The procedures developed by the Laboratory for Advanced Therapy Technologies (LTTA) exploit innovative technologies for the evaluation of the immunomodulatory and antioxidant properties of specific compounds contained in foods, nutraceuticals and other products, such as cosmetics and fillers, by investigating their effects on i) cellular viability, proliferation and metabolism ii) secretion of inflammatory mediators iii) oxidative stress.
Vaccinium floribundum berries, rich in polyphenols and with known antioxidant properties
The assays developed by LTTA are based on innovative molecular and cellular biology technologies that generate reliable and reproducible results on the beneficial / cytotoxic effects of the analyzed compounds in a relatively short time. The advantage of using these cellular systems is constituted by their simplicity and robustness, providing a rapid tool for a preliminary screening on the properties of novel compounds to identify the most promising to be further characterized in in vivo and human studies.
These procedures could be particularly useful for companies interested in evaluating
1) the effectiveness of nutraceuticals, functional foods or other compounds in promoting beneficial effects on health, directly modulating immunological parameters and / or through the action of bioactive metabolites obtained from fermentation processes; 2) biocompatibility and the potential pro-inflammatory effect of cosmetic products and / or hyaluronic acid-based fillers.
RAW 264.7 murine macrophage cells are used as an in vitro model for the evaluation of the pro- or anti-inflammatory and immunomodulatory activity of the compounds of interest.
Evaluation of the immunomodulatory and antioxidant activity of i) Papaya extracts fermented with probiotics or yeasts and ii) Vaccinium floribundum berry extracts fermented or not with probiotics
The immunomodulatory activity is assessed through
i) the analysis of the expression levels of specific mediators of inflammation and immune response such as iNOS (inducible nitric oxide synthase) and TNFα (tumor necrosis factor α) by quantitative RT-PCR in RAW 264.7 mouse macrophage cell line, after treatment with the compound of interest
ii) the Griess assay in the culture medium in RAW 264.7 following treatment with the compound of interest.
The cytotoxic analysis are based on the evaluation of cell proliferation (by the Alamar Blue and Trypan Blue assay) and on the determination in the cell culture medium of the lactate dehydrogenase (LDH) activity released by damaged or dead cells. The antioxidant capacity is evaluated through the ORAC assay (oxygen radical absorption capacity) and in pheochromocytoma embryonic brain cells through the quantification of intracellular levels of reactive oxygen species (ROS).
Results:
1) Formulations of Papaya fermented with probiotics show stronger immunomodulatory properties than those fermented with yeast as they promote more efficiently the expression levels of iNOS and TNFα;
2) Berries extracts fermented with probiotic exhibit more beneficial antioxidant and immunomodulatory properties than berry extracts not fermented.
Incos Cosmeceutica Industriale
The results obtained on Papaya led to the production of a food supplement based on fermented Papaya with probiotics, currently on sale (http://www.lafarmaciadelleerbe.it/papaya-fermentata).
A patent application will be filed shortly
La Farmacia delle Erbe Fermented Papaya was fermented with a selected mix of 3 different probiotic ferments, to which a precious prebiotic fiber was added