Effect of tetrahedral framework nucleic acids on the reconstruction of tendon-to-bone injuries after rotator cuff tears.

Clinicians and researchers have always faced challenges in performing surgery for rotator cuff tears (RCT) due to the intricate nature of the tendon-bone gradient and the limited long-term effectiveness. At the same time, the occurrence of an inflammatory microenvironment further aggravates tissue damage, which has a negative impact on the regeneration process of mesenchymal stem cells (MSCs) and eventually leads to the production of scar tissue. Tetrahedral framework nucleic acids (tFNAs), novel nanomaterials, have shown great potential in biomedicine due to their strong biocompatibility, excellent cellular internalisation ability, and unparalleled programmability. The objective of this research was to examine if tFNAs have a positive effect on regeneration after RCTs. Experiments conducted in a controlled environment demonstrated that tFNAs hindered the assembly of inflammasomes in macrophages, resulting in a decrease in the release of inflammatory factors. Next, tFNAs were shown to exert a protective effect on the osteogenic and chondrogenic differentiation of bone marrow MSCs under inflammatory conditions. The in vitro results also demonstrated the regulatory effect of tFNAs on tendon-related protein expression levels in tenocytes after inflammatory stimulation. Finally, intra-articular injection of tFNAs into a rat RCT model showed that tFNAs improved tendon-to-bone healing, suggesting that tFNAs may be promising tendon-to-bone protective agents for the treatment of RCTs.

The infection of Aeromonas hydrophila activated Multiple programmed cell death pathways in red blood cells of Clarias fuscus.

In contrast to mammalian red blood cells (RBCs), Osteichthyes RBCs contain a nucleus and organelles, suggesting the involvement of more intricate mechanisms, particularly in the context of ferroptosis. In this study, we utilized RBCs from Clarias fuscus (referred to as Cf-RBCs) as a model system. We conducted RNA-seq analysis to quantify gene expression levels in Cf-RBCs after exposure to both Aeromonas hydrophila and lipopolysaccharides. Our analysis unveiled 1326 differentially expressed genes (DEGs) in Cf-RBCs following 4 h of incubation with A. hydrophila, comprising 715 and 611 genes with upregulated and downregulated expression, respectively. These DEGs were further categorized into functional clusters: 292 related to cellular processes, 241 involved in environmental information processing, 272 associated with genetic information processing, and 399 linked to organismal systems. Additionally, notable changes were observed in genes associated with the autophagy pathway at 4 h, and alterations in the ferroptosis pathway were observed at 8 h following A. hydrophila incubation. To validate these findings, we assessed the expression of cytokines (DMT1, TFR1, LC3, and GSS). All selected genes were significantly upregulated after exposure to A. hydrophila. Using flow cytometry, we evaluated the extent of ferroptosis, and the group incubated with A. hydrophila for 8 h exhibited higher levels of lipid peroxidation compared with the 4-h incubation group, even under baseline conditions. An evaluation of the glutathione redox system through GSSG/GSH ratios indicated an increased ratio in Cf-RBCs after exposure to A. hydrophila. In summary, our data suggest that A. hydrophila may induce ferroptosis in Cf-RBCs, potentially by triggering the cystine/glutamate antiporter system (system XC-), while Cf-RBCs counteract ferroptosis through the regulation of the glutathione redox system. These findings contribute to our understanding of the iron overload mechanism in Osteichthyes RBCs, provide insights into the management of bacterial diseases in Clarias fuscus, and offer potential strategies to mitigate economic losses in aquaculture.

Characterization of effects of chitooligosaccharide monomer addition on immunomodulatory activity in macrophages.

The present study aimed to investigate the effects of five chitooligosaccharide monomers of different molecular weights on immunomodulatory activity in macrophage-like RAW264.7 cells. The incubation of various chitooligosaccharide monomers enhanced phagocytosis and pinocytosis activity toward Staphylococcus aureus and Escherichia coli in RAW264.7 cells. The incorporation of chitooligosaccharide monomers significantly boosted the generation of reactive oxygen species and reactive nitrogen species, as well as the release of inflammatory cytokines. To further explore the mechanism of inflammation regulated by chitooligosaccharide, the activation inhibitors of NF-кB (CAPE) and TLR-4 (TAK-242) were utilized, the determination data demonstrated that chitobiose suppressed the expression of inflammatory cytokines and NF-кB p65. In addition, the investigation results revealed that the presence of the mannose receptor inhibitor (mannan) suppressed chitohexaose-induced phagocytic activity and inflammatory cytokines. These results suggested that the five distinct chitooligosaccharide monomers had inconsistent effects, the chitobiose and chitohexaose exhibiting the best biological activity in activating RAW264.7 cells, promoting cell proliferation, and increasing non-specific immunity.

The antibacterial activity of erythrocytes from Clarias fuscus associated with phagocytosis and respiratory burst generation.

It is widely known that red blood cells (RBCs) are responsible for respiration and the transport of gas. However, recent reports have also described the immune properties of RBCs, therefore creating new understanding for the functionality of RBCs. However, little is known about the immunological role of RBCs in bony fish. In this study, we used RBCs from Clarias fuscus as a model and demonstrate that these cells exhibited phagocytic ability with both latex beads and bacteria. Scanning electron microscopy and transmission electron microscopy provided visual confirmation of the phagocytotic process in RBCs. In addition, we used flow cytometry and fluorescence microscopy to analyse the rate of phagocytosis in RBCs. We found that RBCs exhibited stable phagocytotic ability with latex beads ranging from 0.5 to 1.0 µm in size. In response to bacterial stimulation, RBCs produced reactive oxygen species (ROS) and nitric oxide synthase (NOS), which are harmful to bacteria. RBCs also have an antioxidant system. Under conditions of oxidative stress, the expression levels of antioxidant enzymes, and particularly those of superoxide dismutase（SOD） increased significantly. Our results show that the erythrocytes of bony fish are phagocytic and also produce ROS which are toxic to bacteria. In addition, RBCs have an antioxidant system that removes excess ROS production to protect cells from oxidative damage.

Molecular and functional characterization of mitochondrial manganese superoxide dismutase from Macrobrachium rosenbergii during bacterial infection.

Superoxide dismutases (SODs) are the main antioxidant enzymes involved in alleviating oxidative stress. Although mitochondrial manganese SOD (mMnSOD) has been reported to be correlated with the immune response in crustaceans, its biological properties and role in the immune response remain unclear. Here, we cloned the Macrobrachium rosenbergii mMnSOD (MrmMnSOD), analyzed its activity and expression pattern under Staphylococcus aureus and Vibrio parahaemolyticus infection, and further explored its possible mechanism during antibacterial immune response. The results showed that both enzyme activity and the expression of MrmMnSOD were significantly up-regulated by bacterial infection. MrmMnSOD knockdown made the prawn susceptible to Vibrio infection, which increased the mortality rate and the number of bacteria in haemocytes. The bacterial agglutination assay confirmed that MrmMnSOD decreases bacterial abundance via agglutination. Overall, this work identified antibacterial function of MrmMnSOD in the immune response. In addition to contributing to immunological theory, these findings aid disease prevention and control in crustacean aquaculture.

The Antibacterial Activity of Erythrocytes From Goose (Anser domesticus) Can Be Associated With Phagocytosis and Respiratory Burst Generation.

In the lumen of blood vessels, there are large numbers of erythrocytes, which are approximately 95% of the total blood cells. Although the function of erythrocytes is to transport oxygen in the organism, recent studies have shown that mammalian and teleost erythrocytes are involved in the immune response against bacterial infections. However, the immune mechanisms used by avian erythrocytes are not yet clear. Here, we demonstrated that erythrocytes from goose have the ability to phagocytose as well as conduct antimicrobial activity. Firstly, we revealed the phagocytosis or adhesion activity of goose erythrocytes for latex beads 0.1-1.0 µm in diameter by fluorescence microscopy, and scanning and transmission electron microscopy. The low cytometry results also proved that goose erythrocytes had a wide range of phagocytic or adhesion activity for different bacteria. Followed, the low cytometry analysis data further explored that the goose erythrocytes contain the ability to produce reactive oxygen species (ROS) and inducible nitric oxide synthase (iNOS) in response to bacterial stimulation, and also up-regulated the expression of NOX family includes NOX1 and NOX5. Finally, we also found that goose erythrocytes showed a powerful antibacterial activity against all the three bacteria, meanwhile the stimulation of three kinds of bacteria up-regulated the expression of inflammatory factors, and increased the production of antioxidant enzymes to protect the cells from oxidative damage. Herein, our results demonstrate that goose Erythrocytes possess a certain phagocytic capacity and antioxidant system, and that the antimicrobial activity of erythrocytes can occurred through the production of unique respiratory burst against foreign pathogenic bacteria, which provides new clues to the interaction between bacteria and avian erythrocytes.