The combination of mesenchymal stem cell- and hepatocyte-derived exosomes, along with imipenem, ameliorates inflammatory responses and liver damage in a sepsis mouse model.

Aim Sepsis is a medical emergency with no definitive treatment. Animal experiments have confirmed the therapeutic characteristics of exosomes in reducing inflammation and tissue damage. The study investigates the effect of MSC and hepatocyte-derived exosomes along with imipenem in controlling systemic and local (liver) inflammation in a mouse model of sepsis. MAIN METHODS: To induce sepsis in C57BL/6 mice, the Cecal Ligation and Puncture (CLP) model was used. The mice were given various treatments, including imipenem, MSC-derived exosomes, hepatocyte-derived exosomes, and a mixture of exosomes. Blood and liver samples were collected and analyzed for cell blood count, liver enzymes, NO levels, cytokine concentrations, and bacterial presence. The percentages of TCD3 + CD4+/CD8+ and Treg in the spleen and mesenteric lymph nodes were also assessed using flow cytometry. The pathological changes were assessed in the liver, lung, and heart tissues. In addition, the cytokine content of exosomes was measured by ELISA. KEY FINDINGS: Our results demonstrated that MSC-derived exosomes+imipenem could control systemic and local inflammation and increase the TCD4+ and Treg populations. Hepatocyte-derived exosomes+imipenem reduced inflammation in the liver and increased the TCD8+ and Treg populations. The mixture of exosomes+imipenem had the best function in reducing inflammation, maintaining all T lymphocyte populations, reducing liver damage, and ultimately increasing the survival rate. SIGNIFICANCE: The mixture of exosomes derived from MSCs and hepatocytes, along with imipenem, in the inflammatory phase of sepsis could be a promising therapeutic strategy in sepsis treatment.

Study of immunomodulatory effects of mesenchymal stem cell-derived exosomes in a mouse model of LPS induced systemic inflammation.

BACKGROUND: Sepsis is a debilitating systemic inflammation that resulted from infection or injury. Despite many advances in treatment, the resulting mortality rate has remained high due to increasing antibiotic resistance and aging communities. The present study investigated the effects of stem cell-derived exosomes in a mouse model of LPS-induced systemic inflammation. MATERIALS AND METHODS: To induce sepsis, the LPS model was used. Mice were divided into three groups: normal, patient group (LPS + PBS), and treatment group (LPS + exosome). The treatment group received an intravenous exosome 1 h after induction of the model. Patient and treatment groups were sacrificed at 4, 6, 24, and 48 h after induction of the model, and their tissues were isolated. Blood samples were taken from animal hearts to perform biochemical and immunological tests. The study results were analyzed using Graph Pad Prism software version 9. RESULTS: Mesenchymal stem cell-derived exosomes decreased serum levels of ALT and AST liver enzymes, decreased neutrophil to lymphocyte ratio (NLR), and improved kidney, liver, and lung tissue damage at 4, 6, and 24 h after model induction. At 24 h, the exosomes were able to reduce serum urea levels. This study revealed decreased levels of inflammatory cytokines such as IL-6, IL-1ß, and TNF-α after exosome injection. CONCLUSION: Our findings suggest that treating mice with stem cell-derived exosomes can ameliorate the destructive effects of inflammation caused by sepsis by reducing inflammatory factors and tissue damage.

Genetic characterization and phylogenetic of Anaplasma capra in Persian onagers (Equus hemionus onager).

Anaplasma spp. are among the most recognized arthropod-borne infectious agents. Although the novel A. capra has been isolated from wildlife, livestock, and hard ticks from many parts of the world, there is no report regarding the identification of this pathogen from equines and little is known about the epidemiology of A. capra in Equidae. In this study, A. capra was identified in two out of ten blood specimens of wild onagers (Equus hemionus onager) during a routine health check-up in Semnan, Iran by light microscopy and molecular analyses while other pathogens were not detected. First, inclusions on RBC's were observed in two blood smears by light microscopy. Then, the blood specimens of both animals were analyzed by realtime-PCR for Anaplasma, Ehrlichia, and Theileria infections. A 1400 bp sequence of 16S rRNA belonging to Anaplasmataceae and 874 bp fragment for groEL gene for A. capra were amplified in Anaplasma positive samples and sequenced. Preliminary BLAST analysis of sequenced fragments showed high homology to A. capra strains in GenBank database. Finally, nested PCR and restriction enzyme fragment length polymorphism techniques confirmed the pathogen as A. capra. To the best of our knowledge, this study has reported the occurrence of A. capra in wild onagers for the first time and suggests that equines could be infected with this pathogen and act as reservoirs for A. capra.

Study of the Effect of Topical Ozonated Oil on Healing of Calvarial Defect.

BACKGROUND: Fracture healing is one of the important issues in medicine and veterinarian medicine. A new technique should be detected with fewer side effects which improves the speed of the healing. This study was conducted to evaluate the effect of topical ozonated oil on the healing of bone defects created in rat calvarium. METHODS: Defects of 7 mm were induced with a trephine in the calvaria of 45 male Sprague-Dawley rats. The animals were divided into 3 groups, and the defects in each group were filled with ozonated oil, olive oil, or left empty as a control. The animals were euthanized at 3 different time points (14, 28, and 42 days post-operation). Histological and serological (alkaline phosphatase) assessments were carried out on 14, 28, and 42 days post-operation. RESULTS: On 14 and 28 days post-operation, in the edges of the defect, there was a significant difference between the ozonated oil group and olive oil as well as control groups (P ≤ 0.05). However, in the center of the defect, there were no statistically significant differences between the 3 groups (P > 0.05). On day 42, in the edges and center of the defect, there was a significant difference between the ozonated oil group and olive oil as well as control groups statistically (P ≤ 0.05). CONCLUSION: The results of this study indicate the potential efficacy of ozonated oil as a bone substitute in a rat calvarial defect model.

Capecitabine-loaded anti-cancer nanocomposite hydrogel drug delivery systems: in vitro and in vivo efficacy against the 4T1 murine breast cancer cells.

In this work, nanocomposite hydrogel drug delivery systems based on polyvinyl alcohol and montmorillonite loaded with the capecitabine, as an anti-cancer drug, were developed for oral administration. The gel fraction and swelling ability of the prepared nanocomposite hydrogels were experimentally measured. In vitro release kinetics of capecitabine in nanocomposite hydrogel drug delivery systems were studied. In vitro flow cytometry assay was utilized to exhibit the anti-cancer activity of the prepared nanocomposite hydrogel drug delivery systems against 4T1 cancer cell line. The anti-tumor efficacy of the nanocomposite hydrogel drug delivery systems was also studied in vivo on animal models. The results showed that the amount of montmorillonite incorporated into the nanocomposite hydrogel drug delivery systems could be recognized as a key parameter to adjust the values of the gel fraction, swelling and capecitabine release rate in a manner which by increasing the montmorillonite content, the gel fraction is increased while the swelling and drug release rate are decreased. The flow cytometry results demonstrated the better anti-cancer activity of the capecitabine-loaded nanocomposite hydrogel drug delivery systems as compared with the pure capecitabine. The in vivo assays indicated that the administration of nanocomposite hydrogel drug delivery systems had a significant effect on the reduction of the tumor growth in animal models as compared with pure capecitabine administration. In general, the prepared nanocomposite hydrogel drug delivery systems exhibited a suitable efficacy against 4T1 cancer cell line both in vitro and in vivo and they could be considered as promising candidates for controlled release of anti-cancer drugs in chemotherapy with enhanced therapeutic effects.