Artemisin and human endometrial-derived stem cells improve cognitive function and synaptic plasticity in a rat model of Alzheimer disease and diabetes.

Alzheimer disease (AD) is a common form of dementia associated with loss of memory and disruption of synaptic plasticity. There is a strong correlation between the pathophysiological features of AD and diabetes, including induction of oxidative stress, inflammation, and abnormality in blood vessels. Considering the brain's limited capacity to repair damage and the potential of stem cell-derived neural cells in the repair of neurodegenerative disease, we investigated the effects of artemisinin and TSP­1­human endometrial-derived-derived stem cells (TSP­1­hEDSCs) on the cognitive function and synaptic plasticity in AD-diabetes rats. The authors previously showed that artemisinin and TSP­1­hEDSCs suppressed oxidative stress and inflammation in AD-diabetes rats. Thrombospondins-1 (TSPs-1) is a glycoprotein that inhibits angiogenesis. AD and diabetes were induced using streptozotocin. Synaptic plasticity and learning and memory function were studied using the Morris water maze and electrophysiological test, respectively. Streptozotocin increased traveled swimming distance and escape latency in the morris water maze test, decreased the percent time spent in the target quadrant, inhibited the long-term potentiation (LTP), and increased the blood glucose levels. Simultaneous or separate administration of artemisinin and TSP­1­hEDSCs decreased the blood levels of glucose and improved cognitive tasks and synaptic plasticity by considerably reducing traveled swimming distance and escape latency, increasing the percent time spent in the target quadrant, and retrieval of the LTP; therefore, they could be utilized as an adjunct treatment for AD treatment. These results may be due to a decrease in oxidative stress and inflammation.

Effect of eugenol on lipid profile, oxidative stress, sex hormone, liver injury, ovarian failure, and expression of COX-2 and PPAR-α genes in a rat model of diabetes.

BACKGROUND: Diabetes is among the leading causes of reproductive system failure and infertility in both women and men. Inflammation and oxidative stress have a main role in the development of diabetes. Eugenol or clove oil is a phenolic monoterpenoid with antioxidant and anti-inflammatory properties. Here, the effects of eugenol on diabetes features and ovarian function were investigated. METHODS AND RESULTS: Streptozotocin-induced diabetes rats were treated with 12 and 24 mg/kg of eugenol for 4 weeks. The biochemical and histological assay was done to evaluate the effects of eugenol on ovary and pancreas function, liver injury, oxidative status, sex hormones, lipid profile, and mRNA levels of cyclooxygenase-2 (COX-2) and peroxisome proliferator-activated receptor alpha (PPAR-α) genes. Streptozotocin increased levels of serum glucose, total cholesterol, triglyceride, low-density lipoprotein, aspartate transaminase, alanine transaminase, alkaline phosphatase, malondialdehyde, pancreas necrosis and inflammation, COX-2 expression, ovarian cystic, and anovulation. It decreased the levels of insulin, high-density lipoprotein, Superoxide dismutase, estradiol, progesterone, testosterone, luteinizing hormone, follicle-stimulating hormone, and PPAR-α expression. Eugenol administration ameliorated diabetes features through the improvement of lipid profile, oxidative status, insulin and glucose levels, sex hormone levels, liver markers, COX-2 and PPAR-α expression, and pancreas histology. It had no effect on ovarian cystic and follicular development. CONCLUSIONS: Therefore, eugenol may be useful for ameliorating some adverse features of diabetes and used as an adjunct treatment or protective agent accompany by other chemicals in diabetes patients.

The effect of hydroalcoholic extract of Cichorium intybus leaf on aryl hydrocarbon receptor expression in the testis of Wistar rats exposed to cigarette smoke.

Objective: Cigarette smoke (CS) contains compounds such as reactive oxygen species (ROS). Oxidative stress caused by excessive ROS eventually leads to germ cell apoptosis and male infertility. The leaves of Cichorium intybus (chicory) are rich in natural antioxidants, but their protective effects on the adverse effects of CS on testicular tissue have not been studied. Materials and Methods: 24 Wistar rats were classified into four groups: control, extract: treatment with chicory extract (200 mg/kg body weight/day) for 13 weeks, smoke: exposed to CS for 13 weeks, and smoke + extract: exposed to CS and treated with the C. intybus extract. Histological and biochemical analyses and apoptosis assay were done, and Ahr, and Cyp1a1 expression was determined. Results: Treatment with C. intybus compensated for the reduction of Sertoli cells, spermatogonia, spermatocytes, and spermatids caused by CS. Chicory extract reduced free radicals and improved antioxidant status. The lowest and highest percentage of apoptotic cells was observed in the extract and smoke groups, respectively, while simultaneous treatment with C. intybus extract led to a significant reduction of apoptotic cells. The mean Ahr levels in the control, extract, smoke and smoke + extract groups were 1.00±0.57, 1.93±0.25, 5.98±0.42, and 0.62±0.22, respectively (p˂0.05). The mean levels of Cyp1a1 expression in the control, extract, smoke and smoke + extract groups were 1.00±0.31, 2.28±0.65, 5.55±0.40, and 0.21±0.23 (p˂0.05). Conclusion: The C. intybus extract probably affected Cyp1a1 expression by downregulation of Ahr. These led to a decrease in free radicals and apoptosis, and an improvement in antioxidant status.

Effects of empagliflozin on the expression of kisspeptin gene and reproductive system function in streptozotocin-induced diabetic male rats.

One of the main health concerns of diabetes is testicular dysfunction and impairment of reproductive function and sperm quality which can cause male infertility. kisspeptin is a hypothalamic neuropeptide hormone that is involved in the regulation of energy metabolism, gonadotrophin-releasing hormone (GnRH), and reproductive function. In the present study, the therapeutic effects of empagliflozin (sodium-glucose co-transporter 2 inhibitors) on kisspeptin expression along with reproductive function were investigated in diabetic male Wistar rats. Diabetes was induced by a single dose injection of 60 mg/kg streptozotocin. Empagliflozin in doses of 10 and 25 mg/kg body weight was used for 8 weeks. Serum samples, testis, epididymis, and pancreas tissues were collected at the end of the experiments. Lipid profiles, oxidative stress markers, blood hormones, expression of kisspeptin along with pathological alterations of the testis were assayed using real-time PCR, biochemical, and histological technics. Data have shown that empagliflozin improved hyperglycemia, reproductive impairment, oxidative stress condition, and histopathological alterations of pancreatic and testis tissues in diabetic animals. It improved the serum levels of sex hormones, insulin, leptin, and the expression of kisspeptin in the testes tissues. Spermatogenesis is also improved in treated animals. Data indicated that the administration of empagliflozin can ameliorate symptoms of diabetes. It probably has promising antidiabetic potential and may improve the male infertility of diabetic subjects. To our knowledge, this is the first experimental evidence for the potential impact of empagliflozin on kisspeptin expression in diabetic male rats.

Neuroprotective effects of nerolidol against Alzheimer's disease in Wistar rats.

Alzheimer's disease (AD) is the most common type of cognitive disorder in an elderly population associated with the accumulation of amyloid plaques and neurofibrillary tangles. Nerolidol is assumed to have neuroprotection effects. This study aimed to investigate the therapeutic effects of nerolidol on the Aß-induced model of AD in rats. Hippocampal injection of Aß was used to induce AD. Animals were randomly divided into control, sham (received PBS as Aß solvent), AD, DNPZ (AD + donepezil, 4 weeks); NRD-50 (AD + nerolidol, 50 mg/kg, 4 weeks), NRD-100 (AD + nerolidol, 100 mg/kg, 4 weeks; Prot (rats which received 100 mg/kg nerolidol for two weeks before Aß administration), and Solv (AD + sunflower oil as nerolidol solvent, 4 weeks) groups. All rats were subjected to a memory behavioral passive avoidance test by shuttle box. Thioflavin-S staining was performed to confirm Aß plaque formation and measured using ImageJ analyzing program. BDNF and CREB-1 expressions were analyzed by immunohistochemistry assay. Aß induced AD by Aß plaques formation and increasing step-through latency time. It reduced the expression of BDNF and CREB-1 protein. Administration of nerolidol or donepezil improved these features by decreasing Aß and increasing BDNF and CREB-1 expression and latency time. Nerolidol is likely to provide protection against AD. It may prevent dementia through the mediation of BDNF-CREB-1 expression and cholinergic nerve cells restoring. It seems that the administration of nerolidol before the onset of the disease will be more effective than after.

Therapeutic Effects of Eugenol in Polycystic Ovarian Rats Induced by Estradiol Valerate: A Histopathological and A Biochemical Study.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common type of endocrinopathy in women which is accompanied by androgens elevation, insulin resistance, and metabolic dysfunction. Eugenol is a phenolic component of clove oil that has an antioxidant, anti-inflammatory, and anti-diabetic activity. The present study aimed to evaluate the therapeutic effects of eugenol on the PCOS models of rats. MATERIALS AND METHODS: In this experimental study, thirty adults female Wistar rats weighing between 180 and 200 g were used. Estradiol valerate-induced PCOS rats (4 mg/rat) were treated with eugenol (12 and 24 mg/kg) for 28 days. The effects of eugenol were studied on levels of glucose, lipid profile, liver enzymes, reproductive hormones, oxidative stress, and the expression of cyclooxygenase-2 (Cox-2) and peroxisome proliferator-activated receptor alpha (Ppar-α) genes, using biochemical analysis of blood and histopathological evaluation of ovaries.
Results: Estradiol valerate-induced PCOS resulted in the formation of cystic follicles in the ovaries, hyperinsulinemia, hyperglycemia, hyperlipidemia, hyperandrogenism, and anovulation. It altered the Cox-2 and Ppar-α gene expression and increased oxidative stress and activities of liver enzymes. Eugenol treatment improved the PCOS-associated endocrine and metabolic disorder and histopathological alterations, mostly through antioxidant, anti-diabetic, anti hyperlipidemic, and anti-androgenic properties. It showed beneficial effects on serum glucose, serum insulin, fat profile, reproductive hormones, liver activity, oxidative stress, expression of Cox-2 and Ppar-α genes, as well as restoration of normal ovulation in the PCOS animals.
Conclusion: Eugenol could represent a promising natural product to prevent PCOS or reduce its symptoms.

Effects of active, inactive, and derivatives of Akkermansia muciniphila on the expression of the endocannabinoid system and PPARs genes.

This study aimed to investigate the effects of active and heat-inactivated forms of Akkermansia muciniphila, bacterium-derived outer membrane vesicles (OMVs), and cell-free supernatant on the transcription of endocannabinoid system (ECS) members, including cannabinoid receptors 1 and 2 (CB1 and CB2), fatty acid amide hydrolase (FAAH), and peroxisome proliferator-activated receptors (PPARs) genes (i.e., α, ß/δ, and δ) in Caco-2 and HepG-2 cell lines. After the inoculation of A. muciniphila in brain heart infusion enriched medium, OMVs and cell-free supernatant were extracted. For the investigation of the effects of bacteria and its derivatives on the expression of ECS and PPARs genes, the aforementioned cells were treated by active and heat-inactivated bacteria, OMVs, and cell-free supernatant. Quantitative real-time polymerase chain reaction analysis revealed that both forms of the bacterium, bacterial-derived OMVs, and cell-free supernatant could affect the expression of CB1, CB2, FAAH, and PPARs genes (i.e., α, ß/δ, and δ) significantly (P < 0.05). Considering the engagement of the aforementioned genes in metabolic pathways, it might be suggested that both forms of the bacterium, OMVs, and cell-free supernatant might have the potential to serve as a probiotic, paraprobiotic, and postbiotic candidate to prevent obesity, metabolic disorders, and liver diseases.

Therapeutic Effect of P-Cymene on Lipid Profile, Liver Enzyme, and Akt/Mtor Pathway in Streptozotocin-Induced Diabetes Mellitus in Wistar Rats.

Diabetes is a serious public health problem in low- and middle-income countries. There is a strong link between hyperglycemia, oxidative stress, inflammation, and the development of diabetes mellitus. PI3K/Akt/mTOR is the main signaling pathway of insulin for controlling lipid and glucose metabolism. P-cymene is an aromatic monoterpene with a widespread range of therapeutic properties including antioxidant and anti-inflammatory activity. In the present study, the antidiabetic effects of p-cymene were investigated. Diabetes was induced using streptozotocin in male Wistar rats. The effects of p-cymene and metformin were studied on levels of glucose (Glu), lipid profile, liver enzymes, oxidative stress, and the expression of Akt, phospho-Akt, and mTOR (mammalian target of rapamycin) proteins, using biochemical, histological, and immunohistochemical analysis. Data have shown that p-cymene can improve serum levels of Glu, total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-c), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL), alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA), and the expression of mTOR, Akt, and phospho-Akt protein in diabetic animals. These results suggest that p-cymene has hypoglycemia, hypolipidemia, and antioxidant properties. It can regulate Akt/mTOR pathway and reduce hepatic and pancreas injury. It can be suggested for diabetes management alone or simultaneously with metformin.

Molecular and biochemical modifications of suspension-cultured tobacco cell walls after exposure to alternative gravity.

The plant cell wall is a flexible physical barrier surrounding the cell which functions in growth and differentiation, signaling, and response to environmental stimuli including the Earth gravity force. In the present study, structural and molecular modifications of cell wall components of cultured tobacco (Nicotiana tabacum cv. Burley 21) cells under alternative gravity conditions induced by 7 days exposure to 2-D clinostat have been investigated. In comparison with the control group, clinorotation significantly increased biomass but reduced the total amounts of wall and the contents of cellulose, pectin, uronic acidic, and xyloglucan. Gene expression of H+-ATPase was not changed but of expansin A reduced in clinostat-treated cells. However, the gene expression and activity of xyloglucan endotransglycosylase/hydrolases (XTH; EC 2.4.1.207) and endo-(1,4)-ß-D-glucanase (EGase; EC 3.2.1.4), the amount of arabinogalactan proteins (AGP), and the expression of wall-associated kinase (WAK) gene significantly increased by clinorotation. Altered gravity also reduced the activity of polyphenol oxidase and covalently bound peroxidase. The results suggest that altered gravity promoted orchestrated changes of wall-modifying genes and proteins which reduced its stiffness and enhanced cell expansion and division potential.

Effects of artemisinin and TSP­1­human endometrial­derived stem cells on a streptozocin­induced model of Alzheimer's disease and diabetes in Wistar rats.

Alzheimer's disease (AD) is an age­associated dementia disorder characterized by Aß plaques and neurofibrillary tangles. There is a strong link between cerebrovascular angiopathy, oxidative stress, inflammation, and glucose metabolism abnormalities with the development of AD. In this study, we investigated the therapeutic influences of artemisinin and TSP­1­human endometrial­derived stem cells (TSP­1­hEDSCs) on the streptozocin­induced model of AD and diabetes in rats. Hippocampal and intraperitoneal injections of streptozocin were used to induce AD and diabetes in male Wistar rats, followed by intranasal administration of a single dose of TSP­1­hEDSCs and intraperitoneal administration of artemisinin for 4 weeks. Hematoxylin together with eosin staining was performed for demonstrating Aß plaque formation and for analyzing the influence of treatments on the pyramidal cells in the hippocampus. Biochemical analysis was used to assay the serum levels of glucose, MDA, ROS, and TAC. The expression of TNF­α was measured using real­time PCR. Streptozocin induced AD and diabetes via Aß plaque formation and increasing blood glucose levels. It also increased the levels of ROS, MDA, and TNF­α and decreased the levels of TAC. Simultaneous or separate administration of artemisinin and TSP­1­hEDSCs ameliorated this influence by considerably reducing Aß plaque formation in the hippocampus, reducing glucose, MDA, ROS, and TNF­α levels, and increasing TAC levels. It appears that artemisinin and TSP­1­hEDSCs improve the adverse features of AD in a rat model of AD and diabetes. Therefore, artemisinin and TSP­1­hEDSCs could be utilized as an adjunct treatment, as well as a protective agent, in AD patients.

Evaluating the Differential Effects of Valproic Acid on Wharton's Jelly Mesenchymal Stem Cells.

Purpose: The histone deacetylases (HDAC) inhibitor, valproic acid (VPA), is a common antiepileptic drug and is attractive for its broad range of therapeutic effects on many diseases. It has been employed as an inducer of pluripotency in some cultured cells. Conversely, VPA has also been employed as an inducer of in vitro differentiation in many other cells. Therefore, we employed WJMSCs as a cellular target to evaluate the differential effects of of VPA on potency state and differentiation level of Wharton's Jelly mesenchymal stem cells (WJMSCs) in various concentrations and different culture mediums. Methods: The isolated WJMSCs were cultured in DMEM (MSC medium). According to previous protocols, WJMSCs were treated with 0, 0.5 and 1 mM VPA in MSC or embryonic stem cell (ESC) medium and 2 mM VPA in neural differentiation medium. Real-time polymerase chain reaction (PCR) and western blot analysis were performed for evaluating the expression of pluripotency markers. MTT and caspase assays were also performed on VPA-treated cells. Results: The expression of pluripotency markers and the viability of the WJMSCs - determined by MTT assay - were significantly increased after 0.5 mM VPA treatment in ESC medium. A 2 mM VPA treatment in neural differentiation medium significantly diminished the expression of pluripotency markers and the viability of WJMSCs. Conclusion: According to our results, both VPA concentration and the medium context can influence VPA effects on WJMSCs. The differential effects of VPA on WJMSCs can reflect its wide range of effects in the treatment of various diseases.

Energy saving and improvement of metabolism of cultured tobacco cells upon exposure to 2-D clinorotation.

Studies have confirmed that on the ground, the plant cells must expend energy to maintain positional homeostasis against gravity. Under microgravity conditions, such energy may be saved for other process such as biosynthesis of beneficial metabolites for growth. This hypothesis was examined on a cell line of tobacco (Nicotiana tabacum cv. Burley 21). The cells were continuously treated with 2-D clinostat for 1 week. Exposure to clinorotation conditions increased biomass and total protein. Total content of soluble sugar also increased which may provide more precursors for Krebs cycle and adenosine triphosphate (ATP) production. In the case of 2-D clinorotation, the expression and activity of glutamate producing enzyme, glutamate dehydrogenase (GDH) increased, whereas the activity of glutamate decarboxylase (GAD) decreased. Regarding the role of GAD in initiation of gamma amino butyric acid (GABA) shunt, it is plausible that under clinorotation condition, the tobacco cells directed their metabolism toward saving energy for Krebs cycling and more production of ATP rather than shifting to side paths such as GABA shunt. Improvement of radical scavenging enzymes activity and increase of the contents of phenolic compounds and certain peroxide neutralizing amino acids, e.g., His, Pro, Ser, and Asp under clinorotation conditions decreased membrane lipid peroxidation and maintained the growth potential of tobacco cells.

Thymoquinone ameliorates some endocrine parameters and histological alteration in a rat model of polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common form of the endocrine disease which is associated with metabolic dysfunction. PCOS and type 2 diabetes mellitus are related in multiple aspects and are similar in many pathological features. Anti-diabetic effects of Nigella sativa and protective effects of it on reproductive system have been suggested in some reports. OBJECTIVE: The aim of current study was to evaluate the effects of thymoquinone, the main components of Nigella sativa, on PCOS model of rats. MATERIALS AND METHODS: Intraperitoneal injection of estradiol valerate for 25 days was used to induce PCOS in Wistar rats, followed by intraperitoneal administration of 8 and 16 mg/kg thymoquinone for 30 days. Rats were divided into 5 groups; control, sham or PCOS, experiment-1 (PCOS and 8 mg/kg thymoquinone), experiment-2 (PCOS and 16 mg/kg thymoquinone), and metformin (PCOS and metformin administration, 100 mg/kg) groups. All of the animals were subjected to serum biochemical analysis of blood and histopathological study of ovaries. RESULTS: Estradiol valerate induced PCOS while administration of thymoquinone recovered it. The body weight, ovarian morphology, and ovulation had been improved and the serum biochemical parameters including glucose, triglyceride, total cholesterol, low-density lipoprotein, high-density lipoprotein, luteinizing hormone, and follicle stimulating hormone were reversed after thymoquinone intervention. CONCLUSION: Our data suggest that thymoquinone has improvement effects on an ovarian function and ovulation in the PCOS rat model. Therefore, thymoquinone and Nagilla sativa could be used as a protective agent and as an adjunct treatment in PCOS patients.

Thymoquinone recovers learning function in a rat model of Alzheimer's disease.

OBJECTIVE: Alzheimer's disease is a neurodegenerative disorder characterized by accumulation of amyloid beta in the hippocampus. In recent decades, herbal medicine has been widely used to treat many neurodegenerative disorders,as in comparison to conventional drugs, herbal remedies exert minimal side effects. Here, the effects of thymoquinone, as the main active component of Nigella sativa, on passive avoidance memory in rat model of Alzheimer's disease, were evaluated. MATERIALS AND METHODS: Hippocampal injection of amyloid beta (Aß) was used to induce Alzheimer's disease in male Wistar rats, followed by intra peritoneal administrations of 5 and 10 mg/kg thymoquinone on a daily basis for 4 weeks. Animals were subjected to fear learning behavior in passive avoidance test and histopathological analysis of the hippocampus was done. Shuttle box test was used to evaluate the condition studying memory. Thioflavin-S and Hematoxylin and Eosine staining were done to confirm Aß plaque formation and to evaluate the effect of thymoquinone on the pyramidal cells in the hippocampal CA1 region. RESULTS: Amyloid beta caused cognitive dysfunction reflected by increasing initial and step-through latency along with plaque formation and degeneration of pyramidal cells in the hippocampus. Thymoquinone administration ameliorated this effect by significant reductions in plaque formation in CA1 region of the hippocampus and increased latency time. It also increased the number of surviving neurons in the hippocampus. CONCLUSION: It seems that thymoquinone improved learning function in a rat model of Alzheimer's disease. Thus, thymoquinone could be possibly used as an anti-neurodegenerative agent for protecting hippocampal neurons against neurotoxic effects of Aß in patients with Alzheimer's disease.

Construction of scaffolds composed of acellular cardiac extracellular matrix for myocardial tissue engineering.

High rates of mortality and morbidity stemming from cardiovascular diseases unveil extreme limitations in current therapies despite enormous advances in medical and pharmaceutical sciences. Following myocardial infarction (MI), parts of myocardium undergo irreversible remodeling and is substituted by a scar tissue which eventually leads to heart failure (HF). To address this issue, cardiac patches have been utilized to initiate myocardial regeneration. In this study, a porous cardiac patch is fabricated using a mixture of decellularized myocardium extracellular matrix (ECM) and chitosan (CS). Results of rheological measurements, SEM, biodegradation test, and MTT assay showed that the scaffold composed of 3.5% (w/w) CS and 0.5% ECM has the best potential in providing cardiac progenitor cells (CPCs) with a suitable microenvironmental condition for both attachment and growth. This study demonstrates that the fabricated scaffold is capable of transmitting both mechanical and chemical cues that is native to myocardial tissue and supports efficient growth of the CPCs.

Simulated Microgravity Condition Alters the Gene Expression of some ECM and Adhesion Molecules in Adipose Derived Stem Cells.

Adipose- derived stem cells (ADSCs) are widely used for tissue engineering and regenerative medicine. The beneficial effects of ADSCs on wound healing have already been reported. Remodeling of extracellular matrix (ECM) is the most important physiological event during wound healing. ECM is sensitive to mechanical stresses and the expression of its components can be therefore influenced. The aim of this study was to investigate the effect of simulated microgravity on gene expression of some ECM and adhesion molecules in human ADSCs. After isolation and characterization of ADSCs, cells were exposed to simulated microgravity for 1, 3 and 7 days. Real-time PCR, fluorescence immunocytochemistry, and MTT assay were performed to evaluate the alterations of integrin subunit beta 1 (ITGB1), collagen type 3 (ColIII), matrix metalloproteinase-1 (MMP1), CD44, fibrillin (FBN1), vimentin (VIM) genes, and ColIII protein levels as well as cells viability. Microgravity simulation increased the expression of ITGB1, ColIII, MMP1, and CD44 and declined the expression of FBN1 and VIM genes. ColIII protein levels also increased. There were no significant changes in the viability of cells cultured in microgravity. Since the high expression of ECM components is known as one of the fibroblast markers, our data suggest that pretreatment of ADSCs by simulated microgravity may increase their differentiation capacity towards fibroblastic cells. Microgravity had not adversely affected the viability of ADSCs, and it is likely to be used alone or in combination with biochemical inducers for cell manipulation.

A Comparative Study of 660 nm Low-Level Laser and Light Emitted Diode in Proliferative Effects of Fibroblast Cells.

Introduction: In recent years, low-power lasers have been widely used in medicine. With the introduction of affordable light emitted diode (LED), clinical application of LED light has become more and more popular. However, some researchers believe that due to lack of coherence of the LED light, it can be different in terms of biological effects, in comparison with laser. In this study, the biological effects of low-level laser (LLL) to those of LED light are compared and discussed. Methods: Human skin fibroblast cell line Hu02 was irradiated with LLL and LED light with a wavelength of 660 nm, power output of 35 mW and in continuous mode and the control group was not irradiated. The biological effects were compared through analysis of cell proliferation, production of reactive oxygen species (ROS) within the cell and rate of cell division. Results: Our findings showed that production of ROS within the cell was linearly increased both in the LED and laser light irradiated cells. However, laser light is more incremental in comparison to LED light. The MTT results showed that laser light at low energy density (less than 5 J/cm2) increased the rate of cell proliferation after 24 hours. Although, the rate of cell division was increased in energy density of 1 J/cm2 compared to the control group, this increase was not statistically significant. Discussion: The findings indicated that the coherence properties of laser light provided more energy for the cells, and in a constant energy density, laser light created more oxidative stresses in comparison with LED light.

Structural Changes of Human Serum Albumin (HSA) in Simulated Microgravity.

BACKGROUND: Nowadays, the biological effects of microgravity have been the subject of various experimental researches. Microgravity has been confirmed to affect biological systems. Furthermore, as a result of improvement in space technology for instance a manned mission to the moon, probabilities for human exposed to microgravity have incremented undoubtedly. OBJECTIVES: The purpose of this study was to investigate the probable biological effects of microgravity on the human serum albumin (HSA) structure after 3 and 24 h exposure. It is worth mentioning that this is the first effort to investigate the structural alternations of HSA under simulated microgravity condition in biophysico-chemical terms thru different spectroscopic instruments. METHODS: 2D clinostat was utilized for simulating microgravity. The UV-Vis, intrinsic and extrinsic fluorescence, dynamic light scattering (DLS) and circular dichroism (CD) spectra of 3.76 µM HSA in Tris-HCl buffer (pH 7.4, 0.1 M) and 3.76 µM HSA in Tris-HCl buffer (pH 7.4, 0.1 M) kept at simulated microgravity for 3 and 24h were verified. RESULTS: The UV-Visible, near-UV-CD and intrinsic fluorescence spectroscopy represented that microgravity can remarkably change the tertiary structure of HSA. Additionally, the ANS affinity for HSA incremented when the protein was exposed to simulate microgravity compared to unexposed HSA, which may possibly have appeared attributable to expansion of the structure of simulated HSA. Fluorescence quenching by acrylamide demonstrated higher stern-volmer constant for exposed HSA. The results of zeta potential and dynamic light scattering (DLS) experiments depicted that simulated microgravity cause raise in the surface charge and size of HSA. Far-UV CD data demonstrated that simulated microgravity did not perturb the secondary structures of the protein. CONCLUSION: Collectively, our results suggest that HSA after 24 h exposure to microgravity can exhibit a molten globule (MG) structure. This is the first report to demonstrate the molten globule state formation in microgravity condition. Results from this study could give knowledge to understand the role of gravity on protein folding process. In addition, this finding could help to find out safe limits for astronauts and space travelers and to develop adequate countermeasures against any harmful effects of microgravity.

Expression pattern of neurotrophins and their receptors during neuronal differentiation of adipose-derived stem cells in simulated microgravity condition.

OBJECTIVES: Studies have confirmed that microgravity, as a mechanical factor, influences both differentiation and function of mesenchymal stem cells. Here we investigated the effects of simulated microgravity on neural differentiation of human adipose-derived stem cells (ADSCs). MATERIALS AND METHODS: We have used a fast rotating clinostat (clinorotation) to simulate microgravity condition. Real-time PCR and flow cytometry analysis were used to evaluate the regulation of neurotrophins, their receptors, and neural markers by simulated microgravity and their impact on neural differentiation of cells. RESULTS: Our data revealed that simulation microgravity up-regulated the expression of MAP-2, BDNF, TrkB, NT-3, and TrkC both before and after neural differentiation. Also, the neural cells derived from ADSCs in microgravity condition expressed more MAP-2, GFAP, and synaptophysin protein in comparison to the 1G control. CONCLUSION: We showed that simulated microgravity can enhance the differentiation of mesenchymal stem cells into neurons. Our findings provide a new strategy for differentiation of ADSCs to neural-like cells and probably other cell lineages. Meanwhile, microgravity simulation had no adverse effects on the viability of the cells and could be used as a new environment to successfully manipulate cells.