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Ghasemi-Mobarakeh L.,Islamic Azad University at Najafabad | Prabhakaran M.P.,National University of Singapore | Morshed M.,Isfahan University of Technology | Nasr-Esfahani M.H.,Royan Institute for Animal Biotechnology | And 5 more authors.
Journal of Tissue Engineering and Regenerative Medicine | Year: 2011

Among the numerous attempts to integrate tissue engineering concepts into strategies to repair nearly all parts of the body, neuronal repair stands out. This is partially due to the complexity of the nervous anatomical system, its functioning and the inefficiency of conventional repair approaches, which are based on single components of either biomaterials or cells alone. Electrical stimulation has been shown to enhance the nerve regeneration process and this consequently makes the use of electrically conductive polymers very attractive for the construction of scaffolds for nerve tissue engineering. In this review, by taking into consideration the electrical properties of nerve cells and the effect of electrical stimulation on nerve cells, we discuss the most commonly utilized conductive polymers, polypyrrole (PPy) and polyaniline (PANI), along with their design and modifications, thus making them suitable scaffolds for nerve tissue engineering. Other electrospun, composite, conductive scaffolds, such as PANI/gelatin and PPy/poly(ε-caprolactone), with or without electrical stimulation, are also discussed. Different procedures of electrical stimulation which have been used in tissue engineering, with examples on their specific applications in tissue engineering, are also discussed. Copyright © 2011 John Wiley & Sons, Ltd.


Ostadsharif M.,Islamic Azad University at Khorasgan | Ghaedi K.,Royan Institute for Animal Biotechnology | Ghaedi K.,University of Isfahan | Hossein Nasr-Esfahani M.,Royan Institute for Animal Biotechnology | And 5 more authors.
Differentiation | Year: 2011

Peroxisomal matrix protein is mainly expressed in heart, skeletal muscle, and brain tissues. To study the expression of peroxisomal protein (PEP) during neurogenesis, we employed mouse embryonic carcinoma cells (P19) and embryonic stem cells (mESCs) as an in vitro model for neural differentiation by retinoic acid (RA) induction. Expression pattern of PEP was investigated under distinct steps of differentiation by RT-PCR and real-time PCR. The results revealed that expression of PEP transcripts was markedly increased after the RA treatment at embryoid body and neural stages. Therefore, we concluded that PEP might be involved in the early process of neurogenesis, which needs further verification. © 2010 International Society of Differentiation.


Bassiri F.,Royan Institute for Animal Biotechnology | Bassiri F.,Islamic Azad University at Dāmghān | Tavalaee M.,Royan Institute for Animal Biotechnology | Shiravi A.H.,Islamic Azad University at Dāmghān | And 4 more authors.
Human Reproduction | Year: 2012

background: Intracytoplasmic sperm injection (ICSI) is the primary treatment for male infertility. However for this procedure, with the exception of visual morphological selection, there is no standardization for sperm selection. Recently, the hypo-osmotic swelling test (HOST) has been proposed to potentially select sperm with intact membranes. The aim of this study is to evaluate the ability of this technique to select functional sperm in terms of apoptosis and morphology, as well as nuclear integrity. methods: A total of 20 semen samples were randomly collected from men who attended the Andrology Unit of the Isfahan Fertility and Infertility Center. Semen samples were washed and exposed to hypotonic conditions, before being fixed and simultaneously assessed for membrane integrity as well as abnormal morphology, DNA fragmentation and protamine deficiency by using Papanicolaou, TUNEL and CMA3 staining techniques, respectively. The remaining semen samples were washed with calcium buffer and stained by Annexin V, then exposed to hypotonic conditions before being assessed for early apoptosis along with membrane integrity. results: HOST grade 'd', followed by grade 'c', showed the highest percentages of healthy sperm, whereas sperm of HOST grade 'g' in which anomalies in terms of apoptosis, abnormal head morphology or nuclear immaturity or membrane damage, were most frequently observed in the samples assessed. conclusions: Integration of HOST into the sperm selection procedure may provide a valuable tool for selection of functional sperm required for ICSI. According to this study, insemination of HOST grade 'g' sperm should be avoided during ICSI. © The Author 2012.


Kuentz P.,University of Strasbourg | Kuentz P.,Center Hospitalier University | Vanden Meerschaut F.,Ghent University | Elinati E.,University of Strasbourg | And 10 more authors.
Human Reproduction | Year: 2013

STUDY QUESTION: Does DPY19L2 status influence intracytoplasmic sperm injection (ICSI) outcomes with or without assisted oocyte activation (AOA)?SUMMARY ANSWERDPY19L2 mutations have no major impact on ICSI outcomes in globozoospermic patients.WHAT IS KNOWN ALREADYGlobozoospermia is a rare and severe teratozoospermia characterized by round-headed spermatozoa lacking an acrosome. Recently, it has been shown that DPY19L2 mutations can be found in a vast majority of, but not all, globozoospermic patients (66.7%). These patients suffer from primary infertility due to a sperm-related oocyte activation deficiency secondary to the absence of an acrosome that can be overcome by the application of AOA.STUDY DESIGN, SIZE, DURATIONCohort study, retrospective, 34 patients, 83 cycles.MATERIALS, SETTING, METHODSClinical and biologic data were collected from 29 patients mutated for DPY19L2 and 5 non-mutated patients. In total, 35 ICSI cycles using AOA and 48 conventional ICSI cycles were included in the analysis. Patients were divided into groups according to whether or not they were mutated for DPY19L2 and whether or not they received AOA. MAIN RESULTS AND THE ROLE OF CHANCE: Regardless of the presence of a DPY19L2 mutation, the fertilization rates with AOA are restored to normal when compared with conventional ICSI in our cohort of globozoospermic patients. Also, when performing ICSI plus AOA, both mutated and non-mutated cases have similar positive hCG rates, ongoing pregnancy rates and live birth rates per transfer. On the contrary, the fertilization rate in globozoospermic patients using conventional ICSI is correlated with the presence of a DPY19L2 mutation, with slightly better, although still very low, fertilization rates in patients carrying a DPY19L2 mutation. Nevertheless, when performing conventional ICSI, both mutated and non-mutated cases have similar very low positive hCG rates, ongoing pregnancy rates and live birth rates per transfer.LIMITATIONSA limitation of this study is the low number of included non-mutated cases. WIDER IMPLICATIONS OF THE FINDINGS: We propose a pathway for the clinical management of globozoospermic patients depending on the phenotype that includes several diagnostic and therapeutic steps. STUDY FUNDING/COMPETING INTEREST(S): None. © 2013 The Author.


Asgari V.,Royan Institute for Animal Biotechnology | Hosseini S.M.,Royan Institute for Animal Biotechnology | Ostadhosseini S.,Royan Institute for Animal Biotechnology | Hajian M.,Royan Institute for Animal Biotechnology | And 4 more authors.
Molecular Reproduction and Development | Year: 2012

Oocyte vitrification and assisted oocyte activation have increasingly important roles in assisted reproductive technology. Yet, an important area of concern with matured oocyte cryobiology is that elements of oocytes intimately involved in metaphase-II arrest may be modified by cryopreservation. By comparing different cellular characteristics of unvitrified, vitrified-warmed, and unvitrified-activated oocytes, the present study investigated how vitrification-warming process may affect developmental competence of in vitro-matured sheep oocytes following parthenogenetic activation. Structural, ultrastructural, and molecular analyses indicated that the characteristics of vitrified-warmed oocytes vastly differed from fresh oocytes, instead resembling unvitrified-activated oocytes. For unvitrified oocytes, the highest blastocyst yield (41.8±0.6%) was achieved using the maximum ionomycin concentration (5μM), and importantly, the duration of ionomycin treatment was not of utmost importance at this concentration. In contrast, the maximum blastocyst yield of vitrified-warmed oocytes (28.4±1.4%) was achieved with a minimal duration of ionomycin treatment (1min), and further extending the duration dramatically reduced developmental potential of vitrified-warmed oocytes. These results suggested that vitrified-warmed oocytes may need an activation protocol different from unvitrified oocytes. In this respect, unvitrified oocytes were more sensitive to the concentration rather than the duration of ionomycin treatment when compared with vitrified oocytes, which were sensitive to the treatment duration. These results may provide a platform to improve the potential applications of vitrified oocytes in medicine and agriculture. © 2012 Wiley Periodicals, Inc.


Taghipour Z.,Royan Institute for Animal Biotechnology | Taghipour Z.,Isfahan University of Medical Sciences | Karbalaie K.,Royan Institute for Animal Biotechnology | Kiani A.,Royan Institute for Animal Biotechnology | And 6 more authors.
Stem Cells and Development | Year: 2012

Regarding both the neural crest origin and neuronal potential of stem cells from human exfoliated deciduous teeth (SHED), here, we assessed their potential in addition to neural induced SHED (iSHED) for functional recovery when transplanted in a rat model for acute contused spinal cord injury (SCI). Following transplantation, a significant functional recovery was observed in both groups relative to the vehicle and control groups as determined by the open field locomotor functional test. We also observed that animals that received iSHED were in a better state as compared with the SHED group. Immunohistofluorescence evaluation 5 weeks after transplantation showed neuronal and glial differentiation and limited proliferation in both groups. However, myelin basic protein and chondroitin sulfate proteoglycan NG2-oligodendrocyte markers-were increased and glial fibrillary acidic protein-astrocyte marker-was decreased in the iSHED group in comparison with the SHED group. These findings have demonstrated that transplantation of SHED or its derivatives could be a suitable candidate for the treatment of SCI as well as other neuronal degenerative diseases. © 2012 Mary Ann Liebert, Inc.


Tavalaee M.,Royan Institute for Animal Biotechnology | Deemeh M.R.,Royan Institute for Animal Biotechnology | Arbabian M.,Royan Institute for Animal Biotechnology | Nasr-Esfahani M.H.,Royan Institute for Animal Biotechnology | And 2 more authors.
Journal of Assisted Reproduction and Genetics | Year: 2012

Purpose: Although, at present, the selection of sperm prior to ICSI is based on motility and morphology, undetectable anomalies, and more importantly damaged DNA are overlooked. In this regard, novel sperm selection procedures have gained much interest. For instance, sperm has been selected by Magnetic-Activated Cell Sorting (MACS) based on early apoptotic marker, the externalization of phosphatidylserine (EPS). Review of the literature has revealed that the efficiency of this technique has been mainly evaluated post Density Gradient Centrifugation (DGC). Therefore, there is a need to prove the efficiency of this technique independent of DGC. In addition, considering the fact that DGC induces EPS due to capacitation and acrosome reaction, therefore, the role of MACS before DGC(MACS-DGC) and MACS after DGC (DGC-MACS) should be assessed. Methods: Semen samples from fifteen infertile men were divided into three separate fractions: control, DGC, and MACS. To carry out DGC-MACS, DGC samples were further divided into two fractions and MACS was carried on the second fractions. Similarly to carry out MACS-DGC, the MACS samples were further divided into two fractions and DGC was carried on the second fractions. Percentages of sperm with normal morphology, DNA fragmentation, protamine deficiency, EPS and caspase-3 activity were determined in each fraction. Results: DGC is more efficient than MACS in separating intact sperm only in terms of normal morphology, DNA and chromatin integrity but not for active caspase. However, a combination of these procedures was more efficient than a single procedure to separate intact sperm for the aforementioned parameters. Comparison of the combined procedures showed only higher efficiency to separate active caspase in the MACS-DGC group. Conclusion: Based on these results, we propose MACS-DGC rather than DGC-MACS to be implemented in clinical settings. © 2011 Springer Science+Business Media, LLC.


Kiani-Esfahani A.,Royan Institute for Animal Biotechnology | Tavalaee M.,Royan Institute for Animal Biotechnology | Deemeh M.R.,Royan Institute for Animal Biotechnology | Deemeh M.R.,Isfahan Fertility and Infertility Center | And 3 more authors.
Systems Biology in Reproductive Medicine | Year: 2012

The aim of this study was to assess the potential of dihydrorhodamine 123 (DHR123) to measure oxidative stress produced by human spermatozoa. The results were compared with 2', 7'-dichlorodihydrofluorescein diacetate (DCFH-DA) that is routinely used for assessment of H2O2 produced by spermatozoa. Fluorescence intensity and percentage R123 and DCF positive sperm were measured by flow cytometry. The optimal condition for assessment of reactive oxygen species (ROS) produced by sperm with DHR123 was 0.05 M for 1 million sperm per ml for 20 minutes. The results of ROS measurement by DHR123 showed a significant correlation (r0.818, P<0.001) with DCFH-DA staining. Immunofluorescence of sperm stained with DHR123 revealed ROS production in the sperm mid-piece. In addition a significant correlation was observed between oxidant production assessed by DHR123 and semen parameters. Therefore, DHR123 may be considered a suitable probe for estimating oxidants produced by human spermatozoa, and can present heterogeneity in oxidant production between different samples. © 2012 Informa Healthcare USA, Inc.


Nasr-Esfahani M.H.,Royan Institute for Animal Biotechnology
Methods in molecular biology (Clifton, N.J.) | Year: 2013

Sperm selection is an important part of the ICSI procedure. Routinely, sperm selection for ICSI is based solely on sperm morphology and motility. These latter parameters may not be sufficient to select sperm with intact chromatin. Therefore, sperm selected based on sperm functional characteristics may result in the most appropriate sperm for the ICSI procedure. The methodology explained below describes the selection of sperm based on the ability of sperm to bind solid-state hyaluronic acid as its receptor, present on mature sperm with intact chromatin.


Asgari V.,Royan Institute for Animal Biotechnology | Hosseini S.M.,Royan Institute for Animal Biotechnology | Ostadhosseini S.,Royan Institute for Animal Biotechnology | Hajian M.,Royan Institute for Animal Biotechnology | Nasr-Esfahani M.H.,Royan Institute for Animal Biotechnology
Theriogenology | Year: 2011

It is a common practice to rest vitrified-warmed matured oocytes for 1-3 h, as a treatment to recover spindle and cytoskeleton, before commencing a further treatment. Vitrified-warmed matured oocytes, however, are very sensitive and may resume meiosis spontaneously during this recommended rest time. Therefore, the aim of this study was to assess spindle and chromosome status as well as developmental competence of vitrified in vitro matured sheep oocytes activated parthenogenetically, either 0 h (immediately) or 2 h (delayed) after warming. There was no significant effect of post-warming interval on the proportion of degenerated oocytes. Evaluation of chromosomes and meiotic spindle configuration showed that 11.11% of oocytes in the immediate group and 8.82% of oocytes in the delayed group had normal chromosomal alignment on well-structured spindles, compared to non-vitrified group (79.41%). Meanwhile, majority of the chromosomal abnormalities in the immediate and delayed groups were categorized as absent (unobservable) (77.78%) and anaphase II (70.59%), respectively. Oocytes in immediately activated group showed significantly higher blastocyst rate (28.86%) compared to delayed activated group (16.47%). In conclusion, the results suggest that post-warming interval may have important consequence on meiotic progression and parthenogenetic activation of vitrified oocytes. In sheep, it appears that chemical activation without having to await microtubule reorganization improves embryonic development. © 2011 Elsevier Inc.

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