Priprema, karakterizacija i in vitro procjena toksičnosti micela s kapsantinom na modelu stanične linije karcinoma dojke MDA-MB-231

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.Pozadina istraživanja. Karcinom dojke jedan je od najčešćih tipova tumora te je još uvijek glavni uzročnik morbiditeta i mortaliteta među ženama diljem svijeta. U razvijenim zemljama ova multifaktorska bolest predstavlja primarni zdravstveni problem, a u stalnom je porastu u srednje i nisko razvijenim zemljama. Mnoga istraživanja pokazuju da fitokemikalije poput karotenoida suzbijaju rast i potiču apoptozu stanica karcinoma dojke. Nedavno smo poboljšali topljivost kapsantina u vodi postupkom inkapsulacije u novom neionskom surfaktantu, diosgenin polietilenglikol sukcinatu. Stoga je svrha ovoga rada bila pomoću MTT testa ispitati in vitro citotoksičnost micela punjenih kapsantinom, topljivih u vodi, na stanice raka MDA-MB-231. Eksperimentalni pristup. U radu je iz ekstrakta slatke crvene paprike (Capsicum annuum) izoliran hidrofobni karotenoid kapsantin. Iz čvrste disperzije ekstrakta i diosgenin polietilenglikol sukcinata 1000 pripremljene su micele. Ekstrakt kapsantina i micele su okarakterizirani pomoću UV-Vis spektroskopije, visokodjelotvorne tekućinske kromatografije, infracrvene spektroskopije s Fourierovom transformacijom, difrakcije X-zraka, raspodjele veličine čestica, polidisperzije i pretražne elektronske mikroskopije. Ispitali smo učinak ekstrakta kapsantina i micela na preživljavanje i rast trostruko negativnih stanica raka dojke (MDA-MB-231) te citotoksičnost micela. Rezultati i zaključci. Bitno se povećala topljivost kapsantina u inkapsuliranim micelama i time proširila mogućnost njegove primjene za ciljanu isporuku, čime se postižu bolji rezultati u liječenju rezidualnih tumora. Inkapsulirani se kapsantin kontrolirano otpuštao pri simulaciji probave u crijevima (pH=6,8). Predloženi sustav za ciljanu isporuku lijeka s produljenim djelovanjem omogućuje učinkovito i kontrolirano otpuštanje aktivne tvari na mjestu djelovanja. Rezultati pokazuju da se pri inkapsulaciji kapsantina nije promijenila njegova struktura, a niti sastav funkcionalnih skupina. Citotoksičnost micela na stanice MDA-MB-231 (IC50=(3,10±1,09) μg/mL) bila je bitno veća od one ekstrakta kapsantina (IC50=(81,1±1,5) μg/mL). Ekstrakt kapsantina i micele s kapsantinom mogu se upotrijebiti za poticanje apoptoze i povećanje količine reaktivnih kisikovih spojeva u stanicama raka. Novina i znanstveni doprinos. Rezultati prvi put prikazuju citotoksični učinak kapsantina i micela s kapsantinom na stanice raka MDA-MB-231. Kapsantin izoliran iz slatke crvene paprike (Capsicum annuum) imao je izniman citotoksični učinak na trostruko negativne stanice raka MDA-MB-231

Usporedba učinka hranidbe peletima obogaćenih kapsantinom i kapsaicinom iz čilija (Capsicum annuum L.) na pretilost miševa soja C57BL/6J

SUMMARY Research backgroundObesity 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 approachIn 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 conclusionsThe 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 contributionWe 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.Pozadina istraživanja. Pretilost povećava smrtnost i morbiditet jer utječe na razvoj dijabetesa tipa 2, kardiovaskularnih i gastrointestinalnih bolesti, artritisa i nekih tipova karcinoma. Epidemija pretilosti zahtijeva kontinuirano poboljšanje terapeutika radi smanjenja neželjenih nuspojava njihove primjene. Stoga je neophodno ispitati fitokemikalije porijeklom iz hrane, a koje smanjuju pojavu pretilosti. Većina farmakoloških ispitivanja učinaka čilija na pretilost usmjerena su na kapsaicin, a vrlo malo ih se bavi kapsantinom. U prethodnim su se istraživanjima koristili neobloženi kapsaicin i kapsantin. Stoga je svrha ovoga rada bila usporediti utjecaj enteričkih pripravaka kapsaicina i kapsantina na suzbijanje pretilosti miševa hranjenih hranom s velikim udjelom masti. Eksperimentalni pristup. U ovom smo radu ispitali učinak hrane s velikim udjelom masti uz dodatak peleta obogaćenih kapsantinom i peleta kapsaicina dobivenih iz čilija (Capsicum annuum) na pretilost miševa soja C57BL/6J. Miševima je inducirana pretilost unosom hrane s velikim udjelom masti, a zatim su hranjeni peletima. Mjereni su sljedeći parametri: unos hrane, tjelesna masa, pretilost i klinički biomarkeri. Rezultati i zaključci. Miševi hranjeni hranom s velikim udjelom masti imali su veću tjelesnu masu i više bijelog masnog tkiva u usporedbi s miševima koji su hranjeni standardnom hranom. Dodatak peleta obogaćenih kapsantinom i peleta kapsaicina u hranu bitno je reducirao povećanje tjelesne mase miševa. Peleti su bitno (p<0,05) utjecali na biomarkere pretilosti, jer su povećali koncentraciju adiponektina, a smanjili koncentracije leptina, slobodnih masnih kiselina i inzulina u usporedbi s kontrolnim uzorkom (miševi hranjeni zasićenim mastima). Niti u jednoj skupini nije se povećala masa jetre miševa, no bitno se smanjila količina bijelog masnog tkiva. Nakon dodatka peleta obogaćenih kapsantinom u hranu smanjila se količina ingvinalnog masnog tkiva za 37 %, a masnog tkiva epididimisa za 43,64 %. Dobiveni rezultati potvrđuju da se peleti obogaćeni kapsantinom te peleti kapsaicina mogu upotrijebiti za suzbijanje metaboličkih poremećaja, uključujući pretilost, bez štetnih nuspojava. Novina i znanstveni doprinos. Konverzijom kristala u pelete obogaćene kapsantinom povećao se udjel kapsantina u peletima za 50 % i produljila stabilnost peleta pri sobnoj temperaturi za više od godinu dana. Ovaj rad po prvi put potvrđuje mogućnost korištenja kapsantina masenog udjela većeg od 50 % za kontrolu pretilosti