Lutein Encapsulated in PLGA-Phospholipid Nano-Carrier Effectively Mitigates Cytokines by Inhibiting Tumor Necrosis Factor TNF-α and Nuclear Factor NF-κB in Mice Retina.

Lutein, a photo- and thermo-labile macular pigment, prevents the retina from suffering ocular inflammation with its antioxidant and anti-inflammatory activity. However, its biological activity is poor due to poor solubility and bioavailability. Therefore, we developed a PLGA NCs (+PL), (poly (lactic-co-glycolic acid) nanocarrier with phospholipid) to improve the biological availability and bioefficacy of lutein in the retina of lipopolysaccharide (LPS)-induced lutein-devoid (LD) mice. The effect of lutein-loaded NCs with/without PL was studied in comparison with micellar lutein. The induction of inflammation by LPS significantly increased the production of nitrites in the LPS-induced group, revealing higher levels of nitric oxide (NO) in the serum (760%) and retina (891%) compared to the control group. Malondialdehyde (MDA) levels in the serum (93%) and retina (205%) of the LPS-induced group were higher compared to the control group. LPS induction resulted in increased protein carbonyls in the serum (481%) and retina (487%) of the LPS group compared to the control group. Further, to conclude, lutein-PLGA NCs (+PL) effectively down-regulated inflammatory complications in the retina.

miR-7-5p Antagomir Protects Against Inflammation-Mediated Apoptosis and Lung Injury via Targeting Raf-1 In Vitro and In Vivo.

Exacerbated inflammation and apoptosis are considered upstream events associated with acute lung injury (ALI). microRNAs are critical regulators of genes responsible for inflammation and apoptosis and are considered potential therapeutic targets for ameliorating ALI. This study was undertaken to uncover the role of miR-7-5p in LPS-induced lung injury. A LPS-induced inflammation model was established using BEAS-2B cells and C57BL/6 mice. Bioinformatics analysis and the luciferase reporter assay confirmed that Raf-1 is a target of miR-7-5p and that its expression was inversely correlated with expression of proinflammatory markers and miR-7-5p, whereas miR-7-5p inhibition in vitro led to subsequent restoration of Raf-1 expression and prevention of apoptosis. Intranasal (i.n.) administration of antagomir using the C57BL/6 mouse model further confirmed that miR-7-5p inhibition suppresses LPS-induced inflammation and apoptosis via modulating the miR-7-5p/Raf-1 axis. Our findings indicate that blocking miR-7-5p expression by antagomir protects mice from LPS-induced lung injury by suppressing inflammation and activation of mitochondria-mediated survival signalling. In conclusion, our findings demonstrate a previously unknown pathophysiological role of miR-7-5p in the progression of ALI, and targeted i.n. administration of miR-7-5p antagomir could aid in the development of potential therapeutic strategies against lung injury.

Brown algae and barley-based anti-obesity food and its safety in C57BL6 mice.

Brown algae have been considered a potential source of bioactives and used as a dietary supplement to manage obesity and its associated health complications. However, its effective use is limited due to heavy metals and microbial contamination, unawareness of health benefits and limited dietary exploitation. We developed, the Indian brown algae Padina tetrastromatica and barley-based anti-obesity food (AOF) and examined for microbial and heavy metal safety. Additionally, acute [0 (control), 50, 100, 200, 500 g AOF/kg diet] and sub-acute [0, 5, 50 g AOF/kg diet] doses of AOF were fed to C57BL6 mice and toxicity was examined. The physical, locomotory, hematological, biochemical parameters and histopathology were examined. Postprandial plasma and tissue levels of fucoxanthin and its metabolites were analyzed. Feeding AOF did not affect the general behavior, food and water intake, growth or survival of animals. Biochemical indices did not show any differences between AOF-fed and control groups. However, significantly lower levels of plasma cholesterol and triglycerides in groups fed 5 and 50 g of AOF/kg diet were observed. The post-mortem examination revealed no macroscopic/microscopic alteration in the vital organs. Overall, results validate that AOF is a safe and effective dietary supplement (even at higher doses of 500 g AOF/kg) to mitigate obesity. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05483-4.

Astaxanthin ameliorates hyperglycemia induced inflammation via PI3K/Akt-NF-κB signaling in ARPE-19 cells and diabetic rat retina.

Astaxanthin has been reported to possess anti-inflammatory effect but the exact mechanism in protecting the retinal pigment epithelial (RPE) cells is not clear. Hence, we hypothesized that astaxanthin could protect RPE by inhibiting ROS-mediated inflammation. The purpose of this study is to understand the retinal protective mechanism of astaxanthin in modulating hyperglycemia (HG) induced inflammation in ARPE-19 cell and diabetic rat retina. ARPE-19 cells were treated with 30 mM glucose to induce hyperglycemia whereas diabetes was induced in rats with streptozotocin followed by astaxanthin treatment. The level of oxidative stress markers, antioxidant enzyme activity, inflammatory markers (NF-κB, TNF-α, ICAM-1), signaling mediators (PI3K, p-Akt) and nuclear translocation of NF-κB were analyzed in ARPE-19 cells and rat retina. HG-mediated ROS generation and lipid peroxidation were declined upon astaxanthin treatment in ARPE-19 cells. Similarly, astaxanthin treatment found to reduce the elevated levels of nitric oxide, protein carbonyl, and lipid peroxides in diabetic group. Astaxanthin restored the activity of superoxide dismutase, catalase, glutathione peroxidase, and glutathione transferase in serum and retina of diabetic rats. NF-κB, TNF-α, and ICAM-1 levels were higher in HG-treated ARPE-19 cells and diabetic retina compared to control group, whereas astaxanthin treatment lowered their expression. PI3K and p-Akt were higher in high glucose treated ARPE-19 cells and diabetic retina. NAC, LY294002 and PDTC treatment resulted in reduced nuclear translocation of NF-κB and decreased expression of inflammatory markers in HG treated ARPE-19 cells. Thus, we conclude that astaxanthin protected the retinal cells from HG-induced inflammation by modulating NF-κB through ROS-PI3K/Akt signaling cascade.

Turmeric, red pepper, and black pepper affect carotenoids solubilized micelles properties and bioaccessibility: Capsaicin/piperine improves and curcumin inhibits carotenoids uptake and transport in Caco-2 cells.

This study aimed to evaluate the role of spices/spice active principles on physical, biochemical, and molecular targets of bioaccessibility/bioavailability. Carotenoids-rich micellar fraction obtained through simulated digestion of green leafy vegetables (GLV) with individual or two/three combinations were correlated to their influence on bioaccessibility, cellular uptake, and basolateral secretion of carotenoids in Caco-2 cells. Results suggest that carotenoids' bioaccessibility depends on micelles physicochemical properties, which is affected due to the presence of co-treated dietary spices and their composition. Increased bioaccessibility of ß-carotene (BC) and lutein (LUT) is found in GLV (spinach) digested with turmeric (TM) than red pepper (RP) and black pepper (BP). In contrast, enhanced cellular uptake and secretion of BC and LUT-rich triglyceride-rich lipoprotein is observed in the presence of RP and BP compared to the control group. In contrast, TM inhibited absorption, while retinol levels significantly reduced in the presence of TM and RP than BP. Control cells have indicated higher cleavage of ß-carotene to retinol than the spice-treated group. Besides, spice active principles modulate facilitated transport of carotenoids by scavenger receptor class B type 1 (SR-B1) protein. The effect of spices on carotenoids' bioavailability is validated with active spice principles. Overall, carotenoids' bioavailability (cellular uptake and basolateral secretion) was found in the following order of treatments; piperine > capsaicin > piperine + capsaicin > curcumin + capsaicin + piperine > control > turmeric. These findings suggested that the interaction of specific dietary factors, including spice ingredients at the enterocyte level, could provide greater insight into carotenoid absorption. PRACTICAL APPLICATION: Spices/spice active principles play a role in the digestion process by stimulating digestive enzymes and bile acids secretion. Since carotenoids are lipid soluble and have low bioavailability, spice ingredients' influence on intestinal absorption of carotenoids is considered crucial. Hence, understanding the interaction of co-consumed spices on the absorption process of carotenoids may help to develop functional foods/formulation of nutraceuticals to improve their health benefits.

Lactucaxanthin protects retinal pigment epithelium from hyperglycemia-regulated hypoxia/ER stress/VEGF pathway mediated angiogenesis in ARPE-19 cell and rat model.

Hyperglycemia mediated perturbations in biochemical pathways induce angiogenesis in diabetic retinopathy (DR) pathogenesis. The present study aimed to investigate the protective effects of lactucaxanthin, a predominant lettuce carotenoid, on hyperglycemia-mediated activation of angiogenesis in vitro and in vivo diabetic model. ARPE-19 cells cultured in 30 mM glucose concentration were treated with lactucaxanthin (5 µM and 10 µM) for 48 h. They were assessed for antioxidant enzyme activity, mitochondrial membrane potential, reactive oxygen species, and cell migration. In the animal experiment, streptozotocin-induced diabetic male Wistar rats were gavaged with lactucaxanthin (200 µM) for 8 weeks. Parameters like animal weight gain, feed intake, water intake, urine output, and fasting blood glucose level were monitored. In both models, lutein-treated groups were considered as a positive control. Hyperglycemia-mediated angiogenic marker expressions in ARPE-19 and retina of diabetic rats were quantified through the western blot technique. Expression of hypoxia, endoplasmic reticulum stress markers, and vascular endothelial growth factor were found to be augmented in the hyperglycemia group compared to control (P < 0.05). Hyperglycemia plays a crucial role in increasing cellular migration and reactive oxygen species besides disrupting tight junction protein. Compared to lutein, lactucaxanthin aids retinal pigment epithelium (RPE) function from hyperglycemia-induced stress conditions via downregulating angiogenesis markers expression. Lactucaxanthin potentiality observed in protecting tight junction protein expression via modulating reactive oxygen species found to conserve RPE integrity. Results demonstrate that lactucaxanthin exhibits robust anti-angiogenic activity for the first time and, therefore, would be useful as an alternative therapy to prevent or delay DR progression.

Astaxanthin mediated regulation of VEGF through HIF1α and XBP1 signaling pathway: An insight from ARPE-19 cell and streptozotocin mediated diabetic rat model.

Breakdown of outer blood-retina barrier (BRB) has been associated with the pathogenesis of diabetic retinopathy (DR) and diabetic macular edema (DME). Vascular endothelial growth factor (VEGF) might play a detrimental role in the pathogenesis of DME, a major clinical manifestation of DR. In the present study, we investigated the inhibitory mechanism of astaxanthin on VEGF and its upstream signaling pathways under in vitro and in vivo conditions. Astaxanthin has been observed to downregulate VEGF expression under hyperglycemic (HG) and CoCl2 induced hypoxic conditions in ARPE-19 cells. There were compelling pieces of evidence for the involvement of transcription factors like HIF1α and XBP1 in the upregulation of VEGF under HG and hypoxic conditions. Thus, we investigated the role of astaxanthin in the expression and nuclear translocation of HIF1α and XBP1. The activation and translocation of HIF1α and XBP1 induced by HG or CoCl2 conditions were hindered by astaxanthin. Additionally, treatment with HIF1α siRNA and IRE1 inhibitor STF-083010 also inhibited the expression of VEGF induced by HG and CoCl2 conditions. These results indicated that the anti-VEGF property of astaxanthin might be associated with the downregulation of HIF1α and XBP1. Furthermore, astaxanthin mitigated the enhanced migration of retinal pigment epithelial (RPE) cells under DR conditions. As well, astaxanthin protected disorganization of zona occludin-1 (ZO-1) tight junction protein in RPE and reduced HG or hypoxic induced permeability of RPE cells. In streptozotocin-induced diabetic rat model, astaxanthin reduced the expression of HIF1α, XBP1, and VEGF as well as protected the abnormalities in the retinal layers induced by diabetes condition. Thus, astaxanthin may be used as a potential nutraceutical to prevent or treat retinal dysfunction in diabetic patients.

Poly (D, L-lactide-co-glycolide)-phospholipid nanocarrier for efficient delivery of macular pigment lutein: absorption pharmacokinetics in mice and antiproliferative effect in Hep G2 cells.

Lutein has various biological activities, its application in food and pharma industries are limited due to poor aqueous solubility, stability, and bioavailability. To achieve various benefits, lutein-poly (lactic-co-glycolic acid) (PLGA)-phospholipid (PL) nanocapsules were prepared. Lutein-PLGA NCs (+PL) were synthesized, characterized and its bioavailability was studied in vitro and in vivo. The cellular uptake and anti-proliferative activity were analyzed in Hep G2 cells. The mean size and zeta value of lutein-PLGA NCs (+PL) were 140 ± 6 nm and - 44 mV. The amorphous nature of lutein in PLGA NCs (+PL) was confirmed by XRD and DSC. In vitro lutein release kinetics showed an initial burst followed by sustainable release up to 86%. In vitro bioavailability showed 62.7% higher lutein bioaccessibility than lutein in free form. The AUC of lutein after single oral dose of lutein-PLGA NCs (+PL) revealed 3.91-fold (plasma), 2.89-fold (liver), and 3.12-fold (eyes) higher absorption than the control (mixed micelles). The IC50 of lutein-PLGA NCs (+PL) in Hep G2 cells at 72 h was 4.5 µM as opposed to 23.4 µM for lutein in free form. Thus, results reveal that PL added to PLGA NCs helps in enhancing the solubility which in turn resulted in its better bioavailability and bioefficacy.

Fractionation and Characterization of Lycopene-Oxidation Products by LC-MS/MS (ESI)+: Elucidation of the Chemopreventative Potency of Oxidized Lycopene in Breast-Cancer Cell Lines.

Lycopene (LYC) has been correlated with the reduction of certain cancers and chronic diseases. However, the existence and biofunctionality of degraded, oxidized, and biotransformed LYC products in vivo have not been revealed. Therefore, this study aimed to screen and elucidate the potential bioactive lycopene-derived products in breast-cancer and non-cancerous cells. LYC-oxidation or -cleavage products were generated using KMnO4. These oxidation products were separated as fractions I-III by silica column chromatography using gradient solvent systems. Further, LC-MS/MS (ESI)+ was used to elucidate their possible fragmentation patterns and structures. Fraction II showed higher cytotoxicity (IC50 value of 64.5 µM), cellular uptake, and apoptosis-inducing activity in MCF-7 cells. This fraction consists of major peak m/ z 323, identified as apo-8,6'-carotendial. The cytotoxicity-inducing activity may be due to partial ROS generation with mitochondrial dysfunction. Further, the role of apo-8,6'-carotendial in the induction of apoptosis is demonstrated for the first time. These results illustrated that LYC-oxidation derivatives or metabolites are involved in growth inhibition of cancer cells. Exploration of specific oxidized-carotenoid products will give further insight into the field of nutritional biochemistry.

Polymeric chitosan-glycolipid nanocarriers for an effective delivery of marine carotenoid fucoxanthin for induction of apoptosis in human colon cancer cells (Caco-2 cells).

Fucoxanthin (FUCO), a marine carotenoid is photo-, and thermo-labile and poorly bioavailable due to its lipophilicity. Hence, we developed a chitosan (CS) + glycolipid (GL) nanogels (NGs) to increase cellular uptake and anticancer efficacy of FUCO (10 µM) in human colon cells (Caco-2). Effect of FUCO loaded in NGs with/with no GL was studied in comparison with micellar FUCO. Results showed that the cell viability was lower (p < 0.05) in NGs + GL (50.5%) compared to NGs (-GL) (66.5%) and the mixed micelles (72.5%) groups over 48 h exposure. An enhanced reactive oxygen species (ROS) generation was evident in NGs + GL (379.2%) group compared to NGs (-GL) and mixed micelles groups. Further, induction of apoptosis with an increased chromatin condensation and DNA fragmentation as evidenced in DAPI staining and DNA ladder assay were higher in NGs + GL group than other groups. Down-regulation of Bcl-2 (6.6 folds) was higher in NGs + GL group compared to NGs (-GL) (1.94 fold) and mixed micelles (1.19 fold) groups. Higher Bax up-regulation in NGs + GL compared to other groups supports the Bcl-2 down regulation. Mitochondrial membrane polarisation (ΔΨm) was higher in NGs + GL group (2.46 fold) compared to NGs (-GL) (1.91 fold) and mixed micelles (1.26 fold) groups. The cellular FUCO uptake illustrated a positive correlation between its level (pmol/106 cells) in NGs + GL (758.3) and enhanced caspase-3 activity (25.8 folds). This could be the reason for an increased apoptotic activity in NGs + GL group than other groups. Results demonstrate that delivery of FUCO in NGs + GL carrier aids cellular uptake and chemotherapeutic potential of FUCO. Results further demonstrate, for the first time, higher anti-cancer activity of FUCO loaded in NGs + GL and the effect was through ROS generation via a caspase dependent mechanism in Caco-2 cells.

Low-dose doxorubicin with carotenoids selectively alters redox status and upregulates oxidative stress-mediated apoptosis in breast cancer cells.

The combination of carotenoids and doxorubicin (DOX) selectively alters oxidative stress-mediated apoptosis in breast cancer cells. Primarily, cytotoxic efficiency of carotenoids (ß-carotene, BC; lutein, LUT; astaxanthin, AST; or fucoxanthin, FUCO) either with or without a minimal cytotoxic dose of DOX was evaluated in MCF-7 (0.12 µM) and MDA-MB-231 cells (0.28 µM). The higher cell growth inhibition of BC and/or LUT with DOX was selected for testing in further cell-based assays. Low-dose DOX significantly enhanced cytotoxicity in carotenoid (<5 µM)-treated cells compared to high-dose DOX (>1 µM) or carotenoid (20 µM) treatment alone. Depleted glutathione, increased lipid peroxides and increased ROS levels in cells confirmed the cytotoxic effect. Furthermore, mitochondrial dysfunction, cell growth arrest at G0/G1 phase and caspase cascades as well as up- and down-regulated expression levels of related proteins (p21, p27, Bax, p53, Bcl-2, and cyclin D1) revealed the synergistic effect of carotenoid and DOX treatment on ROS-mediated apoptosis. These observations demonstrated increased apoptosis in BC + DOX/LUT + DOX-treated cells due to the pronounced pro-oxidant action. Interestingly, normal breast epithelial cells (MCF 10A) exposed to similar treatments resulted in non-significant cytotoxicity. These newly observed mechanistic differences of anticancer drugs on the mitigation of toxicity with carotenoids may provide insight into the targeting of cancer therapy.

Antidiabetic plant-derived nutraceuticals: a critical review.

Diabetes mellitus (DM) is one of the major health problems in the world, especially amongst the urban population. Chemically synthesized drugs used to decrease the ill effects of DM and its secondary complications cause adverse side effects, viz., weight gain, gastrointestinal disturbances, and heart failure. Currently, various other approaches, viz., diet control, physical exercise and use of antidiabetic plant-derived molecules/foods are advocated to manage DM, as they are economical with fewer or no side effects. This review mainly focuses on antidiabetic plants, chemically characterized plant molecules and plant-based foods in the treatment of DM. Very little science-based evidence is available on the mechanism of action of plant-derived food molecules on the DM targets. Critical DM targets include α-amylase, α-glucosidase, DPP-IV, aldose reductase, PPAR-γ, AMP kinase and GLUT4. In-depth studies carried out on a few of those targets with specific mechanisms of action are addressed in this review. This review may help future researchers in identifying a right plant molecule to treat DM or to develop food formulations for DM management.

Astaxanthin from shrimp efficiently modulates oxidative stress and allied cell death progression in MCF-7 cells treated synergistically with ß-carotene and lutein from greens.

This study investigated the synergistic efficacy of keto-carotenoid astaxanthin (AST, from shrimp) plus hydrocarbon (ß-carotene, BC) and hydroxyl (lutein, L) carotenoids (from greens) on molecular events in MCF-7 cells. MCF-7 cells were treated with either of carotenoid (20 µM, AST or BC or L) separately or the mixture of them (an equimolar concentration of carotenoids mixture, CM) or saponified carotenoid extract from shrimp (SSCE) for 48 h and analyzed cellular uptake, cytotoxicity, and apoptosis. The IC50 and combination-index values of AST co-treatment with a lower concentration of BC and L (5 µM) exhibited enhanced cytotoxicity and oxidative stress as compared with individual carotenoids or SSCE. Further, higher cellular uptake/accumulation of AST along with BC and L found to synergistically induce apoptosis through modulation of cyclin D1, p53, Bax and Bcl-2 expressions by arresting cell cycle at G0/G1 phase. Further, CM or SSCE treatments are unlikely to affect proliferation of normal breast epithelial cells (MCF-10A). The results of selective killing of MCF-7 cells demonstrated a greater insight on the synergistic effect of shrimp AST plus BC and L. It is concluded that consumption of shrimp along with green leafy vegetables helps in combating cancer chemoprevention.

Lactucaxanthin - a potential anti-diabetic carotenoid from lettuce (Lactuca sativa) inhibits α-amylase and α-glucosidase activity in vitro and in diabetic rats.

Intestinal and pancreatic α-amylase and α-glucosidase inhibitors offer an approach to lower the levels of post-prandial hyperglycemia through the control of dietary starch breakdown in digestion. This study hypothesized that lactucaxanthin (Lxn) in lettuce (Lactuca sativa) inhibits the activity of α-amylase and α-glucosidase. In this study, the interaction of Lxn with α-amylase and α-glucosidase in silico and its inhibitory effect on these enzymes were studied using in vitro and STZ-induced diabetic rat models. Lxn was isolated from lettuce with 96% purity confirmed by HPLC and LCMS. The in silico analysis showed that Lxn has a lower binding energy (-6.05 and -6.34 kcal mol-1) with α-amylase and α-glucosidase compared to their synthetic inhibitors, acarbose (-0.21 kcal mol-1) and miglitol (-2.78 kcal mol-1), respectively. In vitro α-amylase and α-glucosidase inhibition assays revealed that Lxn had IC50 values of 435.5 µg mL-1 and 1.84 mg mL-1, but acarbose has values of 2.5 and 16.19 µg mL-1. The in vivo results showed an increased activity for α-amylase and α-glucosidase in the intestine (4.7 and 1.30 fold, p < 0.05) and pancreas (1.3 and 1.48 fold, p < 0.05) of STZ induced diabetic rats compared to normal rats. Whereas the activity decreased (p < 0.05) in the Lxn fed diabetic rats, except for the intestinal α-glucosidase activity (1.69 ± 0.12 PNP per min per mg protein). This was confirmed by the low blood glucose level (239.4 ± 18.2 mg dL-1) in diabetic rats fed Lxn compared to the diabetic group (572.2 ± 30.5 mg dL-1, p < 0.05). Lxn significantly inhibited (p < 0.05) the activity of α-amylase and α-glucosidase and could be of medical and nutritional relevance in the treatment of diabetes.

Lutein attenuates oxidative stress markers and ameliorates glucose homeostasis through polyol pathway in heart and kidney of STZ-induced hyperglycemic rat model.

PURPOSE: Lutein's role on chronic hyperglycemia-induced oxidative stress and associated glucose homeostasis in heart and kidney is limited. Purpose of the study is to investigate the effect of lutein on cardiac and renal polyol pathway enzymes and oxidative stress markers under hyperglycemia-induced oxidative stress condition using streptozotocin (STZ)-injected rat model. METHODS: STZ-induced hyperglycemic (fasting blood glucose ≥11 mM) male Wistar rats were divided into two groups (n = 11/group). Group 1 received micellar lutein (39 nmol/day/rat) and group 2 (negative control) received micelle without lutein for 8 weeks. A separate group (no STZ injected) served as a positive control (n = 11/group). Oral glucose tolerance test (OGTT), biweekly urine glucose and activities of aldose reductase (AR) and sorbitol dehydrogenase (SDH) enzymes were assessed. Activities of antioxidant enzymes and antioxidant level were also evaluated. RESULTS: Lutein-administered hyperglycemic rats showed better glucose tolerance as evidenced with OGTT and biweekly urine glucose when compared to negative control. Activities of AR and SDH were decreased in heart and kidney of lutein-fed hyperglycemic rats. Also, they had significantly (p < 0.05) decreased malondialdehyde levels (66, 34, and 33 %) and increased reduced glutathione level (81, 18 and 92 %) in serum, heart and kidney, respectively. Altered antioxidant enzyme activities such as superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glutathione transferase were also affected in serum, heart and kidney of lutein-fed diabetic group. CONCLUSION: Lutein prevented cardiac and renal injury in STZ-induced hyperglycemic rats due to potential amelioration of altered activities in polyol pathway and oxidative stress markers.

Enhanced cytotoxic and apoptosis inducing activity of lycopene oxidation products in different cancer cell lines.

Currently, upon understanding the metabolomics of carotenoids, it is important to address the key role of carotenoid derived products. In this regard, aim of the study was to elucidate and explore the role of lycopene (LYC) oxidative products generated through autoxidation (AOL) or chemical (KMnO4) oxidation (COL) against proliferation of selected cancer cells. Preliminary, we investigated the effect of LYC on cell viability of various cancer cell lines (PC-3, MCF-7, A431, HepG2, HeLa and A549). Based on the results of LYC treatment on cell cytotoxicity levels, MCF-7, PC-3 and HeLa cell lines were further tested with AOL and COL products. The decreased cell viability with depleted GSH and increased MDA levels were observed when treated with COL products than control, LYC and AOL. In addition, COL products increased ROS levels and percent apoptosis. The typical morphological changes and nuclear condensations showed that COL products have anti-proliferation and apoptosis inducing activity. Based on results, we hypothesized that ROS generation by LYC oxidation products may be one of intermediate step involved in apoptosis. The redox status and therapeutic approach of COL products in modulating ROS and induction of apoptosis in cancer cells were reported for the first time, to our knowledge. To conclude, COL products involves in cancer growth inhibition efficiently than intact LYC and AOL. Hence, there is a great potential for synthesizing or producing such carotenoid oxidation products to augment cancer complication.

Chitosan-glycolipid nanogels loaded with anti-obese marine carotenoid fucoxanthin: Acute and sub-acute toxicity evaluation in rodent model.

Fucoxanthin (FUCO) is a light- and heat-sensitive marine xanthophyll carotenoid, present in brown algae that render physiological properties as anti-oxidants. In this study, nanoencapsulation is an approach adopted to improve bioavailability of FUCO by using ionic-gelation method with polymeric chitosan (CS) dispersed in glycolipid (GL), as a carrier. Further, the aim was to investigate adverse effect of acute and sub-acute toxicity of chitosan nanogels (CS-NGs) loaded with FUCO+GL in rats. In the acute toxicity study, FUCO was fed to rats at doses of 0.1, 1, 10 and 100 mg/kg body weight (BW). In the sub-acute toxicity study, FUCO was fed at doses of 1 and 10 mg/kg BW for 28 days. In both the studies, no mortality and abnormalities in gross morphology were observed. Acute toxicity study revealed that the LD50 of FUCO in CS-NGs is higher than 100 mg/kg BW. No postprandial plasma levels of FUCO were detected. However, fucoxanthinol (FUOH), a hydrolytic metabolite of FUCO was detected in a dose dependent manner (P < 0.01). Compared to the control group(s), no dose-related toxic effects of CS-NGs with FUCO + GL were found in haematological, histopathological, plasma biochemical indices, etc. The no-observed-adverse-effect level (NOAEL) for CS-NGs with FUCO + GL in rats was 10 mg/kg/day. To conclude, no apparent adverse effect of CS-NGs with FUCO + GL demonstrating CS could be a promising polymer matrix for safe delivery of FUCO. This is the first study to demonstrate the safety assessment of CS-NGs with FUCO + GL.

Role of different vehicles in carotenoids delivery and their influence on cell viability, cell cycle progression, and induction of apoptosis in HeLa cells.

The objective of the present study was to determine the role of different vehicles in carotenoids delivery and their influence on cell viability, cell cycle progression and induction of apoptosis in HeLa cells. Cells (5 × 10(3)) were treated with different concentrations (25-100 µM) of ß-carotene (BC) or lutein (L) or astaxanthin (AST) dissolved in 0.5% of tetrahydrofuran (THF), dimethylsulfoxide (DMSO), and fetal bovine serum (FBS), respectively. The effect of delivery vehicle on carotenoids uptake, cytotoxicity, oxidative status, cell cycle distribution, and apoptosis was examined after 48 h of incubation. The results shown that, cell viability reduced significantly in a dose- and time-dependent manner irrespective of carotenoid delivered in vehicles. Cellular uptake of BC delivered in THF was higher by 49.1, 29.7% and L delivered through THF was higher by 41.7 and 37.5% than DMSO and FBS, respectively. While, AST delivered through DMSO was higher by 36.1 and 43.7% than the THF and FBS, respectively. In case of cells treated either with BC or L delivered through THF and AST in DMSO decreased the glutathione and increased the malondialdehyde levels. The net increase in the G 2/M phase percentage of cell cycle progression was observed in carotenoid-treated cells. The % induction of apoptosis by BC or L delivered with THF and AST in DMSO was higher than other treated groups. In conclusion, choice of suitable vehicle for specific carotenoids delivery is essential that in turn may influence on cell proliferation and cell-based assays.

Biodegradable Poly (Lactic-co-Glycolic Acid)-Polyethylene Glycol Nanocapsules: An Efficient Carrier for Improved Solubility, Bioavailability, and Anticancer Property of Lutein.

Lutein bioavailability is limited because of its poor aqueous solubility. In this study, lutein-poly (lactic-co-glycolic acid) (PLGA)-polyethylene glycol (PEG) nanocapsules were prepared to improve the solubility, bioavailability, and anticancer property of lutein. The scanning electron microscopy and dynamic light scattering examination revealed that the nanocapsules are smooth and spherical with size ranging from 80 to 500 nm (mean = 200 nm). In vitro lutein release profile from nanocapsules showed controlled sustainable release (66%) up to 72 h. Aqueous solubility of lutein nanocapsules was much higher by 735-fold than the lutein. Fourier transform infrared spectroscopy analyses showed no chemical interaction among PLGA, PEG, and lutein, indicating possible weak intermolecular forces like hydrogen bonds. X-ray diffraction revealed lutein is distributed in a disordered amorphous state in nanocapsules. Postprandial plasma kinetics (area under the curve) of an oral dose of lutein from nanocapsules was higher by 5.4-fold compared with that of micellar lutein (control). The antiproliferative effect of lutein from nanocapsules (IC50 value, 10.9 µM) was higher (43.6%) than the lutein (IC50 value, 25 µM). Results suggest that PLGA-PEG nanocapsule is an efficient carrier for enhancing hydrophilicity, bioavailability, and anticancer property of lipophilic molecules such as lutein.

Quality characteristics and lutein bioavailability from maize and vegetable-based health food.

Health food (ready-mix) was prepared from maize and vegetables a source of lutein (L) and zeaxanthin (Z) and studied for its quality characteristics (moisture sorption isotherm, sensory, microbiological, chemical composition, and storage stability) on storage at varying temperatures for 3 months and L+Z bioavailability in mice. Results revealed a decrease in the L+Z level (4.70, 9.24, and 13.85%) of ready-mix stored at 4, 27, and 37°C, respectively. Critical relative humidity and critical moisture content of the product was 64% and 12.24%, respectively. The product is well accepted and was not affected adversely during storage. L+Z bioavailability from ready-mix in mice was higher in plasma (29.4%), liver (58.7%), and eye (14.6%) than control (mice received diet with purified L). To conclude, L+Z in the ready-mix is stable and more bioavailable than control. These findings may help in understanding the importance of simple food processing to improve L bioavailability under its deficient condition among an elderly population.