Ciencias Exactas y Ciencias de la Salud

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Immunostimulant activity of aggregates of phenolic compounds and polysaccharides extracted from Jalapeño pepper seeds (Capsicum annuum L.)
(Instituto Tecnológico y de Estudios Superiores de Monterrey, 2024-05) Fortiz De Ita, Jorge; Gutierrez Uribe, Janet Alejandra; emimmayorquin; Antunes Ricardo, Marilena; Rocha Pizaña, Maria del Refugio; School of Engineering and Sciences; Campus Puebla; Cortes Ferre, Hector Emmanuel
Thanks to previous studies, the use of enzyme-assisted extraction (EAE) has been demonstrated as one of the best techniques to extract phenolic compounds specifically from pepper seeds. Likewise, the anti-inflammatory activity of these compounds has been studied and how the use of enhancers benefits the extraction and enhances the properties of the extracts. Therefore, the present research corresponds to a continuation of these findings, to be able to demonstrate how ferulic acid as an enhancer is ideal for improving extraction, extracting better quality phenolic compounds, promoting the generation of aggregates to these phenols and also enhancing the properties of the same, also testing how ultrafiltration helps us separate these aggregates to be able to compare their immunostimulant activities and mainly to be able to demonstrate the immunostimulant properties of these extracts. The use of ferulic acid as an enhancer was shown to reduce the amount of total soluble carbohydrates (TSC) and that, when passing through ultrafiltration, it achieved the lowest loss of suspended solids (6%) and the presence of more ppm of phenolic compounds in the samples. With this enhancer, we verified the value of using this phenolic acid in the extraction of phenolic compounds. The findings in particle size gave us the conclusion of the formation of small polyphenols and polysaccharides that manage to pass through the ultrafiltration pores and achieve particle sizes even larger (JPSC = 893.8 nm and JPSF = 1039 nm) than those of the aggregates at times 120 and 180 minutes. The characterization of free phenols served to demonstrate the disappearance of phenolic acids in the filtered compounds and also to quantify vanillin (NF-JPSC = 14.66 – 1.28 μg/g, NF- JPSF = 38.04 – 0 μg/g, F-JPSC = 16.778 – 0 μg/g and F- JPSF = 27.474 – 0 μg/g), which is decreased as the hydrolysis time increases, resulting in the formation of an aggregate of this phenolic compound with sucrose, which is the most abundant sugar in these extracts and which was also characterized and demonstrated that it disappears when it is filtered and only fructose remains as the most abundant sugar. Finally, it was verified that the immunostimulant activity is higher in the concentrated extracts (2x) and in those without filtering, achieving a production of nitric oxide (NOx) of up to 327.76%, and comparing different concentrations of solids in suspension (800, 400, 200 , 100, 50, 25, 12.5 and 6.25 ug/mL) with the minimum concentration being the one with the most immunostimulating activity and also being the least cytotoxic (cell viability = 143.81%) in the 2x sample at 120 minutes of hydrolysis, this being the best of all the samples analyzed.
Biological Validation of Isorhamnetin Glycosides Loaded Nanofibers as a Drug Release System
(Instituto Tecnológico y de Estudios Superiores de Monterrey) García Valderrama, Elsy Jazmín; 889465; 889465; 889465; Gutierrez Uribe, Janet Alejandra; Mamidi, Narsimha; School of Engineering and Sciences; School of Engineering and Sciences; Campus Monterrey; Antunes Ricardo, Marilena
Nanofibers have emerged as a potential novel platform due to its physicochemical properties for healthcare application. Nanofibers advantage rely in their high specific surface area-tovolume ratio and porosity. Their peculiar assembly allow cell accommodation, nutrient infiltration, gas exchange, waste excretion, high drug release rate and stable structure. Different strategies have been reported to enhance physical and mechanical characteristics of nanofibers and increase their production yield rate. Herbal bioactive compounds have raised special interest due to their prominent beneficial properties in health care. Nanofiber properties let these systems to serve as bioactive compounds carriers to generate functional matrixes. Opuntia ficus-indica (L.) Mill, is commonly consumed in Mexico due to its beneficial properties since ancient times for the treatment of diabetes, hypertension, metabolic syndrome, cardiovascular and neurodegenerative diseases, hypercholesterolemia and asthma. Additionally, O. ficus-indica isorhamnetin glycosides have clinical potential for the treatment of topical injuries. The following project assessed the release kinetics and the in vitro anti-inflammatory potential of isorhamnetin glycosides loaded to nanofibers with different concentration of gelatin (25% and 30%). Isorhamnetin glycosides were extracted from the plant using acetic acid 80% (v:v) and the resultant solution was mixed with gelatin to create a polymeric solution. Isorhamnetin glycosides loaded nanofibers were fabricated through the ForcespinningTM technique and crosslinked with glutaraldehyde (GTA) vapors. Isorhamnetin glycosides within the nanofibers were quantified by HPLC-UV. SEM and FTIR analysis of nanofibers were used to elucidate nanofibers morphology and chemical interactions between the functional groups of the compounds. Alike, the in vitro drug release assays revealed the role of polymer concentration and chemical interactions between particles to enhance the release of isorhamnetin glycosides. 25% GL+NP and 30% GL+NP nanofibers displayed a release percentage of 37.15% and 62.98%, respectively. Additionally, celular uptake assays demonstrated the feasibility of glycosides to cross human dermal fibroblasts (HDFa cell line) cell membrane according the glycosylation pattern of the flavonoids. Cell proliferation was enhanced by the presence of the matrix and no significant effect on cell viability was observed at the concentration of total isorhamnetin glycosides used in the experiments. Nitric Oxide (NO), Cyclooxigenase-2 (COX-2), tumor necrosis- alpha (TNF- alpha) and Interleuikin-6 (IL-6) were monitored. Isorhamnetin glycosides loaded to nanofibers remarkably decrease the production of NO, 30%GL+NP nanofiber reached a NO inhibition of 93.8% at 8h of isorhamnetin release. Also, they inhibited the production of COX-2, TNF- alpha and IL-6. Among the two nanofibers, 30% GL+NP nanofiber displayed the most adequate physicochemical characteristics such as diameter, non-bead formation and prevalence of non-covalent bonding for ease compound´s release to act as a controlled release system a potential anti-inflammatory system in topical injuries than 25% GL+NP nanofiber.

Ciencias Exactas y Ciencias de la Salud

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