Cardiac aorta-derived extracellular matrix scaffold enhances critical mediators of angiogenesis in isoproterenol-induced myocardial infarction mice

An incapability to improve lost cardiac muscle caused by acute ischemic injury remains the most important deficiency of current treatments to prevent heart failure. We investigated whether cardiomyocytes culturing on cardiac aorta-derived extracellular matrix scaffold has advantageous effects on cardiomyocytes survival and angiogenesis biomarkers’ expression. Ten male NMRI mice were randomly divided into two groups: (1) control (healthy mice) and (2) myocardial infarction (MI)-induced model group (Isoproterenol/subcutaneously injection/single dose of 85 mg/kg). Two days after isoproterenol injection, all animals were sacrificed to isolate cardiomyocytes from myocardium tissues. The fresh thoracic aorta was obtained from male NMRI mice and decellularized using 4% sodium deoxycholate and 2000 kU DNase-I treatments. Control and MI-derived cardiomyocytes were seeded on decellularized cardiac aorta (DCA) considered three-dimensional (3D) cultures. To compare, the isolated cardiomyocytes from control and MI groups were also cultured as a two-dimensional (2D) culture system for 14 days. The cell viability was examined by MTT assay. The expression levels of Hif-1α and VEGF genes and VEGFR1 protein were tested by real-time PCR and western blotting, respectively. Moreover, the amount of VEGF protein was evaluated in the conditional media of the 2D and 3D systems. The oxidative stress was assessed via MDA assay. Hif-1α and VEGF genes were downregulated in MI groups compared to controls. However, the resulting data showed that decellularized cardiac aorta matrices positively affect the expression of Hif-1α and VEGF genes. The expression level of VEGFR1 protein was significantly (p ≤ 0.01) upregulated in both MI and healthy cell groups cultured on decellularized cardiac aorta matrices as a 3D system compared to the MI cell group cultured in the 2D systems. Furthermore, MDA concentration significantly decreased in 3D-cultured cells (MI and healthy cell groups) rather than the 2D-cultured MI group (p ≤ 0.015). The findings suggest that cardiac aorta-derived extracellular scaffold by preserving VEGF, improving the cell viability, and stimulating angiogenesis via upregulating Hif-1α, VEGF, and VEGFR1 in cardiomyocytes could be considered as a potential approach along with another therapeutic method to reduce the complications of myocardial infarction and control the progressive pathological conditions related to MI.

Proposition of a Simplified Protocol and New Parameter Introduction in NMRI Mice Anhedonia Induction

Introduction: A broad spectrum of research involving stress and stress protocols has long proven that a point of anhedonia, social defeat and learned helplessness can be achieved and observed – with anhedonia being a clinical symptom of Anxiety, Depression and Bipolar Affective Disorder. Aim: The aim of this study is the development of a simplified protocol for anhedonia induction in NMRI male mice in order to shorten the period of mice suffering and decrease complexity of the procedure for other researchers and introduction of new parameter in order to achieve better standardization of results. Materials and Methods: 21 male NMRI mice were introduced to 2 different stress protocols (one found in literature and one simplified) where cognitive- behavioral status was tested using the Sucrose Preference Test, Open Field Test, Grooming Pattern and histological examination of adrenal glands, and to propose a new protocol for fellow researchers. Results: Results observed include the successful induction of anhedonia proven by Sucrose Preference Tests, Barbering effect and microhemorrhage of the adrenal glands. Simplified protocol showed superiority compared to the one found in literature. Conclusion: Simplified protocol showed higher efficiency and reduced amount of work during testing phase. Introduction of NMI as a new parameter during behavioral evaluation resulted in better standardization of measured SPT values that incorporates common knowledge of mammal physiology. A big developmental step was the introduction of the Normalised Mass Index to even out mass fluctuations and differences in basal metabolism, which we recommend to other researchers and institutions.

The Effect of Different Doses of Mesobuthus eupeus (Scorpionida: Buthidae) Scorpion Venom on the Production of Liver Necrosis in Nmri Mice

Background: Scorpion venom has a variety of different components considerably. Some of these compounds are pro-teins such as Phospholipase A2 which is one of the most important. Use of scorpion venom for the treatment of any disease requires an initial study to determine the therapeutic dose or safe dose. Therefore, due to the necessity of study-ing scorpion venom, it is of special importance to study the effects of its dose response in animal tissues. Methods: To determine the inflammatory effects of scorpion’s venom (Mesobuthus eupeus), 50 Nmri mice with an average weight of 24±7g were selected for investigation in two experiments. In first-round 25 of them were divided into 5 groups and were exposed to different doses of venom injection paralleling the control group. Single-injection of vari-ous doses on 25 mice was performed and results were compared. Results: There was a significant differences between the test and control groups (in most groups). Liver necrosis was one of the important symptoms in this study, the severity of which was measured and statistically analyzed. Conclusion: It was determined that 0.05ppm is a safe dose and sub-lethal doses can use for the investigation of thera-peutic effects of venom on cancer, diabetes, dermatitis, and so on.

Protective Effects of Curcumin on Sperm and Stereological Parameters in Testes of Formaldehyde-Exposed NMRI Mice: An Experimental Study

Background and Aims: Formaldehyde (FA) exposure is an important cause of cellular injury and oxidative damage in testis, leading to infertility. This study aimed to assess the protective effects of curcumin on sperm and stereological parameters in testes from formaldehyde-exposed NMRI mice. Materials and Methods: At 6-8 weeks of age, 24 adult male NMRI mice weighing 30-35 g were categorized into four groups (n=6) based on the treatment they received: Group І (control) received no treatment, group ΙΙ received FA (10 mg/kg), group ΙΙΙ received FA (10 mg/kg) and curcumin (100 mg/kg), and group IV (Solvent) received dimethyl sulfoxide (0.2 ml/day). Materials were administered intraperitoneally for 35 days. After excision, epididymis tissues were placed in 1 mL aliquots of Ham’s F10 medium at 37˚C for 20 min and were then used in analyses of sperm parameters. Testes were fixed and stained with Hematoxylin & Eosin to investigate stereological indices. We also determined lipid peroxidation levels using malondialdehyde assays. Results: Mean sperm parameters (count, motility, viability, and morphology) differed significantly between groups ΙΙ and ΙΙΙ (p≤0.001). Stereological indices, including Leydig and spermatogonia cell numbers and surface-to-volume ratios of seminiferous tubules were significantly higher in group ΙΙΙ than in group ΙΙ (p≤0.001) . Finally, malondialdehyde levels in group III were significantly lower than in group II (p=0.001). Conclusions: The data showed that the curcumin, as an antioxidant, reduced FA-induced damage in sperm parameters and stereological indices in mice testes.