RESULTS
SCIENTIFIC REPORT -Phase 2_ 2022
In vitro studies for iron oxide nanoparticles carried by probiotics, cellular uptake and cytotoxicity assessment.
Activity 2.1. In vitro cytotoxicity studies on normal stomach and intestinal cells; To establish therapeutic doses, cytotoxicity studies are performed on normal, stomach and intestinal cells- MTT Method The obtained Nano Capsules were used for in vitro studies. At first, proliferation assays were performed by the standard MTT colorimetric assay. Cell viability was determined after exposure of cells to the treatment for 12, 24 and 48 hours.
The results of the MTT test of cell proliferation and microchondrial activity on Hs-27, CaCo2 and DDL-1 cell lines treated with nanoparticles obtained by the solvothermal method
Activity 2.2. LDH activity detection - test LDH.;
In correlation with the cell proliferation assay, LDH leakage was determined to demonstrate the cytotoxicity of the compound on the tumor cell line. The treatment was applied to cells and increased LDH activity correlates with increased cell wall damage and LDH leakage into the extracellular space, thus acting as an indicator of cell membrane integrity.
Lactate dehydrogenase activity in treated cells
Activity 2.3. Investigating the type of cell death - Caspase activity. . The investigation into the type of cell death – was carried out by evaluating the activity of caspases, which is a crucial indicator and mediator of programmed cell death (apoptosis).
Effects of treatment (150 µg Fe/mL) on cell lines: Hs27, Caco2, DDL-1. EB/AO staining of cells with and without treatment for 24 h. Cells were observed with an inverted fluorescence microscope
Scheme showing the cytotoxicity of iron oxide nanoparticles at the cellular level (source_original)
Activitya 2.4. Dissemination of results - conference, scientific articles;
- A patent application titled "Characterization and method of obtaining iron oxide nanoparticles with Lactobacillus fermentum" was submitted - A00699/13.10.2022
- An international book chapter was published in the Elsevier publishing house.
- Participated in 4 conferences, including 3 international and one national, with 3 oral presentations and a poster.
- S-a depus o cerere de brevet intitulat “Caracterizarea şi procedeul de obţinere a nanoparticulelor de oxid de fier cu Lactobacillus fermentum” – A00699/13.10.2022
Conference Probiotics, prebiotics & novel foods, nutraceuticals for human and gastrointestinal nutrition and health
September 12-14, 2021
Urban University of Rome
