Decellularization with triton X-100 provides a suitable model for human kidney bioengineering using human mesenchymal stem cells.

AIMS: Regeneration of discarded human kidneys has been considered as an ideal approach to overcome organ shortage for the end-stage renal diseases (ESRDs). The aim of this study was to develop an effective method for preparation of kidney scaffolds that retain the matrix structure required for proliferation and importantly, differentiation of human adipose-derived mesenchymal stem cells (hAd-MSCs) into renal cells. MAIN METHODS: We first compared two different methods using triton X-100 and sodium dodecyl sulfate (SDS) for human kidney decellularization; followed by characterization of the prepared human renal extracellular matrix (ECM) scaffolds. Then, hAd-MSCs were seeded on the scaffolds and cultured for up to 3 weeks. Next, viability, proliferation, and migration of seeded hAd-MSCs underwent histological and scanning electron microscopy (SEM) assessments. Moreover, differentiation of hAd-MSCs into kidney-specific cell types was examined using immunohistochemistry (IHC) staining and qRT-PCR. KEY FINDINGS: Our results indicated that triton X-100 was a more effective detergent for decellularization of human kidneys compared with SDS. Moreover, attachment and proliferation of hAd-MSCs within the recellularized human kidney scaffolds, were confirmed. Seeded cells expressed epithelial and endothelial differentiation markers, and qRT-PCR results indicated increased expression of platelet and endothelial cell adhesion molecule 1 (PECAM-1), paired box 2 (PAX2), and E-cadherine (E-CDH) as markers of differentiation into epithelial and endothelial cells. SIGNIFICANCE: These observations indicate the effectiveness of decellularization with triton X-100 to generate suitable human ECM renal scaffolds, which supported adhesion and proliferation of hAd-MSCs and could induce their differentiation towards a renal lineage.

Neuroprotective and long term potentiation improving effects of vitamin E in juvenile hypothyroid rats.

Protective effects of vitamin E (Vit E) on long term potentiation (LTP) impairment, neuronal apoptosis and increase of nitric oxide (NO) metabolites in the hippocampus of juvenile rats were examined. The rats were grouped (n=13) as: (1) control; (2) hypothyroid (Hypo) and (3) Hypo-Vit E. Propylthiouracil (PTU) was given in drinking water (0.05%) during 6 weeks. Vit E (20 mg/ kg) was daily injected (IP). To evaluate synaptic plasticity, LTP from the CA1 area of the hippocampus followed by high frequency stimulation to the ipsilateral Schafer collateral pathway was carried out. The cortical and hippocampal tissues were then removed to measure NO metabolites. The brains of 5 animals in each group were removed for apoptosis study. The hypothyroidism status decreased the slope, 10-90% slope and amplitude of field excitatory post synaptic potential (fEPSP) compared to the control group (P<0.01-P<0.001). Injection of Vit E increased the slope, 10-90% slope and amplitude of the fEPSP in the Hypo-Vit E group in comparison to the Hypo group (P<0.05-P<0.01). TUNEL positive neurons and NO metabolites were higher in the hippocampus of the Hypo rats, as compared to those in the hippocampus of the control ones (P<0.001). Treatment of the Hypo rats by Vit E decreased apoptotic neurons (P<0.01-P<0.001) and NO metabolites (P<0.001) in the hippocampus compared to the Hypo rats. The results of the present study showed that Vit E prevented the LTP impairment and neuronal apoptosis in the hippocampus of juvenile hypothyroid rats.

Renoprotective Effect of Plantago major Against Proteinuria and Apoptosis Induced by Adriamycin in Rat.

OBJECTIVE: Adriamycin (ADR) is an important anti-cancer drug which can cause renal toxicity. Given the known anti-inflammatory and antioxidant effects of Plantago major (P. major), the aim of this study was to determine the effects of hydroalcoholic extract of P. major on ADR- induced nephropathy in rats. METHODS: Fifty male Wistar albino rats were randomly divided into 5 groups including: control, ADR (5 mg/kg), ADR + P. major (600 and 1200 mg/kg) and P. major (1200 mg/kg). The animals were treated with P. major extract for 5 consecutive weeks and ADR was intravenously injected on the 7th day of the study. Urine and serum samples were collected on days 0, 14, 21, 28, and 35 for the measurement of serum cholesterol and albumin levels and urine protein excretion rate. At the end of the study, the left kidneys were removed for apoptosis assessment. RESULTS: Administration of ADR significantly decreased serum albumin level and increased serum cholesterol and urine protein excretion rate as well as, apoptotic cell numbers compared to the control group (P < 0.001) while had no effect on glomerular filtration rate (P > 0.05). Treatment with P. major, in both 600 and 1200 mg/kg doses, increased serum albumin level and decreased serum cholesterol concentration, urine protein excretion rate and as well as the number of apoptotic cell compared to the ADR group (P < 0.001). CONCLUSION: Our results showed that the P. major extract effectively protects against ADR- induced nephropathy by reducing kidney apoptosis and improving renal functioning in rats.

Neurotoxic Effects of Stanozolol on Male Rats' Hippocampi: Does Stanozolol cause apoptosis?

Stanozolol is an anabolic-androgenic steroid which is commonly abused by athletes for improved energy, appearance, and physical size. It has been previously shown to cause changes in behaviour and has various physical effects. Studies have previously been conducted on its neurotoxic effect on the central nervous system (CNS), which are typically psychological in nature. This study was performed to investigate the apoptotic effect of stanozolol on different parts of the rat hippocampus. Sixteen male Wistar rats were divided randomly into two groups (experimental and control). The experimental group received subcutaneous injections of stanozolol (5mg/kg/day) for consecutive 28 days, whereas the control group received saline using the same dosing schedule and administration route. After routine procedures, coronal sections of rat brain were stained with Toluidine blue and TUNEL for pre-apoptotic and apoptotic cell detection, respectively. In order to compare groups, the mean number of TUNEL-positive and pre-apoptotic neurons per unit area were calculated and analysed. Histopathological examination revealed that the mean number of pre-apoptotic and apoptotic neurons in the CA1, CA2, CA3 and DG areas of the hippocampus were significantly increased in the stanozolol treated group. In conclusion, stanozolol abuse may induce pre-apoptotic and apoptotic cell formation in different regions of the hippocampus.

Plantago major protects against cisplatin-induced renal dysfunction and tissue damage in rats.

The aim of the present study was to determine the effect of Plantago major (P. major) on cisplatin-induced kidney injury in the rat. Cisplatin was injected on the 6th day of the experiment. Animals were treated with P. major extract (300, 600, and 1200 mg/kg) and Vitamin E for five days before and two weeks after cisplatin administration. Cisplatin caused a significant decrease in glomerular filtration rate (GFR), urine osmolarity, and urinary excretion rate of potassium, but significant increase in the kidney index and histological damage compared with the control group. Administration of Vitamin E and P. major (300 and 600 mg/kg) significantly increased GFR compared to cisplatin group. Furthermore, urine osmolarity in Vitamin E and P. major (600 mg/kg) groups were significantly elevated compared to the cisplatin group. P. major (600 mg/kg) significantly increased the urinary excretion rate of potassium compared with cisplatin group. Furthermore, all doses of P. major and Vitamin E significantly attenuated the percentage of kidney tissue damage compared to the cisplatin group. However, only P. major (600 mg/kg) and Vitamin E treated rats showed a significant reduction in the kidney index. This study revealed that P. major extract in a dose-dependent manner provides protection against renal damage induced by cisplatin.

Thymoquinone ameliorates renal damage in unilateral ureteral obstruction in rats.

BACKGROUND: It is well established that renin-angiotensin system (RAS) play a central role in pathophysiology of renal damage following unilateral ureteral obstruction (UUO). The present study investigated the effects of thymoquinone and RAS blockade on renal tissue damage and renal expression of angiotensin II after UUO in rats. METHODS: Forty male rats were divided to five groups: Sham-operated group, UUO group and rats with UUO treated with losartan, captopril and thymoquinone. The rats were evaluated two weeks after UUO by measuring renal oxidative stress, apoptosis and the expression of tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein (MCP-1) and angiotensin II. RESULTS: Two weeks after UUO there was a significant increase in the number of renal apoptotic cells and renal malondialdehyde and TNF-α expression. In addition, renal total thiol content and the activity of antioxidant enzymes were significantly decreased in UUO group, compared with the sham group. Also, UUO was associated with upregulation in the expression of angiotensin II and MCP-1 in the obstructed kidney. Losartan, captopril and thymoquinone significantly improved oxidative damage, apoptosis and TNF-α expression and markedly decreased the upregulation of angiotensin II and MCP-1 compared with the UUO group. CONCLUSIONS: The current study suggests that thymoquinone is able to improve the UUO-induced renal tissue damage. These favorable actions of thymoquinone on UUO model in rat are comparable with the well-known RAS inhibitors captopril and losartan.

The effects of exposure to titanium dioxide nanoparticles during lactation period on learning and memory of rat offspring.

Nanoscale titanium dioxide (TiO2), which is massively produced and widely used in living environment, seems to have a potential risk on human health. The central nervous system (CNS) is the potential susceptible target of nanoparticles, but the studies on this aspect are limited so far. The aim of this study was to evaluate the effects of exposure to TiO2 nanoparticles during lactation period on learning and memory of offspring. Lactating Wistar rats were exposed to TiO2 nanoparticles (100 mg/kg; gavage) for 21 days. The Morris water maze and passive avoidance tests showed that the exposure to TiO2 nanoparticles could significantly impair the memory and learning in the offspring. Therefore, the application of TiO2 nanoparticles and the effects of their exposure, especially during developmental period on human brain should be cautious.

Maternal exposure to titanium dioxide nanoparticles during pregnancy; impaired memory and decreased hippocampal cell proliferation in rat offspring.

Titanium dioxide nanoparticles (TiO2-NPs) are massively produced in the environment, and because of their wide usage, they are a potential risk of damage to human health. TiO2-NPs are often used as additives for paints, papers, and foods. The central nervous system (CNS), including hippocampal regions, is potentially susceptible targets for TiO2-NPs. This study aimed to determine the effects of exposure to TiO2-NPs during pregnancy on hippocampal cell proliferation and the learning and memory of offspring. Pregnant Wistar rats received intragastric TiO2-NPs (100 mg/kg body weight) daily from gestational day (GD) 2 to (GD) 21. Animals in the control group received the same volume of distilled water via gavage. After delivery, the one-day-old neonates were deeply anesthetized and weighed. They were then killed and the brains of each group were collected. Sections of the brains from the rat offspring were stained using Ki-67 immunolabeling and the immunohistochemistry technique. Some of the male offspring (n=12 for each group) were weaned at postnatal day (PND21), and housed until adulthood (PND60). Then the learning and memory in animals of each group were evaluated using passive avoidance and Morris water maze tests. The immunolabeling of Ki-67 protein as a proliferating cell marker showed that TiO2-NPs significantly reduced cell proliferation in the hippocampus of the offspring (P<0.05). Moreover, both the Morris water maze test and the passive avoidance test showed that exposure to TiO2-NPs significantly impaired learning and memory in offspring (P<0.05). These results may provide basic experimental evidence for a better understanding of the neurotoxic effects of TiO2-NPs on neonatal and adult brains.

The effect of PTZ-induced epileptic seizures on hippocampal expression of PSA-NCAM in offspring born to kindled rats.

BACKGROUND: Maternal epileptic seizures during pregnancy can affect the hippocampal neurons in the offspring. The polysialylated neural cell adhesion molecule (PSA-NCAM), which is expressed in the developing central nervous system, may play important roles in neuronal migration, synaptogenesis, and axonal outgrowth. This study was designed to assess the effects of kindling either with or without maternal seizures on hippocampal PSA-NCAM expression in rat offspring. METHODS: Forty timed-pregnant Wistar rats were divided into four groups: A) Kind+/Seiz+, pregnant kindled (induced two weeks prior to pregnancy) rats that received repeated intraperitoneal (i.p.) pentylenetetrazol, PTZ injections on gestational days (GD) 14-19; B) Kind-/Seiz+, pregnant non-kindled rats that received PTZ injections on GD14-GD19; C) Kind+/Seiz-, pregnant kindled rats that did not receive any PTZ injections; and D) Kind-/Seiz-, the sham controls. Following birth, the pups were sacrificed on PD1 and PD14, and PSA-NCAM expression and localization in neonates' hippocampi were analyzed by Western blots and immunohistochemistry. RESULTS: Our data show a significant down regulation of hippocampal PSA-NCAM expression in the offspring of Kind+/Seiz+ (p = 0.001) and Kind-/Seiz+ (p = 0.001) groups compared to the sham control group. The PSA-NCAM immunoreactivity was markedly decreased in all parts of the hippocampus, especially in the CA3 region, in Kind+/Seiz+ (p = 0.007) and Kind-/Seiz+ (p = 0.007) group's newborns on both PD1 and 14. CONCLUSION: Our findings demonstrate that maternal seizures but not kindling influence the expression of PSA-NCAM in the offspring's hippocampi, which may be considered as a factor for learning/memory and cognitive impairments reported in children born to epileptic mothers.

Effects of maternal nicotine exposure on expression of laminin alpha 5 in lung tissue of newborn.

Maternal smoking has been clearly demonstrated to be associated with increased health problems in infants and children. Nicotine is the chemical substance with high level of toxicity. It crosses through the placenta and accumulates in the developing organs of fetus. Previous investigation indicated that maternal nicotine exposures induce decreased fibronectin expression in lung parenchyma. In this study, the effect of maternal nicotine exposure on laminin expression of the newborn mice lungs has been evaluated. Female pregnant Balb/C mice were divided randomly in to four groups as fallow: Experimental group 1 (Exp D1); was received 3 mg kg(-1) nicotine intra peritoneal injection (IP) from gestational day 7 (GD7) to the last day of pregnancy, Experimental group 2 (Exp D14); was received 3 mg kg(-1) nicotine from GD7 to postnatal day 14, Groups 3 and 4; as sham control groups (Sha-Con) were received the same volume (3 mg kg(-1)) of normal saline parallel to experimental groups. At the end of exposure times, all of newborns were anesthetized; their lungs were removed and prepared for immunohistochemical method and real-time polymerase chain reaction. The finding indicated that laminin alpha 5 (Lama5) mRNA expressions in the lung of newborn in the nicotine treated Exp D1 decreased by 0.63 fold but increased in Exp D14 by 1.57 fold comparing to Sh-Con groups. Lama5 immunoreactivity was not similar in different parts of the lungs including alveoli and bronchiole, having a significant increase in the experimental groups in contrast to the Sh-Con groups. However, increase in immunoreactivity observed more in Exp D14. Immunoreactivity intensity in small vessels of all experimental groups was not significantly different. These data also indicate that maternal nicotine exposure may induce abnormal laminin expression which may cause defects in lung function during life time.