The Effects of Hydrogen-Rich Water on Skin Wound Healing in Dogs

Background: Hydrogen-rich water (HRW) has been shown to be one of underlying therapeutic strategies regarding wound treatments. This study explored the effects of drinking HRW on skin wound healing in dogs. Eight circular wounds were analyzed in each dog. The experimental group was treated with HRW twice daily, while the control group was provided with distilled water (DW). The wound tissues of dogs examined histopathologically. The fibroblasts, inflammatory cell infiltration, the average number of new blood vessels, and the level of malondialdehyde (MDA) and superoxide dismutase (SOD) activity in the skin homogenate of the wound was measured using the corresponding kits. The expression of Nrf-2, HO-1, NQO-1, VEGF, and PDGF were measured by real-time fluorescence quantitative method. Results: We observed that HRW wound had significant average healing rate and faster average healing time. Histopathological results showed that the average number of blood vessels and the average thickness of epidermis were significantly different from the DW group. The MDA levels were higher in the DW group than in the HRW group but the SOD levels were higher in the HRW group than in the DW group. The results of qRT-PCR showed that the expression of each gene was significantly different between the two groups. Conclusions: The results showed that HRW treatment could promote skin wound healing in dogs, accelerate wound epithelization, reduce inflammatory reaction, stimulate the expression of cytokines related to wound healing, and shorten wound healing time.

Study of the intensity of microbial contamination and nonspecific immunity under the experimental diabetes after using different ways of fixing the wound edges

Diabetes mellitus and its complications are one of the most important causes of death. As a consequence of hyperglycemia in patients with diabetes there is an increased risk of concomitant diseases, and one of the important consequences of diabetes is impaired ability to repair. Healing disorders in diabetes are the result of complex pathophysiology involving vascular, neuropathic, immune and biochemical componentsAim of research: was to analyze changes in inflammation markers, microbial contamination and nonspecific immunity in skin homogenate of rats with diabetes after using sutures and skin glue.Material and research methods. Experimental studies were performed on 130 white outbred adult male rats, weight from 240 to 320 g. Diabetes mellitus was induced by using streptozotocin (Sigma, USA) (intraperitoneally - 65 mg / kg) with previous (15 minutes) injection of nicotinamide (intraperitoneally - 230 mg / kg). On the background of obesity, which was caused by 4 weeks of keeping animals on a high-fat diet. The development of diabetes mellitus 2 was confirmed by determining the concentration of glucose in the blood using a glucometer BAYER Contour Next (Germany). Animals of all groups (I-IV) under thiopental anesthesia (40 mg / kg body weight of rats) were made full-layer rectilinear incisions, 2 cm long, in the anterior-lateral region of the abdomen. All animals were divided into 4 experimental groups: І group (30 rats) – healthy rats, wound edges were sutured with Vicryl 5/0 thread (ETHICON, Inc. and Johnson & Johnson company (USA); ІI group (30 rats) – healthy rats, fixing of the wound edges was performed by applying skin glue Dermabond (ETHICON, Inc. and Johnson & Johnson company (USA)); ІII group (30 rats) – rats with diabetes mellitus, wound edges were sutured with Vicryl 5/0 thread; IV group (30 rats) – Rats with diabetes mellitus, fixing of the wound edges was performed by applying skin glue Dermabond. For control, all the results were compared with those of intact animals (10 rats). Animals were removed from the experiment on 3, 7, 28 days after surgery under thiopental anesthesia (90 mg / kg body weight of rats).The degree of contamination of the wound with opportunistic and pathogenic microflora determined by the level of urease activity using Nessler's reagent. Lysozyme activity was determined by bacteriolytic method, using as a substrate a suspension of bacteria Micrococcus lysodeicticus.Results. The obtained results prove that the wound process under conditions of streptozotocin diabetes mellitus in rats leads to a significantly greater contamination of the wound with pathogenic and opportunistic microflora and a decrease in nonspecific immunity, compared with healthy animals. Also, different effects of suture materials on the indicators of microbial contamination and immune protection were found: significantly larger deviations from the norm were found in groups of rats, where we used surgical sutures to fix the wound edges. When comparing the intensity of microbial contamination and nonspecific immunity in the long term (28 days) in the homogenate of the skin of rats with diabetes mellitus and the use of skin glue, the level of urease was 27.3% lower and the level of lysozyme 7.0% lower than in III group where we used surgical sutures.Changes in the activity of the antimicrobial enzyme lysozyme allow to assess the state of nonspecific immunity in the studied tissues. Many authors note a decrease in lysozyme activity in various bioliquids and tissues in diabetes mellitus. The results of our studies prove a significant decrease in the activity of lysozyme in the homogenate of postoperative skin wounds of rats with diabetes.Conclusion. Wound process in rats with experimental diabetes mellitus leads to reliable contamination of the wound with pathogenic and opportunistic microflora, as evidenced by increased urease activity, and reduced nonspecific immunity, characterized by reduced lysozyme activity in the homogenate of postoperative wound/scar tissue in relation to intact animals during all the terms of observation.In the remote period (28 days) in the skin homogenate of rats with diabetes where skin glue was used for wound closure, the level of urease was 27.3% lower and the level of lysozyme was 7.0% lower than that of rats with diabetes, where surgical sutures were used.

FEATURES OF FREE SKIN LIPID HOMOGENATES IN RATS AT DIFFERENT WAYS OF WOUND CLOSURE

According to different authors from 4 % to 22 % of patients who applied to medical institutions have abnormal scars. The aim of our research was to identify the characteristics of oxidative stress in skin homogenate of rats with different methods of wound closure. Analysis of all indicators allows to assert that intensity of free radical oxidation in the skin of the animals after applying skin glue is much lower compared to the animals after suturing. In furtherresearch is planned to analyze indicators of free radical oxidation system antioxidation protection and reparative ability of the skin when used sutures and biological glue in rats with the diabetes.

Cortisol and catecholamine responses to social context and a chemical alarm signal in juvenile lake sturgeon, Acipenser fulvescens

We examined potential benefits of group living in juvenile lake sturgeon (Acipenser fulvescens Rafinesque, 1817) and whether those benefits applied equally to associations with conspecific and allospecific individuals. In the first experiment, one focal lake sturgeon was placed with either seven size-matched conspecifics, allospecifics, or in isolation and given a 30 s air exposure. In the second experiment, one focal lake sturgeon was placed with either seven conspecifics or in isolation and was exposed to a conspecific skin homogenate as a potential alarm substance or a control of distilled water. Cortisol, norepinephrine, and epinephrine levels were measured before and after exposure in each experiment via an indwelling cannula in the caudal sinus. In the first experiment, lake sturgeon held with allospecifics and in isolation had a significantly longer norepinephrine response. Furthermore, plasma norepinephrine concentration in the focal lake sturgeon in the allospecific treatment was significantly higher than the other treatments following air exposure. In the second experiment, there was no behavioural or hormonal response to the introduction of conspecific skin homogenate, yet isolated fish had elevated baseline norepinephrine and cortisol levels. Finally, comparison of baseline levels of all three hormones between two ages of juveniles indicated an effect of age on baseline epinephrine levels. Our findings confirm the potential physiological benefit of group living in lake sturgeon and suggest an ontogenetic shift in both the behavioural response to potential stressors and the baseline hormonal levels that may influence fitness.