Avicenna Journal of Medical Biotechnology
Year:2016 | Volume:8 | Issue:3|Papers Count:8

At present, Iran has been known one of the up-warding countries in the world in regenerative medicine using stem cells therapy. In fact, the outcomes of some clinical trials on stem cell therapy of myocardial infarction, vitiligo, decompensated cirrhosis, and osteoarthritis narrate the feasibility of stem cell-based therapy for treatment of human diseases 1, 2. However, in a similar manner with global configuration, the commercialization and translation of tissue engineering products into clinical phase has been restricted. It might be due to weak collaboration of different specialties for technology transfer of the multidisciplinary projects of tissue engineering field into clinical phase. Basic tissue engineers mostly prefer elegant studies, whereas physicians have tendency to solve medical problems with products indicating efficiency, easy to use, and cost benefit. Actually, a surgeon encountered with a dilemma between a partially effective tissue-engineered product that is both expensive and difficult to apply and a more traditional approach may choose the latter option.

Background: Nowadays, highly specific aptamers generated by cell SELEX technology (systematic evolution of ligands by exponential enrichment) are being applied for early detection of cancer cells. Prostate Specific Membrane Antigen (PSMA), over expressed in prostate cancer, is a highly specific marker and therefore can be used for diagnosis of the prostate cancer cells. The aim of the present study was to select singlestranded DNA aptamers against LNCap cells highly expressing PSMA, using cell–SELEX method which can be used as a diagnostic tool for the detection of prostate cancer cells.Methods: After 10 rounds of cell-SELEX, DNA aptamers were isolated against PSMA using LNCaP cells as a target and PC-3 cell lines for counter SELEX. Five DNA aptamers with more than 70% affinity were selected up on flow cytometry analysis of positive clones.Results: Dissociation constants of two selected sequences (A12-B1) were estimated in the range of 33.78±3.77 and 57.49±2.214 pmol, respectively. Conserved secondary structures of A12 and B1 sequences suggest the necessity of these structures for binding with high affinity to native PSMA. Comparison of the secondary structures of our isolated aptamers and aptamer A10 obtained by protein SELEX showed similar stem-loop structures which could be responsible for the recognition of PSMA on LNCap cell surface.Conclusion: Our results indicated that selected aptamers may turn out to be ideal candidates for the development of a detection tool and also can be used in targeted drug delivery for future smart drugs.

Background: Malignant melanoma is a highly aggressive malignant melanocytic neoplasm which resists against the most conventional therapies. Sea cucumber as one of marine organisms contains bioactive compounds such as polysaccharide, terpenoid and other metabolites which have anti-cancer, anti-tumor, anti-inflammatory and antioxidant properties. The present study was designed to investigate the anticancer potential of saponin extracted from sea cucumberHolothuria leucospilata alone and in combination with dacarbazine on B16F10 melanoma cell line.Methods: The B16F10 cell line was treated with different concentrations of saponin (0, 4, 8, 12, 16, 20mg/ml), dacarbazine (0, 1200, 1400, 1600, 1800, 2000 mg/ml) and coadministration of saponin-dacarbazine (1200 da+8 sp, 1200 da+4 sp) for 24 and 48hr and the cytotoxic effect was examined by MTT, DAPI, acridine orange/propodium iodide, flow cytometry and caspase colorimetric assay.Results: The results exhibited that sea cucumber saponin, dacarbazine, and coadministration of saponin-dacarbazine inhibited the proliferation of melanoma cells in a dose and time dependent manner with IC50 values of 10, 1400 and 4+1200 mg/ml, respectively. Morphological observation of DAPI and acridine orange/propodium iodide staining documented typical characteristics of apoptotic cell death. Flow cytometry assay indicated accumulation of IC50 treated cells in sub-G1 peak. Additionally, saponin extracted induced intrinsic apoptosisvia up-regulation of caspase-3 and caspase-9.Conclusion: These results revealed that the saponin extracted from sea cucumber as a natural anti-cancer compound may be a new treatment modality for metastatic melanoma and the application of sea cucumber saponin in combination with dacarbazine demonstrated the strongest anti-cancer activity as compared with the drug alone.

Background: Sporadic Alzheimer’s Disease (SAD) is caused by genetic risk factors, aging and oxidative stresses. The herbal extract ofRosa canina (R. canina), Tanacetum vulgare (T. vulgare)and Urtica dioica (U. dioica) has a beneficial role in aging, as an anti-inflammatory and anti-oxidative agent. In this study, the neuroprotective effects of this herbal extract in the rat model of SAD was investigated.Methods: The rats were divided into control, sham, model, herbal extract -treated and ethanol-treated groups. Drug interventions were started on the 21st day after modeling and each treatment group was given the drugs by intraperitoneal (I.P.) route for 21 days. The expression levels of the five important genes for pathogenesis of SAD including Syp, Psen1, Mapk3, Map2 and Tnf- a were measured by qPCR between the hippocampi of SAD model which were treated by this herbal extract and control groups. The Morris Water Maze was adapted to test spatial learning and memory ability of the rats.Results: Treatment of the rat model of SAD with herbal extract induced a significant change in expression of Syp (p=0.001) and Psen1 (p=0.029). In Morris Water Maze, significant changes in spatial learning seen in the rat model group were improved in herbal-treated group.Conclusion: This herbal extract could have anti-dementia properties and improve spatial learning and memory in SAD rat model.

Background: Protein aggregation is one of the important, common and troubling problems in biotechnology, pharmaceutical industries and amyloid-re-lated disorders.Methods: In the present study, the inhibitory effects of some carbohydrates (alginate, b-cyclodextrin and trehalose) on the formation of nano-globular aggregates from normal (HSA) and glycated (GHSA) human serum albumin were studied; when the formation of aggregates was induced by the simultaneous heating and addition of dithiotheritol. For the investigations, the biophysical methods of UV-vis spectrophotometry, circular dichroism spectroscopy, transmission electron microscopy and tensiometry were employed.Results: The effect of inhibitory mechanism of these inhibitors on the aggregation of HSA and GHSA was expressed and compared together.Conclusion: The results showed that the nucleus formation step of the aggregation process of HSA and GHSA was different in the presence of alginate (compared toβ-cyclodextrin and trehalose). The inhibition efficiencies of the carbohydrates on the aggregate formation of HSA and GHSA were different, arising from the differences in the hydrophobicities of HSA and GHSA, and also, the differences between HSA- and GHSA-carbohydrate interactions.

Background: Niche cells, regulating Spermatogonial Stem Cells (SSCs) fate are believed to have a reciprocal communication with SSCs. The present study was conducted to evaluate the effect of SSC elimination on the gene expression of Glial cell line-Derived Neurotrophic Factor (GDNF), Fibroblast Growth Factor 2 (FGF2) and Kit Ligand (KITLG), which are the main growth factors regulating SSCs development and secreted by niche cells, primarily Sertoli cells.Methods: Following isolation, bovine testicular cells were cultured for 12 days on extracellular matrix-coated plates. In the germ cell-removed group, the SSCs were removed from thein vitro culture using differential plating; however, in the control group, no intervention in the culture was performed. Colony formation of SSCs was evaluated using an inverted microscope. The gene expression of growth factors and spermatogonia markers were assessed using quantitative real time PCR.Results: SSCs colonies were developed in the control group but they were rarely observed in the germ cell-removed group; moreover, the expression of spermatogonia markers was detected in the control group while it was not observed in the germ cellremoved group, substantiating the success of SSCs removal. The expression ofGdnf andFgf2 was greater in the germ cell-removed than control group (p<0.05), whereas the expression ofKitlg was lower in the germ cell-removed than control group (p< 0.05).Conclusion: In conclusion, the results revealed that niche cells respond to SSCs removal by upregulation of GDNF and FGF2, and downregulation of KITLG in order to stimulate self-renewal and arrest differentiation.

Background: This study was aimed to assess the effects of angiotensin II (Ang II) supplementation to the In Vitro Maturation (IVM) and In Vitro Culture (IVC) media of vitrified-warmed ovine oocytes on their developmental competence and expression of Na+/K+/ATPase in resulting embryos.Methods: The slaughterhouse-derived immature oocytes (n=1069) were randomly distributed into four experimental groups: groups I and II) IVM/IVF and IVC of fresh and vitrified oocytes without angiotensin supplementation (Control-Fresh and Control-Vit groups, respectively); group III) IVM of vitrified oocytes in the presence of Ang II followed by IVF/IVC (Vit-IVM group); and group IV) IVM/IVF of vitrified oocytes followed by IVC wherein the embryos were exposed to Ang II on day 4 of IVC (Vit-D4 group).The embryos were immunostained with primary antibodies against Na+/K+/ATPase a1 and b1 subunits.Results: In Vit-IVM and Vit-D4 groups, the rates of expanded and total blastocysts on day 7 as well as the proportion of blastocysts on day 8 were increased. The expression of Na+/K+/ATPase a1 and b 1 subunits were positively influenced by the addition of Ang II on day 4 (Vit-D4 group).Conclusion: The addition of Ang II to the IVM and IVC media could improve blastocysts formation in vitrified sheep oocytes. This improvement might be related to the greater expression of Na+/K+/ATPase a1 and b 1 subunits when Ang II was added during IVC.

Background: Acromegaly is associated with increased morbidity and mortality related to cardiovascular diseases. Leptin (LEP) and Leptin Receptor (LEPR) gene polymorphisms can increase cardiovascular risks. The aim of this study was to investigate association between the frequencies of LEP and LEPR gene polymorphisms and subclinical atherosclerosis in acromegalic patients.Methods: Forty-four acromegalic patients and 30 controls were admitted to study.The polymorphisms were identified by using polymerase chain reaction from peripheral blood samples. The levels of systolic and diastolic blood pressure, BMI, fasting plasma glucose, fasting insulin, IGF-I, GH, IGFBP3, leptin, triglyceride, carotid Intima Media Thickness (cIMT) and HDL and LDL cholesterol concentrations were evaluated.Results: There was statistically significant difference between the LEPR genotypes of acromegalic patients (GG 11.4%, GA 52.3%, and AA 36.4%) and controls (GG 33.3%, GA 50%, and AA 16.7%) although their LEP genotype distribution was similar. In addition, the prevalence of the LEPR gene G and A alleles was significantly different between patients and controls. No significant difference was found among the G (-2548) A leptin genotypes of groups in terms of the clinical parameters. cIMT significantly increased homozygote LEPR GG genotype group compared to AA subjects in patients.But the other parameters were not different between LEPR genotypes groups of patients and controls.Conclusion: It can be said that the LEPR gene polymorphism may affect cIMT in patients.The reason is that LEPR GG genotype carriers may have more risk than other genotypes in the development of subclinical atherosclerosis in acromegaly.