Thesis title: TECNICHE AVANZATE IN ENDOSCOPIA PER LA MEDICINA DI PRECISIONE
In recent years, diagnostic endoscopy and regenerative medicine have emerged as two rapidly evolving fields within the broader context of precision medicine. This doctoral work integrates these two dimensions, digital diagnostics and three-dimensional cell biology, with the aim of improving the morphological and molecular characterization of biliopancreatic diseases and developing autologous cellular models to support translational research.
Chapter 1 analyzes the combined use of endoscopic ultrasound-guided fine-needle biopsy (EUS-FNB) and digital fluorescence confocal microscopy (VivaScope) as an integrated approach for real-time diagnosis of biliopancreatic lesions. The introduction of VivaScope enabled the immediate visualization of high-quality digital histological images, reducing reporting times and enhancing the diagnostic accuracy of EUS-FNB sampling. The results confirm that the combination of EUS-FNB and VivaScope represents an effective diagnostic platform, capable of merging morphological precision with rapid evaluation, thus paving the way for new models of digital pathology applied to endoscopic practice.
Chapter 2 explores a regenerative medicine and cellular modeling approach through the generation of duodenal organoids derived from submucosal gland stem cells (DSGSCs) obtained via deep endoscopic biopsies. Two protocols for isolation and three-dimensional culture were compared: one based on enzymatic–mechanical digestion (Method 1) and another on cold mechanical dissociation (Method 2). Both methods successfully produced viable and proliferating organoid structures, though with substantial differences in growth kinetics, morphology, and transcriptional profiles. RNA-seq analysis revealed that organoids generated with Method 1 exhibited a multipotent and secretory phenotype, characterized by overexpression of genes associated with submucosal and glandular progenitor cells (SOX9, GATA6, KRT7, KRT19, MUC5AC), whereas those obtained with Method 2 displayed a more intestinal and lineage-committed profile, with higher expression of crypt cell markers (LGR5, OLFM4). Overall, organoids derived from duodenal biopsies represent a functional and stable model of human endodermal epithelium, suitable for studies in physiology, toxicology, personalized pharmacology, and regenerative cell therapy.
In conclusion, the results of this research highlight how the integration between digital endoscopic diagnostics and three-dimensional organoid biology represents a new frontier in precision medicine. The interaction between advanced clinical techniques and innovative experimental models enables improvements in diagnostic accuracy, a deeper understanding of disease pathophysiology, and the foundation for increasingly personalized approaches to patient care.