Treatment with quercetin increases Nrf2 expression and neuronal differentiation of sub ventricular zone derived neural progenitor stem cells in adult rats.

BACKGROUND: The presence of neural precursor stem cells (NPSCs) in some parts of the adult brain and the potency of these types of cells with a therapeutic viewpoint, has opened up a new approach for the treatment and recovery of the defects of central nervous system (CNS). Quercetin, as an herbal flavonoid, has been extensively investigated and shown to have numerous restoratives, inhibitory, and protective effects on some cell-lines and disorders. The purpose of this study is to simultaneously investigate the effect of quercetin on the expression of the nuclear factor erythroid 2-related factor 2 (Nrf2) gene and the effect on the proliferation and differentiation of NPSCs derived from the subventricular zone (SVZ) of the brain of adult rats. METHODS AND RESULTS: The cell obtained from SVZ cultured for one week and treated with quercetin at the concentrations of 1, 5, and 15 µM to evaluate the Nrf2 expression, proliferation and differentiation of NSCs after one week. Cellular and genetic results was performed by RT-PCR, MTT assay test, quantification of images with Image-J and counting. The results indicated that the quercetin increases expression of Nrf2 at concentration above 5 µM. Also differentiation and proliferation rate of NSCs is affected by various concentrations of quercetin in a dose-dependent manner. CONCLUSION: These findings confirmed the dose-dependent effect of quercetin on proliferation and differentiation of cell. In addition, quercetin increased the expression of Nrf2 gene. By combining these two effects of quercetin, this substance can be considered an effective compound in the treatment of degenerative defects in CNS.

Effects of N-acetyl-cysteine supplementation on sperm quality, chromatin integrity and level of oxidative stress in infertile men.

BACKGROUND: Infertile men have higher levels of semen reactive oxygen species (ROS) than fertile men. High levels of semen ROS can cause sperm dysfunction, sperm DNA damage and reduced male reproductive potential. This study investigated the effects of supplementation with N-acetyl-cysteine (NAC) on the sperm quality, chromatin integrity and levels of oxidative stress in infertile men. METHODS: The study was carried out in the unit of ACECR Infertility Research Center, Qom, Iran. The patients consisted of 50 infertile men with asthenoteratozoospermia who received NAC (600 mg/d) orally for 3 months, after which they were compared with pre-treatment status. Semen was analyzed according to WHO (2010), followed by the assessment of protamine content [chromomycin A3 (CMA3)] and DNA integrity [terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)]. Oxidative stress markers, i.e. total antioxidant capacity (TAC) and malondialdehyde (MDA), as well as hormonal profile (LH, FSH, Testosterone and Prolactin) were determined by ELISA kit. RESULTS: After NAC treatment, patients' sperm count and motility increased significantly whereas abnormal morphology, DNA fragmentation and protamine deficiency showed significant decreases compared to pre-treatment levels (P < 0.05). Hormonal profile improvement was associated with lowered FSH and LH levels and increased amount of testosterone (P < 0.05). TAC significantly increased and MDA decreased with an inverse significant correlation between TAC and MDA (P < 0.05). CONCLUSION: NAC oral supplementation may improve sperm parameters and oxidative/antioxidant status in infertile males.

Probing the Interaction of Newly Synthesized Pt(II) Complex on Human Serum Albumin Using Competitive Binding Site Markers.

Considering the importance of pharmacology and the influence of drugs on biological materials, the effects of a newly designed and synthesized platin complex (2,2'-Bipyridine-3,3'-dicarboxylic acid, oxalato Platinum(II), as an antitumor drug was tested on the structure of blood carrier protein of human serum albumin (HSA) using various spectroscopic techniques including UV-visible, fluorescence, and circular dichroism at 25 and 37 °C. Results of the fluorescence measurements revealed that adding the complex caused reduction in intrinsic fluorescence emission of HSA resulted from dynamic quenching of HSA. The number of binding sites and binding constants were calculated at both temperatures of 25 and 37 °C. In addition, in order to identify the complex's binding site on HSA employing spectroscopy, the competitive studies were followed using warfarin, digitoxin and ibuprofen as site markers of Sudlow sites I, II and III. Competitive binding test results have shown that Pt(II) complex bind on the warfarin binding site (or Sudlow sites I) on HSA. Besides, a reduction in thermal stability for HSA was observed in the presence of the newly designed Pt(II) complex.

Protective Effect of Cerium Oxide Nanoparticles on Human Sperm Function During Cryopreservation.

The generation of reactive oxygen species during cryopreservation of human sperm has negative effects on the consistency of the thawed sperm. The antioxidant properties of cerium oxide nanoparticles (CeO2NPs) may be useful for reducing cryodamage in thawed sperm. This research was conducted to determine the effects of CeO2NPs on the quality and function of human sperm after thawing. Samples of semen obtained from 20 normozoospermic individuals were allocated to the following four groups: fresh, frozen control (sperm not treated with CeO2NPs), and those exposed to 0.1 µg/mL CeO2NPs (CeO2-0.1), 1 µg/mL CeO2NPs (CeO2-1), and 5 µg/mL CeO2NPs (CeO2-5). Sperm parameters of motility, viability, membrane integrity, DNA fragmentation, protamination, malondialdehyde (MDA) levels, mitochondria membrane potential, and morphology were evaluated after the freezing-thawing process. The results showed that 0.1 µg/mL CeO2NPs significantly (p < 0.05) improved the following human sperm parameters after thawing: progressive (44.6% ± 1.14% vs. 36.2% ± 1.24%) and total motility (60.9% ± 2.5% vs. 51.3% ± 2.5%), viability (67.9% ± 1.5% vs. 58.1% ± 1.5%), membrane functionality (66.1% ± 1.85% vs. 55.4% ± 1.85%), DNA integrity (30.8% vs. 24.04%), and protamination (69.85% ± 2.09% vs. 57.2% ± 2.09%) compared with the frozen control group. We observed the lowest MDA levels in the CeO2-0.1 (3.06 ± 0.25 nmol/mL), CeO2-1 (3.1 ± 0.25 nmol/mL), and CeO2-5 (3.08 ± 0.25 nmol/mL) groups compared with the frozen control group (3.72 ± 0.25). Different concentrations of CeO2NPs did not significantly change sperm normal morphology and mitochondria activity (p < 0.05).

Astaxanthin ameliorates the impairment consequence of prenatal bacterial lipopolysaccharide exposure in adult male offspring NMRI mice.

In this study, the potential effects of astaxanthin (AST) were investigated on preventing the prenatal LPS-induced injures in mothers and adult male offspring of NMRI mice. Pregnant mice were randomly divided into four groups: 1. Saline + vehicle; 2. Saline + AST: received astaxanthin (4 mg/kg for 3 days, ip) on 11-13 gestation days; 3. LPS + vehicle (LPS-treated group): injected with LPS (20 µg/kg, sc) on gestation day 11; 4. LPS + AST: administrated LPS and astaxanthin on gestation days 11 and 11-13, respectively. In each group, maternal care behaviors and TNF-α serum levels were examined until weaning of male offspring at 23 days. At 60 days old, male pups underwent analysis of body weight and length, serum gonadotropins and testosterone hormone levels, sperm quality, gonadal and brain tissues morphologies, and the expression of SOX9 and GnRH genes by real-time PCR. Serum TNF-α level increased significantly in mothers treated with LPS, while AST reduced it. In adult male offspring, serum hormone levels, sperm quality, and the number of spermatocytes and Leydig cells in the testes improved when AST was administrated. According to histological studies of the brain, neurons in the LPS-treated group were smaller and less active, whereas neurons in the LPS + AST group were larger, more numerous, and more active. LPS significantly reduced GnRH expression, while AST induction improved its expression. AST administration during pregnancy prevented the adverse effects of prenatal exposure to LPS, presumably through its genomic and non-genomic effects, in adult male offspring.

Novel electrospun conduit based on polyurethane/collagen enhanced by nanobioglass for peripheral nerve tissue engineering.

Peripheral nerve injury can significantly affect the daily life of individuals with impaired nerve function and permanent nerve deformity. One of the most common treatments is autograft transplantation. Tissue engineering is one of the efficient methods to regenerate injured nerves using scaffolds, cells, and growth factors. Conduits, which are produced by a variety of techniques, could be used as an alternative treatment for patients with damaged nerves. The electrospinning technique is one of the most important and widely used methods for generating nanofiber conduits from biocompatible polymers. In this study, using the electrospinning method, three different conduits, including polyurethane (PU), polyurethane/collagen (PU/C), and a new conduit based on polyurethane + collagen + nanobioglass (PU/C/NBG), were prepared. The characteristics of these three types of conduits were evaluated by SEM, XRD, and various experiments, including porosity, degradation, contact angle, DMTA, FTIR, MTT, and DAPI staining. The results of MTT and DAPI assays revealed the safety of conduits and proper cell attachment. Overall, the results obtained from various experiments showed that the novel PU/C/NBG conduit has better mechanical properties in terms of porosity, hydrophilicity, and biocompatibility in comparison with PU and PU/C conduits and could be a suitable candidate for peripheral nerve regeneration and axonal growth due to its repair potential.

Nanobioglass enhanced polyurethane/collagen conduit in sciatic nerve regeneration.

The main purpose of neural tissue engineering and regenerative medicine is the development of biological substitutions to preserve, improve, and regenerate the damaged functions of tissues and organs. Three novel conduits, including polyurethane (PU), polyurethane/collagen (PU/C), and polyurethane/collagen/nano-bio glass (PU/C/NBG), were fabricated by the electrospinning technique. After confirming the suitability of conduits in the in-vitro environment, conduits were surgically sutured in a 10-mm gap in the sciatic nerve of a rat to evaluate their role in sciatic nerve reconstruction. After 4, 8, and 12 weeks of surgery, nerve regeneration was assessed by the hot plate test, sciatic functional index, electromyography, histology, and immunohistochemistry against S100, NF200, and CD31 antibodies. The results of various examinations revealed that the PU/C/NBG conduit is significantly more suitable than PU and PU/C conduits in terms of nerve regeneration. However, all three groups of conduits had the potential to be used for nerve regeneration. Overall, this study discovered that the PU/C/NBG conduit is a biocompatible neural conduit, which is a favorable candidate for peripheral nerve regeneration and axonal growth.

Evaluation of Encapsulated Eugenol by Chitosan Nanoparticles on the aggressive model of rheumatoid arthritis.

Chitosan Nanoparticles Eugenol recognizes as a potent antioxidant that can use the first therapeutic chemical to treat rheumatoid arthritis (RA) instead of Methotrexate. The purpose of this study was to investigate the effects of Chitosan Nanoparticles Eugenol as a potent Nano-herbal agent in the healing process of experimental neonatal RA compared to Methotrexate. The neonatal Wistar rats induced rheumatoid arthritis in both genders were divided into sham, control, the treatment receiving Methotrexate, and the second treatment receiving encapsulated Eugenol by Chitosan Nanoparticles groups. Afterward, Malondialdehyde, for assessment of lipid peroxidation as an oxidative stress biomarker by assay kit, FOXO3 protein as an antioxidant up-regulating by western blotting and expression of the TGF-ß and CCL2/MCP-1 genes by real-time PCR evaluation, supported by a cartilage histopathology analysis. Based on these results, Methotrexate and Eugenol encapsulated by Chitosan Nanoparticles, a significant decrease is observed in the serum level of MDA and FOXO3 protein expression in comparison to the control group. Additionally, Nanoparticle herbal agent and Methotrexate has a decreasing effect on the expression of TGF-ß and MCP-1 genes and a significant positive correlation was observed between MCP-1 and TGF-ß. Inflammation, synovial hyperplasia, and pannus formation were extreme in the Collagen Induced Arthritis rats. It can be concluded that Encapsulated Eugenol by Chitosan Nanoparticles and Methotrexate, probably by dint of their immunomodulatory, anti-inflammatory, and antioxidant potential has a protective effect against RA. Nano Eugenol is capable of delivering promising lines results to treat autoimmune diseases such as RA can also be suggested.

Effects of maternal thyroid hormone deficiency on differentiation of mesenchymal stem cells in CSF-exposed neonatal Wistar rats.

Cerebrospinal fluid (CSF) contains growth and neurotrophic factors which regulate proliferation, differentiation, and neurogenesis. Thyroid hormones play a crucial role in the development of the nervous system and hypothyroidism during development of embryos leads to defects in the nervous system. This study aimed to survey the effects of rat neonatal CSF collected from induced hypothyroid mothers on differentiation of bone marrow mesenchymal stem cells (BM-MSCs). We hypothesized that hypothyroidism affected levels of growth factor in CSF. To induce hypothyroidism, pregnant Wistar rats received methimazole at the third day of gestation. BM-MSCs were obtained from rat femurs and tibias and cultured in medium. CSF was collected from the cisterna magna of newborn rats, and cells were subsequently exposed to CSF with concentrations of 5,7, and 10 /100 (v/v) for 72 h. MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and real time polymerase chain reaction (RT-PCR) were used to quantify the cell viability and analyze the expression of neural markers, respectively. Our morphological studies showed that treatment with hypothyroidism CSF (HTH-CSF) resulted in a significant decrease in neurite growth and proliferation as compared to normal CSF (N-CSF). RT-PCR analysis also showed a significant decrease in expression of neural markers (i.e., Nestin, Neurod-1, NeuN) in cells treated with HTH-CSF as compared with the N-CSF group. The most effective concentration of CSF for BM-MSC differentiation was 5% (V/V). Our results showed a significant decrease in differentiation of BM-MSCs in the presence of neonatal CSF of hypothyroid mothers compared with neonatal CSF of healthy mothers. Thus, thyroid hormones are essential in neural development and hypothyroid defects can affect development of the neonatal brain.

In vivo tumor-suppressing and anti-angiogenic activities of a recombinant anti-CD3ε nanobody in breast cancer mice model.

Aim: Achievements in cancer immunotherapy require augmentation of a host's anti-tumor immune response for anti-cancer modality. Materials & methods: Different concentrations of recombinant anti-CD3 nanobody were administered at predetermined time intervals during a 24-day treatment period and then expression of angiogenic biomarkers including VEGFR2, MMP9 and CD31, as well as tumor cell proliferation marker ki67, was determined in tumor sections by immunohistochemistry. Furthermore, expression of cytokines was examined in peripheral blood of mice. Results: Based on our results, administration of nanobody could reduce biomarker expression in tumor sections. Tumor growth was also delayed and survival rate was increased in response to nanobody treatment. Moreover, expression of pro-inflammatory cytokines was reduced. Conclusion: In conclusion, we demonstrated that administration of nanobody could effectively suppress angiogenesis as well as tumor growth.

Enhancement of osteogenic differentiation of adipose-derived stem cells by PRP modified nanofibrous scaffold.

Recent developments in bone tissue engineering have paved the way for more efficient and cost-effective strategies. Additionally, utilization of autologous sources has been considered very desirable and is increasingly growing. Recently, activated platelet rich plasma (PRP) has been widely used in the field of bone tissue engineering, since it harbours a huge number of growth factors that can enhance osteogenesis and bone regeneration. In the present study, the osteogenic effects of PRP coated nanofibrous PES/PVA scaffolds on adipose-derived mesenchymal stem cells have been investigated. Common osteogenic markers were assayed by real time PCR. Alkaline phosphate activity, calcium deposition and Alizarin red staining assays were performed as well. The results revealed that the highest osteogenic differentiation occurred when cells were cultured on PRP coated PES/PVA scaffolds. Interestingly, direct application of PRP to culture media had no additive effects on osteogenesis of cells cultured on PRP coated PES/PVA scaffolds or those receiving typical osteogenic factors. The highest osteogenic effects were achieved by the simplest and most cost-effective method, i.e. merely by using PRP coated scaffolds. PRP coated PES/PVA scaffolds can maximally induce osteogenesis with no need for extrinsic factors. The major contribution of this paper to the current researches on bone regeneration is to suggest an easy, cost-effective approach to enhance osteogenesis via PRP coated scaffolds, with no additional external growth factors.