Behaviour of the Biological Adhesives TachoSil®, GelitaSpon®, and a New Elastic Cyanoacrylate (Adhflex®) in Experimental Renal Trauma and Wound Healing.

BACKGROUND: Renal injuries are relatively frequent in abdominal trauma. In some cases, adhesives and sealants can be used to repair and preserve injured organs. This paper describes the behaviour of three biomaterials - TachoSil®, GelitaSpon®, and a new elastic cyanoacrylate (CyA), Adhflex® - in standardized experimental renal injuries. METHODS: Ninety male Wistar rats (300-350 g) were used. A Stiefel Biopsy Punch (8 mm diameter, 3 mm depth) was used to create injuries to the anterior kidney to evaluate wound healing. The animals were divided into five groups: (1) sham (n = 3); (2) control (n = 6), untreated, standard punch injury created on the anterior left kidney; (3) TachoSil® (n = 27), punch injury treated with TachoSil®; (4) GelitaSpon® (n = 27), punch injury treated with GelitaSpon®, and (5) Adhflex® (n = 27), punch injury treated with the new elastic CyA adhesive. The parameters studied were bleeding time, peritoneal adhesions, and histopathological evaluation of wound healing on days 2, 6, and 18, including measurements of the gap between wound edges, inflammatory reaction (CD68), and vascular neoformation (CD31). RESULTS: The bleeding time was significantly shorter (27.7 ± 12.9 s) in the Adhflex® group than in the control (135.8 ± 11.6 s; p < 0.01), TachoSil® (77.5 ± 7.4 s; p < 0.05), and GelitaSpon® (82.5 ± 14.4 s; p < 0.05) groups. The incidence of intraperitoneal adhesions in the animals treated with Adhflex® was 3.6 times higher than in the non-treated group. It was also higher (p < 0.04) than in the groups treated with TachoSil® and GelitaSpon®. The time point with the largest gap between the wound edges and most abundant granulation tissue was at day 6. The largest gap after 18 days was reported for the Adhflex® adhesive. With regard to the markers CD31 and CD68, Adhflex® showed the largest areas 2 days after surgery, but no differences were found after 6 and 18 days versus the other treatments. The expression of the immunomarkers on the renal samples treated with Adhflex® was consistent with a normal healing process. CONCLUSIONS: In this experimental model of renal injuries, the new elastic CyA (Adhflex®) resulted in the shortest bleeding time. It offers rapid sealing of the bleeding produced by renal injuries, fixation to adjacent tissues, and reduced occurrences of relapse. The evolution of the scarring is similar to other procedures. Given that traumatic renal injuries are always an emergency due to haemorrhage, Adhflex® might offer additional benefits over conventional treatment methods in human clinical practice.

RP-HPLC-DAD determination of the differences in the polyphenol content of Fucus vesiculosus extracts with similar antioxidant activity.

Significant quantities of bioactive compounds have been found in the chemical composition of seaweeds. This source of natural antioxidants such as polyphenols appears to attenuate lipid peroxidation caused by oxidative stress, preventing the harmful effects of a number of injuries including ischemia-reperfusion (I/R). Conventional extraction (CE) has been used for years as a traditional method for obtaining bioactive components from seaweeds. However, recent studies highlight ultrasonic-assisted extraction (UAE) as an alternative and more eco-friendly technique. Therefore, the two methods were optimised and compared to obtain a Fucus vesiculosus extract (FVE) with high antioxidant activity and polyphenol content. The highest antioxidant activity was obtained after 1 h at 25 °C for conventional extraction, and after 5 min at 35 °C for ultrasonic-assisted extraction. Higher concentrations of polyphenols were obtained with the optimal conditions in conventional extraction (13.61 mg PGE/g seaweed), but no significant differences were observed between the antioxidant activity obtained with UAE (89.33%) and CE (89.74%). The characterization of the polyphenols present in both optimised extracts was carried out and compared with reverse-phase high-performance liquid chromatography coupled to a diode array detector (HPLC-DAD). The following compounds were identified: phloroglucinol, gallic acid, catechin, vanillic acid, epicatechin, protocatechuic acid, rutin, gentisic acid, chlorogenic acid, caffeic acid, coumaric acid and ferulic acid. RP-HPLC-DAD results also showed higher concentrations of polyphenols in optimised extracts with CE. Consequently, CE was found to be more effective than UAE in providing extracts with higher concentrations of polyphenols, but UAE constitutes an efficient and more eco-friendly methodology for obtaining a FVE with the highest antioxidant activity.

Combination Therapy of Allopurinol and Dantrolene and Its Role In The Prevention of Experimental Ischemia Reperfusion Injury Of The Small Intestine.

BACKGROUND: The effect of different drugs on ischemia and reperfusion (I/R; induced oxygen free radical damage) was examined in small bowel tissue because the intestine is extremely sensitive to this pathology. Different drugs (allopurinol and dantrolene) can remove oxygen free radicals or inhibit the mechanisms leading to their generation, thus reducing mucosal lesions. We investigated the protective potential of combination therapy in the intestine against I/R damage. METHODS: Forty-eight male Wistar rats were separated into 8 groups: one sham (control), one I/R (ischemia 60 min + reperfusion at 24 h), and 6 groups treated with allopurinol, dantrolene, or combination therapy. The grade of injury in the small bowel was established by the lipid peroxidation (MDA) and antioxidant enzymatic activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in tissue samples. Moreover, the collected samples were subjected to histological study. RESULTS: Combination therapy preserved normal enzymatic levels compared to the I/R groups (p < 0.05) for all parameters studied. The animals treated with combination therapy showed less severe small bowel damage than I/R group in accordance with the histological results. CONCLUSIONS: Results obtained in the experimental process indicate that the administration of antioxidants protects against intestinal damage by I/R. Overall, combination therapy may protect intestinal tissue from I/R injury.

The effect of adhesives on inflammatory immune-markers during renal injury healing.

Renal injury is common in abdominal trauma. Adhesives and sealants can be used to repair and preserve damaged organs. We describe the effect of three biomaterial treatments (TachoSil, GelitaSpon, and Adhflex) on injured renal tissue. Renal traumatic injuries were experimentally induced in male Wistar rats (n = 90) using a punch. Animals were divided into five groups: (1) sham noninjured (n = 3) and punch injury groups; (2) nontreated (n = 6); (3) TachoSil (n = 27); (4) GelitaSpon (n = 27); and (5) Adhflex (n = 27). Wound healing was evaluated 2, 6, and 18 days postinjury by inflammatory cytokines response, histopathological evolution of lesions, inflammatory reaction markers (CD68), and vascular neoformation (CD31). The TachoSil group showed the least inflammatory reaction among the three treated groups, which showed similarly low inflammatory reaction 18 days postinjury. Ciliary neurotrophic factor, soluble intercellular adhesion molecule-1, L-selectin, thymus chemokine, and TIMP metallopeptidase inhibitor 1 expression peaked between 2 and 6 days postinjury. TachoSil promoted the highest cytokine expression. The Adhflex group had the highest CD31 inflammatory immune-marker levels at 2 and 6 days postinjury, but there was a similar decrease in CD31 levels in all three groups at 18 days postinjury. The results show that all three sealant treatments induced a normal healing process with the typical pattern of proinflammatory cytokine and immune-marker expression. Each tested sealant substance could be suitable treatment for renal lacerations. The findings of this study indicate that Adhflex® elastic cyanoacrylate does not induce an adverse inflammatory reaction, and therefore, could be considered as one of the first-line treatments for renal injuries. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 1444-1455, 2018.

The effect of biological sealants and adhesive treatments on matrix metalloproteinase expression during renal injury healing.

BACKGROUND: Renal injuries are relatively common in cases of abdominal trauma. Adhesives and sealants can be used to repair and preserve damaged organs. Using a rat model, this study explores the activity of different matrix metalloproteinases (MMP) during the healing of renal injuries treated by two biological adhesives (TachoSil and GelitaSpon) and a new synthetic elastic cyanoacrylate (Adhflex). METHODS: Renal traumatic injuries were experimentally induced in 90 male Wistar rats by a Stiefel Biopsy Punch in the anterior aspect of the left kidney. Animals were divided into five groups: 1, sham non-injured (n = 3); 2, non-treated standard punch injury (n = 6); 3, punch injury treated with TachoSil (n = 27); 4, punch injury treated with GelitaSpon (n = 27); and, 5, punch injury treated with Adhflex (n = 27). Wound healing was evaluated 2, 6, and 18 days after injury by determining the expression of MMPs, and the histopathological evolution of lesions. FINDINGS: Histologically, the wound size at 6 days post-injury was larger in Adhflex-treated samples than in the other treatments, but the scarring tissue was similar at 18 days post-injury. Only the MMPs subtypes 1, 2, 8, 9, and 13 were sufficiently expressed to be quantifiable. Both time since injury and treatment type had a significant influence on MMPs expression. Two days after injury, the expression of MMP8 and MMP9 was predominant. MMP2 expression was greater 6 days after injury. The Adhflex-treated group had a significantly higher MMPs expression than the other treatment groups at all healing stages. CONCLUSIONS: All three sealant treatments induced almost similar expression of MMPs than untreated animals indicating a physiological healing process. Given that all renal trauma injuries must be considered emergencies, both biological and synthetic adhesives, such as Adhflex, should be considered as a treatment options.

Antioxidant potential of Himanthalia elongata for protection against ischemia-reperfusion injury in the small bowel.

BACKGROUND: Seaweed has been associated with the prevention and/or treatment of various diseases related to oxidative stress because of its antioxidant activity. We investigated the protective potential of extract of Himanthalia elongata against ischemia-reperfusion (I/R) injury in the intestine of rats. METHODS: Seventy-two (72) male Wistar albino rats were randomly assigned into 12 groups as follows: sham, I/R only, I/R plus vehicle at 3 time points, and I/R plus extract at 3 time points. The degree of intestinal injury was determined by oxidative stress using lipid peroxidation, superoxide dismutase, catalase, and glutathione peroxidase after mesenteric ischemia-reperfusion. A histological study was also performed. RESULTS: The algae extract helps to maintain normal enzymatic levels because, for all the studied parameters, groups treated with the extract showed significant differences (P < .05) compared with the I/R groups, and there were no differences compared with the sham group. The histological study showed that damage to the intestinal mucosa was less severe in animals treated with extract of H elongata after up to 24 hours of reperfusion compared with the I/R group. CONCLUSION: These results suggest that the extract of H elongata can protect intestinal tissue against ischemia-reperfusion injury.

Pentoxifylline in liver ischemia and reperfusion.

Pentoxifylline is a methylxanthine compound which was first filed in 1973 and registered in 1974 in the United States by Sanofi-Aventis Deustchland Gmbh for the treatment of intermittent claudication for chronic occlusive arterial disease. This methylxanthine was later discovered to be a phosphodiesterase inhibitor. Furthermore, its hemorheological properties and its function as an inhibitor of inflammatory cytokines, like TNF-α, allowed researchers to study its effects in organ ischemia and reperfusion and transplantation. Although this drug has demonstrated beneficial effects, the mechanisms by which Pentoxifylline exerts a protective effect are not fully understood. This paper focuses on reviewing the literature to define the effect of Pentoxifylline when used in liver ischemia and reperfusion injury. Our research shows different animal models in which Pentoxifylline has been used as well as different doses and time of administration, as the ideal dose and timing have not yet been ascertained in liver ischemia and reperfusion. In conclusion, Pentoxifylline has shown positive effects in liver ischemia and reperfusion injury, and the main mechanism seems to be associated with the inhibition of TNF-α.

Allopurinol in renal ischemia.

Allopurinol is a xanthine oxidase inhibitor and antioxidant free radical scavenger which facilitates the protection of ischemic organs in part via this mechanism of action. The accumulation of free radicals during ischemia and reperfusion is in great manner overcome by inhibitors of xanthine oxidase and by the development of endogenous antioxidants. The ischemic lesion generates a well-established inflammatory response with the subsequent production of inflammatory molecules characteristically present at the first stages of the injury. Inflammatory cytokines, chemokines, adhesion molecules, and other cellular and molecular compounds are consequently produced as the lesion sets in. Under these conditions, allopurinol diminishes the effect of inflammatory mediators during the ischemic inflammatory response. This study reviews the literature associated with allopurinol and renal ischemia making special emphasis on the best dose and time of administration of allopurinol regarding its protective effect. It also defines the most accepted mechanism of protection on ischemichally damaged kidneys.

Pentoxifylline protects the small intestine after severe ischemia and reperfusion.

OBJECTIVES: Pentoxifylline, a methylxanthine derivative with significant hemorheologic properties, is used for claudication in patients with peripheral vascular disease, and experimentally for ischemic injury to organs because of its antioxidant and antiinflammatory effects. We used a rat model of severe small intestinal ischemia and reperfusion to determine the ability of pentoxifylline in improving survival, molecular response, and pathological protection. MATERIALS AND METHODS: We used 6 groups of male Wistar rats (n=25 each). The superior mesenteric artery was occluded for 120 minutes. Laboratory and tissue studies were done on 5 animals, 1 hour after reperfusion, and animal survival was assessed at 7 days. There were 2 control groups that received normal saline, either before ischemia or during reperfusion. The 4 treated groups received pentoxifylline 1 or 10 mg/kg at the same times mentioned above. Laboratory studies included measuring serum lactic acid dehydrogenase, tumor necrosis factor-α, interleukin-1ß, and interleukin-6.Intestinal tissue malondialdehyde and myeloperoxidase in small intestine tissue also were measured. Histology and laser vascular blood flow at baseline and reperfusion were obtained, and survival was determined 7 days after ischemia. RESULTS: A significant survival benefit in the animals treated with 10 mg/kg of pentoxifylline at reperfusion was noted. This coincided with a reduction in biochemical markers of cell damage - specifically, serum lactic acid dehydrogenase, and tissue malondialdehyde, ischemia, and reperfusion. Additionally, we saw decreased levels of tumor necrosis factor-α, interleukin-1ß, and interleukin-6. Improved postreperfusion blood flow shown by laser Doppler technology also was seen in the treated groups. Histologically, we observed less neutrophil infiltration in the intestine of ischemic-treated rats. Also seen in the control animals were increased necrotic lesions in the microvilli with a higher presence of lysozyme in the Paneth cells. Survival was significantly better at 7 days (70% vs 40%) when we compared the pentoxifylline group treated at reperfusion (10 mg/kg) to the ischemic controls. CONCLUSIONS: Pentoxifylline had a significant protective effect on severely ischemic bowel when administered during reperfusion at a dosage of 10 mg/kg. Better survival, improved histology, and molecular response should urge consideration of the consideration of applying these findings in some general surgery and transplant conditions.