutmeg were 14:0 (79.2%), 18:1 n-9 (7.4%) and 16:0 (6.1%), and in particular the unsaturated fatty acids 18:1 n-9 averaged 32.96 µg/mg of oil. The level of myristicin in the nutmeg essential and fixed oils was also directly quantified by reversed HPLC-DAD. Moreover, the essential oil obtained from nutmeg, as well as myristicin, showed a significant in vitro inhibitory effect on the growth of a colon cancer cell line (undifferentiated Caco-2 cells). PRACTICAL APPLICATION: In this study, the chemical characterization and the anticancer activity of nutmeg oils obtained by supercritical extraction with carbon dioxide were investigated. This is important for their potential application in food and pharmaceutical industries.

Chemical composition and effect on intestinal Caco-2 cell viability and lipid profile of fixed oil from Cynomorium coccineum L.

Cynomorium coccineum L. is a non-photosynthetic plant, spread over Mediterranean countries, amply used in traditional medicine. We investigated the composition and effect on intestinal Caco-2 cell viability and lipid profile of fixed oil obtained from dried stems of the plant. Oil isolation has been performed by supercritical fractioned extraction with CO2. 13C NMR spectroscopy has been used to study the molecular composition of oil lipids; fatty acid composition was identified using GC and HPLC techniques. The fixed oil was composed mainly by triacylglycerols and derivates. The main fatty acids were 18:1 n-9 (38%), 18:2 n-6 (20%), 16:0 (15%), and 18:3 n-3 (10.8%). The oil showed a significant in vitro inhibitory effect on the growth of colon cancer undifferentiated Caco-2 cells. Moreover, cell viability, lipid composition, and lipid peroxidation were measured in intestinal epithelial cells (differentiated Caco-2 cells) after 24 h incubation with fixed oil. The oil did not show a toxic effect on colon epithelial cell viability but induced a significant change in fatty acid composition, with a significant accumulation of the essential fatty acids 18:3 n-3 and 18:2 n-6. The results showed remarkable biological activity of Maltese mushroom oil, and qualify it as a potential resource for food/pharmaceutical applications.