Estela Sasso-Cerri

Correction on: by de Oliveira SA, da Silva AAS, Hinton BT, Gomes GF, Cunha TM, Cerri PS, Sasso-Cerri E. SARS-CoV-2 exploits steroidogenic machinery, triggers lipid metabolism for viral replication and induces immune response in Leydig cells of K18-hACE2 mice. Front Cell Infect Microbiol. 2025 May 27;15:1538461. doi: 10.3389/fcimb.2025.1538461.In the published article, information was missing regarding the ethical documentation and source of the human tissue sample used in Figure 1F.A correction has been made to the Ethics Statement section: "This study utilizes a human testicular tissue photomicrograph (Figure 1) obtained from a histological slide provided by IPC Medicina Diagnóstica (Araraquara, SP, Brazil). Ethics committee approval was not required according to local legislation (Brazilian National Health Council Resolution CNS 466/2012) because the material consists solely of a de-identified (anonymous) archival sample, with no possibility of identifying the donor."The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated.

Background: Pro‐inflammatory cytokines, TNF and IL‐1B, are essential for testicular homeostasis. Diacerein, an anti‐inflammatory drug, inhibits these cytokines, impairing M2 macrophages and Leydig cells (LCs). However, its impact on Sertoli cells (SCs) and M1 (CD68) macrophages remains unknown. Objective: To investigate the impact of diacerein‐induced cytokine downregulation on SC integrity and the testicular immune profile. Materials and Methods: Male rats were treated with diacerein (100 mg/kg) or saline for 30 days. The seminiferous epithelium was evaluated by light and transmission electron microscopy. The number of spermatocytes and Ki‐67+ cells was quantified. SC structural (actin, vimentin, claudin‐1, connexin‐43) and functional (transferrin, Ldha) proteins, and the interstitial immunolocalization of macrophage inhibitory factor (MIF), CD68, CD45, IL‐1B, iNOS, NF‐κB, and karyopherin were assessed, alongside immune response genes ( Tnfa, Il1b, Nfkb1, Nos2, Tgfb1 ) and nitrite levels. TUNEL method was also performed. Results and Discussion: TNF and IL‐1B were downregulated; steroidogenesis and IL‐1B production by LCs decreased, coupled with reduced MIF levels. SCs exhibited electron‐dense cytoplasm, disrupted filaments, and decreased blood–testis barrier proteins. Transferrin and Ldha expression were reduced, underlying reduced spermatocyte counts, arrested proliferation, and germ cell death. TUNEL‐positive apoptotic bodies (probably from LCs) were found within macrophages. Despite increased CD45+ and CD68+ macrophages, a noninflammatory phenotype was observed due to NF‐κB cytoplasmic sequestration and weak karyopherin expression, corroborating Tgfb1 upregulation and reduced Nos2 expression and nitrite levels. Conclusion: Basal cytokines are essential for SC structure and function. Despite testicular damage, diacerein shifted the immune response toward a homeostatic and immunotolerant profile, avoiding a self‐destructive inflammatory process.

We examined SARS-CoV-2 infection in the epididymis of K18-hACE2 mice, focusing on the cauda region. Thirty mice were divided into control (CG), 2-day infection (2D), and 5-day infection (5D) groups. The animals were intranasally infected with SARS-CoV-2. The 5D group showed a significant increase in IL-6, TNF-a, and IL-1b, high mRNA levels of Ifna, Ifnb, and Ifng, and infiltration of CD8⁺ T cells. SARS-CoV-2 directly infects smooth muscle cells, leading to atrophy of the smooth muscle layer. The infection impaired the basement membrane and downregulated blood-epididymis barrier-related genes Ocln and Jam-a. Clear cells exhibited increased apical V-ATPase and immunoexpression of hACE2, spike, and nucleocapsid. Interestingly, the infected groups showed a greater number of juxtaposed clear cells with PCNA- and Ki67-positive nuclei. The findings also revealed a newly identified NF-kB-driven inflammatory-proliferative response in clear cells. By combining in vivo and in silico analysis of a parallel LPS-stimulation RNA-seq dataset, we uncovered significant co-enrichment of the TNF-a_NF-kB pathway alongside cell cycle pathways (G2/M, E2F). The protein-protein interaction network identified key nodes linking the inflammatory NF-kB pathway to the cell cycle machinery. This in silico model was confirmed in vivo, demonstrating co-localization of phosphorylated NF-kB and PCNA in the CC rows. Alterations in spermatozoa, including decreased PNA labeling, ultrastructural changes, and reduced Crisp1 mRNA, were also observed. Therefore, SARS-CoV-2 infection causes structural and functional changes in the epididymal cauda, leading to smooth muscle cell death and basement membrane disruption. Overall, the results suggest that SARS-CoV-2 infection induces dysfunction in the cauda, a microenvironment essential for sperm maturation and storage. Lay Summary SARS-CoV-2, the virus that causes COVID-19, can reach many organs beyond the lungs, including the male reproductive tract. We studied how this virus affects epididymis, a sperm- carrying tube where sperm are mature and stored, in a genetically modified mouse model that expresses the human ACE2 receptor, which allows SARS-CoV-2 infection. We demonstrated that infection triggers a strong local inflammatory response and impairs the muscular wall and spermatozoa. The virus also infects specialized clear cells that control the epididymal fluid pH where sperm are stored. Following infection, clear cell groups showed proliferation and possible acidification of the sperm storage microenvironment. All these combined changes suggest that the normal environment required for sperm maturation is disrupted. Our findings raise concern that severe COVID-19 may have lasting effects on male fertility and highlight the epididymis as an important but overlooked target organ of this infection.

Objective: To longitudinally investigate the effect of non-surgical periodontal therapy (NSPT) on the transcriptional and translational levels of Superoxide Dismutase (SOD) and Insulin Receptor Substrate type 2 (IRS2) in individuals with Type 2 Diabetes Mellitus (T2DM) and Periodontitis (P). Methods: This clinical study was registered at the Brazilian Clinical Trials Registry (ReBEC-RBR-5m3yxmb). Saliva, peripheral blood mononuclear cells (PBMCs), and gingival biopsies were collected from 156 individuals, distributed into five groups, each with at least 30 participants: T2DM_poorly_controlled+P, T2DM_well_controlled+P, T2DM_without_P, Periodontitis, and Control. Systemic levels of messenger RNA (mRNA) of Superoxide Dismutase 1 (SOD1) and IRS2 were measured using real-time polymerase chain reaction at baseline, 90, and 180 days after NSPT. SOD enzymatic activity in Saliva and IRS-2 immunohistochemistry in gingival biopsies were also assessed. Results: Higher SOD1 mRNA levels were observed in Control individuals at baseline. In contrast, higher IRS2 mRNA levels were detected in individuals with Periodontitis at baseline, followed by a significant reduction over time. A significant positive longitudinal correlation was identified between IRS2 and SOD1 gene expression in the groups without T2DM, indicating potential functional interaction between the molecules. Salivary SOD enzymatic activity was lower in individuals from the T2DM_poorly_Controlled+P and T2DM_well_Controlled+P groups. SOD concentration (U/g) normalized to the total protein content was higher in the saliva of individuals with Periodontitis. T2DM+P and Periodontitis groups showed extensive inflammatory infiltrate in the gingival biopsies, with predominant IRS-2 immunopositive cells in the T2DM+P groups, independently of the metabolic control. Conclusions: This study shows that non-surgical periodontal therapy (NSPT) is followed by longitudinal changes in IRS2 and SOD1 expression at the mRNA and protein levels in individuals with T2DM+P (poorly/well controlled) and periodontitis, reinforcing the clinical relevance of periodontal treatment in the systemic context of T2DM.

Objectives: We evaluated the effect of diacerein, an anti-inflammatory drug, on the activity and survival of alveolar bone osteoblasts in rats with periodontitis. Methods: The rats with periodontitis received diacerein (PDG) or saline solution (PSG) for 7, 15 and 30 days. In gingiva samples, Nfkb1 and Bmp2 gene expressions were evaluated, and maxillae were processed for light and transmission electron microscopy. Results: In PDG, the tumor necrosis factor alpha (TNF-α) and interleukin-1 beta (IL-1β) immunoexpression decreased in parallel with the increase in alkaline phosphatase (ALP) and bone area over time. At 15 and 30 days, Nfkb1 expression decreased in PDG compared to PSG, whereas at 30 days, the Bmp2 expression was greater in PDG than in PSG. Immunofluorescence for IL-10, an anti-inflammatory cytokine, was greater in PDG than in PSG at 15 and 30 days. In PSG, the significant increase in the number of TUNEL-positive osteoblasts was accompanied by the presence of osteoblasts with condensed chromatin nuclei or caspase-3-immunolabelled osteoblasts. In contrast, the number of TUNEL-positive osteoblasts was significantly lower in PDG than in PSG specimens at all time points. Conclusions: Therefore, the diacerein-induced TNF-α and IL-1β inhibitory effect caused Nfkb1 downregulation and, hence, prevented apoptosis in osteoblasts. The increased ALP activity and IL-10 in PDG indicate that diacerein mitigates periodontitis impact on alveolar bone in rat molars.

During the COVID-19 pandemic, the prevalence of death in men was higher than in women. Using transgenic mice expressing the human angiotensin-converting enzyme 2 (hACE2), we demonstrated that SARS-CoV-2 infects Leydig cells and uses its steroidogenic machinery for replication. This study investigates the impact of SARS-CoV-2 in the seminiferous epithelium of K18-hACE2 mice, focusing on the immune response, junctional proteins, and spermatogenesis. The seminiferous tubules (STs) and epithelial (EA) areas were measured. The number of Sertoli cells (SCs), spermatocytes, and damaged ST was quantified. Ultrastructural analysis was performed under transmission electron microscopy. Angiotensin II levels and immunolocalization of hACE2, spike, and nucleocapsid were evaluated. TUNEL and immunoreactions for Ki-67, TNF-α, INF-γ, iNOS, NF-κB, and Conexin-43 were performed and correlated with Jam-α, Stat1, Stat3, and iNOS expressions. hACE2, spike, and nucleocapsid immunolabeling were detected in the epithelium along with high angiotensin II levels in the infected mice. The infection caused a significant reduction in ST, EA, spermatocytes, SCs, Ki-67+ cells, Cx43 immunoexpression, and Jam-a expression. In the epithelium, TNF-α, IFN-γ, iNOS, and nuclear NF-κB immunolabeling increased along with Stat1 upregulation. These findings, combined with the increased epithelial hACE2 and high angiotensin II levels, confirm epithelial responsiveness to the infection and explain the spermatogenic failure and impaired junctional proteins. The presence of viral particles, increased TNF-α immunolabeling, and apoptotic features in Sertoli cells suggests that these sustentacular cells are targets for viral infection in the epithelium, and, due to their extensive projections and ability to phagocytize dying infected germ cells, they may disseminate the viruses throughout the epithelium.

This study investigated how estrogen deficiency and streptozotocin‐induced diabetes mellitus (DM) affect rat gingival mucosa, focusing on autophagy and NLRP3 inflammasome expression. Twenty rats were ovariectomized (OVX) or SHAM‐operated (SHAM) and divided into SHAM, OVX, SHAM‐DM, and OVX‐DM groups. After 7 weeks, rats were euthanized, and maxillae containing gingival mucosa were processed for histology and immunohistochemistry for autophagy (Beclin‐1, LC3B) and inflammasome (NLRP3, IL‐1β) markers. Publicly available data were analyzed to assess these markers in gingival tissue from periodontitis patients. Epithelial and stratum corneum thickness increased in OVX, SHAM‐DM, and OVX‐DM groups. SHAM‐DM and OVX‐DM rats showed more keratinocytes with large vesicular nuclei and prominent nucleoli. The highest numbers of mast cells, especially degranulated ones, occurred in OVX‐DM. Increased Beclin‐1, LC3B, NLRP3, and IL‐1β expression was detected in the lamina propria and gingival epithelium, particularly in OVX‐DM. These markers showed positive correlations. Bioinformatics revealed increased LC3B, NLRP3, and IL1B transcription in periodontitis patients’ gingiva. Positive correlations were found between BECN1 and LC3B , BECN1 and NLRP3 , LC3B and NLRP3 , and NLRP3 and IL1B ; BECN1 and IL1B correlated negatively. These findings suggest that increased autophagy and NLRP3 inflammasome activation contribute to gingival mucosa damage in estrogen‐deficient diabetic rats and in periodontitis patients.

The aim of this study was to evaluate whether the experimental sealers, composed of tricalcium silicate (CaSi) and CaSi supplemented with calcium hypochlorite (CaSi‐H), stimulate tissue repair. The tissue response of experimental materials was compared with commercially available materials Bio‐C sealer (BC sealer) and BioRoot (BROOT). Polyethylene tubes filled with materials or left empty (controls) were implanted in rats for 7, 15, 30 and 60 days. The number of fibroblasts, mast cells, collagen content, interleukin‐10 (IL‐10) and fibroblast growth factor‐1 (FGF‐1) immunoexpression were evaluated. Data were analysed with two‐way anova and Tukey's test, and linear regression. The fibroblast–FGF correlation was estimated using the Pearson correlation coefficient. In all groups, the number of fibroblasts and collagen content increased significantly over time. At all‐time points, CaSi‐H and control groups showed the highest values of fibroblasts, whereas the lowest values were observed in BROOT. At 60 days, no significant differences were detected among CaSi, CaSi‐H, BC sealer and controls. At all‐time points, the capsules of CaSi and CaSi‐H exhibited the highest values of FGF‐1 and IL‐10 immunoexpression. CaSi and CaSi‐H induced the formation of fibrous capsules, indicating that these sealers stimulate connective tissue repair.

This study aimed to develop and validate a dual murine model integrating a high-fat diet (HFD) and a single streptozotocin (STZ) dose to induce diabetes mellitus (DM), alongside periodontitis (Perio) induced by ligature placement and oral inoculation with Porphyromonas gingivalis (P. gingivalis). The goal was to mimic human pathological conditions, creating a physiologically relevant environment to study the interplay between DM and Perio. A total of 128 six-week-old male C57BL/6J mice were randomly divided into four groups: Control, DM, Perio, and DM-P. DM was induced by HFD and STZ injection, and Perio by ligature placement and P. gingivalis infection. Evaluations occurred at baseline and days 7, 14, and 21. Alveolar bone loss was assessed by micro-computed tomography, and inflammation was examined histologically. DM mice showed elevated glucose levels and insulin resistance. Perio and DM-P groups experienced significant bone loss compared with Control and DM groups. The morphometric analysis revealed abundant inflammatory cells and reduced collagen fibers in Perio and DM-P groups, especially at day 7. This dual murine model successfully replicated the key features of DM and Perio, maintaining overall health of the animals, and good tolerability by those subjects to the stress of both interventional procedures.