Enrico Gugliandolo

Joint inflammation and extracellular matrix degradation are major contributors to the development of equine osteoarthritis and reduced athletic performance. Alpha-2-macroglobulin is a naturally occurring broad-spectrum inhibitor of proteases and inflammatory mediators that has emerged as a promising orthobiologic therapy. This study investigated the anti-inflammatory and matrix-protective effects of alpha-2-macroglobulin-enriched plasma in complementary in vitro and ex vivo equine models. Primary equine chondrocytes and synoviocytes isolated from healthy horses were stimulated with interleukin-1β or lipopolysaccharide and treated with alpha-2-macroglobulin-enriched plasma or matched platelet-poor plasma. In parallel, an equine tendon explant model challenged with collagenase was used to evaluate extracellular matrix preservation. Alpha-2-macroglobulin-enriched plasma significantly reduced the production of tumor necrosis factor-α, interleukin-6, and matrix metalloproteinase-13, downregulated interleukin-1β gene expression, and partially restored collagen type II expression in inflamed joint cells compared with platelet-poor plasma. In tendon explants, treatment preserved tissue architecture, reduced collagen degradation, and improved histological indices of matrix integrity under proteolytic challenge. These findings demonstrate that alpha-2-macroglobulin-enriched plasma exerts anti-inflammatory, anti-catabolic, and matrix-protective effects in experimental equine tissues. The results provide mechanistic support for the development of alpha-2-macroglobulin-based orthobiologic therapies as a complementary strategy for the management of synovitis and early osteoarthritis in horses.

The journal retracts the article titled “Aggravation of TGFβ1-Smad Pathway and Autoimmune Myocarditis by Fungicide (Tebuconazole) Exposure”

Chronic joint disease is a major cause of lameness and reduced performance in sport horses and is characterized by persistent synovial inflammation and protease-mediated matrix degradation. This exploratory prospective pilot study investigated clinical outcomes and synovial biomarker changes following intra-articular administration of an α-2-macroglobulin plasma-derived preparation. Twenty client-owned sport horses were observed in the treatment group (n = 10) or a comparison group (n = 10) and monitored for up to 180 days under field conditions. Clinical outcomes were assessed longitudinally, while synovial fluid was analyzed at baseline and 30 days post-treatment only in treated horses. Mixed-effects analysis showed significant group × time interactions for American Association of Equine Practitioners (AAEP) lameness score, flexion test response, and joint effusion. Treated horses showed early and sustained improvement in clinical scores, whereas minimal changes were observed in the comparison group. At 30 days, treated horses exhibited consistent within-subject reductions in synovial total protein, total nucleated cell count, polynuclear cell percentage, pro-inflammatory cytokines (PGE₂, TNF-α, IL-6, IL-1β), matrix metalloproteinases (MMP-9, MMP-13), sulphated glycosaminoglycans, and neurogenic mediators (NGF, Substance P). These findings indicate a coherent pattern of clinical improvement associated with parallel changes in synovial biomarkers in treated horses. However, as longitudinal biomarker data were not collected in the comparison group, these observations should be interpreted as exploratory and do not establish causality. The observed findings support the rationale for further investigation of protease-targeted approaches in equine joint disease.

Background: Orthobiologics such as platelet‐rich plasma (PRP) and alpha‐2‐macroglobulin (A2MG) are increasingly used for equine musculoskeletal injuries. However, their composition and safety with repeated allogeneic intra‐articular administration remain poorly investigated. Objectives: To characterise orthobiological preparations with or without 1 h incubation at 37°C (PRP, PRP + Inc, A2MG, A2MG + Inc) and evaluate immunological safety following repeated intra‐articular administration (autologous and allogeneic) of PRP combined with A2MG + Inc in healthy horses. Study Design: In vivo experiments. Methods: This was a two‐phase study with biochemical characterisation followed by repeated intra‐articular injections in four healthy horses. Blood was processed to produce PRP or A2MG, with or without incubation, and characterised using haematology, biochemistry and enzyme‐linked immunosorbent assay. For immunogenic assessment, healthy horses received three intra‐articular injections of combined PRP and A2MG + Inc at 2‐week intervals (Days 0, 14 and 28) (allogeneic n = 3, autologous n = 1). Blood and synovial fluid were collected on Days 0, 14, 28 and 42. Blood was analysed for haematology, biochemistry, CD4/CD8 ratio and SAA, while synovial fluid was analysed for leukocyte counts and immunoglobulins. Results: Biochemical characterisation demonstrated effective removal of blood cells from all orthobiologics. PRP‐based formulations were enriched in platelets and growth factors, whereas A2MG preparations had higher alpha‐2‐macroglobulin and lower growth factor levels. Repeated intra‐articular administration of PRP and A2MG + Inc caused no clinically relevant changes in systemic or synovial immune cells, lymphocyte subsets, or acute‐phase markers, indicating no immunological activation. Main Limitations: Small sample sizes, the limited scope of characterised compositions and the use of combined PRP and A2MG + Inc for which only individual components were characterised. Conclusions: PRP may promote repair via growth factors, and A2MG is proposed to modulate immunity and catabolism. Repeated intra‐articular injections of PRP with A2MG + Inc caused no systemic or synovial immune response, with no differences between autologous and allogeneic use.

Background: Canine mammary carcinoma (CMC) is characterised by a chronic inflammatory microenvironment resembling human breast cancer; however, the upstream regulatory mechanisms driving this phenotype remain unclear. The endocannabinoid system (ECS) has emerged as a potential modulator of inflammation and tumour biology. This study investigated the role of the ECS in CMC and evaluated the anti‐inflammatory effects of cannabidiol (CBD). Methods: Primary cell cultures were established from surgically excised CMC tissues, with matched normal mammary epithelium used as controls. Basal mRNA expression of ECS‐related receptors (CB1, CB2, transient receptor potential vanilloid 1 [TRPV1], G‐protein‐coupled receptor 55 [GPR55] and peroxisome proliferator‐activated receptor alpha [PPAR‐α]) and inflammatory mediators (COX‐1, COX‐2, interleukin [IL]‐4, IL‐6, IL‐33, IL‐17A, tumour necrosis factor‐alpha [TNF‐α] and LCN2) was assessed by reverse transcription quantitative polymerase chain reaction. Cytokine secretion (IL‐6, IL‐8, TNF‐α and IL‐17A) was quantified by enzyme‐linked immunosorbent assay. Cell viability assays were performed to determine the 24‐h IC ₅₀ of CBD (32 µM), and sub‐cytotoxic concentrations (3, 10 and 20 µM) were subsequently applied for 24 h. Results: Canine mammary carcinoma‐derived cells exhibited significant overexpression of ECS receptors (CB1, CB2, TRPV1, GPR55 and PPAR‐α) compared to normal controls. These cells also showed increased secretion of pro‐inflammatory cytokines, including IL‐6, IL‐8, TNF‐α and IL‐17A. Treatment with CBD at 10–20 µM significantly downregulated key inflammatory genes, particularly COX‐2, IL‐6 and TNF‐α, and reduced corresponding cytokine release without compromising cell viability. Conclusion: The ECS is upregulated in CMC and appears to contribute to the inflammatory tumour microenvironment. Cannabidiol effectively attenuates this inflammatory phenotype at sub‐cytotoxic concentrations, supporting its potential as a therapeutic agent in CMC.

The journal retracts the article titled “Ultramicronized Palmitoylethanolamide and Paracetamol, a New Association to Relieve Hyperalgesia and Pain in a Sciatic Nerve Injury Model in Rat”

The journal retracts the article “Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Inhibitor as a Novel Therapeutic Tool for Lung Injury”

Fumonisin B1 (FB1), a mycotoxin commonly found in contaminated food and feed, has been increasingly implicated in neurotoxicity, although its mechanisms remain poorly understood. This study investigates the neurotoxic potential of FB1 in human SH‐SY5Y neuroblastoma cells, both undifferentiated and RA‐differentiated, and evaluates the protective effect of bromelain, a natural proteolytic enzyme with antioxidant and anti‐inflammatory properties. Cells were exposed to 50 μM FB1 for 24 h, with or without co‐treatment with bromelain (10 or 50 μg/mL). FB1 significantly reduced cell viability and triggered reactive oxygen species (ROS) production, mitochondrial membrane depolarization, lipid peroxidation, inflammatory cytokine release (IL‐6 and TNF‐α), and apoptosis, particularly in differentiated cells. Bromelain co‐treatment attenuated these effects in a dose‐dependent manner, preserving mitochondrial function, reducing oxidative and inflammatory markers, and lowering apoptotic cell death. These findings emphasize the neurotoxic risk posed by FB1 and highlight bromelain as a promising multi‐target protective agent. Identifying effective countermeasures against environmental neurotoxins is crucial for public health and disease prevention.

Background: Bovine mastitis is inflammation of the mammary gland mainly caused by bacterial infections, with relevant economic costs and implications related to antibiotic resistance. In light of the increasing demand for sustainable therapies, this study evaluated the anti‐inflammatory and antioxidant effects of snail secretion filtrate (SSF) from the species Helix Aspersa Muller in an in vitro model of bovine mastitis. Methods: Bovine mammary cells (MAC‐T) stimulated with lipopolysaccharide (LPS) were used to induce the inflammatory process. The cells were then treated with SSF to analyse its effects on oxidative stress, production of inflammatory cytokines (tumour necrosis factor‐α, interleukin‐6 and interleukin‐1β), expression of enzymes associated with inflammation such as inducible nitric oxide (iNOS) and cyclo‐oxygenase‐2 (COX‐2) and cytoprotection as heme‐oxygenase 1 (HO‐1). Results: Snail secretion filtrate significantly reduced the levels of reactive oxygen species and the production of pro‐inflammatory cytokines induced by LPS. Furthermore, it positively modulated the expression of iNOS and COX‐2, reducing their levels and increasing the expression of HO‐1, suggesting a cytoprotective action. Conclusions: Snail secretion filtrate was effective in reducing inflammation and oxidative stress in an in vitro model of bovine mastitis. The results suggest the potential use of SSF as a natural and sustainable remedy, in line with the One Health approach.

This study examines the occurrence of bacteria resistant to antibiotics and heavy metals in Terra Nova Bay, a coastal area of the Ross Sea in Antarctica that is increasingly recognised as vulnerable to human influence. During the 37th Italian Antarctic Expedition (2021–2022), researchers collected seawater, sediment, and fish samples from the notothenioid species Trematomus bernacchii to evaluate microbial resistance in an environment once considered largely pristine. Fifty heterotrophic bacterial isolates were obtained and tested against twenty-eight antibiotics, revealing a notable presence of multidrug resistance. These multidrug-resistant isolates were then assessed for their tolerance to eight heavy metal salts to understand whether resistance traits extended beyond antimicrobials. Twelve isolates showing resistance to both antibiotics and metals were selected for further genetic screening, targeting key resistance genes linked to tetracycline, vancomycin, sulphonamides, and other antimicrobial classes. The detection of multiple resistance genes in genera such as Pseudomonas, Pseudoalteromonas, and Psychrobacter indicates that both natural selective pressures and local, human-related contamination may be shaping resistance patterns in this region. Overall, the study demonstrates that even remote Antarctic marine ecosystems can host bacteria with complex resistance profiles. While these ecosystems are largely isolated, human activities such as scientific research, tourism, and the introduction of pollutants may contribute to the dissemination of antibiotic resistance genes, raising important ecological and potential public health considerations regarding the spread of resistance in polar environments.