Roberta Di Pietro

Satellite cells (SCs) are essential for skeletal muscle regeneration, but their function declines with aging, often associated with increased pro-apoptotic signaling. This study investigated the impact of in vitro serum starvation—as a model of acute microenvironmental and nutrient stress—on the apoptosis and differentiation potential of human SCs from young and aged donors. SCs were isolated from the Vastus Lateralis of young and aged subjects and cultured in serum-free medium for up to 72 h. We assessed apoptosis through Annexin V/PI staining, TUNEL assays, and caspase activity measurements, while transcriptional profiles were analyzed via RT-PCR. Aged SCs displayed a significantly higher susceptibility to stress-induced apoptosis compared to young controls, marked by the early upregulation of CASP9 and FOXO1. While typical nucleosomal DNA fragmentation was absent, we observed the activation of caspase-3 after 72 h of starvation. In aged cells, activated caspase-3 co-localized with myogenin and extranuclear DNA at sites of nuclear remodeling. Notably, treatment with a pan-caspase inhibitor (z-VAD-fmk) prevented the formation of micronuclei and myotubes, further highlighting a non-apoptotic role for these enzymes. Aged SCs also showed a distinct cell cycle profile characterized by an enlarged G0/G1 phase and altered expression of CDK and CCNB1 genes. Our findings suggest that in human aged SCs, caspase enzymes serve a dual role: mediating a heightened stress response and facilitating the nuclear remodeling necessary for myogenic differentiation. These results clarify how intrinsic aging shapes the response of muscle stem cells under severe environmental and metabolic resource deprivation.

Biomaterials play a central role in tissue engineering and regeneration by providing scaffolds that support cell adhesion, proliferation and differentiation while modulating the surrounding microenvironment. They represent promising alternatives to traditional surgical approaches that may lead to complications or tissue damage, and their performance is influenced by chemical composition, mechanical behavior, architecture and interfacial properties, all of which can be precisely tuned through advanced fabrication and surface modification strategies. This review synthesizes evidence from a comprehensive literature search across major scientific databases, focusing on highly cited studies and available clinical data, and examines natural and synthetic biomaterials, their biological responses, functional characteristics, and surface modification methods. Emphasis is placed on Cold Atmospheric Plasma (CAP), which selectively modifies the outermost nanolayer of materials, enhancing hydrophilicity, functional group density, protein adsorption and overall cell–material interactions, as well as improving drug loading capacity. The review also considers stem cell interactions with biomaterials and emerging applications of artificial intelligence (AI) for predicting performance and guiding material optimization. Overall, the analysis highlights that natural matrices provide intrinsic bioactivity, synthetic polymers offer tunable mechanics and degradation profiles, and composite systems integrate these advantages. Advances in technologies such as electrospinning and 3D/4D printing enable precise control over architecture, supporting cell colonization and vascularization. Collectively, developments in CAP treatments and AI-driven design strategies are strengthening the regenerative potential of biomaterials and advancing their clinical translation.

Background: The present case report study aims to describe, from both clinical and histological aspects, the bone healing pattern in a patient under oral bisphosphonates therapy. Case Presentation: an 82-year-old female patient has been under oral nitrogen bisphosphonates therapy for two years. She underwent a tooth extraction. After four months, two bone biopsies were harvested, during standard implant drilling procedures. The first one corresponded to the healed alveolar socket of the previously extracted tooth (specimen A), while the second one corresponded to the bone ridge that was edentulous before starting the bisphosphonates therapy (specimen B). Morphometric and histologic analyses were performed. Results: In both, the bone resulted vital and no evidence of empty lacunae was detected. A reduction in the haversian canal diameter was observed in specimen B. The present case report highlights histological findings suggesting that patients undergoing oral bisphosphonates therapy may be eligible for surgical therapy. A pre-operative careful anamnesis and the observance of international guidelines for treating patients taking bisphosphonates are mandatory. Conclusions: These preliminary results will be used to plan a large clinical study in order to better understand the influence of bisphosphonates on the bone healing process.

The Board of Italian Society of Histochemistry has dedicated significant effort to identifying and contacting prestigious researchers to serve as keynote and invited speakers for this important event as well as Italian societies like GIC (Italian Society of Cytometry), GISN (Italian Group for the Study of Neuromorphology) and SIBS (Italian Society of Experimental Biology) for organizing joint sessions. Furthermore, all member societies of the IFSHC have been actively reached out to define and expand the Congress program. A particular mention must go to the Histochemical Society and to the Society for Histochemistry for their active contribution in organizing sessions. With a focus on ensuring a high-caliber scientific program, the team has successfully formulated a comprehensive agenda that includes plenary sessions, workshops, thematic panels, and poster sessions addressing the latest advancements in histochemistry and cytochemistry applied to medicine, biology, nanotechnology and artificial intelligence. Professor Pavel Hozak, PhD, President, International Federation of Societies for Histochemistry and CytochemistryProfessor Roberta Di Pietro, MD, President, Italian Society of Histochemistry and the Local Organizing Committee

Satellite cells (SC) proliferation and differentiation play a crucial role in skeletal muscle regeneration after damage and adaptation in response to hypertrophic stimuli. Skeletal muscle ageing affects SC proliferation and differentiation and rely on an enhanced expression of several pro-apoptotic factors. As previous researches demonstrated a modified expression of some genes associated with SC antioxidant and repair activity due to age, in the present project we aimed at assessing the occurrence of apoptotic features in human SCs. Cells were collected from Vastus Lateralis of 9 young (27.3+2.0 years old) and 9 old (71.1+1.8 years old) subjects and cultured in serum-free medium to be collected at 4-24-48 and 72 h. Apoptosis was investigated with Annexin V/PI staining, terminal deoxynucleotidyl transferase (TdT)-mediated nick-end labelling (TUNEL) technique, flow cytometry analysis of Caspase-8 activity and RT-PCR detection of transcripts of apoptotic genes. Interestingly, CAS9 and FOXO1 genes were found upregulated in aged SCs at all-time points tested. Furthermore, we showed the activation of caspase-3 enzyme after 72 h starvation in culture with a nice co-localization with myogenin and TUNEL-positive micronuclei in the elderly. Interestingly, after treatment for 24 h with a broad pan-caspase inhibitor Cbz-Val-Ala-Asp- fluoromethyl-ketone (z-VAD-fmk) no micronuclei neither myotube formation were revealed, suggesting the involvement of caspase enzymes in the nuclear remodelling occurring during myotube formation.

Pancreatic cancer, most frequently as ductal adenocarcinoma (PDAC), is the third leading cause of cancer death. Clear-cell primary adenocarcinoma of the pancreas (CCCP) is a rare, aggressive, still poorly characterized subtype of PDAC. We report here a case of a 65-year-old male presenting with pancreatic neoplasia. A histochemical examination of the tumor showed large cells with clear and abundant intracytoplasmic vacuoles. The clear-cell foamy appearance was not related to the hyperproduction of mucins. Ultrastructural characterization with transmission electron microscopy revealed the massive presence of mitochondria in the clear-cell cytoplasm. The mitochondria showed disordered cristae and various degrees of loss of structural integrity. Immunohistochemistry staining for NADH dehydrogenase [ubiquinone] 1 alpha subcomplex, 4-like 2 (NDUFA4L2) proved specifically negative for the clear-cell tumor. Our ultrastructural and molecular data indicate that the clear-cell nature in CCCP is linked to the accumulation of disrupted mitochondria. We propose that this may impact on the origin and progression of this PDAC subtype.

Extracellular vesicles (EVs) are secreted from many tumors, including glioblastoma multiforme (GBM), the most common and lethal brain tumor in adults, which shows high resistance to current therapies and poor patient prognosis. Given the high relevance of the information provided by cancer cell secretome, we performed a proteomic analysis of microvesicles (MVs) and exosomes (EXOs) released from GBM-derived stem cells (GSCs). The latter, obtained from the brain of GBM patients, expressed P2X7 receptors (P2X7Rs), which positively correlate with GBM growth and invasiveness. P2X7R stimulation of GSCs caused significant changes in the EV content, mostly ex novo inducing or upregulating the expression of proteins related to cytoskeleton reorganization, cell motility/spreading, energy supply, protection against oxidative stress, chromatin remodeling, and transcriptional regulation. Most of the induced/upregulated proteins have already been identified as GBM diagnostic/prognostic factors, while others have only been reported in peripheral tumors. Our findings indicate that P2X7R stimulation enhances the transport and, therefore, possible intercellular exchange of GBM aggressiveness-increasing proteins by GSC-derived EVs. Thus, P2X7Rs could be considered a new druggable target of human GBM, although these data need to be confirmed in larger experimental sets.

Pancreatic cancer (PC) is the seventh leading cause of cancer-related death. PC incidence has continued to increase by about 1% each year in both men and women. Although the 5-year relative survival rate of PC has increased from 3% to 12%, it is still the lowest among cancers. Hence, novel therapeutic strategies are urgently needed. Challenges in PC-targeted therapeutic strategies stem from the high PC heterogeneity and from the poorly understood interplay between cancer cells and the surrounding microenvironment. Signaling pathways that drive PC cell growth have been the subject of intense scrutiny and interest has been attracted by necroptosis, a distinct type of programmed cell death. In this review, we provide a historical background on necroptosis and a detailed analysis of the ongoing debate on the role of necroptosis in PC malignant progression.

Trop-2 proteolytic processing in cancer cells exposes epitopes that were specifically targeted by the 2G10 antibody. We sought additional recognition of Trop-2 within difficult-to-reach, densely packed tumor sites. Trop-2 deletion mutants were employed in immunization and screening procedures, and these led to the recognition of a novel epitope in the N-terminal region of Trop-2, by the 2EF antibody. The 2EF mAb was shown to bind Trop-2 at cell–cell junctions in MCF-7 breast cancer cells, and in deeply seated sites in prostate cancer, that were inaccessible to benchmark anti-Trop-2 antibodies. The 2EF antibody was shown to inhibit the growth of HT29 colon tumor cells in vitro, with the highest activity at high cell density. In vivo, 2EF showed anticancer activity against SKOv3 ovarian, Colo205, HT29, HCT116 colon and DU-145 prostate tumors, with the highest impact on densely packed tumor sites, whereby 2EF outcompeted benchmark anti-Trop-2 antibodies. Given the different recognition modes of Trop-2 by 2EF and 2G10, we hypothesized the effective interaction of the two mAb in vivo. The 2EF mAb was indeed demonstrated to enhance the activity of 2G10 against tumor xenotransplants, opening novel avenues for Trop-2-targeted therapy. We humanized 2EF by state-of-the-art CDR grafting/re-modeling, yielding the Hu2EF for therapy of Trop-2-expressing tumors in patients.

Pancreatic cancer (PC) is the seventh leading cause of cancer-related death. PC incidence has con-tinued to increase by about 1% each year in both men and women. Although the 5‐year relative survival rate of PC has increased from 3% to 12%, it is still the lowest among cancers. Hence, novel therapeutic strategies are urgently needed. Challenges in PC-targeted therapeutic strate-gies stem from the high PC heterogeneity and from the poorly understood interplay between cancer cells and the surrounding microenvironment. Signaling pathways that drive PC cell growth have been the subject of intense scrutiny and interest has been attracted by necroptosis, a distinct type of programmed cell death. In this review, we provide a historical background on necroptosis and a detailed analysis of the ongoing debate on the role of necroptosis in PC malignant progression.