Teresa Barra

Background: Inflammatory bowel disease (IBD) is a chronic inflammatory condition, with therapy-resistant patients undergoing surgery and a high risk of developing colorectal cancer. Novel therapeutic approaches have shown limited efficacy in IBD treatment, highlighting the need for safer and more personalized strategies. The potential of natural compounds to modulate inflammation suggests their use as a potential adjunct therapy for IBD patients. Methods: Intestinal epithelial cells organoids (IECOs) were derived from IBD and non-IBD tissues from IBD patients, and levels of inflammation markers and epithelial barrier permeability were assayed using qRT-PCR, WB, IF and leaking assays in the presence of Ozoile, an extra virgin olive oil enriched in ozonides. The Luciferase-based IBD-like organoid platform was generated for preliminary screening of anti-inflammatory drugs. Results: In this study, we showed that IBD-ECOs recapitulate tissue architecture and pathological state. We showed that Ozoile has anti-inflammatory and epithelial barrier modulatory effects and that the Luciferase IBD-like organoid model is sensitive to anti-inflammatory compounds. Conclusions: Using IECOs, the specific anti-inflammatory and regenerative properties of Ozoile were assessed. Notably, our study highlights the potential of an IBD-like organoid platform to use in high-throughput screenings for rapid selection of anti-inflammatory drugs.

Combination therapy has shown promise in treating aggressive cancers by using several drugs simultaneously to target different biological pathways, with the added benefit of potentially reducing toxicity. Given the critical role of mitochondrial dysfunction in tumor progression, targeting mitochondrial metabolism represents a promising therapeutic avenue. In this study, we developed a mitochondria-targeted nanofiber based on self-assembling peptides, engineered to co-deliver two complementary therapeutic agents for glioblastoma treatment. The nanofiber carries 1,3-bromopyruvate (BrP), a glycolysis inhibitor, and temozolomide, an alkylating chemotherapeutic, conjugated via a matrix metalloproteinase-9 (MMP-9)-responsive linker for controlled, on-demand release. To enhance selectivity for glioblastoma cells, the nanofiber surface was functionalized with the targeting peptide falGea binding specifically to EGFRvIII, commonly overexpressed in tumor cells, and gH625, a cell-penetrating peptide known to facilitate the blood-brain barrier (BBB) transport. The nanofibers were comprehensively characterized for their aggregation behavior, structural stability, and morphology. Mitochondrial targeting and functional effects were evaluated by using isolated rat brain mitochondria. Therapeutic efficacy was assessed in U-87 MG glioblastoma cells cultured in both 2D and 3D systems. Additionally, BBB permeability was examined by using a dynamic 3D in vitro BBB model, demonstrating the transport-enhancing role of gH625. These findings support the potential of multifunctional, mitochondria-targeted nanofibers as an effective platform for glioblastoma therapy, offering both precision targeting and enhanced drug delivery across the BBB.

Parkinson’s disease (PD) is an aggressive and devastating age-related disorder. Although the causes are still unclear, several factors, including genetic and environmental, are involved. Except for symptomatic drugs, there are not, to date, any real cures for PD. For this purpose, it is necessary develop a model to better study this disease. Neuroblastoma cell line, SH-SY5Y, differentiated with retinoic acid represents a good in vitro model to explore PD, since it maintains growth cells to differentiated neurons. In the present study, SH-SY5Y cells were treated with 1-methyl-4-phenylpyridinium (MPP⁺), a neurotoxin that induces Parkinsonism, and the neuroprotective effects of pituitary adenylate cyclase-activating polypeptide (PACAP), delivered by functionalized liposomes in a blood–brain barrier fluid dynamic model, were evaluated. We demonstrated PACAP neuroprotective effects when delivered by gH625-liposome on MPP⁺-damaged SH-SY5Y spheroids.

The blood–brain barrier (BBB) selectively protects the central nervous system (CNS) from external insults, but its function can represent a limit for the passage of therapeutic molecules. Numerous in vitro models of the BBB have been realized in order to study the passage of drugs for neurodegenerative diseases, but these in vitro models are not very representative of the physiological conditions because of a limited supply of oxygen and nutrients due to static conditions. To avoid this phenomenon, we used a millifluidic bioreactor model that ensures a circulation of the medium and, therefore, of the nutrients, thanks to the continuous laminar flow. This dynamic model consists of a double-culture chamber separated by a membrane on which brain endothelial cells are cultured in order to evaluate the passage of the drug. Furthermore, in the lower chamber, SH-SY5Y were seeded as 3D spheroids to evaluate the drug passage through these cells. As nanodelivery system, we used liposomes functionalized with viral fusion peptide to evaluate the passage of a neuroprotective agent, pituitary adenylate cyclase-activating polypeptide (PACAP), through the dynamic in vitro model of the BBB. We showed that our nanodelivery system, made of functionalized liposomes and loaded with specific molecules, efficiently crosses the in vitro fluid-dynamic model of the BBB. Our findings represent an important step for further experimental investigations on PACAP administration as a therapeutic agent by an enhanced drug delivery system. Our results can improve the diffusion of good practice in neuroscience laboratories, helping to spread the 3R rules.