Cell and Tissue Journal

Abstract  
Aim: Tissue engineering is a new approach to regeneration and repair lost or damaged tissues. The aim of this study is to design and manufacture polycaprolactone (PCL) randomly electrospun nanofiber scaffold for use in regenerative medicine.
Material and Methods: Human adipose derived stem cells (hADSCs) were isolated (from superficial layer of abdominal fat), cultured (in DMEM/Ham'sF12 medium) and characterized (flow cytometry for CD29, CD73, CD34, CD105 and CD45). Electrospinning was used to produce PCL nanofiber scaffolds, scanning electron microscopy (SEM) was used to investigate the binding, penetration and morphology of hADSCs, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was used to determine the toxicity of scaffolds.
Results: Flow cytometry showed extensive expression of the CD29, CD73 and CD105 (positive) and very low expression of the CD34 and CD45 (negative) in hADSCs. The results of MTT assay, showed the viability and proliferation of hADSCs which were seeded on the nanofiber scaffold. Microscopic photographs of SEM, showed hADSCs attached to PCL nanofiber scaffolds and migrating.
Conclusion: The results of this study showed that electrospun PCL nanofiber scaffolds are suitable for implantation, binding and propagation of hADSCs.

Abstract Aim: The toxic effects of cadmium exposure are mainly due to the production of oxygen free radicals, reduction of cellular antioxidants and oxidative stress, which can lead to cell component destruction, DNA damage, apoptosis and ultimately tissue damage. In this study, the protective effects of N-acetylcysteine, as a substance with antioxidant properties, in cadmium-exposed Wistar mice were investigated by liver histology and measuring the expression of effective genes in apoptosis and cell proliferation.
Material and Methods: Mice weighing approximately 150-200 g were classified into three treatments including, G1) control treatment, G2) cadmium recipient treatment, and G3) concomitant cadmium and N-acetylcysteine treatment and for four weeks received the desired. Then liver tissue samples were taken for histopathological examination and expression of eif4e and mad1 genes.
Results: The results of this study showed that cadmium exposure resulted in serious damage to rat liver tissue, including central artery hyperemia, increased number of inflammatory cells and inflammation in the liver parenchyma, while the use of N-acetylcysteine significantly reduced the mentioned injuries. Also, the use of N-acetylcysteine resulted in a significant reduction in the expression of eif4e and mad1 genes by 2.14 and 2.27 times, compared with mice that received only cadmium.
Conclusion: The results of this study suggest that N-acetylcysteine as an antioxidant can play an important role in preventing tissue damage due to oxidative stress and preventing the induction of cellular apoptosis.

Abstract Aim: The aim of this study was to collect and identify Fusarium of Elegans and Martiella-Ventricosum sections related to cucurbits plant in Tehran province and evaluation the efficiency of ITS-RDNA RFLP polymorphisms as a molecular marker in assessment of relationships between these Fusaria. Materials and Methods: During the cropping season, sampling from cucurbit field was done from the root, crown, stem up to height of 15 cm, as well as the soil (rhizosphere). Fungal isolates were cultured on PPA culture medium, purified by single sporulation method, and then identified based on morphological characteristics. Genomic DNA of fungi was extracted. The genomic DNA of Fusarium isolates was extracted and ITS-rDNA region was amplified with ITS1 and ITS4 primers. PCR products were digested with SmaI, BglΠ and MboI restriction enzymes and then loaded on 2.5% agarose gel. Data analyses were performed by NTSYS-PC V2.02 software. At first, a data matrix of the presence (1) or absence (0) of each band was drawn for each isolate. Then using the SM similarity coefficient, the genetic distance matrix was produced and the similarity dendogram of isolates was drawn based on the SAHN coefficient and the UPGMA method. Results: A total of 95 Fusarium isolates belonging to species of Fusarium solani and Fusarium oxysporum were identified. Of which, 45 isolates belonging to F. solani and 50 isolates were identified as F. oxysporum. In this research, based on morphological characteristics, F. redolens isolates were not differentiated from F. oxysporum isolates and both were placed under F. oxysporum group. PCR reproduction of ITS region result in fragments in size of 550±25 bp in F. oxysporum isolates, and 575±25 bp in F. solani isolates. Bgl Π enzyme had no cleavage site in ITS products in both species, neither in F. solani nor in F. oxysporum. SmaI enzyme had one cleavage site in F. solani isolates and produced two fragments (350bp and 230bp) but had no cleavage site in F. oxysporum isolates. The MboI enzyme in some F. oxysporum isolates produced four fragments (180bp, 160bp, 130 bp and 90 bp), so these isolates called as Fusarium redolens. MboI enzyme in others Fusarium oxysporum isolates produced two bands (320 bp and 190 bp). MboI enzyme had no restriction site in F. solani isolates. Conclusion: It seems that the use of rDNA-ITS RFLP marker is a suitable method to differentiate Fusarium species belonging to section Elegance from those of Martiella-Ventricosum sections. It is more efficient method for studying diversity within and between species in section Elegans.

Abstract Aim: Nowadays, much attention is paid to the effects of natural factors on physiological and pathological processes in human body. In this regard, lack of oxygen or hypoxia is one of the crucial biological factors involved in various physiological processes such as wound healing and pathological processes such as cancer. Moreover, it is very important to find natural compounds affecting the characteristics and functions of cells. Thymoquinone (TQ) is a natural compound derived from certain plants such as Nigella Sativa. It has many biopharmacological effects, including anti-bacterial, anti-oxidant, anti-inflammatory, anti-diabetic, anti-aging, anti-cancer, etc. Given the biopharmacological properties of TQ and the importance of hypoxia as an important factor affecting physiological and pathological processes, this study was designed to investigate the effect of TQ under cobalt (II) chloride-mediated hypoxia on breast cancer and wound healing by evaluating the expression of SOX2, CDK4, c-MET, and DNMT1 genes in a breast cancer cell line (MCF7) and a normal fibroblastic cell line (HDF) that treated with these compounds.
Materials and Methods: In the present study, after the cultivation of MCF7 and HDF cell lines, each of the cells were divided into two groups. The treatment group was treated simultaneously with 500 ng/ml of TQ and 100 μM of cobalt (II) chloride for 24 h and the control group was only treated with cobalt (II) chloride. After incubation time, total RNA extraction, DNase I treatment, and cDNA synthesis were carried out and finally, the expression of target genes was examined by real-time PCR assay. In this study, relative threshold method was used to determine the amount of gene expression changes, and SPSS software and Student's t-test statistical method were used to find the significance of gene expression changes in the treated groups compared to the controls.
Results: The results showed that simultaneous treatment of MCF7 cells with TQ and cobalt (II) chloride significantly (P < 0.05) reduced the expression of CDK4, c-MET, and DNMT1 genes at about 4.35-, 1.89-, and 2.08-fold, respectively, compared to the control group. However, the treatment of MCF7 cells caused a limited increase in the expression of SOX2 at about 1.14-fold, which was not significant according to the significance level of ≥ 1.5. Moreover, simultaneous treatment of HDF cells with TQ and cobalt (II) chloride significantly increased c-MET gene expression by about 1.86-fold. In addition, the treatment of HDF cells caused a slight increase in the expression of CDK4 at about 1.26-fold, which was not significant according to the significance level of ≥ 1.5. Also, the expression of SOX2 and DNMT1 genes has decreased at about 1.28- and 1.32-fold in the treatment group compared to the control group, which were as not significant according to the significance level of  ≥ 1.5.
Conclusion: Overall, it can be concluded that TQ under cobalt (II) chloride-mediated hypoxia may inhibit breast cancer by inhibiting the expression of genes involved in proliferation and migration. In addition, due to the important role of fibroblasts in the wound healing process, TQ may help wound healing under hypoxic conditions by increasing the migration potential of fibroblast cells.

Abstract Nanotechnology is an emerging branch of science that is widely used in various fields including medicine. Nanoparticles (NPs) are produced by various compounds in size, shape, and different chemical properties, which can be used in a variety of biological and biomedical applications. Metal NPs are used widely in many fields and have various properties making them appropriate for use in medical applications.
Zinc is an essential trace element for almost all living organisms. Due to possessing a significant role in versatile biological processes, including fetal development, natural growth, wound healing, metabolism, immunity, cognitive functions, sperm generation, bone mineralization, neurological, and enzymatic processes.
In recent decades, zinc oxide NPs have been one of the most popular types of NPs with numerous biological applications due to their biocompatibility and low toxicity and are used in a wide range of commercial applications including applications in many different industries such as pharmaceuticals, textiles, dyes, rubber, tissue engineering, antibacterial agents, and anti-cancer. Nowadays, significant scientific interest has been directed toward the application of nanomaterials in the modulation of stem cell proliferation and differentiation for further application in regenerative medicine. Zinc is one of the most plentiful trace metals in the human body and was reported to be essential for the regeneration of bone. ZnO-NPs exhibit attractive antibacterial properties. Particular emphasis was given to bactericidal and bacteriostatic mechanisms with a focus on the generation of reactive oxygen species (ROS). ROS has been a major factor for several mechanisms including cell wall damage due to ZnO-localized interaction, enhanced membrane permeability, internalization of NPs due to loss of proton motive force, and uptake of toxic dissolved zinc ions.  ZnO NPs present certain cytotoxicity in cancer cells and induce cancer cell death via the apoptosis signaling pathway. This autophagy induction was positively correlated with the dissolution of ZnO NPs in lysosomes to release zinc ions, and zinc ions released from ZnO NPs were able to damage lysosomes, leading to impaired autophagic flux and mitochondria.
ZnO nanostructures, featuring antimicrobial activity, osteogenesis, and angiogenesis, have been also combined with additive manufacturing technologies with the final aim of designing novel advanced hybrid scaffolds for tissue engineering.
Zinc deficiency is positively correlated with diabetes and may also affect the progress of Type 2 diabetes. Several zinc complexes have been synthesized and proven to be effective in rodent models of diabetes.
ROS are generated by metal oxide nanoparticles which considerably help in fibroblast proliferation. The interlinkage of the fibroblast cells and zinc oxide nanoparticles was impacted by the surface area and particle size of the nanoparticles. Zinc oxide nanoparticles dressing increases apoptosis, bacteria clearance, platelet activation, tissue necrosis, re-epithelialization, tissue scar formation, debris removal, angiogenesis, and stem cell activation through wound healing. Nanoparticle-based drug delivery systems (DDS) can overcome the aforementioned limitations by releasing the drugs in a slow and sustained manner and delivering them to the desired area of the body system. This article provides an overview of some of the research relating to the use of zinc oxide nanoparticles in biological sciences.

Abstract Aim: One of main challenges in worldwide is increasing rate of depression and sexual dysfunction associated with antidepressant drug consumption. Regarding the role of Sertoli cells in spermatogenesis, this study investigated the effect of duloxetine on viability, apoptosis and expression of Bax and Cx43 (Connexin 43) expression in Sertoli cells.
Material and Methods: TM4 Sertoli cells were cultured in DMEM/F12 medium containing 2.5% FBS, 5% horse serum and 1% penicillin-streptomycin. Cells were treated with different doses of duloxetine (3.75, 7.5, 15, 30, 60 µg/ml) for 24 to 72 hours. MTT assay was performed to evaluate cell viability. The rate of apoptosis was measured by flow cytometry and RT-qPCR was performed to evaluate of Bax (proapoptotic gene) and Cx43 (essential for spermatogenesis) genes.
Results: Duloxetine reduced cell survival in a dose and time-dependent manner. On the basis of