Cell Biology, General Biochemistry, Genetics and Molecular Biology, Health, Toxicology and Mutagenesis, Multidisciplinary
22
Scopus Publications
Scopus Publications
Acetamiprid exerts sex-specific effects on adipose tissue of subjects with severe obesity Giulia Zanchi, Alessia Tammaro, Valentina Monteleone, Rosaria Varì, Carmela Santangelo, et al. Frontiers in Toxicology, 2026 Background Neonicotinoid pesticides, including acetamiprid (ACE), are widely used in agriculture and pose increasing concerns due to their persistence in the environment and potential human exposure mainly through diet. Available evidence suggests that ACE may disrupt adipocyte function and promote metabolic dysfunctions such as obesity; however, there is limited research on how ACE negatively affects adipose tissue (AT) in men and women. This study utilizes an ex vivo translational model to clarify the sex-specific effects of ACE on AT metabolic and inflammatory profile of a vulnerable human substrate, such as the visceral AT of subjects with severe obesity. Methods Twenty-four subjects with severe obesity (11 men and 13 women) undergoing bariatric surgery were recruited from St. Andrea University Hospital (Rome, Italy). Visceral adipose tissue biopsies were collected and either treated with ACE or left untreated for further gene and protein expression analysis by RT-qPCR and Western blot, respectively. In addition, adipocytokines secretion, reactive oxygen species production, and free fatty acid release were measured in adipose tissue culture media using commercial or in house assays. Results Our findings demonstrate that ACE induces distinct sex-dependent alterations in lipid metabolism, Adipokines regulation, and inflammatory pathways. Specifically, it significantly lowers PPARγ gene expression but raises protein levels, particularly in men. Free fatty acid release increases and Hormone Sensitive Lipase (HSL) drops in both sexes, while Lipoprotein Lipase (LPL) decreases only in women. ACE also promotes inflammation mainly in women, increasing TNF-α, NF-κB, and reactive oxygen species. Conclusion These results show that the neonicotinoid ACE worsens AT dysfunction via inflammatory and metabolic pathways in a sex-specific way, likely leading to different risks of obesity-related complications. Overall, these findings provide a mechanistic basis for understanding the toxicological risk of neonicotinoids, highlighting the importance of sex-specific assessment in evaluating metabolic risks of environmental pesticide exposure.
Screening of Estrogenic and Antiestrogenic Effects of Estradiol, Bisphenol A, and Fulvestrant Using 2D and 3D Breast Cancer Cell Systems With a Luciferase Reporter Gene Assay Valentina Monteleone, Katia Barbaro, Innocenzi Elisa, Marcoccia Daniele, Sofia Bisegna, et al. Journal of Applied Toxicology, 2026 Endocrine‐disrupting chemicals (EDCs) like bisphenol A (BPA) pose health risks by interfering with hormones. This study develops and utilizes in vitro 2D and 3D cell models to evaluate the estrogenic and antiestrogenic properties of compounds. Human breast cancer cell lines T47D and MCF7, stably transfected with a luciferase reporter gene (ERE‐LUC), were first compared in 2D. Due to the significantly higher sensitivity and responsiveness observed in the T47D line during preliminary 2D screenings, this cell line was exclusively selected for the development of the 3D spheroid model. Cells were treated with 17 β ‐estradiol (E2), BPA, and Fulvestrant (FUL) to assess cell viability and luciferase activity. In 2D models, T47D ERE‐LUC cells showed higher responsiveness than MCF7 ERE‐LUC, which failed to show significant luciferase induction with E2. In the 3D T47D model, cells exhibited significant and robust changes in luciferase activity in response to E2 and BPA, highlighting the enhanced fidelity of 3D cultures in replicating tissue conditions compared to their 2D counterparts. The study highlights the effectiveness of 3D models over 2D in evaluating estrogenic activity. Specifically, the 3D T47D ERE‐LUC system serves as a superior, sensitive, and reliable platform for screening EDCs, offering benefits in cost, data speed, and reduced in vivo reliance.
Gender differences in eating habits and lifestyles of young students: observational evaluation from MaestraNatura program Annalisa Silenzi, Letizia Giona, Giulia Zanchi, Alessia Tammaro, Valentina Monteleone, et al. Frontiers in Nutrition, 2025 Introduction Unhealthy diet and sedentary life represent the main risk factors for chronic NonCommunicable Diseases. Investing in children’s health education leads to benefits for their whole lifespan and the whole of society. MaestraNatura (MN) is an education program addressed to increase knowledge and skills about food and nutrition in students from primary and first-level secondary school. The main objective of the present paper was to evaluate eating habits and the adherence to the principles of the Italian dietary guidelines (IDG) of 8–14 years old children to identify possible influences of gender and parents on their eating choices and behaviors. Therefore, the potential benefits of MN on promoting changes in lifestyle and dietary patterns were also evaluated. Methods Questionnaires on eating habits and physical activity were completed by 1,037 students and 220 parents from primary and secondary schools from 6 Italian regions. One hundred fifty-eight questionnaires were filled in by a subgroup of secondary schools’ students before and after two consecutive years of MN program. Differences between groups in the responses to questions were assessed by Chi-squared ( χ 2 ) tests. Differences in the IDG adherence score were assessed by analysis of variance (ANOVA), with gender (males, M vs. females, F) and/or type of school as between-subjects’ factors, as appropriate. Results The questionnaires showed an average degree of adherence to the principles of the IDG for children with a similar distribution in M and F. Analysis of individual responses revealed differences between F and M in daily consumption of vegetables (F > M), water (M > F), and in weekly consumption of nuts (F > M) and commercial cookies (M > F). Furthermore, M are more engaged in physical exercise compared to F. Results from a subgroup of students attending MN for two consecutive years showed increases in the consumption of water, vegetables, fruit and legumes, and a decrease in the intake of commercial snacks, as well as an increase in physical activity. Conclusion Italian children evidence gender differences in eating behaviors. Specific nutrition programs are needed to raise children’s awareness of the importance of a healthy lifestyle and to correct eating habits.
Evaluation of estrogenic and anti-estrogenic activity of endocrine disruptors using breast cancer spheroids: a comparative study of T47D and MCF7 cell lines in 2D and 3D models Katia Barbaro, Elisa Innocenzi, Valentina Monteleone, Daniele Marcoccia, Annalisa Altigeri, et al. Frontiers in Toxicology, 2025 IntroductionThe estrogenic and anti-estrogenic effects of endocrine disruptors were examined in vitro using two-dimensional 2D and three-dimensional 3D estrogen receptor-positive T47D and MCF7 human breast cancer cells.MethodsThe in vitro model system was used to test the plasticizer Bisphenol A (BPA), a known endocrine disruptor (EDs) with estrogen-like action, aga inst 17β-Estradiol (E2), the endogenous nuclear estrogen receptor (nERs) ligand, and the anti-estrogenic drug Fulvestrant (FUL). Spheroid formation and gene expression of estrogen-regulated markers (pS2 and TGFβ3) both in 2D and 3D cultures were used to establish the dose-dependent cellular effects of these substances, evaluated cell viability either by separately treating with the individual substances or in co-treatment.ResultsBPA exhibited a dose-dependent estrogenic activity in both 2D and 3D cultures, significantly influencing cell proliferation and gene expression of estrogen-regulated markers (pS2 and TGFβ3). In contrast, FUL displayed anti-estrogenic properties, effectively inhibiting the proliferative effects of E2, thereby highlighting the complex interactions between these compounds and the nERs pathways in human breast cancer cells.DiscussionOur findings indicate that E2 and BPA significantly increase pS2 expression while decreasing TGFβ3, and that FUL co-treatment reverses these effects. Therefore, the in vitro model system could serve to observe the cell-mediated effects caused by the interaction of EDs with nERs. Through the use of these in vitro model systems - 2D and especially 3D, the latter of which allow better emulation of complex physiological and pathological processes occurring in vivo, the effects caused by EDs on nERs pathways can be detected and studied under various conditions. This approach performs as a preliminary screening tool to identify estrogenic substances, offering the potential to reduce reliance on in vivo experiments and contributing to improved environmental and health risk assessments.
Silver Containing Antimicrobial Coatings on Innovative Ti-30Nb-5Mo β-Alloy Prepared by Micro-Arc Oxidation for Biomedical Implant Applications Giovana Collombaro Cardoso, Katia Barbaro, Pedro Akira Bazaglia Kuroda, Angela De Bonis, Roberto Teghil, et al. Coatings, 2024 Micro-arc oxidation (MAO) is a versatile surface-modification method that promotes higher wear and corrosion resistance, osseointegration, and biological activity to titanium alloys’ surfaces. This study aimed to modify the surface of a recently developed metastable β Ti alloy, which exhibits more favorable mechanical properties for implant applications compared to some commercial Ti alloys, by incorporating Ag into the coatings to introduce a bactericidal function to the surface. The Ti-30Nb-5Mo alloy, with lower elastic modulus, was treated by the MAO method using electrolyte solutions containing calcium acetate, magnesium acetate, β-glycerol phosphate, and varied concentrations of silver nitrate (1.5 mM, 2.5 mM, and 3.5 mM). With an increase in the concentration of silver ions in the electrolyte, the galvanostatic period during the MAO process decreased from 1.7 s to 0.5 s. The Ca/P ratio increased from 0.72 up to 1.36. X-ray diffraction showed that the MAO coatings were formed by rutile and anatase TiO2 main phases and calcium phosphates. X-ray photoelectron spectroscopy analysis detected the presence of amorphous Nb2O5, CaCO3, and MgCO3, and metallic and oxide forms of Ag. The increase in Ag in the electrolyte decreased the coating thickness (from 14.2 μm down to 10.0 μm), increased the contact angle (from 37.6° up to 57.4°), and slightly increased roughness (from 0.64 μm up to 0.79 μm). The maximum inhibition of Enterococcus faecalis, Pseudomonas aeruginosa, and Candida albicans strains growth was of 43%, 43%, and 61%, respectively. The Ag did not negatively affect the differentiation of adipose-tissue-derived mesenchymal stem cells. Therefore, the treatment of the surface of the innovative Ti-30Nb-5Mo alloy by the MAO method was effective in producing a noncytotoxic porous coating with bactericidal properties and improved osseointegration capabilities.
Regenerative treatment of canine osteogenic lesions with Platelet-Rich Plasma and hydroxyapatite: a case report Katia Barbaro, Giorgio Marconi, Elisa Innocenzi, Annalisa Altigeri, Alessia Zepparoni, et al. Frontiers in Veterinary Science, 2024 IntroductionThis study examined the efficacy of a therapy based on a combination of Platelet Rich Plasma and hydroxyapatite nanoparticles in a severe clinical case involving a young Rottweiler with a complex spiral fracture of the tibia.MethodFollowing a worsening of the lesion after traditional surgical intervention, the subject was treated with the combined therapy. X-rays were taken at the following stages: immediately post-surgery, four weeks post-surgery, and 10 days post-treatment. Fracture gap and callus density measurements were obtained using ImageJ analysis, allowing for a detailed quantitative assessment of bone regeneration over time.ResultsPost-operative radiographs indicated a clinical worsening of the fracture, revealing an increased fracture gap due to bone loss. However, significant improvements were observed ten days following the treatment, with a marked reduction in fracture gaps and increased callus density. These results demonstrated a notable acceleration in bone healing and callus formation compared to typical recovery times for similar lesions.ConclusionThe method showed potential for enhancing osteogenic regeneration, facilitating faster healing of serious orthopedic injuries compared to traditional methods.
Arthroscopic Treatment of a Subchondral Bone Cyst via Stem Cells Application: A Case Study in Equine Model and Outcomes Fernando Canonici, Daniele Marcoccia, Pamela Bonini, Valentina Monteleone, Elisa Innocenzi, et al. Biomedicines, 2023 Subchondral bone cysts in horses represent one of the main causes of lameness that can occur in different anatomical locations. The study describes the treatment in regenerative therapy of the intracystic implantation of adipose tissue mesenchymal stromal cells (AMSCs) included in platelet-rich plasma (PRP). The ability of AMSCs to differentiate in osteogenic cells was tested in vitro and in vivo. Given the aim to investigate the application of AMSCs in bone defects and orthopedic pathologies in horses, a four-year-old male thoroughbred racing horse that had never raced before was treated for lameness of the left hind leg caused by a cyst of the medial femoral condyle. The horse underwent a new surgery performed with an arthroscopic approach in which the cystic cavity was filled with AMSCs contained in the PRP. Radiographs were taken 3, 5, and 10 months after the surgery to assess the development of newly regenerated bone tissue in the gap left by the cyst. Twelve months after the operation and after six months of regular daily training, the horse did not show any symptoms of lameness and started a racing career. According to the study, the use of AMSCs and PRP suggests promising benefits for treating subchondral bone cysts.
Articular Cartilage Regeneration by Hyaline Chondrocytes: A Case Study in Equine Model and Outcomes Fernando Canonici, Cristiano Cocumelli, Antonella Cersini, Daniele Marcoccia, Alessia Zepparoni, et al. Biomedicines, 2023 Cartilage injury defects in animals and humans result in the development of osteoarthritis and the progression of joint deterioration. Cell isolation from equine hyaline cartilage and evaluation of their ability to repair equine joint cartilage injuries establish a new experimental protocol for an alternative approach to osteochondral lesions treatment. Chondrocytes (CCs), isolated from the autologous cartilage of the trachea, grown in the laboratory, and subsequently arthroscopically implanted into the lesion site, were used to regenerate a chondral lesion of the carpal joint of a horse. Biopsies of the treated cartilage taken after 8 and 13 months of implantation for histological and immunohistochemical evaluation of the tissue demonstrate that the tissue was still immature 8 months after implantation, while at 13 months it was organized almost similarly to the original hyaline cartilage. Finally, a tissue perfectly comparable to native articular cartilage was detected 24 months after implantation. Histological investigations demonstrate the progressive maturation of the hyaline cartilage at the site of the lesion. The hyaline type of tracheal cartilage, used as a source of CCs, allows for the repair of joint cartilage injuries through the neosynthesis of hyaline cartilage that presents characteristics identical to the articular cartilage of the original tissue.
