Benefits of bone marrow mesenchymal stem cells compared to their conditioned medium in valproic acid-induced autism in rats.

BACKGROUND: Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by repetitive behaviors, a limited range of activities, and deficiencies in social communications. Bone marrow mesenchymal stem cells (BM-MSCs), which secrete factors that stimulate surrounding microenvironment, and BM-MSCs conditioned medium (BM-MSCs-CM), which contains cell-secreted products, have been speculated to hold potential as a therapy for ASD. This study aimed to compare the therapeutic effects of BM-MSCs and BM-MSCs-CM on behavioral and microglial changes in an animal model of autism induced by valproic acid (VPA). METHODS AND RESULTS: Pregnant Wistar rats were administered by VPA at a dose of 600 mg/kg at 12.5 days post-conception. After birth, male pups were included in the study. At 6 weeks of age, one group of rats received intranasal administration of BM-MSCs, while another group received BM-MSCs-CM. The rats were allowed to recover for 2 weeks. Behavioral tests, quantitative real-time polymerase chain reaction (qRT-PCR), and immunohistochemistry were performed. Both BM-MSCs and BM-MSCs-CM administration significantly improved some behavioral deficits. Furthermore, these treatments notably reduced Iba-1 marker associated with microgliosis. Additionally, there was a significant reduction in the expression of pro-inflammatory cytokines IL-1ß and IL-6, and an increase in the levels of the anti-inflammatory cytokine IL-10 in rats administered by BM-MSCs and BM-MSCs-CM. CONCLUSIONS: Post-developmental administration of BM-MSCs and BM-MSCs-CM can ameliorate prenatal neurodevelopmental deficits, restore cognitive and social behaviors, and modulate microglial and inflammatory markers. Results indicated that the improvement rate was higher in the BM-MSCs group than BM-MSCs-CM group.

Retinoic acid loaded with chitosan nanoparticles improves spermatogenesis in scrotal hyperthermia in mice.

OBJECTIVE: High temperatures can trigger cellular oxidative stress and disrupt spermatogenesis, potentially leading to male infertility. We investigated the effects of retinoic acid (RA), chitosan nanoparticles (CHNPs), and retinoic acid loaded with chitosan nanoparticles (RACHNPs) on spermatogenesis in mice induced by scrotal hyperthermia (Hyp). METHODS: Thirty mice (weighing 25 to 30 g) were divided into five experimental groups of six mice each. The groups were as follows: control, Hyp induced by a water bath (43 °C for 30 minutes/day for 5 weeks), Hyp+RA (2 mg/kg/day), Hyp+CHNPs (2 mg/kg/72 hours), and Hyp+RACHNPs (4 mg/kg/72 hours). The mice were treated for 35 days. After the experimental treatments, the animals were euthanized. Sperm samples were collected for analysis of sperm parameters, and blood serum was isolated for testosterone measurement. Testis samples were also collected for histopathology assessment, reactive oxygen species (ROS) evaluation, and RNA extraction, which was done to compare the expression levels of the bax, bcl2, p53, Fas, and FasL genes among groups. Additionally, immunohistochemical staining was performed. RESULTS: Treatment with RACHNPs significantly increased stereological parameters such as testicular volume, seminiferous tubule length, and testicular cell count. Additionally, it increased testosterone concentration and improved sperm parameters. We observed significant decreases in ROS production and caspase-3 immunostaining in the RACHNP group. Moreover, the expression levels of bax, p53, Fas, and FasL significantly decreased in the groups treated with RACHNPs and RA. CONCLUSION: RACHNPs can be considered a potent antioxidative and antiapoptotic agent for therapeutic strategies in reproductive and regenerative medicine.

Efficacy of dental pulp-derived stem cells conditioned medium loaded in collagen hydrogel in spinal cord injury in rats: Stereological evidence.

Spinal cord injury (SCI) causes histological alterations which in turn affects functional activity. Studies have demonstrated that dental pulp-derived stem cells conditioned medium has beneficial effects on the nervous system. Besides, collagen hydrogel acts as a drug releasing system in SCI investigations. This research aimed to evaluate effects of dental pulp-derived stem cells conditioned medium loaded in collagen hydrogel in SCI. After culturing of Stem cells from human exfoliated deciduous teeth (SHEDs), SHED-conditioned medium (SHED-CM) was harvested and concentrated. Collagen hydrogel containing SHED-CM was prepared. The rats were divided into five groups receiving laminectomy, compressive SCI with or without intraspinal injection of biomaterials (SHED-CM and collagen hydrogel with or without SHED-CM). After 6 weeks, histological parameters were estimated using stereological methods. The total volume of preserved white matter and gray matter (p < 0.05) as well as the total number of neurons and oligodendrocytes in the rats received SHED-CM loaded in collagen hydrogel were significantly higher, and also lesion volume and lesion length were significantly lower (p < 0.05) compared to those of the other injured groups. In conclusion, intraspinal administration of SHED-CM loaded in collagen hydrogel leads to neuroprotection, proposing a cell-free therapeutic approach in SCI.

Beneficial Effects of Olive Oil on the Rats' Cerebellum: Functional and Structural Evidence.

BACKGROUND: Olive oil is a food additive and used in many biological studies as a solvent for other chemicals, including drugs. AIM: The present study aimed to investigate the effects of olive oil on rats' cerebellum structure and motor function. MATERIALS AND METHODS: Male rats were randomly divided into two groups orally receiving distilled water and olive oil (1 ml/kg/day). At the end of week 4, motor function was assessed in the rotarod test. The cerebellum was removed for stereo-logical assessment. Data were analyzed using a two-way repeated measures ANOVA for rotarod test and Kruskal-Wallis and Mann-Whitney U test for quantitative histological parameters. RESULTS: Performance of the olive oil-treated rats in fixed and accelerating speed rotarod was better and their riding time (endurance) was greater compared to the control group (p<0.05). However, no significant difference was found between the two groups regarding the total volume of the cerebellar hemisphere, its cortex, and deep cerebellar nuclei. The total number of the Purkinje, Bergman, and Golgi of the granular layer as well as neurons of the deep cerebellar nuclei was 26 - 36% higher in the olive oil-treated rats than in the distilled water treated group (p<0.03). CONCLUSION: The study findings suggest that olive oil has neuroprotective effects on the cerebellum and induces better performance of the rats in the rotarod.

Sensory and Motor Behavior Evidences Supporting the Usefulness of Conditioned Medium from Dental Pulp-Derived Stem Cells in Spinal Cord Injury in Rats.

STUDY DESIGN: Experimental animal study. PURPOSE: This study aimed to assess effects of conditioned medium (CM) of dental pulp-derived stem cells loaded in collagen hydrogel on functional recovery following spinal cord injury (SCI). OVERVIEW OF LITERATURE: SCI affects sensory and motor functions, and behavioral recovery is the most essential purpose of therapeutic intervention. Recent studies have reported that CM from dental pulp-derived stem cells has therapeutic benefits. In addition, collagen hydrogel acts as a drug delivery system in SCI experiments. METHODS: Stem cells from human exfoliated deciduous teeth (SHEDs) were cultured, and SHED-CM was harvested and concentrated. Collagen hydrogel containing SHED-CM was prepared. The rats were divided into five groups receiving laminectomy, compressive SCI with or without intraspinal injection of biomaterials (SHED-CM), and collagen hydrogel with or without SHED-CM. Basso, Beattie, and Bresnahan (BBB) scoring, inclined plane, cold allodynia, and beam walk tests were performed for 6 weeks to assess locomotor, motor, sensory, and sensory-motor performances, respectively. RESULTS: Scores of the rats receiving SHED-CM loaded in collagen hydrogel were significantly better than those of the other injured groups at 1-week post-injury for BBB, 2 weeks for inclined plane, 2 weeks for cold allodynia, and 4 weeks for beam walk tests (p <0.05). The differences remained significant throughout the study. CONCLUSIONS: Intraspinal administration of SHED-CM loaded in collagen hydrogel leads to improved functional recovery and proposes a cell-free therapeutic approach for SCI.

Efficacy of Curcumin in the Modulation of Anxiety Provoked by Sulfite, a Food Preservative, in Rats.

Sulfites are used as food preservatives and excessive sulfite might disturb the body systems including the brain. Curcumin shows protective effects on the nervous system toxicity. The present study aimed to evaluate the protective role of curcumin in sulfite-induced anxiety in rats. Male rats were divided into five groups. The rats in groups I to V received distilled water (vehicle of sulfite, 1 mL/d), olive oil (vehicle of curcumin, 1 mL/d), curcumin (100 mg/kg/d), sulfite (25 mg/kg/d), and sulfite+curcumin, respectively, by daily gastric gavage for 8 weeks. At the end of 8 weeks the rats were tested in the elevated plus-maze for anxiety. The results showed that concomitant treatment of curcumin during sulfite consumption prevented the reduction of the time spent in the open arm and entrance to the open arm (the indexes of anxiety). Besides, an increase was found in motor activity of the rats in the sulfite+curcumin group compared to the sulfite-treated animals. Exposure of sulfite in rats can induce anxiety, and curcumin can act as an anti-anxiety agent.

Protective role of curcumin against sulfite-induced structural changes in rats' medial prefrontal cortex.

OBJECTIVES: Sodium metabisulfite as a food preservative can affect the central nervous system. Curcumin, the main ingredient of turmeric has neuroprotective activity. This study was designed to evaluate the effects of sulfite and curcumin on the medial prefrontal cortex (mPFC) using stereological methods. METHODS: Thirty rats were randomly divided into five groups. The rats in groups I-V received distilled water, olive oil, curcumin (100 mg/kg/day), sodium metabisulfite (25 mg/kg/day), and sulfite + curcumin, respectively, for 8 weeks. The brains were subjected to the stereological methods. Cavalieri and optical disector techniques were used to estimate the total volume of mPFC and the number of neurons and glial cells. Intersections counting were applied on the thick vertical uniform random sections to estimate the dendrites length, and classify the spines. Non-parametric tests were used to analyze the data. RESULTS: The mean mPFC volume, neurons number, glia number, dendritic length, and total spines per neuron were 3.7 mm(3), 365,000, 180,000, 1820 µm, and 1700 in distilled water group, respectively. A reduction was observed in the volume of mPFC (∼8%), number of neurons (∼15%), and number of glia (∼14%) in mPFC of the sulfite group compared to the control groups (P < 0.005). Beside, dendritic length per neuron (∼10%) and the total spines per neuron (mainly mushroom spines) (∼25%) were reduced in the sulfite group (P < 0.005). DISCUSSION: The sulfite-induced structural changes in mPFC and curcumin had a protective role against the changes in the rats.

Curcumin prevents the structural changes induced in the rats' deep cerebellar nuclei by sodium metabisulfite, a preservative agent.

OBJECTIVE: To evaluate the the possible neurotoxic effects of sulfite and the protective potential of curcumin on the deep cerebellar nuclei using stereological methods. METHODS: The rats were randomly divided into five experimental groups (n=6): Group I: distilled water, Group II: Olive oil, Group III: Curcumin (100 mg/kg/day), Group IV: Sodium metabisulfite (25 mg/kg/day), and Group V: Sodium metabisulfite+curcumin. At the end of 56 d, the right cerebellar hemispheres were removed and assigned to stereological studies. The total volume and total neuron number of deep cerebellar nuclei were assessed using Cavalieri and optical disector methods, respectively. RESULTS: The data showed ∼20% and ∼16% decrease was respectively observed in the total volume and the total neuron number of the deep cerebellar nuclei of the sulfite-treated rats in comparison to the distilled water group (P<0.04). However, no significant change was observed in the total volume and neuronal number of the deep cerebellar nuclei in sulfite+curcumin-treated rats and curcumin played a protective role against sulfite. Curcumin or its vehicle (olive oil) did not induce any significant changes. CONCLUSIONS: Curcumin, the main part of the turmeric, could prevent the structural changes induced in the deep cerebellar nuclei by sodium metabisulfite, a preservative agent, in rats.

Curcumin and sertraline prevent the reduction of the number of neurons and glial cells and the volume of rats' medial prefrontal cortex induced by stress.

Chronic stress induces morphological changes in the neurons of several brain regions, including medial prefrontal cortex (mPFC). This region is involved in variety of behavioral tasks, including learning and memory. Our previous work showed that stress impaired function. The present work extends the earlier work to study mPFC in stressed and non-stressed rats with or without sertraline or curcumin treatments using stereological methods. Sertraline is a selective serotonin reuptake inhibitor and curcumin is the main ingredient of turmeric with neuroprotective effects. In this study, 42 male rats were randomly assigned to seven groups: stress + distilled water, stress + olive oil, stress + curcumin (100 mg/kg/day), stress + sertraline (10 mg/kg/day), curcumin, sertraline, and control groups. After 56 days, the right mPFC was removed. The volume of mPFC and its subdivisions and the total number of neurons and glia were estimated. The results showed ~8%, ~8%, and 24% decrease in the volume of the mPFC and its prelimbic and infralimbic subdivisions, respectively. However, the anterior cingulated cortex remained unchanged. Also, the total number of the neurons and glial cells was significantly reduced (11% and 5%, respectively) in stress (+distilled water or olive oil) group in comparison to the non-stressed rats (P<0.01). However, no significant reduction was observed in the volume of the mPFC and its subdivisions as well as the total number of the neurons and glial cells in stress + sertraline and stress + curcumin groups in comparison to the non-treated stressed rats (P<0.01). The result indicated that treatment of rats with curcumin and sertraline could prevent the stress-induced changes in mPFC.

Protective effects of curcumin and sertraline on the behavioral changes in chronic variable stress-induced rats.

Behavioral characteristics of the animal models and humans are impaired in chronic stress. The present study aimed to evaluate and compare the protective effects of sertraline and curcumin on stress-induced learning and memory impairment, anxiety and anhedonia in rats. Male rats were divided into seven groups: stress+water, stress+olive oil, stress+curcumin (100 mg/kg/day), stress+sertraline (10 mg/kg/day), curcumin, sertraline, and control groups. The rats were exposed to chronic variable stress for 56 days. At the end of 40 days and while the previous treatments were continued, the rats were tested in the eight radial maze, elevated plus maze, and sucrose consumption for learning and memory, anxiety, and anhedonia, respectively. In comparison to the non-stressed group, stress+water and stress+olive oil groups revealed a significantly lower percent of correct choices and higher reference and working memory errors during learning and retention phases (p<0.001). In addition these stress groups showed a significant lower percent of the open arms time and open arms entries in the elevated plus maze and consuming less sucrose solution. In addition, the stress+curcumin and stress+sertraline groups showed a better performance in the evaluated parameters of the radial arm maze, elevated plus maze, and sucrose consumption tests. It appears that curcumin and sertraline have the similar effectiveness on behavioral changes in chronic variable stress-induced rats.

Curcumin, the main part of turmeric, prevents learning and memory changes induced by sodium metabisulfite, a preservative agent, in rats.

Sodium metabisulfite is used as a disinfectant, antioxidant, and preservative agent in the food, beverage, and drug industries. Neurons are highly sensitive to sulfite toxicity. Curcumin is the main part of turmeric and has neuroprotective effects on a variety of nervous system damages. The present study aimed to investigate the possible protective role of curcumin in learning and memory after exposure to sulfite in rats. The rats were divided into five groups receiving distilled water (solvent of the sulfite), olive oil (solvent of the curcumin), sodium metabisulfite (25 mg/kg/day), curcumin (100 mg/kg/day), and sulfite + curcumin. All the animals received daily gavages for 8 weeks. At the end of the 8(th) week, learning and memory were assessed in a partially-baited eight arm radial maze. The animals treated with sulfite showed fewer correct choices and more reference and working memory errors during the learning phase, at the end of the learning phase, and during the retention testing (p<0.001). The study results demonstrated that sulfite-exposure was associated with impaired learning and memory in rats. Adding curcumin to the rat nutrition plays a protective role in learning and memory after exposure to sulfite.

Curcumin can prevent the changes in cerebellar structure and function induced by sodium metabisulfite in rat.

Sulfites are used as anti-microbial and anti-oxidant agents in the food and pharmaceutical industries. Curcumin, a flavonoid, is an Asian spice that shows neuroprotective activities. The current study aimed to stereologically assess the rats' cerebellar cortex and rotarod performance following sulfite exposure and determine the possible neuroprotective potential of curcumin. The rats were divided into five groups: distilled water, olive oil, curcumin (100 mg/kg/day), sodium metabisulfite (25 mg/kg/day), and sodium metabisulfite+curcumin. At 56 days after treatment, rotarod performance was tested, and then the cerebellum was removed for stereological analysis. The study results revealed 31%, 36%, 19% and 24% decrease in the total volume of the cerebellum, cortex, the total number of the Purkinje cells and length of the nerve fibers in the cortex per Purkinje, respectively in the sodium metabisulfite-treated rats compared to the distilled water group (p<0.01). The pre-trained animals on the rotarod apparatus were tested first on the fixed speed rotarod protocol followed by the accelerating rotarod protocol two days later. The results showed a significant decrease in the latency to fall in both test in sulfite-treated rats. The sulfite effects on the structural parameters and rotarod performance were significantly protected by the concomitant curcumin treatment (p<0.001). Sulfite can induce structural and functional changes in the rats' cerebellum and concomitant curcumin prescription plays a neuroprotective role.