Magnetic Fe3 O4 @graphene oxide improves the therapeutic effects of embryonic stem cells on acute liver damage.

OBJECTIVE: Acute liver failure is usually associated with inflammation and oxidation of hepatocytes and has high mortality and resource costs. Mesenchymal stem cell (MSCs) has occasionally been reported to have no beneficial effect due to poor transplantation and the survival of implanted cells. Recent studies showed that embryonic stem cell (ESC)-derived MSCs are an alternative for regenerative medicine. On the other hand, graphene-based nanostructures have proven useful in biomedicine. In this study, we investigated whether magnetic graphene oxide (MGO) improved the effects of ESC-MSC conditioned medium (CM) on protecting hepatocytes and stimulating the regeneration of damaged liver cells. MATERIALS AND METHODS: To provide a rat model of acute liver failure, male rats were injected intraperitoneally with carbon tetrachloride (CCl4 ). The rats were randomly divided into six groups, namely control, sham, CCl4 , ESC-MSC-CM, MGO and ESC-MSC-CM + MGO. In the experimental groups, the rats received, depending on the group, 2 ml/kg body weight CCl4 and either ESC-MSC-CM with 5 × 106 MSCs or 300 µg/kg body weight MGO or both. Symptoms of acute liver failure appeared 4 days after the injection. All groups were compared and analysed both histologically and biochemically 4 days after the injection. Finally, the results of ESC-MSC-CM and MSC-CM were compared. RESULTS: The results indicated that the use of MGO enhanced the effect of ESC-MSC-CM on reducing necrosis, inflammation, aspartate transaminase, alanine aminotransferase and alkaline phosphatase in the CCl4 -induced liver failure of the rat model. Also, the expression of vascular endothelial growth factor and matrix metalloproteinase-9 (MMP-9) was significantly upregulated after treatment with MGO. Also, the results showed that the ESC-MSC-CM has more efficient effective compared to MSC-CM. CONCLUSION: Magnetic graphene oxide improved the hepatoprotective effects of ESC-MSC-CM on acute liver damage, probably by suppressing necrosis, apoptosis and inflammation of hepatocytes.

Two Novel Compounds with Tri-aryl Structures as Effective Anti-Breast Cancer Candidates In-vivo.

Prognosis of metastatic breast cancer is very poor which urges the necessity to develop novel potential drug candidates. We assessed two compounds with tri-aryl structures (A and B) for their potency to reduce primary breast tumor growth and lung metastasis in 4T1 mice model. MTT assay, 4T1 mammary mouse model, and immunohistochemistry experiments were used in this study. In-vitro results exhibited an anti-proliferative effect for compounds A and B towards MDA-MB-231 cancer cells. Our in-vivo results displayed that administered compounds A and B could suppress the size of the primary tumor and the number of lung metastatic foci in 4T1 BALB/c mice model. Histopathological analysis revealed that treatment of both compounds resulted in necrosis. Our findings provide new evidence that compound B may be promising for slowing the growth of tumor along with metastatic foci via COX-2 independent pathway.

Topical hydrogel containing Fumaria vaillantii Loisel. extract enhances wound healing in rats.

BACKGROUND: Fumaria species (Fumariacea) has traditionally been used in wound healing in Iranian folk medicine. However, with the discovery of newer agents, its use has faded off into total obscurity. This study explored the wound healing potential of a gel containing 10% Fumaria vaillantii Loisel through topical application of total extract in a model of excisional as well as incisional wound healing in albino Wistar rats. METHODS: Rats were anesthetized, and excisional skin wound was established using a sterilized surgical scissors. The animals were then treated with 10% F.vaillantii topical gel formulation along with the gel base. The treatments were administered once a day after the injury for 21 days. For topical treatment, the hydrogel was formulated and evaluated for chemical and physical characteristics. Histopathological analysis with hematoxylin and eosin (H&E) was used for microscopic examination of the skin tissues on 21-day-old sections of excision wound. To verify collagen formation, hydroxyproline determination was performed 21 days post wound healing. Breaking strength was determined in a 10-day-old incision wound by the uniaxial tensile test. RESULTS: Topical administration of F.vaillantii gel formulation significantly enhanced skin wound closure on the 6th post-wounding day compared to both gel base and the negative control, indicating an accelerated wound healing process, while a significant difference was observed on 10th and 14th post -wound days in F.vaillantii treatment compared to the negative control groups. Gel formulation prepared with a 10% F. vaillantii extract exhibited a response in terms of wound epithelialization, angiogenesis and number of hair follicles at wound area better than the gel base on the 21st post-wound day. Application of gel base produced further advantages by increasing hydroxyproline content and collagen fiber thickness. Our results on incision wound model were supported by histopathological data indicating the role of gel base in the enhancement of breaking strength. CONCLUSION: Traditional use of Fumaria species in the skin diseases was justified in this study by revealing the increase in wound healing activity after hydrogel containing F. vaillantii total extract administration.

A novel hydrazide compound exerts anti-metastatic effect against breast cancer.

BACKGROUND: There are currently a number of barriers hindering the successful treatment of breast cancer, including the metastatic spread of cancer cells. In looking for new anticancer agents, we reported two novel hydrazide derivatives with anti-cancer activity in human breast cancer cells. The current study aims to explore the therapeutic potential of the most effective one, N'-((5-nitrothiophen-2-yl)methylene)-2-(phenylthio)benzohydrazide (compound B), on metastatic breast cancer, which is resistant to available chemotherapeutics. METHODS: 4T1 mammary carcinoma cells were inoculated into the fat pad mammary of 5-7-week-old female BALB/c mice and then the effective compound was intraperitoneally administered for 4 weeks. Proliferation index and angiogenesis in tumor and lung tissues were examined with immunohistochemistry. In vitro assessments were also carried out to evaluate the effect of the compound on invasion of MDA-MB-231 cells. RESULTS: Our results demonstrated that this effective derivative significantly inhibited invasion of MDA-MB-231 cells in vitro as shown by Matrigel assay and quantitative real-time method for MMP-9 expression after 48 h of treatment. Daily administration of the compound suppressed the growth of primary tumor and its metastasis to lung, which was confirmed by H&E experiment at a dose of 1 mg/kg in a well-known metastatic model of 4T1 breast cancer in syngeneic BALB/c mice. These outcomes were supported by the immunohistochemical examinations of the tumor and lung tissues of mice. Tumors and lungs in mice treated with the effective compound showed a reduced proliferation index and a smaller microvessel density compared to the control. CONCLUSION: This study highlights an anti-metastatic role for a novel hydrazide derivative in both in vitro and in vivo models of breast cancer.

A novel hydrazide compound exerts anti-metastatic effect against breast cancer

BACKGROUND: There are currently a number of barriers hindering the successful treatment of breast cancer, including the metastatic spread of cancer cells. In looking for new anticancer agents, we reported two novel hydrazide derivatives with anti-cancer activity in human breast cancer cells. The current study aims to explore the therapeutic potential of the most effective one, N'-((5-nitrothiophen-2-yl)methylene)-2-(phenylthio)benzohydrazide (compound B), on metastatic breast cancer, which is resistant to available chemotherapeutics. METHODS: 4T1 mammary carcinoma cells were inoculated into the fat pad mammary of 5-7-week-old female BALB/c mice and then the effective compound was intraperitoneally administered for 4 weeks. Proliferation index and angiogenesis in tumor and lung tissues were examined with immunohistochemistry. In vitro assessments were also carried out to evaluate the effect of the compound on invasion of MDA-MB-231 cells. RESULTS: Our results demonstrated that this effective derivative significantly inhibited invasion of MDA-MB-231 cells in vitro as shown by Matrigel assay and quantitative real-time method for MMP-9 expression after 48 h of treatment. Daily administration of the compound suppressed the growth of primary tumor and its metastasis to lung, which was confirmed by H&E experiment at a dose of 1 mg/kg in a well-known metastatic model of 4T1 breast cancer in syngeneic BALB/c mice. These outcomes were supported by the immunohistochemical examinations of the tumor and lung tissues of mice. Tumors and lungs in mice treated with the effective compound showed a reduced proliferation index and a smaller microvessel density compared to the control. CONCLUSION: This study highlights an anti-metastatic role for a novel hydrazide derivative in both in vitro and in vivo models of breast cancer.

Stimulation Effect of Low Level Laser Therapy on Sciatic Nerve Regeneration in Rat.

Introduction: Recent studies showed that low-level laser therapy (LLLT) accelerates the regeneration process of injured peripheral nerve tissue. The objective of this study was investigate the effect of LLLT (780 nm) on regeneration of injured right sciatic nerve of male Wistar rat. Methods: In this research work, the effect of LLLT (780 nm) on the regeneration process and reconstruction of injured peripheral right side sciatic nerve was investigated. Twelve adult male Wistar rats underwent surgery in aseptic conditions under general anesthesia to induce a lesion to their right side sciatic nerve according to standard protocol. Before suturing the location, only the experimental group was treated by laser. The damaged nerve was directly irradiated with (2 J, 100 mW, 40 seconds). The irradiation procedure was terminated in 21 days with little improvement (4 J, 200 mW, 40 seconds) across the skin surface of experimental group. Rats were selected randomly from each group to be sacrificed on different periods and histopathological examination was carried out on the extracted nerves. Results: Significant acceleration of revascularization and angiogenesis of the injury site was observed in the experimental group. Furthermore, a reduction of hemorrhages and increase in blood supply was observed. Also, Wallerian degeneration decreased while higher axonal density compared to the control rats was observed. Moreover, the cross-section analysis of the injured area on the 14th and 21st days as post-surgery showed that the nerve sheath diameter in the lesion area of the experimental group was reduced. While the ratio between thicknesses increased in the control group. Conclusion: The results of the current study suggest that laser phototherapy at 780 nm exactly could accelerate the regeneration process of injured peripheral nerves tissue.

Beneficial effect of pentoxifylline into the testis of rats in an experimental model of unilateral hindlimb ischemia/reperfusion injury.

OBJECTIVE: The objective of the present study was to investigate the role of pentoxifylline (PTX) on remote testicular injury caused by unilateral hind limb ischemia/reperfusion of rats. MATERIALS AND METHODS: Twenty healthy male Wistar rats were allocated randomly into two groups: ischemia/reperfusion (IR group) and ischemia/reperfusion + pentoxifylline (IR+PTX group). Ischemia was induced by placement of a rubber tourniquet at the greater trochanter for 2h. Rats in IR+PTX group received PTX (40 mg/kg IP) before the reperfusion period. At 24h after reperfusion, testes were removed and levels of superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT) and myeloperoxidase (MPO) activity were determined in testicular tissues. Three rats of each group were used for wet/ dry weight ratio measurement. Testicular tissues were also examined histopathologically under light microscopy. RESULTS: Activities of SOD and CAT in testicular tissues were decreased by ischemia/ reperfusion (P<0.05). Significantly increased MDA levels in testicular tissues were decreased by PTX treatment (P<0.05). MPO activity in testicular tissues in the IR group was significantly higher than in the IR+PTX group (P<0.05). The wet/dry weight ratio of testicular tissues in the IR group was significantly higher than in the IR+PTX group (P<0.05). Histopathologically, there was a statistically significant difference between two groups (P<0.05). CONCLUSIONS: According to histological and biochemical findings, we conclude that PTX has preventive effects in the testicular injury induced by hind limb ischemia/reperfusion.

Beneficial effect of pentoxifylline into the testis of rats in an experimental model of unilateral hindlimb ischemia/reperfusion injury

The objective of the present study was to investigate the role of pentoxifylline (PTX) on remote testicular injury caused by unilateral hind limb ischemia/reperfusion of rats.

Melatonin attenuates lung injury in a hind limb ischemia-reperfusion rat model.

OBJECTIVE: This study evaluated the protective antioxidant effect of melatonin on lung injury as a remote organ after skeletal muscle ischemia-reperfusion in rats. METHODS: Thirty male Wistar rats were allocated randomly into three experimental groups: operated with no ischemia (Sham) group, ischemia-reperfusion group and ischemia-reperfusion+melatonin group. Hind limb ischemia was induced by clamping the femoral artery. After 2h ischemia, the clamp was removed and the animal underwent 24h reperfusion. Rats in the ischemia-reperfusion + melatonin group received melatonin (10 mg/kg i.v.), immediately before the clamp was removed. At the end of the trial, animals were euthanized and the lungs were removed for water content determination, histopathological and biochemical studies. RESULTS: In the ischemia-reperfusion + melatonin group, tissues showed less intense histological abnormalities such as neutrophilic infiltration, intra-alveolar hemorrhage and edema compared with the ischemia-reperfusion group. Histopathologically, there was a significant difference (P < 0.05) between the two groups. The lung water content in the ischemia-reperfusion + melatonin group was significantly lower than the ischemia-reperfusion group (P < 0.05). Lung tissue myeloperoxidase (MPO) activity and nitric oxide (NO) level were significantly (P < 0.05) increased by ischemia-reperfusion. The increase in these parameters was reduced by melatonin. Comparing the ischemia-reperfusion+melatonin group with the sham group, no significant increase in all analyzed aspects of the research was observed. CONCLUSIONS: These findings suggest that melatonin has preventive effects in lung tissue injury after transient femoral artery occlusion.

Effects of light-emitting diode (LED) therapy on skeletal muscle ischemia reperfusion in rats.

Low-level laser therapy has been shown to decrease ischemia-reperfusion injuries in the skeletal muscle by induction of synthesis of antioxidants and other cytoprotective proteins. Recently, the light-emitting diode (LED) has been used instead of laser for the treatment of various diseases because of its low operational cost compared to the use of a laser. The objective of this work was to analyze the effects of LED therapy at 904 nm on skeletal muscle ischemia-reperfusion injury in rats. Thirty healthy male Wistar rats were allocated into three groups of ten rats each as follows: normal (N), ischemia-reperfusion (IR), and ischemia-reperfusion + LED (IR + LED) therapy. Ischemia was induced by right femoral artery clipping for 2 h followed by 2 h of reperfusion. The IR + LED group received LED irradiation on the right gastrocnemius muscle (4 J/cm(2)) immediately and 1 h following blood supply occlusion for 10 min. At the end of trial, the animals were euthanized and the right gastrocnemius muscles were submitted to histological and histochemical analysis. The extent of muscle damage in the IR + LED group was significantly lower than that in the IR group (P < 0.05). In comparison with other groups, tissue malondialdehyde (MDA) levels in the IR group were significantly increased (P < 0.05). The muscle tissue glutathione (GSH), superoxide dismutases (SOD), and catalase (CAT) levels in the IR group were significantly lower than those in the subjects in other groups. From the histological and histochemical perspective, the LED therapy has alleviated the metabolic injuries in the skeletal muscle ischemia reperfusion in this experimental model.

Effect of Otostegia persica extraction on renal injury induced by hindlimb ischemia-reperfusion: a rat model.

INTRODUCTION: It is known that ischemia-reperfusion causes remote organ injury as well as local injury. In traditional systems of medicine, many plants have been documented to be useful for the treatment of various disorders including oxidative esters. This study was designed to investigate whether Otostegia persica extraction pretreatment has a protective effect against renal injury induced by hindlimb ischemia-reperfusion. METHODS: Forty male Wistar rats were allocated into five groups as follows: Control, Sham, Otostegia persica, ischemia-reperfusion and ischemia-reperfusion+Otostegia persica groups. Rats in Otostegia persica and ischemia-reperfusion+Otostegia persica groups received Otostegia persica extraction (300 mg/kg) orally 2 days prior to operation. Hindlimb ischemia was induced by clamping the femoral artery for 2 h. After 24 h of reperfusion, blood and urine samples were obtained for kidney function tests and the kidneys were removed for histological analysis and oxidative stress measurement. RESULTS: The decrease in glomerular filtration rate induced by reperfusion was significantly improved by Otostegia persica extraction administration (P<0.05), which resulted in the decrease in serum urea and creatinine concentrations. Urinary creatinine significantly decreased in ischemia-reperfusion group compared to the other groups (P<0.05). Urinary excretion rate, water intake and the ratio of kidney/body weight significantly increased in animals with reperfusion injury as compared with other groups (P<0.05). On histological examination, rats pretreated with Otostegia persica extraction had nearly normal morphology. Skeletal muscle ischemia-reperfusion produced a significant increase in renal tissue malondialdehyde level, while pretreatment with Otostegia persica extraction was associated with a significantly lower malondialdehyde level (P<0.05). Renal tissue catalase and superoxide dismutase activity and glutathione level were significantly (P < 0.05) decreased by hindlimb ischemia-reperfusion. The increases in these parameters were decreased by pretreatment with Otostegia persica extraction. CONCLUSIONS: The results of this study showed that Otostegia persica extraction pretreatment significantly protected the renal injury from skeletal muscle ischemia-reperfusion.

TBHP-induced oxidative stress alters microRNAs expression in mouse testis.

PURPOSE: Reactive oxygen species (ROS) and oxidative stress is one of the main reasons of male infertility. MicroRNAs (miRNAs) regulate multiple intracellular processes. Alterations in miRNAs expression may occur in different conditions and diseases. In this study, the effect of oxidative stress induced by tertiary-butyl hydroperoxide (TBHP) on the expression of candidate miRNAs in mouse testis was investigated. METHODS: After determining median lethal dose (LD50), TBHP was intraperitoneally (ip) injected at the dilution of 1:10 LD50 into the adult male mice for 2 weeks, and then testis tissues were removed in order to assay the ROS level. Total RNA was extracted and the expression of five miRNAs was quantified by reverse transcription-real time polymerase chain reaction (RT-qPCR). RESULTS: The flow cytometry analysis showed a significant increase in ROS level in testis. The expression of three out of five selected miRNAs, including miR-34a, miR-181b and miR-122a, showed some degrees of changes following exposure to oxidative stress. These miRNAs are involved in antioxidant responses, inflammation pathway and spermatogenesis arrest. CONCLUSIONS: In conclusion, TBHP alters the miRNA expression profile of testis which might play a potential role in oxidative and antioxidative responses and spermatogenesis.

WITHDRAWN: Melatonin attenuates lung injury in a hind limb ischemia-reperfusion rat model.

This article has been withdrawn for editorial reasons because the journal will be published only in English. In order to avoid duplicated records, this article can be found at http://dx.doi.org/10.1016/j.rppnen.2014.01.010. The Publisher apologizes for any inconvenience this may cause. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.

On the benefit of Teucrium in murine colitis through improvement of toxic inflammatory mediators.

Regarding the role of free radicals in pathogenesis of inflammatory bowel disease (IBD), we were interested to investigate the effects of Teucrium persicum with approved antioxidant and anti-inflammatory properties in an experimental model of colitis. Immunologic colitis was induced by rectal administration of a mixture of 2,4,6-trinitrobenzene sulphonic acid (TNBS) and ethanol through rubber cannula into rats. Three different doses of Teucrium (100, 200, and 400 mg/kg) were gavaged in a duration of 10 days to rats. Endpoint markers of colitis included macroscopic and microscopic examination of colon tissue and measuring colonic cells concentrations of tumor necrosis factor-alpha (TNF-alpha), interlukin-1beta (IL-1beta), total antioxidant power as ferric reducing antioxidant power (FRAP), myeloperoxidase (MPO), and lipid peroxidation as thiobarbitoric acid-reactive substance (TBARS). Teucrium at all doses improved both macroscopic and histological damages of rats with colitis. Teucrium reduced colonic MPO activity and concentrations of cellular lipid peroxides, TNF-alpha, and IL-1beta, with a concomitant increase in FRAP value in rats with colitis. It is concluded that beneficial effects of Teucrium in experimental colitis is mediated through its antioxidant and anti-inflammatory potentials. Examination of this herbal medicine in patients with IBD as a supplement would further reveal the potential of Teucrium.