Potential therapeutic effect of pomegranate seed oil on ovarian ischemia/reperfusion injury in rats.

OBJECTIVES: The aim of this study is to determine the therapeutic effects of pomegranate seed oil, which is a powerful antioxidant and anti-inflammatory agent, on ovarian-ischemia and reperfusion injury in rats. MATERIALS AND METHODS: Fifty-six female albino Wistar rats were divided into 7 equal groups. Group 1; Sham Operation, Group 2; Ischemia, Group 3; Ischemia + Reperfusion, Group 4; Ischemia + Pomegranate 0,32 ml / kg (IP), Group 5; Ischemia + Pomegranate 0.64 ml / kg, Group 6; Ischemia + Pomegranate 0,32 ml / kg + reperfusion, Group 7; Ischemia + Pomegranate 0,64 ml / kg + reperfusion. Three hours after ischemia and 3 hours after reperfusion, the study was terminated. RESULTS: While NADPH oxidase activity, MDA and TNF-α levels were significantly increased, SOD activity and GSH levels were reduced in ischemia and I/R groups. Low dose pomegranate seed oil application reduced significantly oxidative stress and NADPH oxidase activity in both ischemic and ischemic/reperfusion groups. At the same time, low-dose pomegranate seed oil extract reduced TNF-α levels and significantly increased antioxidant activity. CONCLUSION: PSO demonstrated an important therapeutic effect in the treatment of ovarian ischemia and reperfusion injury.

Astrocyte/neuron ratio and its importance on glutamate toxicity: an in vitro voltammetric study.

The purpose of this study was to clarify the relationship between neuron cells and astrocyte cells in regulating glutamate toxicity on the 10th and 20th day in vitro. A mixed primary culture system from newborn rats that contain cerebral cortex neurons cells was employed to investigate the glutamate toxicity. All cultures were incubated with various glutamate concentrations, then viability tests and histological analyses were performed. The activities of glutamate transporters were determined by using in vitro voltammetry technique. Viable cell number was decreased significantly on the 10th day at 10(-7) M and at 10(-6) M glutamate applications, however, viable cell number was not decreased at 20th day. Astrocyte number was increased nearly six times on the 20th day as compared to the 10th day. The peak point of glutamate reuptake capacity was about 2 × 10(-4) M on the 10th day and 10(-3) M on the 20th day. According to our results, we suggested that astrocyte age was important to maintain neuronal survival against glutamate toxicity. Thus, we revealed activation or a trigger point of glutamate transporters on astrocytes due to time since more glutamate was taken up by astrocytes when glutamate transporters on the astrocyte were triggered with high exogenous glutamate concentrations. In conclusion, the present investigation is the first voltammetric study on the reuptake parameters of glutamate in vitro.

The in vitro protective effect of salicylic acid against paclitaxel and cisplatin-induced neurotoxicity.

Paclitaxel (PAC) and cisplatin (CIS) are two established chemotherapeutic drugs used in combination for the treatment of various solid tumors. However, the usage of PAC and CIS are limited because of the incidence of their moderate or severe neurotoxic side effects. In this study, we aimed to assess the protective role of salicylic acid (SA) against neurotoxicity caused by PAC and CIS. For this purpose, newborn Sprague Dawley rats were decapitated in sterile atmosphere and primary cortex neuron cultures were established. On the 10th day SA was added into culture plates. PAC and CIS were added on the 12th day. The cytotoxicity was determined by using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Oxidative alterations were assessed using total antioxidant capacity and total oxidative stress assays in rat primary neuron cell cultures. It was shown that both concentrations of PAC and CIS treatments caused neurotoxicity. Although SA decreased the neurotoxicity by CIS and PAC, it was more effective against the toxicity caused by CIS rather than the toxicity caused by PAC. In conclusion it was clearly revealed that SA decreased the neurotoxic effect of CIS and PAC in vitro.

Genotoxic and oxidative damage potentials in human lymphocytes after exposure to terpinolene in vitro.

Terpinolene (TPO) is a monocyclic monoterpene found in the essential oils of various fir and pine species. Recent reports indicated that several monoterpenes could exhibit antioxidant effects in both human and animal experimental models. However, so far, the nature and/or biological roles of TPO have not been elucidated in human models yet. The aim of this study was to investigate the genetic, oxidative and cytotoxic effects of TPO in cultured human blood cells (n = 5) for the first time. Human blood cells were treated with TPO (0-200 mg/L) for 24 and 48 h, and then cytotoxicity was detected by lactate dehydrogenase (LDH) release and [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] (MTT) assay, while DNA damage was also analyzed by micronucleus assay, sister chromatid exchanges assay and 8-oxo-2-deoxyguanosine (8-OH-dG) level. In addition, biochemical parameters [total antioxidant capacity (TAC) and total oxidative stress (TOS)] were examined to determine oxidative effects. The results of LDH and MTT assays showed that TPO (at concentrations greater than 100 mg/L) decreased cell viability. In our in vitro test systems, it was observed that TPO had no genotoxicity on human lymphocytes. Again, TPO (at 10, 25, 50 and 75 mg/L) treatment caused statistically important (p < 0.05) increases of TAC levels in human lymphocytes without changing TOS levels. In conclusion, TPO can be a new resource of therapeutics as recognized in this study with its non-genotoxic and antioxidant features.

Novel pHEMA-gelatin SPHs as bone scaffolds in dynamic cultures.

The effectiveness of poly(2-hydroxyethyl methacrylate)-gelatin superporous hydrogels (pHEMA-gelatin SPHs) was investigated for bone tissue engineering. The cell culture studies were performed with preosteoblastic MC3T3-E1 cells. Dynamic culture conditions were provided using 100 ml spinner flask rotating at 50 rpm. According to the results of mitochondrial activity test (1-3-[4,5-dimethylthiazol-2-yl]-diphenyltetrazolium bromide), there is no significant difference between proliferation behavior of cells cultured under static and dynamic conditions during 28 days. Observations by scanning electron microscopy and confocal laser scanning microscopy showed that, cells attached well onto the scaffolds and spread through the pores for both culture conditions. However, it was found that, calcium deposition and alkaline phosphatase activity in the scaffolds cultured under dynamic conditions were higher than that of static conditions. The expression of osteogenic differentiation markers, i.e. collagen I and osteopontin, based on real-time reverse transcriptase-polymerase chain reaction demonstrated increased responses under the spinner flask culture conditions. The combination of dynamic culture conditions in spinner flask with the use of superporous pHEMA-gelatin scaffolds enhanced the outcomes related to bone tissue engineering.

Novel scaffolds based on poly(2-hydroxyethyl methacrylate) superporous hydrogels for bone tissue engineering.

In this study, poly(2-hydroxyethyl methacrylate) (pHEMA)-based superporous hydrogels were synthesized by radical polymerization of 2-hydroxyethyl methacrylate (HEMA) in the presence of a gas blowing agent, sodium bicarbonate. These hydrogels are: pHEMA, pHEMA-gelatin, glycerol phosphate (GP) cross-linked pHEMA-gelatin, glutaraldehyde (GA) cross-linked pHEMA-gelatin superporous hydrogels (SPHs) and pHEMA-hydroxyapatite (HA) superporous hydrogel composites (SPHCs). The hydrogels have a structure of interconnected pores with pore sizes of approx. 500 µm. Although the extent of swelling decreased when gelatin and HA were incorporated to the pHEMA structure, the time to reach the equilibrium swelling (approx. 20 s) was not affected so much. In the presence of gelatin and cross-linkers, mechanical properties significantly improved when compared with pHEMA SPH. Among all the synthesized hydrogels, pHEMA-HA SPHC showed great improvement in mechanical strength and its elastic modulus value was 0.027±0.002 N/mm(2). Osteogenic activities of pHEMA-based scaffolds were examined by preosteoblastic MC3T3-E1 cell-culture studies. The mitochondrial activity test (MTT) showed that gelatin-containing scaffolds stimulated cell proliferation compared with other scaffolds, while alkaline phosphatase levels (ALP) and mineralization were found highest for the GP cross-linked pHEMA-gelatin SPH. However, pHEMA SPH and pHEMA-HA SPHC did not support cell proliferation and also differentiation. In conclusion, pHEMA-gelatin SPH and GP cross-linked pHEMA-gelatin SPH can be considered as potential scaffolds for bone tissue-engineering applications.