Capsanthin-Loaded Micelles: Preparation, Characterization and in vitro Evaluation of Cytotoxicity Using MDA-MB-231 Breast Cancer Cell Line.

Research background: Breast cancer is one of the most common cancers and remains a major cause of morbidity and mortality among women worldwide. In developed countries, breast cancer as a multifactorial disease is a major health concern, and its incidence is constantly rising in low and middle-income countries. Numerous studies have demonstrated that phytochemicals such as carotenoids inhibit breast cancer growth and induce apoptosis. We recently enhanced the solubility of capsanthin in water by encapsulating it in diosgenin polyethylene glycol succinate, a novel non-ionic surfactant. Thus, this study aims to evaluate the cytotoxicity of water-soluble capsanthin-loaded micelles in MDA-MB-231 cells in vitro through tetrazolium dye MTT assay. Experimental approach: In the current study, capsanthin, a hydrophobic carotenoid, is extracted from sweet red pepper (Capsicum annuum). Capsanthin-loaded diosgenin polyethylene glycol succinate 1000 (cap-DPGS-1000) micelles were prepared from capsanthin extract (cap) and diosgenin polyethylene glycol succinate 1000 (DPGS-1000) using the solid dispersion method. The capsanthin extract and cap-DPGS-1000 micelles were characterized by UV-visible spectroscopy, high-performance liquid chromatography (HPLC), Fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), particle size distribution, polydispersity, and scanning electron microscopy (SEM). The effects of capsanthin extract and cap-DPGS-1000 micelles on a human triple-negative breast cancer cell line (MDA-MB-231) were tested to check the cell viability, proliferation and cytotoxicity of the micelles. Results and conclusions: The solubility of encapsulated cap-DPGS-1000 micelles in water is greatly enhanced and leads to an increased scope for localized drug delivery, a better delivery option for treating residual cancerous tumours. The encapsulated capsanthin showed a sustained release in simulated intestinal fluid (pH=6.8). Our research proposes a sustained drug delivery system that ensures effective and controlled release to the affected site. The characterization data revealed no change in the structure and functional groups in the encapsulated capsanthin. The IC50 value of the cap-DPGS-1000 micelles against MDA-MB-231 breast cancer cells was (3.10±1.09) µg/mL, which is much lower than of capsanthin extract ((81.1±1.5) µg/mL). Capsanthin extract and capsanthin-loaded micelles are promising drug candidates to induce apoptosis and increase reactive oxygen species (ROS) in cancer cells. Novelty and scientific contribution: The result shows the cytotoxic effect of capsanthin and capsanthin-loaded micelles on MDA-MB-231 cell line for the first time. Capsanthin from sweet red pepper (Capsicum annuum) showed remarkable cytotoxic effect on the triple-negative MDA-MB-231 cell line.

Comparison of the Anti-Obesity Effect of Enriched Capsanthin and Capsaicin from Capsicum annuum L. Fruit in Obesity-Induced C57BL/6J Mouse Model.

Research background: Obesity increases mortality and morbidity due to its impact on type 2 diabetes, cardiovascular and gastrointestinal diseases, arthritis and certain cancers. The epidemic of excessive mass and obesity require constant research to improve therapies without undesirable side effects. Therefore, exploring the anti-obesity phytochemicals from food sources is essential. Most pharmacological studies of the anti-obesity potential of Capsicum annuum have been directed towards capsaicin and very few towards capsanthin. However, these studies utilized uncoated capsaicin and capsanthin. This study aims to compare the anti-obesity effects of enteric-coated capsaicin and capsanthin in a high-fat diet-induced obesity in animal model. Experimental approach: In this study, we investigated the anti-obesity properties of capsanthin-enriched pellets and capsaicin pellets derived from red chili fruit (Capsicum annuum) on high-fat diet (HFD)-induced obesity in C57BL/6J mice. First, the animals received HFD to induce their obesity. Animals were supplemented orally with pellets. The food intake, body mass, obesity and clinical biomarkers were assessed. Results and conclusions: The mice fed with HFD gained body mass and white adipose tissue mass compared to the mice that consumed a normal diet. The oral administration of capsanthin-enriched pellets and capsaicin pellets significantly reduced the body mass gain. These pellets have a statistically significant (p<0.05) impact on obesity biomarkers by increasing adiponectin and decreasing leptin, free fatty acid and insulin concentrations relative to HFD control. There was no change in the liver mass in all groups, but there was a significant decrease in white adipose tissue amounts. Inguinal adipose tissue amount was reduced by 37.0% and that of epididymal adipose tissue by 43.64% after treatment with capsanthin-enriched pellets. These results suggest that capsanthin-enriched pellets and capsaicin pellets may be useful in combating metabolic diseases, including obesity, without adverse effects. Novelty and scientific contribution: We increased the content of capsanthin for more than 50% in capsanthin-enriched extract and extended the room temperature stability for more than one year by converting the crystals into capsanthin-enriched pellets. This study breaks new ground by examining the potential of capsanthin >50% in the management of obesity for the first time.

Capsanthin from Capsicum annum fruits exerts anti-glaucoma, antioxidant, anti-inflammatory activity, and corneal pro-inflammatory cytokine gene expression in a benzalkonium chloride-induced rat dry eye model.

Dry eye disease (DED) is a complex ocular surface inflammatory disease. Its occurrence varies widely over the world, ranging from 5% to 34%. The use of preservatives, specifically benzalkonium chloride, in the ocular drops worsens the DED conditions. Furthermore, the Covid-19 pandemic increased screen time and the use of face masks and shields. As a result, the number of people suffering from dry eye disease (DED) has increased significantly in recent years. The main objective of our study is to find a solution to manage the dry eye disease (DED) preferably from natural source without any adverse events. In this study, the beneficial effects of capsanthin from Capsicum annum (CCA) were evaluated on benzalkonium chloride (BAC)-induced dry eye disease (DED) in Albino Wistar rats. Oral supplementation of CCA resulted in a statistically significant decrease in intraocular pressure (IOP) (p < .0001), increase in tear break-up time (TBUT) (p < .01), decline in Schirmer test results (p < .01), and decrease in corneal surface inflammation (p < .01). Capsanthin ameliorated in reducing oxidative stress by increasing serum antioxidant levels such as glutathione peroxidase (GPX), nitric oxide (NO), and lactoferrin (LTF) and inhibiting matrix metalloproteinases 2 and 9 (MMP2 and MMP9) (p < .0001). Capsanthin treatment significantly inhibited the expression of inflammatory cytokines, tumor necrosis factor-alpha (TNF-α), interleukins (IL-2, IL-4, IL-6), and pro-inflammatory mediator, matrix metalloproteinase-9 (MMP9). Furthermore, the lacrimal gland expressed vascular cell adhesion molecule (VCAM-1), and prostaglandin-endoperoxide synthase 2 (PTGS2) was suppressed by CCA treatment. PRACTICAL APPLICATIONS: Benzalkonium chloride (BAC), a preservative widely used in the topical ocular drug delivery system (ODDS), causes undesirable effects such as dry eye disease as well as ameliorating intraocular pressure leading to optical nerve damage and irreversible vision loss. Capsanthin from Capsicum annum (CCA) can be used to treat symptoms related to dry eye disease such as inflammation, eye irritation, visual disturbance, ocular discomfort with potential damage to the ocular surface. The CCA may be beneficial in the treatment of glaucoma, an elevated intraocular pressure. Capsanthin from C. annum can be useful in managing DED by increasing tear break-up time (TBUT), declining in Schirmer test results and decreasing in corneal surface inflammation.

Ninety-day repeated oral toxicity study of saponified Capsicum annum fruit extract with 50% capsanthin in Sprague-Dawley Rats with a 28-day recovery period.

A ninety-day oral toxicity study of saponified Capsicum annum fruit extract with 50% (w/w) capsanthin (SCFE-50 C) was performed by oral gavage administration to male and female Sprague-Dawley (SD) rats at doses of 0, 500, 1000 and 2000 mg/kg BW/day for a period of ninety consecutive days. To assess the reversal of toxicity, the treatment phase was followed with a twenty-eight-day recovery period. The treatment with SCFE-50 C in both male and female SD rats showed no mortality, and no treatment-related toxicologically significant changes were observed in any groups. No significant differences between treated and control groups were found in feed consumption, body weight gain, individual organ weights, ocular examination, clinical chemistry or blood biochemistry. The necroscopy and histopathology examination did not reveal any clinically significant changes in male and female rats from the 2000 mg/kg BW/day group. According to this study, the no observable adverse effect level (NOAEL) for saponified Capsicum annum fruit extract with 50% (w/w) capsanthin (SCFE-50 C) administered by oral gavage for 90-days is > 2000 mg/kg BW/day in SD rats.

Extraction of capsanthin from Capsicum annum L fruits and its effect on carbomer-induced intraocular pressure in Albino Wistar rats.

The present study was aimed to explore the antiglaucoma activity of capsanthin enriched fraction (CEF) of Capsicum annum L fruits against carbomer-induced experimental glaucoma in Albino Wistar rats. CEF was orally administered to carbomer-induced glaucomatous rats, and pilocarpine 2% eye drops were used as a standard drug. Intraocular pressure (IOP) levels were determined after oral administration of a low, medium, and a high dose of CEF (20, 40, and 80 mg/kg bwt) in glaucomatous rats. In rats with elevated IOP in both eyes, oral administration of CEF resulted in a significant reduction in IOP (p < .05) even at a low dose of 20 mg/Kg body weight. There were no treatment-related changes in histopathology, hematology, and clinical chemistry parameters. Thus, CEF when administered orally in IOP-bearing rats successfully reduced IOP without any adverse effects. PRACTICAL APPLICATIONS: Capsanthin enriched fraction can be used to prevent permanent vision loss due to age-related macular diseases and high intraocular pressure. The intraocular pressure reduction action of capsanthin can be useful in the treatment of glaucoma. The medication available to treat glaucoma are topical drugs, and for the first time, we proved the oral supplementation of capsanthin from a food source can reduce the intraocular pressure in rats.

Effectiveness of Cucumis sativus extract versus glucosamine-chondroitin in the management of moderate osteoarthritis: a randomized controlled trial.

PURPOSE: Osteoarthritis (OA) is an age-related disease caused by the wear and tear of the joints. Presently, there is no known cure for OA, but its management involves the use of high doses of pain killers and antiinflammatory agents with different side and dependency effects. Alternative management strategies involve the use of high doses of glucosamine-chondroitin (GC). This study was carried out to evaluate the efficacy of Q-Actin™, an aqueous extract of Cucumis sativus (cucumber; CSE) against GC in the management of moderate knee OA. PATIENTS AND METHODS: Overall, 122 patients (56 males and 66 females) aged between 40 and 75 years and diagnosed with moderate knee OA were included in this randomized double-blind, parallel-group clinical trial that took place in three different centers. The 180 day intervention involved two groups of 61 participants in each: the GC group, which received orally the generally prescribed dose of 1,350 mg of GC twice daily and the CSE group, which received orally10 mg twice daily of CSE. The Western Ontario McMaster Universities Osteoarthritis Index (WOMAC), Visual Analog scale, and Lequesne's Functional Index were used to evaluate pain, stiffness, and physical function of knee OA in participants at baseline (Day 0) and on Days 30, 60, 90, 120, 150, and 180. RESULTS: In the CSE group, the WOMAC score was decreased by 22.44% and 70.29% on Days 30 and 180, respectively, compared to a 14.80% and 32.81% decrease in the GC group. Similar trends were observed for all the other pain scores. No adverse effect was reported during the trial period. CONCLUSION: The use of 10 mg CSE, twice daily, was effective in reducing pain related to moderate knee OA and can be potentially used in the management of knee pain, stiffness, and physical functions related to OA.