The protective effect of Ozone on the mice testicular damage induced by methotrexate.

OBJECTIVE: Methotrexate (MTX) is widely administered for the treatment of various cancers. However, MTX induces male reproductive toxicity. In the current study, the effect of ozone therapy (OT) on reducing the toxic effects of MTX in the mouse testicles has been investigated. METHODS: Twenty-four mice were divided into four groups: control, OT (4 mg/kg ozone), MTX (20 mg/kg), and MTX + OT. Testosterone levels, histological changes, and oxidative stress biomarkers were assessed to evaluate the protective effects of OT. RESULTS: The results demonstrated that MTX disrupted germinal epithelium, reduced serum testosterone levels, and enhanced oxidative stress in testicular tissue. However, treatment with OT attenuated these adverse effects. OT effectively restored the levels of antioxidant enzymes, such as catalase (CAT), glutathione (GSH), and superoxide dismutase (SOD). OT reduced lipid peroxidation, as indicated by decreased malondialdehyde (MDA) levels. OT preserved normal spermatogenesis, improved morphometric parameters, and reduced histological changes by MTX. Moreover, OT effectively restored testosterone levels. CONCLUSIONS: OT protects against MTX-induced testicular damage by suppressing oxidative stress.

Therapeutic Utilization of Zinc Supplementation Concurrent with Ozone Therapy Ameliorates Diabetic Foot Ulcer and Attenuates Serum Level of C-reactive Protein- A Case Report Study.

Non-healing diabetic foot ulcer (DFU) is one of the main complications in diabetic patients. This case reported a 65-year-old male with a neuropathic ulcer in the right foot came to Ahwaz Wound Clinic after the wound had not healed with routine treatments. In addition to the routine treatment program, we used tropical ozone therapy and autohemotherapy (blood ozone therapy) for 2 months. Zinc supplementation (50 mg) was also administered daily during the treatment. The DFU was clearly healed with diminishing inflammation and wound closing, and there were no side effects. Additionally, the C-reactive protein level was obviously decreased during the treatment indicating effective suppression of infection. This way indicates a helpful new intervention approach to the treatment of DFU.

Single intratesticular injection of blood-serum-derived exosomes can potentially alleviate testopathy following testicular torsion.

BACKGROUND: Testicular torsion (TT) is an acute inflammatory process leading to male infertility. Today, anti-inflammatory effects of exosomes derived from blood serum are used in various laboratory procedures. In the present study, the anti-inflammatory effects of blood-serum-derived exosomes in treatment of acute inflammation following TT in mice were evaluated. MATERIALS AND METHODS: Eighteen male mice were grouped as healthy control, TT, and TT + exosome. TT was induced surgically, and exosomes were extracted from blood serum and administrated by a single intratesticular injection (10 IU). Malondialdehyde (MDA) and Griess assays were used to evaluate the level of oxidative stress. Sperm indices, testosterone (Tes), and apoptotic gene expression (p-53, Bcl2, and Caspase-3) were also assessed. H&E and immunohistochemistry (IHC) stainings were used for histopathological investigations. Data analysis was applied by SPSS (v.19) software. RESULTS: Oxidative stress and apoptotic genes expression were increased significantly (p < 0.05) in TT group compared with control. Sperm parameters and Tes were significantly increased, and expression of apoptotic genes was significantly reduced in TT + exosome group (p < 0.05). CONCLUSION: Since the blood-serum-derived exosomes have anti-inflammatory features, the intratesticular application of blood-serum-derived exosomes can be used clinically in acute phase of orchitis following TT to inhibit testicular inflammation.

The effects of ozone and melatonin on busulfan-induced testicular damage in mice.

OBJECTIVE: Busulfan is one of the most common chemotherapeutic drugs and has the ability to induce apoptosis in testicular germ cells, which leads to infertility. In this study, the effects of ozone therapy and melatonin were evaluated on testicular disorders induced by busulfan. METHODS: In this study, we divided 24 male mice into four groups: control group, groups treated with busulfan, busulfan/melatonin, and busulfan/ozone. At the end of a 35-day period, blood samples were taken from the mice and their testosterone levels were measured. Both of the mice's testes were removed and weighed, afterwards, each one of them was used for evaluation of morphology by Johnson's score, as well as for measuring the diameter and thickness of seminiferous tubules. The other testis was homogenized for measuring Malondialdehyde (MDA) and antioxidant status using Catalase (CAT), Super Oxide Dismutase (SOD), and Total Antioxidant Capacity (TAC) levels. Epididymis spermatozoa were also used to evaluate motility, morphology, and sperm count. RESULTS: Busulfan significantly reduced the testis quality (weight, sperm parameters, testosterone, CAT, SOD, and TAC levels) and increased MDA and destruction of seminiferous tubules compared to the control group. Ozone and melatonin treatments significantly increased testis quality, sperm parameters, MDA, and antioxidant status, but they did not affect the TAC level. CONCLUSIONS: This study showed that similar to melatonin, ozone can reduce the effect of busulfan toxicity on mice testis. However, further studies are needed to understand the precise mechanism of ozone function on testis.