The effects of melatonin on the concentrations of inflammatory cytokines and proteins, serotonin, cortisol and melatonin in ovariohysterectomised female dogs.

BACKGROUND: Ovariohysterectomy (OHE) induces inflammation and stress in female dogs. The anti-inflammatory effects of melatonin have been reported in several studies. OBJECTIVES: The goal of this study was to assess the effects of melatonin on the concentrations of melatonin, cortisol, serotonin, α-1-acid glycoprotein (AGP), serum amyloid A (SAA), c-reactive protein (CRP), interleukin-10 (IL-10), interleukin-8 (IL-8), interleukin-1ß (IL-1ß) and tumour necrosis factor-α (TNF-α) before and after OHE. METHODS: The total number of animals was 25 and aligned in 5 groups. Fifteen dogs were divided into three groups (n = 5): melatonin, melatonin+anaesthesia and melatonin+OHE and received melatonin (0.3 mg/kg, p.o.) on days -1, 0, 1, 2 and 3. Ten dogs were assigned to the control and OHE groups (n = 5) without melatonin treatment. OHE and anaesthesia were performed on day 0. Blood samples were obtained via jugular vein on days -1, 1, 3 and 5. RESULTS: Melatonin and serotonin concentrations significantly increased in the melatonin, melatonin+OHE and melatonin+anaesthesia groups compared with the control group, while cortisol concentration decreased in the melatonin+OHE group compared with the OHE group. The concentrations of acute-phase proteins (APPs) and inflammatory cytokines significantly increased after OHE. The CRP, SAA and IL-10 concentrations decreased significantly in the melatonin+OHE group compared with the OHE group. The concentrations of cortisol, APPs and proinflammatory cytokines increased significantly in the melatonin+anaesthesia group compared with the melatonin group. CONCLUSIONS: The oral administration of melatonin before and after OHE help controlling the high levels of inflammatory APPs, cytokines and cortisol induced by OHE in female dogs.

Effects of short-term administration of melatonin before gonadectomy on oxidative stress, cortisol and sex hormones in male dogs.

The aim of this study was to investigate gonadectomy stress, steroid hormones and serotonin in male dogs treated with melatonin before gonadectomy. Twenty-five mixed breed adult dogs were divided into five equal groups. The melatonin and melatonin + gonadectomized groups received melatonin treatment (3 mg/10 Kg, PO, TID) the day before gonadectomy; the gonadectomized and anaesthesia groups did not receive melatonin; and the control group just received the melatonin vehicle. Blood sampling was performed before melatonin administration (day -1) and on days 0 (gonadectomy), 1, 3 and 6 after gonadectomy. Superoxide dismutase and glutathione peroxidase concentrations decreased significantly in gonadectomized dogs compared with dogs treated with melatonin before gonadectomy and intact dogs. Gonadectomy led to a significant decrease in catalase concentration in gonadectomized dogs compared with other study groups. Malondialdehyde levels increased significantly in gonadectomized dogs compared with other groups. Melatonin administration before gonadectomy led to decreased malondialdehyde concentration in gonadectomized and intact dogs compared to the control group. Cortisol concentration increased significantly in gonadectomized dogs compared to the control dogs. Serotonin levels decreased in gonadectomized dogs, but melatonin treatment increased serotonin concentration in gonadectomized and intact dogs. Melatonin treatment before gonadectomy suppressed oxidative stress and the cortisol but increased serotonin level.

The effects of melatonin treatment on oxidative stress induced by ovariohysterectomy in dogs.

BACKGROUND: As one of the most common surgeries performed in veterinary medicine, ovariohysterectomy (OHE) can induce oxidative stress in dogs. The antioxidant properties of melatonin have been confirmed in various studies. This study aimed to investigate the effects of melatonin administration on oxidative stress in dogs before and after OHE. In this study, 25 mature female intact dogs were selected and randomly divided into five equal groups: Melatonin (melatonin, no surgery), OHE (no melatonin, surgery), OHE + melatonin (melatonin, surgery), anesthesia+melatonin (melatonin, sham surgery), and control (no melatonin, no surgery) groups. Melatonin (0.3 mg/Kg/day, p.o.) was administrated to the dogs in the melatonin, OHE + melatonin, and anesthesia+melatonin groups on days - 1, 0, 1, 2, and 3 (day 0 = OHE). Blood sampling was performed on days - 1, 1, 3, and 5 of the study. Blood samples were immediately transferred to the laboratory and sera were separated and stored at - 20 °C. Superoxide dismutase (SOD), glutathione peroxidase (GPX), catalase (CAT) and malondialdehyde (MDA) concentrations were measured with commercial kits. RESULTS: The levels of SOD, GPX and CAT were significantly higher in the melatonin and anesthesia+melatonin groups compared to those of the control group at days 3 and 5. The level of antioxidant enzymes significantly decreased in the OHE group compared to that of other groups at days 3 and 5. The administration of melatonin increased the level of antioxidant enzymes in ovariohysterectomized dogs. Ovariohysterectomy significantly increased the concentration of MDA in comparison to that of other groups at day 3. Melatonin administration significantly decreased the level of MDA in melatonin, anesthetized, and ovariohysterectomized dogs at day 3. CONCLUSIONS: Administration of melatonin on day - 1, 0, 1, 2 and 3 modulate the oxidative stress induced by OHE in dogs by increasing antioxidant enzymes concentration and decreasing MDA levels.

Are Stem Cells Derived from Synovium and Fat Pad Able to Treat Induced Knee Osteoarthritis in Rats?

BACKGROUND: Osteoarthritis (OA) is a chronic disease and a significant cause of joint pain, tenderness, and limitation of motion. At present, no specific treatment is available, and mesenchymal stem cells (MSCs) have shown promising potentials in this regard. Herein, we aimed to evaluate the repairing potentials of stem cells derived from the synovium and fat pad in the treatment of OA. METHODS: Twenty-eight male rats (220 ± 20 g, aged 10-12 weeks), were randomly divided into four groups (n = 7): C1: nontreated group, C2: Hyalgan-treated group, E1: adipose tissue-derived stem cell-treated group, and E2: synovial membrane-based stem cell-treated group. Collagenase type II was injected into the left knee; after eight weeks, OA was developed. Then, stem cells were injected, and rats were followed for three months. Afterward, specimens and radiological images were investigated. p value ≤ 0.05 was set as statistically significant. RESULTS: Compared to the C1 group, the E1 and E2 groups showed significantly better results in all six pathological criteria as well as joint space width and osteophytes of medial tibial, medial femoral, and medial fabellar condyles (p ≤ 0.001). Similarly, compared to the C2 group, the E1 and E2 groups had better scores regarding surface, matrix, cell distribution, and cell population viability (p < 0.05). E2 showed considerably higher scores compared to C2 regarding subchondral bone and cartilage mineralization (p < 0.05). The joint space width was similar between the C2 and E groups. CONCLUSION: Treatment of OA with MSCs, particularly synovial membrane-derived stem cells, not only prevented but also healed OA of the knee to some extent in comparison to the Hyalgan and nontreatment groups.

The detection of canine anti-sperm antibody following parenteral immunization of bitches against homogenized whole sperm.

BACKGROUNDS: The development of a canine-specific method of immunocontraception is one of the non-invasive controlling strategies for humanely decreasing the dog population. This study was aimed to investigate the potential of whole sperm in stimulating the immune system and producing specific anti-sperm antibodies (ASAs) in female dogs. Mature, mixed-breed bitches were subcutaneously immunized with high (200 × 106 cells/mL) and low (100 × 106 cells/mL) doses of sperm vaccine, emulsified with Freund's adjuvants. Booster immunizations were given at weeks 1, 2, 4, and 6, and serum samples were collected at days 0, 14, 28, 42, 63, and 84 prior to each immunization. Reproductive tract samples, including vaginal and uterine lavages, were also collected by flushing each section with sterile PBS at the end of the experiment. Canine anti-sperm antibody titer and specificity in sera and genital secretions were measured using an enzyme-linked immunosorbent assay technique. RESULTS: Specific anti-sperm antibodies were detected in the serum of both high and low dose groups and were significantly higher than those observed in the controls. A high dose of sperm induced elevated immune responses over the low dose antigen. Immunization with a high dose of sperm increased the level of ASAs in the uterine secretions and vaginal secretions significantly. Higher ASAs were observed to have transduced to the uterine lumen compared to the vagina. CONCLUSIONS: Based on the results obtained in this study, parenteral immunization with whole sperm can induce a high level of specific antibodies in the serum and genital secretions of female dogs and the response would be dose-dependent.

CONTEXTE: Le développement d'une méthode d'immunocontraception spécifique à la race canine constitue l'une des stratégies non invasives pour réduire humainement la population canine. Cette étude a pour objectif d'évaluer le potentiel du sperme entier à stimuler le système immunitaire des femelles et à produire des anticorps anti-sperme spécifiques (ASA) chez ces dernières. Des chiennes matures de race croisée sont immunisées par voir sous-cutanée avec soit de fortes doses (200 millions de cellules/mL) soit de faibles doses (100 millions de cellules/mL) de vaccin constitué de sperme entier émulsifié avec des adjuvants de Freund. Des vaccinations de rappel sont faites aux semaines 1, 2, 4 et 6, et des échantillons sanguins sont prélevés aux jours 0, 14, 28, 42, 63 et 84 avant chaque immunisation. Des échantillons de l'appareil reproducteur, incluant des lavages vaginaux et utérins, sont recueillis par flushing de chaque section avec du PBS stérile à la fin de l'expérimentation. Le titre et la spécificité des anticorps anti-sperme entier canin dans le sérum et les sécrétions génitales ont été mesurés par la technique de dosage ELISA. RÉSULTATS: Des anticorps anti-sperme entier spécifiques ont été détectés dans le sérum des femelles immunisées tant avec de faibles que de fortes doses, et de façon significativement plus élevé que chez le groupe témoin. Une dose forte de sperme entier induit des réponses immunitaires élevées par rapport à l'antigène à faible dose. L'immunisation avec une forte dose de sperme entier augmente de façon significative le niveau d'ASA dans les sécrétions utérines et dans les sécrétions vaginales. On a observé que les ASA ont plus été transduits vers la lumière utérine que vers la lumière vaginale. CONCLUSIONS: Basée sur les résultats obtenus dans la présente étude, l'immunisation parentérale par du sperme entier peut induire un taux élevé d'anticorps spécifiques dans le sérum et le sécrétions génitales de chiennes ; et la réponse serait dose-dépendante.

Use of ovine acellular peritoneal matrix combined with honey and ovine fetal skin extract in the healing of full-thickness infected burn wounds in a rat model.

Treatment of infected burn wounds remains a challenge in burn units. Silver-sulfadiazine (SSD) is the most commonly used topical antimicrobial agent in managing these wounds. We aimed to accelerate the healing of burn wounds by combined application of ovine acellular peritoneal matrix (OAPM), honey (H), and ovine fetal skin extract (OFSE). Sixty-four standardized burn wounds were created on the dorsum of 16 rats and were subsequently inoculated with Staphyloccocus aureus and Pseudomonas aeruginosa. After 48 hr, the wounds were surgically debrided and received either physiologic saline (control group) or SSD, OAPM+SSD, OAPM+H+SSD, and OAPM+H+OFSE+SSD. The healing wounds were evaluated for size, bacterial counts, histopathology, and biomechanical properties on days 3, 7, 14, 21, and 28 after surgery. All treatments had effectively reduced the level of S. aureus and P. aeruginosa on wounds compared to the control group by day 3 and 7. The wounds treated with combined application of OAPM+H+OFSE+SSD demonstrated considerable inflammation reduction, fibroplasia, complete re-epithelialization, and wound contraction together with significantly lesser scar tissue formation. Treatment with OAPM+H+OFSE+SSD showed superior biomechanical properties of the healing wounds. The findings suggested that the synergistic effect of dressing the wounds with OAPM, H, and OFSE was a very effective approach in accelerating the healing process of the experimentally induced infected full-thickness burn wounds in rats.

The concurrent use of probiotic microorganism and collagen hydrogel/scaffold enhances burn wound healing: An in vivo evaluation.

Treatment of burn wounds is technically demanding and several attempts have been taken to improve wound healing. Silver sulfadiazine antibiotic has been shown to have some beneficial effects on wound healing via reduction in infection. This study was designed to investigate the impact of collagen hydrogel-scaffold dressing with or without topical use of Saccharomyces cerevisiae on cutaneous burn wound healing in rat. Four circular 1cm cutaneous wounds were created in the dorsum back of rats, 48h post-burning. Thirty male rats were divided into the three major groups (1-3, n=10), then the wounds in each group were equally divided into two subgroup treatments (6 treatments), (1) including (a) silver sulfadiazine (SSD) as positive control (PC) and (b) untreated wounds as negative control (NC) (2) including (c) S. cerevisiae and (S.C.) and (d) collagen scaffold (CS), (3) including (e) collagen hydrogel-scaffold (CH-S) and (f) S. cerevisiae with collagen hydrogel-scaffold (CH-S-S). In each group, the animals were euthanized at 12 and 22 days post-injury (DPI) and the skin samples were used for histopathological and biomechanical investigations. Collagen scaffold and hydrogel modulated the inflammation, especially when combined together. Moreover, they increased wound healing, epithelialization and biomechanical performance of wound area and also reduced the scar size. The best results gained when the combination of collagen scaffold and hydrogel were mixed with probiotic. The CH-S biological dressing along with probiotic microorganism (S.C.) significantly increased collagen content compared to the negative controls. Moreover, the CH-S-S treated lesions demonstrated greater ultimate load and stiffness compared to the untreated wounds. In conclusion, application of S. cerevisiae with a bi-phase biological dressing (CH-S) improved the morphological and biomechanical characteristics of the healing burned wounds in rats and the results were comparable to the positive control.

Epididymis ligation: a minimally invasive technique for preparation of teaser rams.

OBJECTIVE: To describe a minimally invasive technique for preparation of teaser rams by needle-assisted ligation through the tail of the epididymis. STUDY DESIGN: Experimental study. ANIMALS: Mature rams (n=6), estrus-induced ewes (2). METHODS; After local anesthesia, epididymis ligation was achieved by restraining the testis distally within the scrotal sac and passing suture through a hypodermic needle inserted between tail of epididymis and distal pole of testis, caudomedial to craniolateral through the scrotum. The needle was removed leaving the suture in place and the testis pushed up dorsally, then the needle was reinserted through the original holes and the suture passed back through the needle, which was withdrawn. This resulted in the suture forming a complete loop around the epididymis. The suture ends were tied ligating the epididymis. Semen was evaluated pre- and postligation. Testes were removed after 30 days for gross and histologic examination. RESULTS: Epididymis ligation was accomplished without postoperative complications. Three weeks after the epididymis ligation, no motile and live spermatozoa were found in ejaculates. From 5 to 28 days after epididymis ligation, attraction to ewes and libido was unchanged and similar to 14 days before ligation. CONCLUSIONS: This novel minimally invasive technique is a simple, alternative method for preparation of teaser rams. CLINICAL RELEVANCE: This method is simply performed, without skin wounds, and minimal postoperative care is needed. The technique should be readily adaptable to other species.