Development of BDNF/NGF/IKVAV Peptide Modified and Gold Nanoparticle Conductive PCL/PLGA Nerve Guidance Conduit for Regeneration of the Rat Spinal Cord Injury.

Spinal cord injuries are very common worldwide, leading to permanent nerve function loss with devastating effects in the affected patients. The challenges and inadequate results in the current clinical treatments are leading scientists to innovative neural regenerative research. Advances in nanoscience and neural tissue engineering have opened new avenues for spinal cord injury (SCI) treatment. In order for designed nerve guidance conduit (NGC) to be functionally useful, it must have ideal scaffold properties and topographic features that promote the linear orientation of damaged axons. In this study, it is aimed to develop channeled polycaprolactone (PCL)/Poly-D,L-lactic-co-glycolic acid (PLGA) hybrid film scaffolds, modify their surfaces by IKVAV pentapeptide/gold nanoparticles (AuNPs) or polypyrrole (PPy) and investigate the behavior of motor neurons on the designed scaffold surfaces in vitro under static/bioreactor conditions. Their potential to promote neural regeneration after implantation into the rat SCI by shaping the film scaffolds modified with neural factors into a tubular form is also examined. It is shown that channeled groups decorated with AuNPs highly promote neurite orientation under bioreactor conditions and also the developed optimal NGC (PCL/PLGA G1-IKVAV/BDNF/NGF-AuNP50) highly regenerates SCI. The results indicate that the designed scaffold can be an ideal candidate for spinal cord regeneration.

Active shrinkage protects neurons following axonal transection.

Trauma, vascular events, or neurodegenerative processes can lead to axonal injury and eventual transection (axotomy). Neurons can survive axotomy, yet the underlying mechanisms are not fully understood. Excessive water entry into injured neurons poses a particular risk due to swelling and subsequent death. Using in vitro and in vivo neurotrauma model systems based on laser transection and surgical nerve cut, we demonstrated that axotomy triggers actomyosin contraction coupled with calpain activity. As a consequence, neurons shrink acutely to force water out through aquaporin channels preventing swelling and bursting. Inhibiting shrinkage increased the probability of neuronal cell death by about 3-fold. These studies reveal a previously unrecognized cytoprotective response mechanism to neurotrauma and offer a fresh perspective on pathophysiological processes in the nervous system.

High-resolution mapping of sensory fibers at the healthy and post-myocardial infarct whole transgenic hearts.

The sensory nervous system is critical to maintain cardiac function. As opposed to efferent innervation, less is known about cardiac afferents. For this, we mapped the VGLUT2-expressing cardiac afferent fibers of spinal and vagal origin by using the VGLUT2::tdTomato double transgenic mouse as an approach to visualize the whole hearts both at the dorsal and ventral sides. For comparison, we colabeled mixed-sex transgenic hearts with either TUJ1 protein for global cardiac innervation or tyrosine hydroxylase for the sympathetic network at the healthy state or following ischemic injury. Interestingly, the nerve density for global and VGLUT2-expressing afferents was found significantly higher on the dorsal side compared to the ventral side. From the global nerve innervation detected by TUJ1 immunoreactivity, VGLUT2 afferent innervation was detected to be 15-25% of the total network. The detailed characterization of both the atria and the ventricles revealed a remarkable diversity of spinal afferent nerve ending morphologies of flower sprays, intramuscular endings, and end-net branches that innervate distinct anatomical parts of the heart. Using this integrative approach in a chronic myocardial infarct model, we showed a significant increase in hyperinnervation in the form of axonal sprouts for cardiac afferents at the infarct border zone, as well as denervation at distal sites of the ischemic area. The functional and physiological consequences of the abnormal sensory innervation remodeling post-ischemic injury should be further evaluated in future studies regarding their potential contribution to cardiac dysfunction.

The Effects of Transcranial Focused Ultrasound Stimulation of Nucleus Accumbens on Neuronal Gene Expression and Brain Tissue in High Alcohol-Preferring Rats.

We investigated the effect of low-intensity focused ultrasound (LIFU) on gene expression related to alcohol dependence and histological effects on brain tissue. We also aimed at determining the miRNA-mRNA relationship and their pathways in alcohol dependence-induced expression changes after focused ultrasound therapy. We designed a case-control study for 100 days of observation to investigate differences in gene expression in the short-term stimulation group (STS) and long-term stimulation group (LTS) compared with the control sham group (SG). The study was performed in our Experimental Research Laboratory. 24 male high alcohol-preferring rats 63 to 79 days old, weighing 270 to 300 g, were included in the experiment. LTS received 50-day LIFU and STS received 10-day LIFU and 40-day sham stimulation, while the SG received 50-day sham stimulation. In miRNA expression analysis, it was found that LIFU caused gene expression differences in NAc. Significant differences were found between the groups for gene expression. Compared to the SG, the expression of 454 genes in the NAc region was changed in the STS while the expression of 382 genes was changed in the LTS. In the LTS, the expression of 32 genes was changed in total compared to STS. Our data suggest that LIFU targeted on NAc may assist in the treatment of alcohol dependence, especially in the long term possibly through altering gene expression. Our immunohistochemical studies verified that LIFU does not cause any tissue damage. These findings may lead to new studies in investigating the efficacy of LIFU for the treatment of alcohol dependence and also for other psychiatric disorders.

Comparison of the efficiencies of intrathecal and intraganglionic injections in mouse dorsal root ganglion.

Background/aim: Dorsal root ganglia (DRG) are structures containing primary sensory neurons. Intraganglionic (IG) and intrathecal (IT) applications are the most common methods used for viral vector transfer to DRG. We aim to compare the efficiencies and pathological effects of IT and IG viral vector delivery methods to DRG, through in vivo imaging. Materials and methods: Mice were divided into four groups of six each: IT, IG, IT-vehicle, and IG-vehicle. Adeno-associated virus (AAV) injection was performed for EGFP expression in IT/IG groups. DRGs were made visible through vertebral window surgery and visualized with multiphoton microscopy. After imaging, spinal cords and DRGs were removed and cleared, then imaged with light sheet microscopy. Results: No neuronal death was observed after IT injection, while the death rate was 17% 24 h after IG injection. EGFP expression efficiencies were 90%-95% of neurons in both groups. EGFP expression was only observed in targeted L2 DRG after IG injection, while it was observed in DRGs located between L1-L5 levels after IT injection. Conclusion: IT injection is a more suitable method for labeling DRG neurons in neurodegenerative injury models. However, when the innervation of DRG needs to be specifically studied, IT injection reduces this specificity due to its spread. In these studies, IG injection is the most suitable method for labeling single DRG neurons.

Lithium promotes long-term neurological recovery after spinal cord injury in mice by enhancing neuronal survival, gray and white matter remodeling, and long-distance axonal regeneration.

Spinal cord injury (SCI) induces neurological deficits associated with long-term functional impairments. Since the current treatments remain ineffective, novel therapeutic options are needed. Besides its effect on bipolar mood disorder, lithium was reported to have neuroprotective activity in different neurodegenerative conditions, including SCI. In SCI, the effects of lithium on long-term neurological recovery and neuroplasticity have not been assessed. We herein investigated the effects of intraperitoneally administered lithium chloride (LiCl) on motor coordination recovery, electromyography (EMG) responses, histopathological injury and remodeling, and axonal plasticity in mice exposed to spinal cord transection. At a dose of 0.2, but not 2.0 mmol/kg, LiCl enhanced motor coordination and locomotor activity starting at 28 days post-injury (dpi), as assessed by a set of behavioral tests. Following electrical stimulation proximal to the hemitransection, LiCl at 0.2 mmol/kg decreased the latency and increased the amplitude of EMG responses in the denervated hindlimb at 56 dpi. Functional recovery was associated with reduced gray and white matter atrophy rostral and caudal to the hemitransection, increased neuronal survival and reduced astrogliosis in the dorsal and ventral horns caudal to the hemitransection, and increased regeneration of long-distance axons proximal and distal to the lesion site in mice receiving 0.2 mmol/kg, but not 2 mmol/kg LiCl, as assessed by histochemical and immunohistochemical studies combined with anterograde tract tracing. Our results indicate that LiCl induces long-term neurological recovery and neuroplasticity following SCI.

Anatomical characterization of vagal nodose afferent innervation and ending morphologies at the murine heart using a transgenic approach.

Heart is an extensively innervated organ and its function is strictly coordinated by autonomic neural circuits. After pathological events such as myocardial infarction (MI), cardiac nerves undergo a structural and functional remodeling contributing to cardiac dysfunction. Although the efferent component of the cardiac nerves has been well described, sensory innervation of the heart has not been defined in detail. Considering its importance, comprehensive description of vagal afferent innervation on the whole heart would enable a better description of autonomic imbalances manifesting as sympathoexcitation and vagal withdrawal in post-ischemic states. To address this issue, we globally mapped the vagal nodose afferent fibers innervating the whole murine heart with unprecedented resolution. By using the Phox2b-Cre::tdTomato transgenic mouse line, we described the detailed distribution and distinct vagal sensory ending morphologies at both the dorsal and ventral sides of the mouse heart. By neural tracing analysis, we quantitated the distribution and prevalence of vagal afferent nerve fibers with varying diameters across dorsal and ventral surfaces of the heart. Moreover, we demonstrated that vagal afferents formed flower spray and end-net-like endings within the atria and ventricles. As distinct from the atria, vagal afferents formed intramuscular array-like endings within the ventricles. Furthermore, we showed that vagal afferents undergo structural remodeling by forming axonal sprouts around the infarct area in post-MI hearts. These findings improve our understanding of the potential effect of vagal afferent remodeling on autonomic imbalance and generation of cardiac arrhythmias and could prospectively contribute to the development of more effective neuromodulatory therapies.

The association of oxidative stress and DNA damage with XRCC1 and XRCC3 polymorphisms in radiology technicians.

Ionizing radiation has widespread use in medicine in the diagnosis and treatment of many medical conditions. Radiology technicians are one group that is occupationally exposed to low doses of radiation. There are questions regarding whether low dose exposure to radiation could have long-term health consequences. Assessing the effect of radiation on genetic material is essential for appraising long-term health results. Hereditary variations in DNA repair genes cause differentiation in individual responses to radiation related health effects. This study aimed to determine oxidative stress and DNA damage, and their relationship to XRCC1 (Arg399Gln) and XRCC3 (Thr241Met) polymorphisms in radiology technicians occupationally exposed to low dose radiation. Peripheral blood samples were collected from 45 radiology technicians and age-matched with 40 healthy control individuals working in office environments. Our results showed that radiology technicians had significantly greater oxidative stress and DNA damage than the control group, and women appeared more susceptible to occupational radiation exposure than men. Individuals with wild-type genotypes for XRCC1 (Arg/Arg) and XRCC3 (Thr/Thr) had less DNA damage. Lower DNA damage levels could be explained by the enhanced capacity to repair low dose radiation induced DNA damage. Further studies are needed to evaluate the role of DNA repair genes in individuals that are occupationally exposed to low dose radiation.

The effects of human amniotic membrane on silicone related capsule formation in rats.

Silicone breast implants are commonly used materials in plastic surgery for breast augmentation and reconstruction and the most severe complication of silicone implants are capsule contraction which occurs in 40% of patients. The aim of our study is to evaluate how the amniotic membrane alters the capsule formation effects of silicone 24 wistar rats were used in the study. We placed a bare silicone block into the left side (Subgroup A) and single layer amniotic membrane coated silicone block into the right side (Subgroup B) of the rats back. The rats were then separated into three groups and in group 1 rats were euthanized after 3 weeks, in group 2 after 12 weeks and in group 3 after 24 weeks. Then capsule thickness, fibroblast and lymphocyte cell counts were evaluated for each sample. In Group 2 and group 3, the capsule thickness in Subgroup B was detected to be statistically significantly lower than that in Subgroup A. In Group 1, 2, and 3, the lymphocyte count in the capsule tissue taken from Subgroup B was lower than Subgroup A but the difference was not statistically significant. In Group 2 and 3, the fibrocyte count detected in the capsule tissue in Subgroup B was found to be statistically significantly lower than Subgroup A. the amniotic membrane was demonstrated to reduce capsule thickness by the antifibrinolytic effect in our study.

Therapeutic effects of metformin for noise induced hearing loss.

OBJECTIVE: This study aimed to investigate the healing effect of metformin on noise induced hearing loss (NIHL) by measuring audiological, biochemical and histological parameters. MATERIALS AND METHODS: 32 rats were divided into four groups (Group 1: Noise, Group 2: Noise + Metformin, Grup 3: Metformin, Grup 4: Control). Broadband noise was applied to Group 1 and Group 2 after basal measurements. Measuring audiological (distortion product otoacoustic emission (DPOAE) and Auditory Brainstem Response (ABR)), biochemical (total antioxidant status (TAS), total oxidant status (TOS), oxidative status index (OSI), DNA damage, IL-1 beta, IL-6, TNF alfa, HSF-1 and COX-2) and histological parameters. RESULTS: Group 2 had significant decreases in ABR thresholds on day 7 and day 14 compared to day 1. DPOAE values of Group 2 on the 7th and 14th days were significantly higher than the post-noise levels. DNA damage, TOS and OSI values of Group 1 were significantly higher than the other groups. The Cox-2 value of Group 1 was higher than all other groups. The HSF-1 value of Group 2 was significantly higher than that of Group 1. In terms of IL-1 Beta, IL-6 and TNF-alpha values, there was no significant difference between groups 2, 3 and 4 and these values were significantly lower than group 1. In histopathological results of our study, no significant difference was found between the groups being exposed to noise and the control group. CONCLUSION: This study showed that early period of Metformin treatment has therapeutic effect on NIHL.

Transfer and Integration of Breast Milk Stem Cells to the Brain of Suckling Pups.

Beside its unique nutritional content breast milk also contains live cells from the mother. Fate of these cells in the offspring has not been adequately described. In this study, we aimed to detect and identify maternal cells in the suckling's blood and the brain. Green fluorescent protein expressing transgenic female mice (GFP+) were used as foster mothers to breastfeed wildtype newborn pups. One week and two months after the birth, blood samples and brains of the sucklings were analyzed to detect presence of GFP+ cells by fluorescence activated cell sorting, polymerase chain reaction and immunohistochemistry on the brain sections and optically cleared brains. The tests confirmed that maternal cells were detectable in the blood and the brain of the pups and that they differentiated into both neuronal and glial cell types in the brain. This phenomenon represents breastfeeding - induced microchimerism in the brain with functional implications remain to be understood.

Different concentrations of mesna application have an effect on the internal ear?

OBJECTIVE: The use of Mesna as a chemical dissector in higher concentrations may reduce the length of time of operation by providing more effective dissection as well as being used in otologic pathologies such as tympanosclerosis. In this study, it was aimed to assess the effect of Mesna on the internal ear, which was applied intra-tympanically in higher concentrations than the conventional use. METHODS: Twenty-four female rats were included in our study. The rats were randomly divided into three groups (Group 1: Mesna 50%, Group 2: Mesna 100%, Group 3: Saline). At the beginning of the study, DPOAE and ABR measurements were carried out on every rat on days 7 and 14. At the end of the study, cochleas of the rats were excised and histopathological assessments were carried out. RESULTS: Basal values and DPOAE and ABR values on day 7 and 14 of Group 1, Group 2, and Group 3 were similar to each other. No significant difference was detected among the three groups in the histopathological assessment carried out at the end of the study. CONCLUSION: It was revealed by audiological and histopathological parameters that the use of Mesna at 50% and 100% concentrations did not create toxicity effects on the internal ear. Mesna would be more effective by being used in higher concentrations in audiological surgeries, that its duration of operation world reduce and could being used in different indications including tympanosclerosis.

Assessment of the effectiveness of cyclosporine nasal spray in an animal model of allergic rhinitis.

The aim of this study is to show if cyclosporine has an antiallergic role in a rat model of ovalbumin-induced allergic rhinitis. The 54 rats were divided into six equal groups. The first group was a negative control group without induced allergic rhinitis; the second group a positive control with induced allergic rhinitis not receiving treatment. The remaining four groups, after induction of allergic rhinitis, received intranasal cyclosporine treatment in doses of 0.05, 0.1, or 0.2% or nasal steroid treatment. In the biochemical examination, on the surface of the tissue tumor necrosis factor (TNF) interferon (IFN), interleukin (IL)-5, IL-13, as well as IL-2, IL-4, IL-17A, and IgE were studied. Histologically, ciliary loss, increase of goblet cells, vascular congestion, and the degree of eosinophil infiltration were rated. In all treatment groups, on average, a significant reduction in all histological and biochemical values was found compared to the positive control group. Comparing each of the three cyclosporine-using groups with the group of nasal corticosteroid did not show any significant difference in the average scores. Cyclosporine nasal drops are effective to be used in an animal model of experimental allergic rhinitis without systemic effects.

Evaluation of ototoxicity of intratympanic administration of Methotrexate in rats.

OBJECTIVE: Methotrexate is a dihydrofolate reductase enzyme inhibitor with very high selectivity, and it is an antiproliferative folic acid antagonist used for the treatment of autoimmune diseases. In this study, our objective was to evaluate the effect of intratympanic Methotrexate application in the inner ear. METHODS: This study was planned as an animal study. This study performed in a tertiary referral center. 24 healthy female rats were used in our study. They were separated into three groups. 0.2 cc intratympanic saline was applied to both ears of Group 1. Paracentesis was applied to the tympanic membrane in both ears of Group 2. 0.2 cc intratympanic Methotrexate was applied to both ears of Group 3. At the beginning of the study, Distortion-product otoacoustic emissions (DPOAE) and Auditory brainstem response (ABR) of all rats were measured and then again on the 5th, 10th and 15th day. Histologic examinations of all groups were compared. RESULTS: There was not any significant difference between basal DPOAE and ABR measurement values of the groups and the results were measured again on the 5th, 10th and 15th day (p > 0.05). There was no difference between the groups in terms of histology. CONCLUSION: The intratympanic Methotrexate injection does not have any ototoxic effect on inner ear. We assume that intratympanic Methotrexate could be used safely on inner ear diseases in which steroid treatment is contraindicated or not effective.

Effect of Energy Drink on Bone Formation in the Expanded Inter-Premaxillary Suture.

Maxillary expansion of the median palatal suture is a common procedure in orthodontics. Even after retention, there is a strong tendency to relapse in the expanded suture. The authors' objectives are to accelerate the bone formation process in the expanded suture and to reduce the required retention time by using an energy drink (ED). Twenty rats were divided into 2 groups (n = 20). The expansion-only group was defined as the control group (Group C). The other group was defined as the expansion-plus-energy drink group (Group ED). In Group ED, ED was administered systemically through oro-gastric tubes after the expansion period. After 5 days of expansion, the springs were removed and replaced with short lengths of rectangular retaining wire. Tooth separation was maintained for 12 days. Histomorphometric analysis showed significant differences between the 2 groups in terms of newly formed bone (P = 0.018) and the bone area (P = 0.007). For the parameters that were investigated, Group ED had better results than Group C. These results show that systemic administration of an ED during the early stages of the orthopedic expansion of the inter-maxillary suture areas can stimulate bone formation and decrease the time required for retention.

Neuroprotective Effects of Chronic Fenofibrate Treatment via Modulating the Immunoreactivity of Cleaved Caspase-3 in Stroke Induced by Transient Middle Cerebral Artery Occlusion Rat Model.

AIM: Current stroke therapies include lipid-lowering drugs, which reduce inflammation and serve to stabilize the atherosclerotic plaque to demonstrate better outcome and neuroprotection. Peroxisome proliferator activated receptors (PPAR) ? regulates lipid homeostasis and is a target of fibrates, which have a neuroprotective function by various mechanisms. In this study, we aimed to evaluate the role of the PPAR? agonist, fenofibrate, in the modulation of cleaved caspase-3 immunoreactivity and at the final infarct volume in an experimental ischemia/reperfusion rat model by induced transient proximal middle cerebral artery occlusion. MATERIAL AND METHODS: A total of 65 male Sprague Dawley rats were allocated into 4 groups; sham (n =5), experiment 1 (n=20), experiment 2 (n=20), experiment 3 (n=20). All experiment groups were divided to 3 subgroups in order to evaluate the final infarct volume at 24th hour (n=5) and the immunoreactivity of cleaved caspase -3 at different time periods [at first hour (n=5), at 6th hour (n=5), at 24th hour (n=5)] after transient middle cerebral artery occlusion (MCAo). At the study, the experiment groups (Experiment 1 and Experiment 2) were received the fenofibrate-diet during 14 days before ischemia procedure. All animals were sacrificed at 24th hours after MCAo. Infarction volumes were calculated from 2,3,5,triphenyltetrozolium chloride (TTC)- stained brain sections. RESULTS: We found that fenofibrate-therapy reduced significantly more body weight than the other experiment groups (p < 0.05). At the time intervals, a decrease of immunoreactivity of cleaved caspase-3 was significantly observed with fenofibrate therapy after MCAo (p < 0.05). Chronic fenofibrate treatment before cerebral ischemia significantly reduced the infarction size after MCAo compared with the other groups (respectively; p = 0.011 and p < 0.000). CONCLUSION: Fenofibrate treatment has neuroprotective effects on middle cerebral artery infarcts.

Protective effect of ß-glucan on acute lung injury induced by lipopolysaccharide in rats.

BACKGROUND/AIM: Lipopolysaccharide (LPS)-induced endotoxemia can cause serious organ damage such as acute lung injury and death by triggering the secretion of proinflammatory cytokines and acute-phase reactants. The goal of this study was to evaluate the effects of ß-glucan on inflammatory mediator levels and histopathological changes in LPS-induced endotoxemia. MATERIALS AND METHODS: Forty-seven male Wistar albino rats were randomly allocated into four groups as follows: control group, LPS group (10 mg/kg LPS), LPS + ß-glucan group (100 mg/kg ß-glucan before LPS administration), and ß-glucan group. Twelve hours after LPS administration, lung and serum samples were collected. Concentrations of IL-6, IL-8, C-reactive protein (CRP), and procalcitonin were measured in the serum at hours 0 (basal) and 12. The severity of lung damage was assessed by an appropriate histopathological scoring system. RESULTS: Serum levels of CRP in the LPS group at 12 h were significantly higher than in the other groups, whereas serum IL-6 levels in the LPS and LPS + ß-glucan groups at 12 h were significantly decreased. The mean histopathological damage score of the LPS group was slightly higher than that of the LPS + ß-glucan group. Moreover, mortality rate was significantly decreased in the LPS + ß-glucan group versus the LPS group. CONCLUSION: ß-glucan reduces endotoxemia-induced mortality and might be protective against endotoxemia-induced lung damage.

Histopathological evaluation of Ankaferd blood stopper use in the rabbit septoplasty model.

OBJECTIVE: The purpose of this study was to assess the use of Ankaferd blood stopper (ABS), an organic hemostatic agent of plant origin, in septoplasty operations, and to determine its effect on nasal septal tissues in the rabbit model. METHOD: The study was performed on 30 New Zealand adult male rabbits each weighing 2500-3500g (average: 3000g). The rabbits were randomly divided into 4 groups, namely, the control group (without septoplasty), the septoplasty+tampon group, the septoplasty+ABS group, and the septoplasty+tampon+ABS group. All animals were sacrificed after two weeks, and the nasal septums were total removed using the lateral rhinotomy technique. The specimens were obtained from similar sites of cartilaginous nasal septum. The sections were stained with hematoxylin-eosin and Mason trichrome stains and studied under the light microscope by the same pathologist who evaluated the mucosal ulcerations, the severity of inflammatory cell infiltration, the mucosal thickness, and the cartilage thickness. The data obtained were statistically analyzed using the Kruskal-Wallis variance analysis and the Mann-Whitney U test. RESULTS: No mucosal ulceration or inflammatory cell infiltration was detected in any of the rabbit groups. There was a statistically significant difference between the groups in terms of mucosal thickness and cartilage thickness (p<0.05). The values of mucosal thickness in the groups mentioned above were 147.7±17.6µm, 205.7±36.7µm, 139.6±14.8µm, and 190.2±17.5µm, respectively. The values of cartilage thickness were 398.2±28.9µm, 546.2±35.3µm, 363.7±24.7µm, and 447.8±28.2µm, respectively. There was no significant difference between the control group and the septoplasty+ABS group in terms of mucosal thickness and cartilage thickness (p>0.05). However, there was a significant increase in nasal mucosal and cartilage thickness in tampon-using groups when compared with the other groups (p<0.05). CONCLUSION: Although nasal tampons provide the contact of mucoperichondrium with the cartilage, they are generally accepted as a discomfort for patients. Ankaferd blood stopper can be used instead of nasal tampons to increase patient comfort.

Effects of caffeic acid phenethyl ester on wound healing in calvarial defects.

OBJECTIVE: The aim of this study is to analyze histologically the effect of CAPE on bone healing of Critical Size Defect (CSD) in rat calvaria. STUDY DESIGN: Thirty-two 3-month-old male rats were used. The animals were randomly divided into four groups. Group A received isotonic saline solution, Group B received CAPE (50 mmol/kg) locally, Group C received CAPE (100 mmol/kg) locally and Group D received CAPE (10 mmol/kg/day i.p. for 28 days) systematically. A 5-mm diameter calvarial defect was created in the right side of the parietal bone without damaging the underlying dura mater. Twenty-eight days after the surgery, all the animals were sacrificed. The original defect area was removed from the animal's calvarium bone en bloc. Beginning at the center of the surgical defect, serial sections of 6 µm thick were cut longitudinally. The sections were stained with hematoxylin and eosin for analysis under a light microscope. The sections were analyzed for the presence of inflammatory infiltrate, connective tissue formation and new bone formation. Computer-assisted histomorphometic measurements were carried out with an automated image analysis system. RESULTS: The total new bone areas were significantly greater in group D than in all groups and group C was statistically insignificant from the other groups (p < 0.05). Group B had a greater, but not statistically significant (p > 0.05), amount of total regenerated bone area than the control group. CONCLUSION: The results indicate that 100 mmol/kg topical and 10 mmol/kg/day systemic application of CAPE increases bone healing, especially with systemic application.

A comprehensive study of myocardial redox homeostasis in naturally and mimetically aged rats.

Age-related myocardial dysfunction has important implications with impaired redox homeostasis. Current study focused on investigation of redox homeostasis and histopathological changes in the myocardium of mimetically (MA), naturally aged (NA), and young control (YC) rats. Chronic D-galactose administration to young male Wistar rats (5 months old) was used to set up experimental aging models. We investigated 16 different oxidative damage biomarkers which have evaluated redox homeostasis of cellular macromolecules such as protein, lipid, and DNA. As a protein oxidation biomarker, advanced oxidation end products, protein carbonyl groups, protein-bound advanced glycation end products, dityrosine, kynurenine, and N-formylkynurenine concentrations in MA and NA rats were found to be significantly higher compared to those in YC rats. On the other hand, the levels of protein thiol groups were not significantly different between groups, whereas lipid peroxidation biomarkers such as conjugated diens, lipid hydroperoxides, and malondialdehyde in MA and NA rats were found to be significantly higher in comparison to those in YCs. For the assessment of oxidative DNA damage, we analyzed eight hydroxy-5'-deoxyguanosine concentrations of MA and NA groups which were higher than YCs. As an antioxidant status in the MA and NA groups, Cu-Zn superoxide dismutase, ferric reducing antioxidant power, and total thiol levels were lower than those in the YCs. Only nonprotein thiol levels were not significantly different. We also observed similar histopathological changes in MA and NA rats. We concluded that the mimetic aging model could be considered as a reliable experimental model for myocardial senescence.