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EgeUn###, Neurodegenerative diseases affect millions of people. Major reasons behind the onset and progression of these diseases are still under investigation. Therefore, any approach that would treat/prevent progression is important. In this study, we aimed to investigate the potential protective effects of Psephellus pyrrhoblepharus (Boiss.) Wagenitz extracts in MPP+-induced dopaminergic cell damage and compare the effectiveness of different extracts (methanol:water (1:1), chloroform and n-hexane). The cells were pretreated with four different concentrations (10, 50, 100, and 200 mu g/ml) of methanol:water (1:1), chloroform and n-hexane extracts of P. pyrrhoblepharus following MPP+ treatment for 12 or 24 h. The changes in cell viability were determined using the MTT assay. Additionally, antioxidant activities and total phenolic/flavonoid contents of the extracts were determined with radical scavenging capacity, Folin-Ciocalteu and aluminum chloride assays, respectively. The extracts at selected concentrations were found to be protective in a dose-dependent manner at 12 and 24 h. Nevertheless, the methanol extract of the plant showed the highest protection both at 100 and 200 mu g/ml (115.13%+/- 3.98, 121.87%+/- 1.66; p < 0.05) against dopaminergic damage at 24 h. The results showed that selected concentrations were not toxic and did not affect cell proliferation rate. Besides, the chloroform extract was found to have higher antioxidant activity than the other extracts (p < 0.05). The total phenolic and total flavonoid contents were found consistent with antioxidant activities. Our findings support the neuroprotective and antioxidant potential of P. pyrrhoblepharus. However, further studies on identifying the presence of chemicals in P. pyrrhoblepharus extracts which are responsible for protection should be carried out to confirm their therapeutic potential.

WOS: 000474396700008, PubMed: 30415440, ObjectiveThe aim of this in vitro study is to evaluate the effects of resveratrol (RES) addition on the cytotoxicity and microtensile bond strength (mu TBS) of different adhesives.Materials and methodsFive self-etching adhesives (G-aenial Bond-GC, Optibond All in One-Kerr, Gluma Self Etch-Kulzer, Clearfil S-3 Bond-Kuraray, and Nova Compo-B Plus-Imicryl) were tested. They were applied to L-929 cell culture by the extract method. In the test groups, 0.5 mu M RES (Sigma-Aldrich) was added into the medium. Cell viability was assessed by MTT assay after 24h. Human extracted third molars were used for mu TBS test (n=7). The adhesives with or without 0.5 mu M RES addition were applied on dentin surfaces. A composite build-up was constructed. Then, the specimens were sectioned into multiple beams with the non-trimming version of the microtensile test and subjected to microtensile forces. Statistical analysis was performed using ANOVA and post hoc Tukey test (p?0.05).ResultsThe extracts of all adhesives decreased the cell viability. However, RES addition increased the cell viability in all groups (p?0.05). RES addition did not cause any decrease in mu TBS values of the adhesives compared to baseline. Optibond All in One showed the highest mu TBS after RES addition. It was followed by Clerafil S-3 Bond and Nova Compo-B Plus. No difference was determined between the Optibond All in One and Clearfil S-3 Bond. There was difference between Optibond All in One and Nova Compo-B Plus (p?0.05).ConclusionRES addition may improve the biocompatibility without causing negative influence on mu TBS of the adhesives.Clinical relevanceRES addition has clinical applicable potential to overcome the adverse biocompatibility of adhesives.

Spinal cord injury (SCI) leads to vascular damage and disruption of blood-spinal cord barrier which participates in secondary nerve injury. Epidermal growth factor (EGF) is an endogenous protein which regulates cell proliferation, growth and differention. Previous studies reported that EGF exerts neuroprotective effect in spinal cord after SCI. However, the molecular mechanisms underlying EGF-mediated protection in different regions of nervous system have not shown yet. In this study, we aimed to examine possible anti-apoptotic and protective roles of EGF not only in spinal cord but also in brain following SCI. Twenty-eight adult rats were divided into four groups of seven animals each as follows: sham, trauma (SCI), SCI + EGF and SCI + methylprednisolone (MP) groups. The functional neurological deficits due to the SCI were assessed by behavioral analysis using the Basso, Beattie and Bresnahan (BBB) open-field locomotor test. The alterations in pro-/anti-apoptotic protein levels and antioxidant enzyme activities were measured in spinal cord and frontal cortex. In our study, EGF promoted locomotor recovery and motor neuron survival of SCI rats. EGF treatment significantly decreased Bax and increased Bcl-2 protein expressions both in spinal cord and brain when compared to SCI group. Moreover, antioxidant enzyme activities including catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPx) were increased following EGF treatment similar to MP treatment. Our experiment also suggests that alteration of the ratio of Bcl-2 to Bax may result from decreased apoptosis following EGF treatment. As a conclusion, these results show, for the first time, that administration of EGF exerts its protection via regulating apoptotic and oxidative pathways in response to spinal cord injury in different regions of central nervous system.

Although hyperbaric oxygen (HBO) treatment following spinal cord injury (SCI) have been studied in terms of neurological function and tissue histology, there is a limited number studies on spinal cord tissue enzyme levels.The effect of HBO treatment in SCI was investigated by measuring superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), nitric oxide synthase (NOS) and nitric oxide (NO) activity in the injured tissue. SCI was induced by applying an aneurysm clip extradurally at the level of T9-T11 vertebrae. Preoperative HBO (preopHBO) treatment was applied for 5days and postoperative HBO (postopHBO) for 7days.In the preopHBO group, a significant decrease was observed in NOS and NO compared to the SCI group. There was a decrease in SOD, NOS and NO in the postopHBO group when compared to the SCI group. In the pre-postHBO group SOD, GPx, NOS and NO decreased significantly. There was a decrease in SOD in postopHBO compared to preopHBO. In the prepostopHBO, SOD decreased significantly compared to that in the preopHBO group. The prepostopHBO presented a significant decrease in GPx compared to postopHBO (p0.05 for all parameters). No significant difference was observed for catalase for all groups. Significant improvement was found in BBB scores for both postopHBO and prepostHBO groups when compared to the SCI group (p0.05).HBO treatment was found to be beneficial following SCI in terms of biochemical parameters and functional recovery in the postoperative period.

CART mRNA and peptides are highly expressed in the anatomical structures composing the hypothalamo-pituitary-adrenal (HPA) axis and sympatho-adrenal system. Anatomical and functional studies suggest that CART peptides may have a role in the regulation of the neuroendocrine and autonomic responses during stress. Our previous study showed that CART peptides increased significantly in the male hypothalamus and amygdala 10 min after the forced swim stress. The present study aimed to examine the effect of forced swim stress on CART peptide expression in selected brain regions, including those where CART peptide expression has not been reported before (frontal cortex, pons, medulla oblongata), as well as in endocrine glands related to stress in male Sprague Dawley rats. A total of 16 (n = 8) animals were used, including control groups. Rats were subjected to forced swim on two consecutive days, and sacrificed on the second day, 2 h after the termination of the stress procedure. Frontal cortex, pons, medulla oblongata, hypothalamus, pituitary and adrenal glands were dissected and homogenized. CART peptide expression in these tissues was measured by Western Blotting and six different CART peptide fragments were identified. Our results showed that forced swim stress elicited region-specific changes in CART peptide expression. CART was upregulated in the frontal cortex, hypothalamus, medulla oblongata and adrenal gland while there was no change in the pons and pituitary. Enhanced CART peptide fragments in these brain regions and adrenal glands may have a role in the regulation of the HPA and sympatho-adrenal axis activity during stress response.

We investigated functional recovery of the lower urinary system in rats with spinal cord injury after transplanting neuronal restricted precursors/glial restricted precursors or neural cells derived from bone marrow stromal cells into the injured area of the spinal cord.A total of 30 rats underwent experimentation in 4 groups, including group 1--sham operation, group 2--spinal cord injury plus neuronal restricted precursor/glial restricted precursor transplantation, group 3--spinal cord injury plus bone marrow stromal cell transplantation and group 4--spinal cord injury control. All rats in the 4 groups were investigated urodynamically and sacrificed on day 28 after transplantation. The cells transplanted into the injured spinal cord underwent histological investigation.Transplanted cells (neuronal and glial restricted precursors, and bone marrow stromal cells) were found to maintain a presence in the injured spinal cord area. Baseline pressure, maximum capacity, mean uninhibited contraction amplitude, mean voiding pressure, voided volume and post-void residual volume were significantly better in groups 2 and 3 than in group 4, while baseline pressure in group 2 was better than that in group 3. We found no significant difference among the groups according to mean uninhibited contraction frequency.Although neuronal/glial restricted precursor transplanted rats seemed to have more improvement, all rats in groups 2 and 3 showed some significant improvement in lower urinary system function. On the other hand, the level of this improvement was far from complete functional recovery.

Results of electrophysiological studies suggest a significant role of the lateral spinal nucleus (LSpN) in the transmission of nociceptive signals. In our study, the presence of Fos immunoreactivity and NADPH-diaphorase positivity was observed in the rat LSpN following noxious peripheral subcutaneous stimulation. Formalin-induced unilateral hindpaw stimulation in the rat caused bilateral NADPH-d reactivity and ipsilateral Fos expression in this nucleus. In the LSpN of the L3-L5 segments of stimulated rats, on average, 4.1 +/- 1.2 NADPH-d-positive, NADPH-d(+), 5.1+1.8 Fos-immunoreactive, Fos(+), and 3.0 +/- 1.1 double-labeled neurons per 25-mu m-thick section were found unilaterally. A close anatomical relationship between NADPH-d(+) processes and Fos(+) cell nuclei in the LSpN was also observed following noxious peripheral stimulation. These neuroanatomical findings support the hypothesis that the LSpN is involved in pain processing and suggest an important role of nitric oxide-mediated signal transduction in this nucleus.

WOS: 000378324000020, PubMed ID: 26981753, The aim of this study was to evaluate whether medium modification improves the odontogenic differentiation of human dental pulp stem cells (DPSC) in vitro and in vivo. DPSC isolated from human impacted third molar teeth were analysed for clusters of differentiation with flow cytometry. Odontogenic differentiation was stimulated by medium modification with the addition of bone morphogenetic protein 2 (BMP2). The expression of dentin sialophosphoprotein, dentin matrix protein 1, enamelysin/matrix metalloproteinase 20 and the phosphate-regulating gene with homologies to endopeptidases on the X chromosome of the cells were analysed with RT-PCR at 7, 14 and 21 days. Then, DPSC were transplanted on the back of immunocompromised mice via a hydroxyapatite tricalcium phosphate scaffold, and the structure of the formed tissue was investigated. The cells were identified as mesenchymal stem cells with a 98.3% CD73 and CD90 double-positive cell rate. The increase in mineralization capacity and expression of human enamel-dentin specific transcripts proportional to the culture period were determined after differentiation. Six weeks after transplantation, an osteo-dentin matrix was formed in the group in which odontogenic differentiation was stimulated, and the odontogenic characteristics of the matrix were confirmed by histological examination and RT-PCR analysis. Odontogenic differentiation of the isolated and characterized human DPSC was improved with medium modification by the addition of BMP2 in vitro and in vivo. The defined medium and applied technique have a potential use for forming reparative dentin in the future, but the effects of the method should be investigated in long-term studies., Ege University's Scientific Research FoundationEge University [2009-Dis-036]; Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK), This study was supported by Ege University's Scientific Research Foundation (2009-Dis-036). We also acknowledge The Scientific and Technological Research Council of Turkey (TUBITAK) for its PhD scholarship support.

We explored the effects of early and late application of botulinum-A toxin (BTX-A) on reservoir function and histological bladder changes in spinal cord injured rats.The study was done in 30 Sprague-Dawley rats randomly allocated into 5 groups. Group 1 of 6 rats underwent sham operation only. Group 2 of 6 rats underwent spinal cord transection. Group 3 of 6 rats underwent spinal cord transection followed by BTX-A application into the detrusor muscle 7 days later. Group 4 of 6 rats underwent spinal cord transection, followed by BTX-A application into the detrusor muscle 28 days later. Group 5 of 6 rats underwent spinal cord transection followed by saline injection into the detrusor muscle after 28 days. Spinal cord injury was created by transecting the cord at the T9 to T10 level. All rats underwent cystometric examination initially and on day 42 before sacrifice. The bladders were removed and examined histologically for fibrosis and hyperplasia.On cystometric examination BTX-A caused an improvement in baseline pressure, and the frequency and amplitude of uninhibited detrusor contractions (p0.001). No significant differences were observed in maximal bladder capacity or urethral opening pressure (p0.05). Histologically BTX-A led to decreased fibrosis and hyperplasia (p0.001). No significant differences were found between histological or cystometric among the groups with respect to receiving BTX-A in the early and late periods (p0.05).BTX-A has a functional and histological healing effect on detrusor hyperreflexia subsequent to spinal cord injury in rats. Although administering BTX-A in the early period had better quantifiable functional and histological outcomes compared to the late period, the difference was not statistically significant.

Traumatic spinal cord injury (SCI) can lead to post-traumatic inflammation, oxidative stress, motor neuron apoptosis, necrosis and autophagy of tissue. To promote and enhance recovery after SCI, recent development of devices and therapeutic interventions are needed. The aim of the present study was to investigate the possible role of C-terminal domain of tetanus toxin (Hc-TeTx) on cell death mechanisms including apoptosis and autophagy following SCI. Thirty five adult rats were divided into five groups (n=7 each) as follows: control, sham, trauma (SCI), SCI+Hc-TeTx and SCI+methylprednisolone groups. The functional neurological deficits due to the SCI were assessed by behavioral analysis using the BBB open-field locomotor test. The alterations in pro-/anti-apoptotic and autophagy related-protein levels were measured by western blotting technique. In our study, Hc-TeTx promotes locomotor recovery and motor neuron survival of SCI rats. Hc-TeTx decreased expression of bax, bad, bak, cleaved caspase-3, Ask1 and autophagy-related proteins including Atg5 and LC3II in brain. Our study provides an evidence that cell death mechanisms play critical roles in SCI and non-toxic peptides may exert protective effect and decrease cell death following SCI.

Aims The aim of this study is to determine the role of nitric oxide (NO) in neuropathic pain and the effect of embryonic neural stem cell (ENSC) transplantation on NO content in rat spinal cord neurons following spinal cord injury (SCI). Main methods Ninety adult male Sprague–Dawley rats were divided into 3 groups (n = 30, each): control (laminectomy), SCI (hemisection at T12–T13 segments) and SCI + ENSC. Each group was further divided into sub-groups (n = 5 each) based on the treatment substance (L-NAME, 75 mg/kg/i.p.; l -arginine, 225 mg/kg/i.p.; physiological saline, SF) and duration (2 h for acute and 28 days for chronic groups). Pain was assessed by tail flick and Randall–Selitto tests. Fos immunohistochemistry and NADPH-d histochemistry were performed in segments 2 cm rostral and caudal to SCI. Key findings Tail-flick latency time increased in both acute and chronic L-NAME groups and increased in acute and decreased in chronic l- arginine groups. The number of Fos (+) neurons decreased in acute and chronic L-NAME and decreased in acute l -arginine groups. Following ENSC, Fos (+) neurons did not change in acute L-NAME but decreased in the chronic L-NAME groups, and decreased in both acute and chronic l -arginine groups. NADPH-d (+) neurons decreased in acute L-NAME and increased in l -arginine groups with and without ENSC transplantation. Significance This study confirms the role of NO in neuropathic pain and shows an improvement following ENSC transplantation in the acute phase, observed as a decrease in Fos(+) and NADPH-d (+) neurons in spinal cord segments rostral and caudal to injury.

Introduction Erectile dysfunction is common among patients with spinal cord injury (SCI). Aim This study aims to investigate the recovery of penile erectile functions of the rats with spinal cord injury (SCI) following transplantation of endogenous neuronal precursors cell (neuronal restricted precursors [NRP]/glial restricted precursors [GRP]) into the injured area of spinal cord. Methods Twenty‐two rats were experimented in three groups. Group 1 (N = 6): Sham; Group 2 (N = 10): SCI + NRP/GRP transplanted in day 9 after operation; Group 3 (N = 6): SCI + culture medium transplanted in day 9 after operation. Analysis of penile reflexes and cavernosal nerve stimulation studies were performed in day 28 after transplantation for each group. All rats in three groups were then sacrificed and the injured regions of spinal cords underwent histological investigation. Main Outcome Measures These results show improvements to some extent in locomotor and erectile functions although these improvements are far from full functional recovery. Results Cavernosal nerve stimulation resulted in significantly higher intracavernosal pressure in Group 3 (SCI) although there was no difference between Group 1 (sham) and Group 2 (SCI + NRP/GRP). Number of clusters was similar between groups. Number of erections was higher in Group 3 (SCI) than Groups 1 and 2, and number of cups was higher in Group 2 (SCI + NRP/GRP) than the other two groups. Number of flips was similar in Groups 1 and 2 but lower in Group 3. Number of long flips was highest in Group 1 and lowest in Group 3. The differences between groups were significant. Conclusion. This study emphasized the healing potential of NRP/GRP transplantation following experimental SCI. However, further experimental and clinical studies are required to advance this treatment modality. Temeltas G, Dagci T, Evren V, and Lekili M. Effects of neuronal and glial restricted precursor cells transplantation on erectile function after experimentally induced spinal cord injury.

WOS: 000268125600014, PubMed ID: 18637713, The oxidative mechanisms of injury-induced damage of neurons within the spinal cord are not very well understood. We used a model of T8-T9 spinal cord injury (SCI) in the rat to induce neuronal degeneration. In this spinal cord injury model, unilateral avulsion of the spinal cord causes oxidative stress of neurons. We tested the hypothesis that apurinic/apyrimidinic endonuclease (or redox effector factor-1, APE/Ref-1) regulates this neuronal oxidation mechanism in the spinal cord region caudal to the lesion, and that DNA damage is an early upstream signal. The embryonic neural stem cell therapy significantly decreased DNA-damage levels in both study groups-acutely (followed up to 7 days after SCI), and chronically (followed up to 28 days after SCI) injured animals. Meanwhile, mRNA levels of APE/Ref-1 significantly increased after embryonic neural stem cell therapy in acutely and chronically injured animals when compared to acute and chronic sham groups. Our data has demonstrated that an increase of APE/Ref-1 mRNA levels in the caudal region of spinal cord strongly correlated with DNA damage after traumatic spinal cord injury. We suggest that DNA damage can be observed both in lesional and caudal regions of the acutely and chronically injured groups, but DNA damage is reduced with embryonic neural stem cell therapy.

Object. Spinal cord injury (SCI) is a complex pathological entity, the treatment of which requires a multipronged approach. One way to integrate different therapeutic strategies for SCI is to develop implantable scaffolds that can deliver therapies in a synergistic manner. Many investigators have developed implantable “bridges,” but an important property of such scaffolds—that is, mechanical compatibility with host tissues—has been neglected. In this study, the authors evaluated the results of implanting a mechanically matched hydrogel-based scaffold to treat SCI. Methods. A nonbiodegradable hydrogel, poly(2-hydroxyethylmethacrylate) (PHEMA), was engineered using thermally initiated free radical solution polymerization. Two groups of 12 adult Sprague—Dawley rats underwent partial cervical hemisection injury followed by implantation of either PHEMA or PHEMA soaked in 1 µg of brain-derived neurotrophic factor (BDNF). Four rats from each group were killed 1, 2, or 4 weeks after induction of the injury. Immunofluorescence staining was performed to determine the presence of scarring, cellular inflammatory responses, gliosis, angiogenesis, and axonal growth in and around the implanted scaffolds. Conclusions. The implanted PHEMA with 85% water content had a compressive modulus of 3 to 4 kPa, which matched the spinal cord. Implanted PHEMA elicited modest cellular inflammatory responses that disappeared by 4 weeks and minimal scarring was noted around the matrix. Considerable angiogenesis was observed in PHEMA, and PHEMA soaked in BDNF promoted axonal penetration into the gel. The authors conclude that mechanically engineered PHEMA is well accepted by host tissues and might be used as a platform for sustained drug delivery to promote axonal growth and functional recovery after SCI.