Hybrid 3D-printed/electrospun scaffolds drive myogenic differentiation of mesenchymal stem cells (MSCs) Silvia Pisani, Stefania Marconi, Valeria Mauri, Beatrice Rossetti, Aleksandra Evangelista, Giovanna Bruni, Marco Benazzo, Ferdinando Auricchio, Bice Conti Scientific Reports, 2026 The development of functional scaffolds that support cell differentiation and tissue regeneration remains a major challenge in regenerative medicine. In this study, we designed and fabricated hybrid scaffolds (HS) composed of poly(lactic-co-caprolactone) (PLA-PCL) by integrating 3D extrusion-based printing and electrospinning techniques. Two distinct 3D-printed architectures were explored-aligned filaments and grid patterns-which were subsequently coated with a layer of electrospun nanofibers to better mimic the anisotropic and hierarchical structure of native skeletal muscle tissue. Bone marrow porcine mesenchymal stem cells (p-MSCs) were seeded onto the scaffolds and cultured under myogenic conditions. To evaluate the progression of myogenic differentiation, we assessed the expression of early and late myogenic markers, MyoD and Myogenin respectively, at 14, 21 and 28 days. A comprehensive physical-mechanical characterization was performed, including morphological analysis, porosity measurements, and uniaxial tensile testing. The results demonstrated that the HS provided a biomimetic microenvironment that supported p-MSC attachment, viability, and differentiation. Notably, the HS aligned architecture enhanced the expression of myogenic markers compared to the grid design, suggesting a role of topographical cues in directing lineage commitment. These findings highlight the potential of dual-fabricated PLA-PCL scaffolds as a promising platform for guiding myogenic differentiation and may serve as a foundation for promoting functional skeletal muscle regeneration.
B7-33 modulates fibrotic signalling in hypertrophic scar fibroblasts: An in vitro study supporting a novel therapeutic strategy Martina Tamburriello, Laura Benedetti, Enrica Chiesa, Silvia Pisani, Ida Genta, Bice Conti, Gabriele Ceccarelli, Rossella Dorati Journal of Drug Delivery Science and Technology, 2026 B7-33 is a synthetic single-chain peptide analogue of relaxin (RLX), the latter known for its powerful organ-protective effects however its use is limited by the complex and costly synthesis. B7-33 reproduces the beneficial effects of RLX without triggering pathways often associated with tumorigenesis. Although explored in several therapeutic areas, its potential in hypertrophic scar (HTS) management had not been investigated. This study evaluated, for the first time, the biocompatibility and antifibrotic efficacy of B7-33 on human hypertrophic scar fibroblasts (HSF) and proposed an electrospun peptide-based dressing as a topical strategy for HTS management. Both normal (NHDF) and HSF maintained > 70% viability after 72h exposure to B7-33 (0.03–0.3 μg/mL). Treatment with 298.7 ng/mL B7-33 significantly modulated profibrotic gene expression within 3 days, showing greater Collagen Type I α-1 Chain (COL1A1) downregulation than reference antifibrotic agents. Poly(L-lactide-co-ε-caprolactone) (PLA-PCL, 70:30) solutions above the entanglement concentration (>7.21% w/v ) were electrospun to obtain B7-33-loaded dressings (1.5% w/w ). The resulting fibres (7.06 ± 1.53 μm) enabled controlled release over 72h (96 ± 3.1%). The peptide retained antifibrotic activity after incorporation, as confirmed by qRT-PCR up to 72h. The B7-33 dressing reduced wound closure in HSF, consistent with α-Smooth Muscle Actin (α-SMA) downregulation, without affecting NHDF behaviour. B7-33 dressing exhibited biocompatibility (89.13 ± 6.56%) and hydrophobic behaviour, both in ultrapure water (108.59 ± 2.16°) and simulated wound fluid (111 ± 1.3°), supporting its suitability for exudate management. These findings support B7-33 as an antifibrotic peptide and the feasibility of electrospun dressings for topical HTS management. • Hypertrophic scars arise from persistent TGF-β/Smad pathway activation • B7-33, a Relaxin analogue, shows antifibrotic properties • B7-33 is biocompatible with normal and hypertrophic human fibroblasts (HSF) • B7-33 modulates pro-fibrotic gene expression and cells proliferation in HSF • Advanced Electrospun Dressing enables controlled B7-33 release for scar management
Models for Training in Pediatric Otologic Surgery: A Systematic Review Elena Carlotto, Serena Cirillo, Stefania Marconi, Silvia Pisani, Mirko Bertozzi, Cesare Chiapperini, Simone Mauramati, Marco Benazzo, Pietro Canzi Children, 2026 Background/Objectives: Temporal bone surgery in children is technically challenging due to their smaller anatomical structures, developmental differences, and the closer proximity of critical neurovascular structures. The limited availability of conventional training materials and pediatric cadaveric specimens has led to greater enthusiasm for simulation-based methods. The aim of this systematic review was to identify existing otologic simulation models and evaluate their anatomical accuracy, teaching effectiveness, and supporting evidence. Methods: In accordance with PRISMA guidelines, the PubMed, Embase, Scopus, and Cochrane Library databases were searched for studies reporting simulation tools for pediatric otologic surgery. Articles describing three-dimensional printed (3DP) models, virtual reality (VR) platforms, cadaver specimens, and animal models were included. Studies focusing on children and providing educational outcomes were selected. The extracted data were synthetized and analytically discussed. Results: Thirteen studies met the inclusion criteria: nine on 3DP models and four on VR environments. No research involving cadavers or animals was identified. 3DP models exhibited consistent anatomical accuracy and notable educational advantages. Five studies used surveys for their evaluations, and three relied on expert observer assessments. The studies including validation analyses showed a high correlation between printed models and computed tomography (CT) images. VR systems supported anatomical reconstruction and segmentation tasks, as well as guided simulation exercises. However, most of the research consisted of feasibility studies with limited participant groups. Conclusions: Simulation-based training with 3DP and VR models could be ethical and accurate methods for obtaining relevant skills in pediatric otologic surgery. The reviewed data suggest that these tools may be suitable as a first-line step within an integrated, multimodal training pathway prior to direct patient contact.
Fe3O4@LDH Hybrids as Drug Delivery Systems for Meloxicam: A Physical–Chemical Characterization and In Vitro Study Marcella Bini, Maria Cristina Mozzati, Deborah Fabris, Vittorio Berbenni, Giovanna Bruni, Lauretta Maggi, Silvia Pisani, Valeria Friuli Applied Sciences Switzerland, 2026 Magnetic nanoparticles represent the next-generation drug delivery systems, enabling drug targeting to specific organs without adverse effects on the body and with a controlled release rate. Their strengths are represented by biocompatibility, low cost, and easy drug loading; some drawbacks are aggregation and poor stability in biological media. In the present work, we synthesized magnetic core–shell structures with a magnetite core coated with layered double hydroxides (LDHs) based on Mg2+ or Zn2+ and Al3+ ions and loaded with meloxicam, a poorly water-soluble anti-inflammatory drug. Several syntheses have been attempted to obtain iron oxides based on the only magnetite phase. The combined use of different characterization techniques allowed us to reveal that the best product, showing the crucial room temperature superparamagnetism and a good level of compositional uniformity, was obtained from co-precipitation in nitrogen flow. The next LDH coating was successful, even if the hybrids showed the occurrence of aggregation. The drug was mainly adsorbed onto the LDH surfaces, as shown by the X-ray diffraction and Infrared Spectroscopy techniques. The loaded meloxicam amount was low, but the subsequent release into simulated body fluid could be prolonged for 4 days. Our study provides a proof of concept about the importance of a thorough characterization of the nanocomposite hybrids and their possible use for tricky drugs, such as those of class II of the Biopharmaceutical Classification System.
Liposomal tobramycin and ceftazidime as advanced nanocarriers against Pseudomonas aeruginosa infections Shafia Tufail, Silvia Pisani, Gabriele Trespidi, Rossella Dorati, Ida Genta, Silvia Buroni, Bice Conti International Journal of Pharmaceutics, 2026 The worrisome decline of antibiotic efficacy against Pseudomonas aeruginosa infections emphasizes the urgent need for new antibiotic delivery techniques. Liposomal formulations of tobramycin and ceftazidime were developedand comprehensively studied utilizing an active loading approach. Both formulations showed nanoscale size (<120 nm), low polydispersity index (<0.3), and spherical morphology, as validated by TEM examination. Tobramycin and ceftazidime encapsulation efficiency was in the rank of 20 %. Drug entrapment exploits both temperature change and electrostatic interaction between the charged drugs and lipids. The latter is demonstrated from Zeta potential shifts after drug integration. Tobramycin-loaded vesicles remained negatively charged (-22 mV), while ceftazidime-loaded vesicles remained positively charged (+22 mV), with magnitudes above ± 20 mV ensuring colloidal stability. In-vitro release experiments indicated temperature-dependent behavior, with rapid tobramycin release at 37 °C (∼71 % in 6 h) and sustained ceftazidime release (∼80 % over 48 h). Both showed slower release at 4 °C, indicating storage stability. Antimicrobial tests against P. aeruginosa PAO1 revealed significant improvements: liposomal tobramycin reduced MIC by 2.78-fold, and ceftazidime by 1.72-fold, compared to free antibiotic. Time-killing studies showed the liposomal formulations extended bactericidal action, which significantly reduced bacterial regrowth after 24 h at MIC values. Stability experiments conducted over three weeks revealed good colloidal stability, with minimal increases in size and PDI while retaining an absolute zeta potential. These findings imply that liposomal encapsulation of aminoglycosides and β-lactams increases antibacterial activity, prolongs therapeutic action, and provides stable nanoscale carriers, indicating their potential as advanced treatments against P. aeruginosa infections.
Advances in Fetal Repair of Spina Bifida Integrating Prenatal Surgery, Stem Cells, and Biomaterials Aleksandra Evangelista, Luigi Ruccolo, Valeria Friuli, Marco Benazzo, Bice Conti, Silvia Pisani Biomedicines, 2026 Spina bifida (SB) is a congenital malformation of the central nervous system (CNS), resulting from incomplete closure of the neural tube (NT) during early embryogenesis. Myelomeningocele (MMC), the most severe form of SB, leads to progressive neurological, orthopedic, and urological dysfunctions due to both NT developmental failure and secondary intrauterine injury (“two-hit hypothesis”). Prenatal repair of MMC has progressed considerably since the Management of Myelomeningocele Study (MOMS, 2011) trial, which showed that open fetal surgery can decrease the need for shunting and improve motor function, although it carries significant maternal risks. To address these limitations, minimally invasive techniques have been developed, with the goal of achieving similar benefits for the fetus while reducing maternal morbidity. Recent research has shifted toward regenerative strategies, integrating mesenchymal stem cells (MSCs), bioengineered scaffolds, and cell-derived products to move beyond mere mechanical protection toward true NT repair. Preclinical studies in rodent and ovine models have shown that amniotic- and placenta-derived MSCs exert neuroprotective and immunomodulatory paracrine effects, promoting angiogenesis, modulating inflammation, and supporting tissue regeneration. Minimally invasive, cell-based interventions such as Transamniotic Stem Cell Therapy (TRASCET), in preclinical rodent models, offer the possibility of very early treatment without hysterotomy, although translation remains limited by the lack of large-animal validation and long-term safety data. In parallel, advances in biomaterials, nanostructured scaffolds, and exosome-based therapies reinforce a regenerative paradigm that may improve neurological outcomes and quality of life in affected children. Ongoing translational studies are essential to optimize these approaches and define their safety and efficacy in clinical settings. This review provides an integrated overview of embryological mechanisms, diagnostic strategies, and prenatal therapeutic advances in SB treatment, with emphasis on prenatal repair, fetal surgery and emerging regenerative approaches.
A pilot evaluation of a 3D bioprinted tumor model for assessment of electroporation-based therapies: This article belongs to the Special Issue: Advanced Strategies in 3D Bioprinting for Disease Modelling Franca Scocozza, Silvia Pisani, Aleksandra Evangelista, Ferdinando Auricchio, Michele Conti, Bice Conti, Marco Benazzo International Journal of Bioprinting, 2026 Head and neck squamous cell carcinomas (HNSCCs) are aggressive malignancies with poor prognosis and limited therapeutic options. Electrochemotherapy (ECT), combining short electric pulses with chemotherapeutic agents to enhance intracellular drug uptake, has shown clinical potential but still requires physiologically relevant in vitro models for protocol optimization and mechanistic studies. Here, we introduce a three-dimensional 3D bioprinted in vitro HNSCC model specifically designed for the assessment of electroporation. Structures were fabricated using a composite hydrogel composed of 8% sodium alginate and 4% gelatin (w/w), crosslinked with calcium chloride at concentrations of 0.5%, 1%, and 2%. Uniaxial compression testing confirmed elastic moduli spanning the physiological tumor stiffness range, with the 1% calcium chloride formulation providing optimal mechanical and handling characteristics (42.96 &plusmn; 19.89 kPa). Hypopharyngeal carcinoma FaDu cells (5&times;106/mL) embedded in three-layer structures (thickness: 1.05 mm) maintained 75&ndash;80% viability for up to 21 days and formed tumor-like spheroids (mean diameter: 303 &plusmn; 113 &mu;m), reflecting native tumor architecture. Electroporation with eight pulses at 200 V for 100 &mu;s efficiently permeabilized the cell membrane, as evidenced by the internalization of propidium iodide, while maintaining high cell viability as confirmed by live/dead analysis. Programmed death-ligand 1 expression was preserved and upregulated in 3D spheroids compared to two-dimensional (2D) controls, supporting the platform&rsquo;s relevance for immuno-oncology studies. Compared to other 3D HNSCC models, our system integrates mechanical tuning, electroporation compatibility, and immune-related biomarker expression, enabling functional validation of electric field-mediated intracellular delivery. This proof-of-concept platform demonstrates structural fidelity, long-term cell viability, and high reproducibility, offering a scalable, human-relevant tool for preclinical optimization of ECT and other electrically based therapies, bridging the gap between conventional 2D cultures and complex in vivo models.
Reconstruction after hypopharyngeal cancer surgery: Comparison of outcomes in pectoralis major, radial forearm, and jejunal flap Simone Mauramati, Fabio Sovardi, Irene Herman, Sergio Carnevale, Giulia Bertino, Antonio Occhini, Pietro Canzi, Giuseppe Trisolini, Alberto Luchena, Antonino Maniaci, Emanuele Robiolio, Rebecca Gelli, Eugenio Errico, Silvia Pisani, Marco Benazzo, Patrizia Morbini Jpras Open, 2025 <h2>Abstract</h2> Reconstruction following hypopharyngolaryngectomy remains a challenge for head and neck surgeons. Several techniques have been described. There is no consensus on the primary preference for hypopharyngeal reconstruction. We compared clinical and oncological outcomes between three different types of reconstruction. We identified patients who had undergone reconstruction of the pharyngeal defect using radial forearm flap (RFF) pectoralis major myocutaneous flap (PMMF) or jejunal free flap (JFF). We revised site and stage of the tumor, postoperative complications, surgical margins, morbidity, recurrence, survival and mortality. Resumption of feeding and hospital stay were also recorded. Primary reconstruction (<i>n</i> = 153) was obtained by JFF in 67 patients (43.8 %) PMMF in 47 cases (30.7 %) and RFF in 39 (25.5 %). Flap necrosis was significantly less frequent in PMMF than in the other flap types (p:0.002). PMMF treated patients were significantly more frequently 65-years or older than JFF and RFF ones (p:0.011 and 0.04). There was no difference in the rate of minor complications in the three groups. Overall survival (OS) was significantly shorter in PMMF than in JFF and RFF (p: 0.008 and 0.035 respectively). Parameters associated with shorter OS were PMMF surgery (p:0.0071) pT4 (p:0.0315) positive surgical margins (p:0.0028) neck metastasis (p:0.0376) ENE (p:0.0054) and a history of previous squamous cell carcinoma (p:0.0130). In our experience JFF patients had a lower rate of complications requiring repair surgery and a shorter hospital stay and feeding tube weaning than RFF. We observed a significantly higher rate of major complications requiring surgical repair in RFF than in the other flap types.
Electrospun Bio-Scaffolds for Mesenchymal Stem Cell-Mediated Neural Differentiation: Systematic Review of Advances and Future Directions Luigi Ruccolo, Aleksandra Evangelista, Marco Benazzo, Bice Conti, Silvia Pisani International Journal of Molecular Sciences, 2025 Neural tissue injuries, including spinal cord damage and neurodegenerative diseases, pose a major clinical challenge due to the central nervous system’s limited regenerative capacity. Current treatments focus on stabilization and symptom management rather than functional restoration. Tissue engineering offers new therapeutic perspectives, particularly through the combination of electrospun nanofibrous scaffolds and mesenchymal stem cells (MSCs). Electrospun fibers mimic the neural extracellular matrix, providing topographical and mechanical cues that enhance MSC adhesion, viability, and neural differentiation. MSCs are multipotent stem cells with robust paracrine and immunomodulatory activity, capable of supporting regeneration and, under proper stimuli, acquiring neural-like phenotypes. This systematic review, following the PRISMA 2020 method, analyzes 77 selected articles from the last ten years to assess the potential of electrospun biopolymer scaffolds for MSC-mediated neural repair. We critically examine the scaffold’s composition (synthetic and natural polymers), fiber architecture (alignment and diameter), structural and mechanical properties (porosity and stiffness), and biofunctionalization strategies. The influence of MSC tissue sources (bone marrow, adipose, and dental pulp) on neural differentiation outcomes is also discussed. The results of a literature search show both in vitro and in vivo enhanced neural marker expression, neurite extension, and functional recovery when MSCs are seeded onto optimized electrospun scaffolds. Therefore, integrating stem cell therapy with advanced biomaterials offers a promising route to bridge the gap between neural injury and functional regeneration.
Assessing liposomal nanocarriers for targeted drug delivery through electroporation Silvia Pisani, Godspower Tochukwu Isaac, Rossella Dorati, Ida Genta, Giulia Bertino, Marco Benazzo, Bice Conti International Journal of Pharmaceutics, 2025 Electroporation (EP) is a technique that temporarily increases cell membrane permeability through high-voltage electrical pulses, facilitating the internalization of hydrophilic drugs. When used in clinics, reversible EP offers significant advantages in drug delivery with minimal systemic toxicity, making it a promising approach in cancer therapy (Electrochemotherapy). However, is still challenging to increase therapeutic efficacy, such as increasing the amount of drug internalized by cells after EP. To address these limitations, integrating nanocarriers-particularly liposomes-into EP-based drug delivery strategies has shown great promise. Due to their structural similarity to cell membranes, liposomes can undergo electroporation without causing irreversible cell damage, enabling localized and controlled drug release at targeted sites. This study preliminary evaluates the effectiveness of positively charged gentamicin sulfate loaded liposomes (GS-Lipo) in enhancing gentamicin sulfate uptake through electroporation. The focus is on liposome behavior under EP, drug release, and cellular internalization. The results reveal a strong interplay between liposomes and EP. While EP minimally affects liposome size (sizes lower than 250 nm before and after EP) and PDI, it significantly enhances intracellular uptake and drug release by creating transient pores in liposomal bilayer, facilitating gentamicin diffusion. In vitro uptake studies performed with fluorescent liposomes and GS-Lipo, confirmed superior performance when combined treatment (EP + GS-Lipo) is used. By optimizing electroporation parameters (160 V, 200 V, and 250 V), this study succeeds in maximizing intracellular drug concentration, with the long-term goal of improving therapeutic outcomes, particularly in cancer treatment.
Emerging and re-emerging infectious disease in otorhinolaryngology F. Scasso, G. Ferrari, G. D. De Vincentiis, A. Arosio, S. Bottero, M. Carretti, A. Ciardo, S. Cocuzza, A. Colombo, B. Conti, A. Cordone, M. DE Ciccio, E. Delehaye, L. Della Vecchia, I. De Macina, C. Dentone, P. Di Mauro, R. Dorati, R. Fazio, A. Ferrari, G. Ferrea, S. Giannantonio, I. Genta, M. Giuliani, D. Lucidi, L. Maiolino, G. Marini, P. Marsella, D. Meucci, T. Modena, B. Montemurri, A. Odone, S. Palma, M. Panatta, M. Piemonte, P. Pisani, S. Pisani, L. Prioglio, A. Scorpecci, L. Scotto di Santillo, A. Serra, C. Signorelli, E. Sitzia, M. Tropiano, M. Trozzi, F. M. Tucci, L. Vezzosi, B. Viaggi Acta Otorhinolaryngologica Italica, 2018
