Optimizing hydrocortisone concentration for skeletal muscles differentiation of ADSCs on PLLA nano-scaffolds

Skeletal muscle is a highly differentiated tissue with very specific functions which has low potential of regeneration. Skeletal muscle injuries especially in athletes almost have lead to muscular dysfunctions and healing may be prolonged for several years. Therefore, working on skeletal muscle differentiation remained an importance in biomedical researches. Adipose derived stem cells [ADSCs] are novel source of mesenchymal stem cells which are an excellent alternative for satellite cells in in-vitro skeletal muscle differentiation. Differentiation potential of ADSCs on both tissue culture plate [TCP] and also on Poly l-lactide acid [PLLA] electrospun fibrous nano-scaffold which now is widely used at tissue engineering investigations has studied in this research. Scanning electron microscopy [SEM] and Tensile test were performed for evaluating scaffold properties. Hydrocortisone has considered a critical factors for skeletal muscle differentiation while, the recommended concentrations of it for inducing myogenesis in stem cells is yet discussing. Statistical analysis of our results from colorimetric MTT assay for various concentrations of hydrocortisone showed that the concentration of 10[-7] mol/L is the optimum dose for myogenic differentiation of murine ADSCs which was used on both TCP and PLLA scaffolds and skeletal myosin fiber formations was confirmed with immunocytochemistry. DAPI staining proved myocytes nuclei and syncytium formations. Our results also showed that ADSCs and PLLA nano-scaffolds are the suitable biomaterials for engineering skeletal muscle tissue

ADSCs on PLLA/PCL hybrid nanoscaffold and gelatin modification: cytocompatibility and mechanical properties

Development of tissue engineering and regenerative medicine has led to designing scaffolds and their modification to provide a better microenvironment which mimics the natural niche of the cells. Gelatin surface modification was applied to improve scaffold flexibility and cytocompatibility. PLLA/PCL aligned fibrous scaffold was fabricated using electrospinning method. ADSCs were seeded after O[2] plasma treatment and gelatin coating of the scaffolds. The morphological and mechanical properties of blends were assessed by Scanning Electron Microscopy [SEM], tensile test and ATR-FTIR. The cells proliferation was evaluated by MTT assay. Based on the results, it is supposed that gelatin coating is a brilliant method of surface modification which significantly increases the mechanical properties of scaffold without any changes on the construction or on the direction of nanofibers which conducts cell's elongation. MTT analysis exhibited that ADSCs attachment, viability and proliferation significantly [p<0.05] increased after gelatin treatment. Gelatin surface modification is a highly beneficial method to improve cytocompatibility, flexibility and mechanical features of the scaffolds which doesn't affect the nanofibers construction. Proliferation of Adipose Derived Stem Cells [ADSCs] as a remarkable source of stem cells was investigated for the first time on PLLA/PCL hybrid scaffold

comparative study of osteogenic differentiation human induced pluripotent stem cells and adipose tissue derived mesenchymal stem cells

Human induced pluripotent stem cells [iPSCs] have been shown to have promising capacity for stem cell therapy and tissue engineering applications. Therefore, it is essential to compare the ability of these cells with the commonly used mesenchymal stem cells [MSC] for bone tissue engineering in vitro. In this experimental study, the biological behavior and osteogenic capacity of the iPSCs were compared with MSCs isolated from human adipose tissue [AT-MSCs] using 3-[4,5-di-methylthiazol-2-yl]-2,5-diphenyltetrazolium bromide [MTT] assay, Alizarin red staining, alkaline phosphatase [ALP] activity measurements, calcium content assay and common osteogenic-related genes. Data were reported as the mean +/- SD. One-way analysis of variance [ANOVA] was used to compare the results. A p value of less than 0.05 was considered statistically significant. There was a significant difference between the rate of proliferation of the two types of stem cells; iPSCs showed increased proliferation compared to AT-MSCs. During osteogenic differentiation, ALP activity and mineralization were demonstrated to be significantly higher in iPSCs. Although AT-MSCs expressed higher levels of Runx2, iPSCs expressed higher levels of osteonection and osteocalcin during differentiation. iPSCs showed a higher capacity for osteogenic differentiation and hold promising potential for bone tissue engineering and cell therapy applications

effects of pyridaben pesticide on the DNA integrity of sperms and early in vitro embryonic development in mice

Pyridaben, a pyridazinone derivative, is a new acaricide and insecticide for control of mites and some insects such as white flies, aphids and thrips. This study was designed to elucidate how pyridaben can affect the sperms' morphological parameters, its DNA integrity, and to estimate the effect of various quantities of pyridaben on in vitro fertilization rate. In this study, 80 adult male Balb/C strain mice were used. Animals were divided into control and two test groups. Control group received distilled water. The test group was divided into two subgroups, viz, high dose [212 mg/kg/day] and low dose [53 mg/kg/day] and they received the pyridaben, orally for duration of 45 days. The spermatozoa were obtained from caudae epididymides on day 45 in all groups. Sperm viability, protamin compression [nuclear maturity], DNA double-strand breaks, and in vitro fertilizing [IVF] ability were examined. The pyridaben treatment provoked a significant decrease in sperm population and viability in epididymides. The data obtained from this experiment revealed that, the pyridaben brings about negative impact on the sperm maturation and DNA integrity in a time-dependent manner, which consequently caused a significant [p<0.05] reduction in IVF capability. Embryo developing arrest was significantly [p<0.05] higher in treated than the control group. Theses results confirmed that, the pyridaben is able to induce DNA damage and chromatin abnormalities in spermatozoa which were evident by low IVF rate

[ effect of fatigue on passive avoidance learning and its interaction with simvastatin drug in male Wistar rats]

The aim of this study was to evaluate the sequence effects of simvastatin and fatigue, specifically the interaction between fatigue effects and passive avoidance learning in simvastatin treated rats. In this experimental study, 40 male Wistar rats weightening 300-350 g were divided into 4 groups: control group: treated with enough tap [n=10], Sham group: rats which were gavaged with distilled water and food [n=10], first experimental [Exp1] and Second experimental [Exp2] group: rats which were gavaged with distilled water and food [n=10] and simvastatin with dose of 1 mg/kg for 35 days [n=10]. Then, both experimental groups were forced to swim in aquariums for 35 days at 9-10 AM to 13-14 PM. At the end of treatment days, passive avoidance learning [PAL] system was considered and monitored. The learning and memory of all groups were studied. After performing behavioral tests, testosterone and cortisol were measured. fatigue decreased learning activity in Exp group one [P<0.001], while simvastatin causes increase in learning actitives in Exp group two. Also, fatigue induced by forced swimming caused significant decrease of testosterone hormone level [P<0.001], increase of cortisole hormone level [P<0.001] in experimental groups in comparison with sham and control groups. Our study showed that fatigue due to long term swimming causes decrease in learning, while simvastatin drug is effective for prevention of decrease of learning and memory in fatigued rats.

Role of serotonergic system on modulation of depressogenic-like effects of silymarine

In the present study, we have investigated the effects of silymarine on depression and the possible role of serotonergic system in these effects. The rats were anesthetized intraperitoneally with ketamine hydrochloride and placed in a Stoelting stereotaxic instrument. A stainless steel guide cannula [22-gauge] was implanted in the third ventricular region. The third ventricular region was infused by means of an internal cannula [27-gauge], terminated 1 mm below the tip of the guide cannula. Forced swimming test was used for evaluating the depression. The results obtained from this study showed that oral administration of silymarin [35, 70, 140 and 280 mg/rat] for two weeks increased the immobility time in forced swimming test, indicating an increase in depression level of the treated rats. Intra-third-ventricle [Intra-TV] infusion of 5HT1A receptor agonist 8-OH-DPAT [25 and 10 ng/rat] decreased the immobility time indicating an anti-depression effect, while injection of 5HT1A receptor antagonist NAN190 [0.25, 0.5 and 1 microg/rat] had no significant effect on immobility time. An effective dose of 8-OH-DPAT [10 ng/rat] co-administered with silymarin [140 and 280 mg/rat] decreased the depressogenic effects of silymarin. These results showed that the depressogenic effects of silymarin may be modulated via 5HT1A receptor of serotonin

In vivo study of the effect of kombucha on the injured skin of adult NMRI mice strain

Kombucha is used for the treatment of several diseases, including wound healing, in traditional medicine. Due to its effective abilities, such as antibiotic activities, detoxification, and its impact on skin luminosity, we attempted to determine its effect on injured skin in in vivo condition. 30 male NMRI strain were randomly assigned in control [injured skin], sham [exposed with sweet tea] and experimental [received Kombucha] groups. A 3mm diameter wound with complete skin thickness was made on the back of each mouse. Treatment with Kombucha was applied three times a day for 18 days. At the end of study, histological study was carried out on the under-treatment tissues. Significant decrease in wound diameter was observed in experimental group compared to control group [P<0.001]. Furthermore, significant increased epidermal thickness and decreased derm and hypoderm thickness were seen in experimental group compared to control group [P<0.05]. Hair follicle diameter was significantly increased in experimental group compared to control group, while skin thickness diameter was in coordination with control group. External use of Kombucha on open wound skin with complete skin thickness effectively accelerates wound-healing procedure and also increases hair follicle diameter and length in in vivo condition

Study the effect of extremely low frequency electromagnetic fields on some blood serum's lipoproteins, liver enzymes and P448/P450 cytochrome enzyme system in NMRI female mice

Arterial diseases are the first causes of death in the developed countries. It seems necessary to investigate the role of enzymes, lipids, proteins and para-oxidation lipids in these diseases. The tribulation of fats is one of the most risk factors which results in atherosclerosis of coronary arteries. In this study, the short time effects of Extremely Low Frequency Electromagnetic Fields with the frequency of 50Hz, on the amount of some blood lipo-proteins, liver enzymes, P448 and P450 cytochrome enzyme system are investigated in the NMRI female mice. The results indicate that the blood serum's levels of lipoproteins decrease significantly, in comparison with control and sham groups [P<0.05]. Experimental groups of 3 and 5 showed significant increase of HDL compare with control and sham groups [P<0.001]. While levels of Aspartate aminotransferase [AST] increased in liver, Alanin aminotransferase [ALT] enzyme levels did not show any significant changes in experimental groups [P<0.01]. In addition, Ames test shows similarity in control and experimental liver extract in the reveres colonies, so the electromagnetic field with intensity of 0.06 militesla [P<0.01], does not disturb the P448/P450 enzyme system

[Effect of vibration stress on the sexually dimorphic nucleus [SDN] of hypothalamus of immature male wistar rat]

The sexually dimorphic nuclei in the preoptic area [SDN-POA] of hypothalamus are a cluster of cells which size, shape and their cellular morphology are related to gender and age. Different stressors affect on hypothalamus nuclei. In this study, effect of vibration stress was studied on endocrine system and sexually dimorphic nucleus [SDN] of hypothalamus. In this experimental study, male immature Wistar rats [15 days old] were divided into 3 groups Control, Experimental E350 and E500. Experimental groups were exposed to vibration stress by shaker with frequency 350mot/min and 500mot/min, 15 minutes daily for 3 weeks. At the end of this period, serum plasmic hormones, such as cortisol, testosterone, estradiol and progesterone, were evaluated. Sections of the brain tissue were prepared for histological and microscopic studies. Cortisol levels increased significantly in experimental E350 [p<0.05] and E500 [p<0.001] groups. Testosterone level decreased significantly in experimental E500 [p<0.01] group. Progesterone increased significantly in experimental E500 [p<0.01]. Estradiol did not show a significant decrease. Microscopic studies of brain showed significant decrease of number and density of sexually dimorphic nuclei in the preoptic area [SDN-POA] of hypothalamus in male immature Wistar rats with vibration stress. Vibration stress induces dysfunction of hypothalamus-pituitary-adrenal axis which can lead to homosexuality and other sexually behavioral disorders

[ effects of chronic use of imatinib on wistar rat fetuses]

Imatinib mesylate selectively inhibits bcr/abl and other non-specific tyrosine kinases and represents a model of targeted therapy for chronic myeloid leukaemia [CML] as well as gastrointestinal stromal tumors [GIST]. This study was designed to evaluate effects of imatinib on pregnancy and development of fetus. In this experimental study, imatinib was administrated orally at doses of 7, 12, 22, 50 and 100 mg/kg/day and control groups received sterile water. The pregnant rats were subdivided into 2 groups. In group one, the pregnant rats were sacrificed on day 18 of gestation and the number alive and dead of foetuses were checked. The brain of fetuses were fixed in formalin and embedded in paraffin for histological studies. Selected slides were stained with hematoxylin and eosin [H and E]. In group two, the fetuses were allowed to become mature. The effect of drug on learning and memory were assessed by a passive avoidance method using shuttle box apparatus. Histological studies revealed no evidence of teratogenic effects of imatinib on development of frontal and parietal bones. Imatinib given in 100 mg/kg dose caused weight decrease [p<0.001] and increase mortality in fetuses [p<0.01]. Administration of imatinib in 7, 12, 22 and 50 mg/kg doses showed statistically significant reduction in learning and memory of fetuses [p<0.05]. Imatinib can decrease development, learning and memory of fetuses. So, it is recommended that women treated with imatinib avoid becoming pregnant

[ role and effect of finasteride on the spermatogenesis, prostate gland and epididyim of mature NMRI mouse in in vivo and in vitro conditions]

Finasteride, a 4-azasteroid compound, is a specific inhibitor of type II 5alpha-reductase that converts testosterone into 5alpha-dihydrotestosterone. In the present study, the effects of human dose of finasteride on the number of spermatogenic cells, seminiferous, prostatic and epididymal tubules diameter and thickness of mature NMRI mouse were investigated, both in in vivo and in vitro conditions. In this experimental study, in in vivo condition, 18 mature NMRI mice were divided into 3 groups of 6: control [without treatment], sham [treatment whit physiologic serum] and experimental [treatment with drug]. Experimental group was received intraperitoneal injection of 5 mg/kg/day finasteride for 7 days. In in vitro condition, testes, prostates and epididyms of 18 mature NMRI mice were divided into 3 groups of 6: control [fixation with bouin without treatment], sham [treatment with physiologic serum] and experimental [treatment with 5 mg/kg/day of finasteride in culture media for 3 days]. Data were analyzed using one way ANOVA and Tukey test by SPSS software. P- value< 0.05 was considered significant. In in vivo condition, treatment with finasteride did not cause significant reduction in the number of spermatogenic cells, seminiferous tubules diameter compared with control group. But, inin vitro condition, significant decrease was observed. In both conditions, the drug could cause significant decrease in prostatic tubules diameter and thickness compared with control group. Diameter and thickness of epididymis tubules were decreased just in in vitro condition. Finasteride can act in long-term treatment, high doses and in in vitro condition better than other conditions