Cell and Tissue Journal

Abstract Aim: According to the importance of coumarin derivatives as an effective medication on cancer cells and various other therapeutic effects, in this study we investigated the effect of a new derivative of coumarin named 3- (tetrazol-5-yl) coumarin on single-stranded DNA by different spectroscopic methods in solution.
Material and Methods: The present study has investigated the effect of 3- (tetrazol-5-il) coumarin on single-stranded DNA in vitro. The findings demonstrates that the rate of single strand DNA absorption enhances by interaction with 3-(tetrazol-5-yl) coumarin at 210 and 260 nm. The fluorescence intensity of single-stranded DNA increases in a concentration-dependent of 3- (tetrazol-5-yl) coumarin, indicating the binding of 3- (tetrazol-5-yl) coumarin to the chromophores in single-stranded DNA.
Results: Binding of 3- (tetrazol-5-yl) coumarin to single-stranded DNA causes a significant increase in ellipticity in circular dichroism of DNA molecules in the regions of 220 and 275 nm which is more positive at 245 nm. The results indicate a stronger binding of 3- (tetrazol-5-l) coumarin to single-stranded DNA, which may be due to the fact that single-stranded DNA may be more available during replication.
Conclusion: The results obtained from the effect of 3- (tetrazol-5-yl) coumarin on single-stranded DNA can provide valuable information to design medications by coumarin derivatives which have more anti-tumor effect and less side effects.

Abstract Aim: The aim of this study is to evaluate the interaction of bacterial inoculation and iron treatment (nano and Fe-chelate) on physiological traits of alfalfa.
Material and Methods:  In this study, effects of inoculation with standard Rhizobium meliloti, effects of different levels of iron (Fe-chelate, 0, 5, 10, 20 and 25 μM Fe₂O₃ nanoparticles) and the interaction of bacterial inoculation and iron treatment were investigated on alfalfa in a factorial experiment in completely randomized design with three replications for 45 days. The measured traits were growth indexes, photosynthetic pigments, protein, proline, antioxidants activity, DPPH(diphenyl-picryl-hydrazyl)-radical scavenging activity percent and elements content.
Results: Rhizobium incoculation alone showed beneficial effects on the alfalfa growth and was caused increasing in growth parameters, pigmants, protein content, potassium and phosphours uptake. However inoculation did not effect on the proline and antioxidant content. Iron treatment had a positive effect on the growth parameters, pigmants, protein content and elemant uptake. Highest values of growth parameters was observed 25μM Fe₂O₃ nanoparticles. The highest values of proline and antioxidants activity were measured in control (0μM nanoparticles). Since this concentration is considered a stress for alfalfa. Negative effects of  0μM nanoparticles decreased in inoculated alfalfa plants with R. meliloti. Indeed rhizobium causes increasing in inoculated plant resistant by reducing stressful conditions.
Conclusion: Rhizobium-alfalfa symbiosis plus iron nanofertilizer can cause increasing in plant resistance to stress, in addition to increase growth of plant. The highest amount of growth parameters, pigmants and protein content was measured in inoculated plant  with Rhizobium meliloti and 10μM nanoparticles.
 
 
 

Abstract

Abstract Aim: Cancer remains a global health problem, with ovarian cancer ranking fifth among cancers affecting women and the leading cause of cancer-related death in women. There are different types pf ovarian cancer, including Epithelial ovarian cancer, Stromal tumors and Germ cell tumors.  Epithelial ovarian cancer is the most common type that includes several subtypes, including serous carcinoma and mucinous carcinoma. To this end, several factors have been identified to increase your risk of ovarian cancer, including older age, inherited gene changes, family history of ovarian cancer, being overweight or obese, postmenopausal hormone replacement therapy, endometriosis, and never having been pregnant. The current strategies to treat ovarian cancer include surgery, chemotherapy, radiotherapy and targeted therapies, and hormone therapy. Scientists continue to investigate the foundational mechanisms involving cancer development, as well as to find new drugs and metabolites having the capacity to control cancer. Alpha-ketoglutarate (AKG), a critical metabolite in the Krebs cycle involved in cellular energy production and the regulation of gene expression. Recent studies have shown that AKG may have the potential to enhance the efficacy of cancer treatments, by modulating the tumor microenvironment and improving the immune response against cancer cells. This study investigates the effect of AKG on ovarian cancer cells.
Material and methods: SKOV3 cells were obtained from Tehran University and cultured in complete culture medium (RPMI, 10% Fetal bovine serum (FBS), and 1% Penicillin-Streptomycin (Pen/Strep)). To find the proper concentration of AKG, SKOV3 cells were cultures in 96-well plate, and treated with different concentration of AKG (range 20 to 220 µM). After 24 and 48 h, the viability of the cells was determined using MTT assay. Based on the results obtained from viability assay, 200 μM of AKG was selected for the next assessments. To evaluate the effect of AKG on SKOV3 cell proliferation using plotting a growth curve, cells were cultured in the presence (200 μM AKG) and absence of AKG, and counted the number of cells every 24h for one week. To determine the population doubling time (PDT), the cells were cultured in the presence (200 μM AKG) and absence of AKG for 72h, then the cell were collected and the number of living cells was counted using Neubauer Chamber. The PDT was calculated using a related standard method. To assess colony formation potential, the SKOV3 cell were cultured in the presence (200 μM AKG) and absence of AKG. After 7 days, the cells were fixed using 10% formalin solution, then the colonies were stained using crystal violet dye, and the number of colonies were counted using inverted microscope. To investigate the effect of AKG on the migration rate of SKOV3 cells, the cells were cultured in complete medium to reach 85% confluence, then treated with mitomycin (10 μM) for 3h, then Created a scratch in the cell monolayer using a sterile pipette tip. the cells were cultured in the presence (200 μM AKG) and absence of AKG for 3 days. The images of the scratch were taken at regular intervals using a microscope, and the closure of the scratch over time was analyzed. To analyze the cell cycle profile, the SKOV3 cell were cultured in the presence (200 μM AKG) and absence of AKG for 48h. Then, the cells were collected and analyzed using a flow cytometry.
Results: Based on the MTT assay, 200 μM AKG was determined as the proper concentration to investigate other biological behaviors of SKOV3 cells. Colony forming assay showed a decrease in the number and size of colonies in the AKG-treated group (P<0.05). In addition, the cell doubling time increased in the treatment group, indicating slower growth rate (P<0.05). Growth curve analysis confirmed reduced cell growth in treated group. Cell cycle analysis showed a higher percentage of treated cells arrested in S and G1 phases. The scratch assay showed slow cell migration and metastasis in the cells treated with 200 µM AKG.
Conclusion: In conclusion, AKG has an inhibitory effect on the proliferation, viability, migration in SKOV3 ovarian cancer cells, highlighting its potential as an adjuvant treatment with existing therapies. More research is necessary to fully investigate the therapeutic effect of AKG in ovarian cancer.
 

Abstract Introduction: St. John's Wort, scientifically known as Hypericum perforatum L., belongs to the family Hypericaceae and is an important medicinal plant widely used today for the treatment of depression. It is one of the best-selling herbal products worldwide. Its medicinal properties are attributed to the secondary metabolites present in its extract, including hypericin, hyperforin, flavonoids, xanthones, and other valuable compounds. To meet the growing demands of the pharmaceutical industry and to obtain high-quality biomass, it is cultivated in various countries. However, plants grown under field conditions often face challenges that may affect their phytochemical composition. Plant tissue culture under controlled conditions can mitigate these issues and serve as an attractive alternative to field cultivation. Today, one of the key aspects of biotechnology in Hypericum perforatum is the enhancement of bioactive molecule content using various approaches, including elicitation in tissue culture.
Aim:This research aims to investigate the callogenesis optimization and morpho-physiological, phytochemical, and biochemical changes in the callus of Hypericum perforatum L. in response to chitosan elicitor under In Vitro culture conditions.
Materials and Methods: After obtaining Hypericum perforatum seeds, the effect of sterilization methods on seed contamination percentage was evaluated using a completely randomized design. Subsequently, a factorial experiment in a completely randomized design was conducted to optimize callus induction using hormonal treatments (E₁ to E₂), explants (E₁, E₂), and different light conditions (L₁, L₂). After preparing the chitosan elicitor at concentrations of 0 (control), 25, 50, and 75 mg, another experiment was carried out in a completely randomized design to apply the elicitor to the seeds. Finally, the effect of the chitosan elicitor on improving morphological, physiological, metabolic, and biochemical traits was examined.
Results: The results of the mean comparison for the effect of sterilization methods showed that treatment C (no sterilization) and S₂ exhibited the lowest and highest levels of contamination percentage, respectively. The mean comparison results for the interaction effect of hormonal treatment explant type, and light conditions indicated that the best hormonal combination for callus induction was H₁E₁L₁. The findings revealed a significant increase in traits such as fresh weight, dry weight, callus volume, flavonoids, phenols, antioxidant activity, anthocyanins, carotenoids, catalase, peroxidase, proline, and chlorophyll a and b at all chitosan concentrations compared to the control. The highest and lowest values for these traits were observed in the 50 mg/L chitosan treatment and the control, respectively. For malondialdehyde content, the highest and lowest levels were associated with the control treatment and the 50 mg/L chitosan treatment, respectively.
Conclusion: Overall, the results of this study demonstrated that the chitosan elicitor improved the growth, physiological, metabolic, and biochemical characteristics of Hypericum perforatum. Among all the concentrations used, the 50 mg/L concentration was the most effective in enhancing these traits.
 

Abstract Aim: Cancer is a global concern and colorectal cancer (CRC( accounts for the second most common cause of cancer related death in the world. Nanotechnology could enhance the effectiveness of chemotherapy as a common therapeutic approach through development of smart nanoparticles (NPs). In this context, theranostic nanoparticles with both imaging and therapeutic potentials are considered as promising platforms in diagnosis and treatment of advanced cancers.
Materials & Methods: Here, we designed and synthesized magnetic mesoporous silica nanoparticles (SPION-MSNs) in which release of 5-fluorouracil (5-FU) at physiological conditions was inhibited with pH-responsive gold gatekeepers. Heterofunctional polyethylene glycol (PEG) polymer was then conjugated onto the outer surface of nanoparticles and non-targeted nanoparticles were successfully synthesized. In order to achieve active and specific targeting, non-targeted nanoparticles were armed with an epithelial cell adhesion molecule (EpCAM) aptamer (Apt) for selective drug delivery of 5-FU to colorectal cancer cells. Finally, the physicochemical properties of NPs including functional groups, surface charge and their size were fully characterized with Fourier transform infrared (FTIR) spectra and dynamic light scattering (DLS) in each step. Moreover, morphology and homogeneity of SPION-MSNs were evaluated using field emission scanning electron microscopy (FESEM), high-resolution transmission electron microscopy (HR-TEM) and atomic force microscopy (AFM). The cumulative release of 5-FU from nanoparticles was compared in buffer solutions with two different pH values (pH 7.4 and 5.4). In the final step, anti-cancer potential and cytotoxicity of free 5-FU, non-targeted, and targeted nanoparticles were assessed on human colorectal adenocarcinoma HT-29 cells and Chinese hamster ovary cells.
Results: Core-shell NPs, SPION-MSNs, were successfully prepared and the FTIR spectra showed specific peaks at the surface of nanoparticles. Obtained results from DLS measurements showed that the synthesized formulation had negative charges with size of 20 nm. Moreover, the morphology of SPION-MSNs indicated spherical shape with uniform distribution. After introducing amine groups, the surface charge was shift to positive and the two bands at 2965 and 1,560 cm⁻¹ in the FT-IR spectrum were appeared and assigned to CH₂-CH₂ and N-H groups, respectively. The results indicated, 5-FU was encapsulated in the open pores of MSNs and the encapsulation efficiency (EE%) and drug loading capacity (LC%) were about 98% and 49%, respectively. The release of 5-FU from NPs showed pH-dependent manner, with an initial rapid release (within 6 h) followed by a sustained release for 96 h at pH 5.4. interestingly, the cumulative release of 5-FU was about 3.9% in neutral medium over 96 h. the results supported the Intelligent release of cargoes from theranostic nanoparticles. At the final step, targeted nanoparticles were successfully synthesized with a final size diameter of 78 nm and negative surface charge. In vitro results demonstrated higher cytotoxicity and anti-cancer property of targeted nanoparticles against EpCAM-positive HT-29 cells as compared to the EpCAM-negative CHO cells, confirming the effectiveness of aptamer as a targeting ligand.
Conclusion: These findings suggest that application of the targeted formulation can be considered as a promising theranostic platform for EpCAM-positive CRC cells. However, further experiments are required before it can be practiced in the clinic.

Abstract Introduction: Cancer is one of the most significant diseases of the present century and ranks as the second leading cause of death worldwide. Its incidence, particularly breast cancer in women, is steadily increasing. This disease arises from uncontrolled cell proliferation and the ability of cancer cells to migrate to healthy tissues, leading to tumor formation and metastasis. The genetic and biological heterogeneity of cancer cells, especially in breast cancer, complicates effective treatment. Conventional therapies such as surgery, chemotherapy, and radiotherapy, although effective in reducing tumor size, are associated with limitations, including damage to healthy cells, drug resistance, and systemic side effects. Therefore, the development of targeted and innovative therapeutic strategies is a priority in cancer research. One promising approach is the use of nanoparticles for targeted drug delivery. Due to their small size, high surface-to-volume ratio, and modifiable surfaces, nanoparticles can efficiently carry drugs and deliver them to target cells. Controlled drug release from nanoparticles reduces uptake by healthy cells and minimizes systemic toxicity. Iron oxide nanoparticles (Fe₃O₄) possess unique magnetic and chemical properties that enable precise guidance to tumor sites and allow real-time monitoring of drug distribution via MRI. Surface modification with biocompatible polymers such as polyethylene glycol (PEG) enhances nanoparticle stability, prolongs systemic circulation, reduces immune clearance, and provides sites for conjugating targeting ligands such as folic acid and glucose. Polylactic acid (PLA), a biodegradable and biocompatible polymer, increases drug-loading capacity and enables sustained and controlled drug release. Combining PLA with PEG and targeting ligands creates multifunctional nanoparticles that are stable, biocompatible, and capable of selectively recognizing cancer cells while minimizing side effects on healthy tissues. Targeting ligands such as folic acid and glucose facilitate selective cellular uptake; folic acid binds to overexpressed receptors on many cancer cells, promoting intracellular drug delivery, while glucose exploits the high metabolic demand of cancer cells, improving the delivery of therapeutic agents or genes.
Aims: Dual-ligand nanoparticles enable multi-pathway targeting, enhancing therapeutic efficacy and potentially overcoming drug resistance. In this study, multifunctional PLA-PEG nanoparticles functionalized with folic acid and glucose and incorporating Fe₃O₄ were developed for targeted drug delivery to triple-negative breast cancer cells (Hs-578T). This system combines magnetic guidance, biocompatibility, controlled drug release, and selective targeting, providing a promising platform for effective and safe breast cancer treatment with reduced side effects compared to conventional therapies.
Materials and Methods: Various chemicals and reagents, including MTT ((3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide)), Ethanol, HCl, NaCl, KH₂PO₄, Chloroform, NH2-PEG-FA, NH2-PEG-Glu, NaOH, and paraformaldehyde were procured from commercial suppliers, while FBS, DMEM, and trypsin-EDTA were used for in vitro studies with Hs-578T breast cancer cells. Multifunctional nanoparticles PPF (PLA-PEG-FA), PPG (PLA-PEG-Glu), and PPGF (PLA-PEG-Glu/FA) were synthesized via the reaction of PLA-acrylate with the respective NH2-PEG derivatives in chloroform under mechanical stirring and mild heating, followed by purification through dialysis and freeze-drying. Methanolic extract of Silybum marianum was prepared by ultrasonication of powdered aerial parts in 80% methanol, followed by centrifugation and filtration. Fe₃O₄ nanoparticles were synthesized via co-precipitation of Fe (III) and Fe (II) salts under nitrogen, followed by gradual addition of Silybum marianum plant extract and NaOH, leading to nanoparticle formation. The particles were collected magnetically, washed, freeze-dried, and stored in dark, dry conditions. Targeted delivery of Curcumin (CUR) was achieved using folic acid and glucose ligands. CUR and Fe₃O₄-OA were incorporated into PPF, PPG, or PPGF matrices using sonication and emulsification in PVA solutions, followed by rotary evaporation to remove chloroform, washing, and filtration to remove unencapsulated drug and large particles. Structural and chemical composition were analyzed using 1H-NMR and FTIR spectroscopy. Morphology was observed via TEM, and average size and zeta potential were measured using dynamic light scattering (DLS). Drug encapsulation efficiency was determined spectrophotometrically by measuring unencapsulated CUR in the supernatant. CUR release kinetics were evaluated in two pH environments: pH=7.4 (normal cells), and pH=4.5 (tumor-like), with periodic sampling and replacement with fresh medium over three days. Cytotoxicity was assessed via the MTT assay, with IC₅₀ values determined after treating Hs-578T cells with varying concentrations of CUR, nanoparticles with CUR, or blank nanoparticles. Apoptosis was analyzed using Annexin V/propidium iodide staining and flow cytometry to distinguish apoptotic and necrotic cells. All experiments were performed in triplicate. Data were analyzed using one-way ANOVA followed by Duncan’s test at a 5% significance level, and normality was verified with the Kolmogorov–Smirnov test. Results are reported as mean ± standard deviation.
Results: Targeted drug delivery has emerged as a promising strategy for treating difficult-to-treat diseases, including cancer, by directing therapeutic agents specifically to diseased tissues while minimizing uptake by normal cells. Biocompatible and biodegradable polymers are essential for efficient delivery systems. PLA, a widely used biodegradable polymer, allows loading and controlled release of hydrophobic drugs like curcumin. However, PLA alone can aggregate in serum due to hydrophobic interactions, leading to immune recognition and clearance. To overcome this, PLA was conjugated with PEG, which enhances nanoparticle circulation time and provides a stealth effect, reducing recognition by the immune system. PEG also improves drug loading and stability, although it is not biodegradable and may accumulate over repeated doses. Magnetic Fe₃O₄ nanoparticles were incorporated to increase targeting efficiency, while folic acid and glucose ligands were used for selective recognition of cancer cells. The synthesized PLA-based nanoparticles demonstrated good biocompatibility with Hs-578T cells, showing no significant cytotoxicity. Drug encapsulation within PLA-PEG nanoparticles allowed controlled and sustained release of curcumin, which was particularly enhanced under acidic conditions mimicking the tumor microenvironment. Notably, nanoparticles containing both folic acid and PEG showed higher drug release compared to those with PEG alone, highlighting the role of folic acid in facilitating curcumin release. This pH-sensitive release profile minimizes drug exposure to normal tissues, potentially reducing side effects. Cytotoxicity assay revealed that curcumin-loaded nanoparticles significantly inhibited Hs-578T cell proliferation compared to free curcumin, with the dual-ligand nanoparticles exhibiting the lowest IC₅₀ values. Flow cytometry analysis demonstrated that these nanoparticles primarily induced apoptosis rather than necrosis, suggesting selective activation of programmed cell death pathways. The enhanced apoptotic effect is likely due to increased cellular uptake and targeted delivery, which facilitates higher intracellular concentrations of curcumin.
Discussion: Overall, the results indicate that PLA-PEG-based nanoparticles, functionalized with magnetic Fe₃O₄ and surface ligands, provide an effective platform for targeted curcumin delivery. These nanoparticles combine biocompatibility, controlled release, tumor-specific accumulation, and apoptosis induction, which collectively enhance therapeutic efficacy while minimizing systemic toxicity. The study underscores the potential of multifunctional nanocarriers as a promising approach for improving the effectiveness of anticancer therapies and offers a foundation for the further development of targeted, stimuli-responsive drug delivery systems.
Conclusion: As a final conclusion of this study, it can be stated that the development of novel drug delivery systems based on PLA-PEG copolymer nanoparticles is a promising and strategic step toward overcoming the therapeutic challenges of intractable diseases such as cancer. This study clearly demonstrated that the design of such nanocarriers not only offers high biocompatibility and safety for normal cells, but also, by taking advantage of the camouflage effect caused by PEG, increases blood circulation time and prevents premature elimination by the immune system. The outstanding feature of this system is the ability to control and intelligently release the drug in response to specific stimuli of the tumor microenvironment, especially higher acidity, which allows for maximum drug delivery to the target tissue and, subsequently, a significant reduction in systemic side effects. In this study, the effect of adding targeting ligands such as folic acid and glucose on the accuracy and efficiency of these nanoparticles in recognizing and binding to cancer cells was also clearly observed. In addition, the results of this study showed that the designed nanoparticles were successfully able to increase the induction of cell apoptosis in Hs-578T cancer cells. These findings strongly support the superiority of this nanotechnology compared to conventional drug formulations, both in increasing therapeutic efficacy and reducing toxicity. Overall, it can be concluded that PLA-PEG-based smart nanoparticles, with their multimodal capabilities, have great potential to become a comprehensive and reliable approach in the next generation of targeted cancer therapies, paving the way for further studies and ultimately effective clinical applications.

Abstract Aim: In the present study, the effects of hydroalcoholic extract of Berberis integrrima root on the induction of differentiation in mesenchymal stem cells derived from adipose tissue of male Wistar rats towards osteoblasts were investigated.
Material and Methods: In this study, stem cells derived from male Wistar rat tissue were isolated and flow cytometry was performed to confirm the surface markers. Stem cells were treated with 10, 20, 40 and 80 μg/ml hydroalcoholic extract of B. integrrima root. The cell toxicity of the extract was evaluated by MTT assay and its differentiated effects was evaluated by alizarin red staining and alkaline phosphatase activity, calcium deposition assay. Results were analyzed using one-way ANOVA at a significance level of P <0.05.
Results: Stem cells markers including CD105, CD44, and CD73 had positive, and CD45 and CD34 had negative expression in these cells. The results of the MTT assay showed Concentrations less than 20 µg/ml do not have significant toxic effects on cells. Higher alkaline phosphatase activity was observed in the treatment group compared to the control group on day 10. The results of Alizarin red staining and measurement of calcium content for 21days showed that this extract in a concentration dependent manner leads to the differentiation of stem cells into osteoblasts.
Conclusion: The findings of this study showed that mesenchymal stem cells derived from adipose tissue of male Wistar rats treated with hydroalcoholic extract of Berberis integrrima root enter osteoblasts in the differentiation pathway.

Abstract

Abstract Aim: The aim of the current work was to investigate the anticancer activities of Zinc Oxide nanoparticles (ZnONPs) through modulation of Glutation peroxidase and Glutationreductase gene expression in breast cancer cells.
Material and Methods: In this investigation, the breast cancer MCF-7 and normal HEK293 cell lines were treated with various concentrations of ZnONPs for overnight. Cell viability was measured using MTT(3-(4, 5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay against cancer and normalcells. The quantity of Glutation peroxidase and Glutationreductase genes compared to GAPDH gene expressions were evaluated using the real time PCR method.
Results:The MTT data showed that ZnONPs significantly decreased the viability of cancer cells compared to normal cells in dose-dependent mode. Moreover, the mRNA levels of Glutation peroxidase and Glutation reductase genes was significantly increased by 2.13±0.07 (p < 0.001) and 1.22±0.05 (p < 0.05) fold, respectively, following treatment withZnONPs.
Conclusion: According to the results of this investigation, the Glutation peroxidase and Glutation reductase gene expression was the key factors of glutathione system in elimination of increasing reactive oxygen species treated with ZnONPs.
 

Abstract Aim: The ovary is one of the tissues which progesterone receptor is expressed in it. The aim of this study was to investigate the effects of antiprostrogen and estrogen on the ovaries and fallopian tubes of ovulated stimulated mice.
Material and Methods: In this study, mice were divided into four groups after stimulation of ovulation and pregnancy. 1) Control 2) Estrogen, 3) Progesterone, 4) Anti-progesterone and estrogen. 4.5 days after pregnancy, mice were killed by cervical vertebral disslocation and the ovaries and fallopian tubes were prepared by H&E and PAS staining for histomorphometrical changes.
Results: The results showed that there were different types of follicles and corpus luteum in the control group. In the progesterone group, the number of corpus luteum increased compared to the control group and in the estrogen group, The corpus luteum decreased and the majority of follicles were primitive or growing. The amount of atretic follicles was higher in the anti-progesterone with estrogen group. The results obtained from the fallopian tubes indicate that progesterone injection reduces the height of the luminal epithelium and the number of ciliated cells. In the estrogen-containing groups, an increase in ciliated cells and epithelial height were observed.
Conclusion: The results of this study showed that progesterone alone could not provide better conditions for fertilization and adding estrogen to progesterone may improve this condition.

Abstract Aim: The aim of this research was to investigate the effects of different plant growth regulators and MS culture medium on the characteristics of callus weight, percentage of callus formation and embryogenesis in anther culture of two cultivars of Sina and Negin cucumbers (Cucumis sativus L.).
Materials and methods: The experiment was conducted as a factorial in a completely randomized design with three replications. In this research, to determine the stage of growth and development of microspores, male flower buds in the sizes of 0.5 to 2.2 cm were taken from mother plants. For dyeing, the anthers were separated from the buds and placed in stocarman dye solution containing one gram of carmen powder and one hundred milliliters of 45% acetic acid for one day. Then the anthers were crushed on the laboratory slide and the microspores inside them were observed under a light microscope. In the callus induction test, the factors include two cucumber cultivars (Sina and Negin), the combination of different concentrations of BAP (0, 0.7, 0.9 and 1.1 mg/liter) and 2,4-D (0, 0.75, 1 and 1.25 mg/liter). 4 anthers were cultured in each petri dish. After placing the anthers in the culture medium, the lids of the petri dishes were closed and sealed with parafilm, and then they were covered with aluminum foil and placed in the growth chamber at a temperature of 25 ± 2 degrees Celsius in the dark for 2 weeks and the environments They were cultivated once every 2 or 3 weeks.
Results: Cytogenetic experiments showed that 1.5 cm buds containing anthers and microspores in the mononuclear stage are suitable for anther culture. The results of the analysis of the variance of the data showed a statistically significant difference at the level of 0.01 between the interaction effects of genotype and culture medium. Anthers cultured in MS base medium and media prepared with different concentrations of BAP did not show a response and were destroyed after one month of cultivation. The average main effect of cultivars on callus fresh weight after several weeks of cultivation showed that the highest callus fresh weight was obtained in Sina variety. The comparison of the average triple interaction effects of cultivar × BAP × 2,4-D for callus fresh weight in this research has shown that Sina cultivar in MS culture medium contains the growth regulator compound 0.7 mg/liter BAP and 1 mg/liter 2,4 -D has the highest callus fresh weight. By examining the interaction effect of cultivar, auxin and cytokinin, it was found that the highest percentage of callus formation was 100% in both cultivars in environments with the combination of BAP (0.7 and 0.9 mg/liter) with 2,4-D (0.75, 1 and 1.25 mg/L) and in Sina cultivar BAP (1.1 mg/L) with 2,4-D (0.75, 1.1 and 1.25 mg/L). However, the lowest percentage of callus formation was observed in Nagin cultivar in the presence of 2,4-D hormone with a concentration of 1 mg/liter with an average of 33.333%.
Conclusion: In general, the use of growth regulator BAP and 2,4-D in MS culture medium induced callus in anther culture of both cucumber cultivars.

Abstract Aim: Nanoparticles are superior to conventional elemental forms due to the novel physicochemical properties that enable them to act as plant growth promoters. Recent research on nanomaterials has been shown both positive and negative effects. Titanium dioxide nanoparticles (TiO₂ NPs) are assayed worldwide in large quantities for various purposes. This study investigated the effect of TiO₂ NPs on some physiological and biochemical traits and the induction of the antioxidant systems in Origanum vulgare L. and Origanum majorana L., in a completely randomized design with three repetitions.
Material and Methods: Surface-sterilized seeds were sown with 2 cm depth in each plastic pots filled with a 1: 1 mix of loamy soil and sand. Until reaching the appropriate physiological age, they were exposed to greenhouse conditions (16 h light/8 h dark photoperiod). Two-month-old plants were foliar sprayed with 10, 60, and 120 mg/L TiO₂ NPs and harvested 14 days after the last treatment. Plants sprayed with distilled water were used as controls. Changes in the photosynthetic pigments (the content of chlorophyll a, chlorophyll b, total chlorophyll, and carotenoid), biochemical properties (the levels of membrane stability index, malondialdehyde, and proline contents), and the activity of antioxidant enzymes (Guaiacol peroxidase, ascorbate peroxidase, polyphenol oxidase, superoxide dismutase, catalase, and glutathione s-transferase) were measured in the collected samples.
Results: The comparison of data means showed that the treatment of 60 mg/L TiO₂ NPs caused the maximum amount of chlorophyll a, total chlorophyll, and carotenoids in both Origanum species. However, the highest content of chlorophyll b in O. majorana was observed in the treatment of 120 mg/L TiO₂ NPs. It is worth mentioning that the negative effect of the concentration of 120 mg/L TiO₂ NPs on the membrane system of two species is statistically significant and remarkable due to the leakage of electrolytes. Although the increase in the concentration of TiO₂ NPs led to fewer levels of malondialdehyde in O. vulgare in comparison with O. majorana; this concentration was found to be effective to the high levels of proline in O. vulgare. Most of the antioxidant enzymes (Guaiacol peroxidase, ascorbate peroxidase, polyphenol oxidase, superoxide dismutase, and glutathione s-transferase) of O. vulgare showed their maximum activity at the concentration of 60 mg/L TiO₂ NPs. However, the lower activity of catalase in O. vulgare, compared to O. majorana was remarkable.
Conclusion: The results of this research show that the treatment of 60 mg/L TiO₂ NPs improves the physiological characteristics of the medicinal plant Origanum, including the increase of photosynthetic pigments, followed by the rate of photosynthesis and the increase of biomass. It is noteworthy that higher levels than the optimal concentrations of TiO₂ NPs can lead to an increase in the levels of ROS and oxidative burst, which leads to a decrease in plant performance. Therefore, the plant response to nanoparticles depends significantly on the concentration and time of application, as well as the size, shape, and surface functionalization of the particles. Finally, O. vulgare is introduced as a more successful species due to the higher activity of antioxidant enzymes when treating TiO₂ NPs.

Abstract Introduction: Oxidative stress exerts destructive effects on sperm cells, leading to sperm dysfunction and male infertility. It occurs when there is an imbalance between the production of reactive oxygen species (ROS) and the cell's antioxidant defense system. ROS are free radicals and peroxides that can damage cells. They can harm the sperm cell membrane, which is rich in polyunsaturated fatty acids, through lipid peroxidation, impairing sperm membrane fluidity. This affects motility and the sperm's ability to fuse with the egg. Additionally, ROS can directly damage sperm DNA, leading to DNA fragmentation and other abnormalities. Oxidative stress can also impair sperm motility by disrupting energy production in the mitochondria. Severe oxidative stress may result in sperm cell death (apoptosis or necrosis), reducing sperm viability. These negative effects ultimately decrease the chances of successful fertilization and a healthy pregnancy.
Cadmium, a pervasive environmental and industrial pollutant, induces significant toxic effects on human health, particularly on the male reproductive system. It is both a naturally occurring element and a widespread environmental pollutant generated from various industrial and agricultural activities. Its presence in soil, water, air, and food exert risks on human health. Exposure to cadmium is strongly linked to increased oxidative stress, which damages cellular components such as lipids, proteins, and DNA within sperm cells. This oxidative damage can lead to impairments in critical sperm parameters, including motility, viability, morphology, and DNA integrity, thereby compromising fertilization potential and contributing to male infertility. The severity of these effects underscores the need for effective strategies to mitigate cadmium-induced sperm damage.
Alpha-lipoic acid (ALA), a naturally occurring organosulfur compound, is produced in the body, obtained through dietary sources, and taken as a supplement or medication. ALA is a potent antioxidant with the unique ability to function in both aqueous and lipid environments. This versatility allows ALA to scavenge a wide range of free radicals and ROS, thereby protecting cells from oxidative damage. Furthermore, ALA can regenerate other endogenous antioxidants, such as glutathione, further amplifying its protective effects. Given ALA’s well-documented antioxidant properties and its potential to mitigate oxidative stress, this study aims to investigate its protective effects on human sperm exposed to cadmium.
Aim: This research will explore the extent to which ALA can counteract the detrimental effects of cadmium-induced oxidative stress on vital sperm parameters. By evaluating the impact of ALA supplementation on sperm motility, viability, morphology, and DNA integrity in the presence of cadmium, this study seeks to determine the potential therapeutic role of ALA in preserving sperm function and improving male fertility outcomes. The results of this investigation will provide valuable insights into the efficacy of ALA as a protective agent against cadmium-induced reproductive toxicity and contribute to the development of strategies for mitigating the harmful effects of environmental pollutants on male reproductive health.
Materials and Methods: In this experimental study, human sperm samples were divided into five groups: 1) spermatozoa at 0 hours, 2) spermatozoa at 180 minutes (control group), 3) spermatozoa treated with cadmium chloride (10 μM) for 180 minutes, 4) spermatozoa treated with ALA (50 μM) + cadmium chloride (10 μM) for 180 minutes and 5) spermatozoa treated with ALA (50 μM) for 180 minutes.
Vital sperm parameters, including motility, viability, plasma membrane and acrosome integrity, mitochondrial membrane potential, and DNA fragmentation, as well as oxidative stress indices (total antioxidant capacity and lipid peroxidation), were examined in different groups. Data were expressed as mean ± standard deviation and analyzed using one-way analysis of variance (ANOVA). Means with p

Abstract

Abstract Aim: The aim of this study was to investigate the effects of inoculation of two species of mycorrhizal fungi on growth, colonization percentage, antioxidant enzymes activity, health and risk index of coriander under cadmium stress.
Material and methods: A pot experiment was conducted in a completely randomized design with 2 factors and 3 replications in the research greenhouse of the Department of Horticultural Science and Landscape Engineering at the Ferdowsi University of Mashhad in 2018. The first factor was cadmium nitrate at 4 levels of 0, 20, 40, and 80 mg kg^-1 soil and the second factor was mycorrhiza application at 3 levels of non-inoculation, inoculation with Glomus mosseae and Glomus intraradices.
Results: The results showed that with increasing cadmium concentration in the soil, the shoot and total biomass of plant, leaf area, number of seeds, 1000 seeds weight, colonization percentage, soluble protein, catalase, ascorbate peroxidase, guaiacol peroxidase, and polyphenol oxidase activities were significantly decreased in coriander. But malondialdehyde content, health and risk index increased. However, the use of mycorrhizal fungi reduced the harmful effects of cadmium in the plant. This resulted in a decrease of 47.1% of the risk and health index and 17.2% of malondialdehyde in the plant.
Conclusion: According to the findings, use of mycorrhizal fungi had a significant effect on the improving of harmful effects of cadmium in coriander, thus improved the risk and health index for consumers. Therefore, using mycorrhizal fungi as a management strategy in polluted areas with this heavy metal is 

Abstract Aim: The aim of this study was to investigate the effect of Zinc oxide nanoparticles on growth and physiological characteristics, rosmarinic acid production and the expression of key genes in the biosynthetic pathway of this compound in lemongrass.
Material and Methods: In this study, 30-day-old seedlings of lemongrass were treated with concentrations of 0, 0.06 and 0.12 mg/l Zinc oxide nanoparticles, and then growth parameters, photosynthetic pigments, proline, Glycine betaine, protein, activity of antioxidant enzymes and production of rosmarinic acid were examined. Also, the expression of key genes in the rosmarinic acid biosynthetic pathway was examined by real-time PCR.
Results: The results indicate that Zinc oxide nanoparticles treatment increased chlorophyll photosynthetic pigments, carotenoids and antioxidant activity of this plant. Also, the highest amount of proline and glycine betaine was obtained at a concentration of 12 mg/L of this treatment. Zinc oxide nanoparticles also increased the expression of rosmarinic acid biosynthetic pathway genes (TAT and 4-Cl) and thus increased this compound.
Conclusion: Based on the results of this study, Zinc oxide nanoparticles can affect the growth and physiological stages of lemongrass and therefore it can be used to increase the production of rosmarinic acid.

Abstract Aim: The use of algae as a food additive has many positive effects such as increasing the quality, health, and greed of food and increasing the level of growth and immunity in aquatic animals. Among the algae with many uses are the members of the red algae branch, which are seen in red due to the large amounts of phycoerythrin and phycocyanin pigments. These algae belong to primary multicellular plants of the marine species, non-flowering without roots, stems, and leaves, and they reproduce sexually. The current research was conducted to replace red algae in the diet in stomach, liver histomorphology, and some growth factors of Nile tilapia, one of the most important commercial species in the world.
Material and Methods: For this purpose, after 8 weeks of preparation, fish were fed diets containing zero, 3%, 6%, and 9% of food weight with red algae with three repetitions in each treatment. After the end of the rearing period, the fishes were anesthetized and after opening the abdominal area while measuring some growth factors, a 0.5 cm tissue sample was taken from the middle part of the stomach and liver and placed in a 10% formalin buffer solution. After going through the routine steps of tissue preparation, including dehydration with alcohol, clarification with xylol, blocking with paraffin, cutting with a thickness of 4-6 microns, and staining, finally, the study was done with a light microscope. SPSS software version 21 was used for statistical analysis of data.
Results: Microscopic and anatomical studies of the stomach and some growth factors of the digestive system showed an increase in the measured factors as percentage of weight gain, especially in the group receiving the diet containing 9% red algae. The results of microscopic studies on the stomach showed that the mucosal lining of the stomach is of a simple type with nuclei near the base. This tissue is stretched from the surface of the mucous membrane to the inner part, so that at the beginning of the stomach, short but deep folds were also observed at different intervals. In the lamina propria of gastric mucosa, simple tubular glands were observed, whose tissue consists of a row of short cubic to cylindrical cells, which have spherical to elongated nuclei and basophils attached to the base of the cells. These cells had strong acidophilic color and covered all the lamina propria. Also liver was completely lobulated and the pancreas was located in the form of an appendix gland inside.
Conclusion: The results of this research showed that in terms of the structural characteristics of the stomach and liver tissue such as mucous glands, epithelium thickness, thickness, mucus height, and anatomical growth of the digestive system, red seaweed powder can be a suitable substitute for fish meal in the diet. Tilapia, along with possible substitutes in other animal and vegetable proteins, can be used to feed this fish, which will eventually reduce the cost and reduce the dependence on fish powder to supply the protein needed by Nile tilapia.

Abstract Introduction: Plant-derived exosome-like nanoparticles (PDENs) are nano-sized vesicles released by plant cells. They contain a lipid bilayer membrane and carry bioactive molecules such as proteins, lipids, nucleic acids, and secondary metabolites. Due to their low toxicity and natural drug delivery potential, they have gained attention for therapeutic applications. Plant extracts are also rich in phenolic compounds and flavonoids with known antioxidant and antimicrobial effects. The rising problem of antibiotic resistance in pathogens like Escherichia coli, Staphylococcus aureus, and Streptococcus pyogenes has created an urgent need for new natural agents.
Aim: This study aimed to isolate and characterize exosomes from ginger, lavender, onion, and lemon, and to compare their antioxidant and antibacterial activities with those of aqueous extracts from the same plants.
Materials and methods: Fresh ginger, lavender, onion, and lemon were used for the preparation of plant extracts and exosome isolation. For aqueous extraction, 2 g of ginger and lavender samples were homogenized with 20 mL of distilled water and incubated in a water bath at 70°C for 90 minutes. The mixtures were centrifuged at 3000 × g for 10 minutes, and the supernatants were filtered and stored at 4°C until analysis. Exosomes were isolated using the Exosun Exosome Isolation Kit (EXOSUN Company). Plant materials were homogenized, filtered, and subjected to differential centrifugation. The resulting supernatants were processed according to the manufacturer's instructions using buffers A and B. Due to the acidic nature of lemon juice, modification of the protocol was required by increasing the concentration of buffer A to facilitate exosome precipitation. Protein concentration of isolated exosomes was determined using the Bradford assay with bovine serum albumin (BSA) as the standard. Exosome morphology was characterized by transmission electron microscopy (TEM). The antioxidant activity of extracts and exosomes was evaluated using the Ferric Reducing Antioxidant Power (FRAP) assay. Samples were analyzed in 96-well microplates, and absorbance was measured at 570 nm. Antibacterial activity was assessed against Escherichia coli, Staphylococcus aureus, and Streptococcus pyogenes using the disk diffusion method. Sterile blank disks were impregnated with plant extracts or exosome preparations at concentrations of 25, 50, and 100 mg/mL and placed on Mueller-Hinton agar plates inoculated with bacterial suspensions adjusted to a 0.5 McFarland standard. Gentamicin and vancomycin served as positive controls. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were determined using the broth microdilution method in 96-well microplates. All experiments were performed in triplicate. Statistical analyses were conducted using SPSS version 27, applying one-way ANOVA followed by Tukey’s post hoc test, with p <0.05 considered statistically significant.
Results: TEM confirmed successful isolation of exosome-like vesicles from all plants. Bradford assay indicated protein-containing nanoparticles. FRAP analysis showed that lavender had the highest antioxidant capacity. Plant extracts generally showed greater antioxidant activity than their corresponding exosomes. Antibacterial testing revealed that exosomes from all four plants had no detectable antibacterial effect. Similarly, aqueous extracts of ginger, lavender, and onion showed no inhibition. However, lemon extract exhibited significant antibacterial activity against all three bacteria. MIC values of lemon extract were 3.12 mg/mL for S. aureus, 6.25 mg/mL for S. pyogenes, and 50 mg/mL for E. coli. Inhibition zones for S. aureus reached 22 mm at 100 mg/mL. Gram-positive bacteria were more susceptible than Gram-negative E. coli.
Discussion:The spread of antibiotic-resistant pathogens has increased the need for new treatments. In this study, exosomes isolated from ginger, lavender, onion, and lemon showed no antibacterial activity, while among aqueous extracts, only lemon extract displayed good antibacterial properties. The lack of activity in aqueous lavender extract compared to alcoholic extracts in previous reports may be due to solvent differences affecting phenolic compound extraction. Aqueous ginger extract also showed no effect, which contrasts with some alcoholic extracts. However, lavender exosomes demonstrated the highest antioxidant capacity among all samples, highlighting their potential for antioxidant applications.
Conclusion: Although plant-derived exosomes from these four species did not show antibacterial effects under the tested conditions, lemon aqueous extract exhibited promising antimicrobial activity, especially against Gram-positive bacteria. Lavender exosomes showed the strongest antioxidant potential. These findings suggest that lemon extract could serve as a natural antibacterial agent, while lavender-derived exosomes and extracts may be valuable sources of natural antioxidants. Further in vivo studies are needed to explore their efficacy and safety.

Abstract Aim: Tissue engineering refers to methods that are based on the use of scaffolds, cells and biologically active molecules to produce tissues with specific functions. The purpose of tissue engineering is to build structures that can regenerate, maintain and improve damaged tissue or the whole organ. Today, by using tissue engineering methods, various natural and synthetic scaffolds have been designed that can be used for nerve grafts. The physical, chemical and biological properties of the scaffold must be similar to the extracellular matrix of the body in order to avoid adhesion, growth and support the differentiation of cells. An ideal neural scaffold should have biodegradability, biocompatibility and proper tensile strength. Recently, the use of polycaprolactan as a suitable biodegradable material has been evaluated in many fields of tissue engineering. Antioxidants are among the substances, which seem to be able to prevent neuronal death by reducing the amount of ROS. Flavonoids include many compounds that have various biological effects in the body. Silymarin (Silybum marianum) is a flavonoid that has many effects, including anti-cancer effects and antioxidant properties. Tragacanth is a known natural polymer that has excellent biological properties such as biodegradability, biocompatibility, antibacterial and wound healing ability. It is obtained from the stems and branches of the Asian species tragacanth. It has outstanding structural stability against heat and acidity. The aim of this study is to produce polycaprolactan/ tragacanth /silymarin nanoscaffolds and to investigate the viability of pc12 cells on the scaffold under oxidative stress. Considering that silymarin has antioxidant properties, the use of polycaprolactan/ tragacanth /silymarin nanoscaffolds can prevent neuropathy of nerve cells.
Material and Methods: Scaffolds used in this research were prepared using the electrophoretic method. For this purpose, an electrospinning machine was used, which is equipped with a rotary collector with a thickness of 70 mm and a width of 50 mm. In order to prepare a polycaprolactan/ tragacanth nanoscaffold and load silymarin on it, a 7% polycaprolactan solution (dissolved in acetic acid), 0.7% by weight tragacanth solution (dissolved in acetic acid) and 0.9% by weight silymarin solution were mixed by a magnetic stirrer for 20 minutes, and in order to make the solution uniform, sodium didecyl sulfate (SDS) with a concentration 1 percent by weight of the solvent was added to the solution and the suspension was homogenized for 20 minutes with an ultrasonic device, then the scaffold was prepared by an electrospinning device. . The nanofibers were collected in a period of 6 hours, the sample collection speed was 1 ml per hour, and the nanofiber samples were collected by rotating at 250 rpm. The distance between the injection needle and the scaffold is 12 cm and this process is done at a voltage of 15 kV. The morphology of the scaffold was evaluated by scanning electron microscope (SEM) and the chemical structure of the scaffold was evaluated by FTIR spectroscopy. To investigate the antioxidant properties of the scaffold, glucose 80 mg/L and H2O2, 150 macro L were used.
Results: Examining the morphology and chemical structure of the scaffold showed the proper porosity of the polycaprolactan/ tragacanth scaffold and the successful loading of silymarin on the scaffold. Evaluation of the oxidant properties of the scaffold after 24 hours of PC12 cell culture on it showed the increase in cell viability on the scaffold and the appropriate antioxidant properties of the scaffold.
Conclusion: The results of this research showed that the enrichment of polycaprolactan/ tragacanth scaffold with silymarin increased the proliferation and survival of PC12 cells under oxidative stress. Therefore, this scaffold can be a suitable candidate for tissue engineering in oxidative stress.

Abstract Introduction: Downstream processes are crucial in pharmaceutical protein manufacturing, bridging drug synthesis and final product formulation. They involve purification strategies to isolate active pharmaceutical ingredients (APIs) from natural or cellular sources, ensuring high purity and potency. Advances in technology and purification methods have significantly impacted the efficiency and quality of pharmaceutical products, particularly for biological drugs like human follicle-stimulating hormone (hFSH) and VHHs antibodies
Aims: To highlight the importance of downstream processes in pharmaceutical protein production, focusing on the purification techniques employed for proteins like human follicle-stimulating hormone and VHHs antibodies, and to demonstrate how technological innovations improve yield, efficiency, and product quality.
Materials and methods: The study emphasizes the use of strategic purification approaches, including advanced chromatographic methods such as recombinant DNA technology for producing hFSH and affinity chromatography utilizing VHHs antibodies. These methods are tailored to preserve the structural and functional integrity of target proteins and enhance purification efficiency.
Results: Innovative purification strategies, especially affinity chromatography with VHHs antibodies, have led to a significant increase in yield (~60%), reduced processing time, and improved purity. These techniques effectively streamline production, reduce costs, and maintain the biological activity of target proteins like hFSH.
Discussion: The integration of cutting-edge technologies—recombinant DNA, affinity chromatography, and sophisticated purification strategies—has transformed downstream processing. These advancements support the shift from natural sourcing to recombinant production, improving efficiency, product quality, and cost-effectiveness, and fostering progress in biopharmaceutical manufacturing.
Conclusion: Refinement of downstream processes, driven by technological innovation, advances the production of vital biological drugs such as hFSH. These developments symbolize a move toward higher standards of quality and efficiency in pharmaceutical manufacturing, showcasing human ingenuity and the ongoing pursuit of excellence in healthcare and biotechnology

Abstract Intoduction: Cancer as one of the most common genetic diseases is the leading cause of death worldwide. Cancer cells undergo various genetic and phenotypic changes to spread and survive. In the early stages, these changes lead to the development of tumor, while at the advanced stages they can provide a suitable pre-metastatic microenvironment in which various uncontrolled events occur including cell proliferation, traversing through the extracellular matrix, and crossing barriers to enter the bloodstream. Extracellular changes in this microenvironment can induce intracellular changes in primary cancer cells that assist in the sustainability and propagation of these cells. Complicated interactions between the external and internal factors result in the establishment of various regulatory networks between different types of carcinogenesis promoting factors. Identification of these modifications plays a critical role in understanding the mechanisms of disease progression, prognosis and management. Text: Various mutations and differential gene expression trigger metastasis of cancer cells by epithelial to mesenchymal transition (EMT) mechanism, among which the role of chromatin structural changes, intracellular signal transduction pathways, regulation of cell cycle and microRNAs, and genomic instability has been reported. The alterations in gene expression patterns of mentioned pathways lead to potential regulatory complications that faced the management of disease progression and response to therapies with problems. Cancer cells provide their requirements by neutralizing biological barriers, modifying the regulation of inhibiting processes of cancer progression, establishing de novo endogenous mechanisms and providing specialized molecular and structural markers, and various combinations of these methods have been demonstrated in different types of cancer. Furthermore, EMT and cancer stem cells (CSCs) have a mutual relationship in which the presence of one assists the occurrence of the other. Altogether,  cancer cells take the advantage of  multiple approaches including  upregulation of main transcription factors such as snail, slug, Foxc2, Twist and ZEB1/2, benefiting the mechanisms of telomere length protection, production of CD133,CD44 and BMI1 biomarkers, mutation in P53 coding gene, Failure in acquiring aging phenotype, mutation in amino acid residue S115 of SIM2S gene and increased genomic instability, enhanced activity of signaling pathways such as NF-κB, TGF-β, Wnt, Notch and Hh, mitotic rounding process, facilitated cell division, epigenetic changes such as acetylation and methylation of histones and dysregulation of miRNAs. Conclusion: EMT plays a crucial role in cancer progression, crossing the cells through the biological and body barriers, and metastasis that are usually associated with poor prognosis of cancer patients. Molecular and cellular changes in the main pathways of cells^, development are considered as the promoting factors of EMT and resulted from the differential expression of genes in EMT compared to the normal phenotype of cells. Advancement in the exploration of these changes and their role in the progression of cancer can remarkably affect the early diagnosis, treatment and management of disease. Aim: In this review, various molecular and cellular mechanisms involved in EMT progression and cancer have been investigated, including signal transduction pathways, structural changes of chromatin and telomeres, up/down-regulation of small non-coding RNAs such as miRNAs, cell cycle regulation and genomic instability.

Abstract Aim:: The renin-angiotensin system (RAS) is an endocrine system essential for regulating blood pressure and fluid balance. Components of this system are expressed not only in various body cells but also in breast cancer cells. Angiotensin-converting enzyme (ACE) inhibitors, which are widely used as antihypertensive agents, have been proposed to have an additional benefit in reducing the risk of certain cancers. Doxorubicin is a commonly used chemotherapy drug, also employed in the treatment of breast cancer. One of the side effects of doxorubicin is its toxicity to cardiac cells. This research aims to investigate the effect of cytotoxicity of four RAS antagonists (enalapril, captopril, valsartan, and losartan) in the presence and absence of doxorubicin against two breast cancer cell lines including MCF-7 and MDA-MB-231. The study’s objective is to determine whether these RAS inhibitors can modulate the cytotoxicity of doxorubicin and their potential impact on breast cancer cell survival.Material and methods:  MTT is one of the colorimetric methods to investigate cell survival. This assay is based on the conversion of the MTT reagent to a formazan product by metabolically active cells, providing a quantitative measure of cell viability. The effect of RAS inhibitors in the presence and absence of doxorubicin on the survival of cancer cells was measured using the MTT test. Chemotherapy drugs, such as doxorubicin, typically reduce cell survival by inducing apoptosis. One of the markers of apoptosis induction in cells is the evaluation of caspase-3/7 activity. The induction of apoptosis in MDA-MB-231 cells treated with the RAS inhibitors was investigated using a caspase-3/7 activity assay kit, which quantifies the enzymatic activity in cell lysates. Results: Enalapril had no significant cytotoxic effect on breast cancer cells at concentrations up to 500 μg/mL. Captopril and losartan were toxic to breast cancer cells. Valsartan had no cytotoxic effect on breast cancer cells and increased cell proliferation over time. The cytotoxic effects of captopril and losartan on breast cancer cells may be attributed to their inhibitory effects on the RAS. The RAS plays a crucial role in regulating blood pressure and fluid balance, but it also has inhibitory effects on tumor growth and progression. By blocking the activity of the RAS with captopril and losartan, the cells may experience reduced proliferation and ultimately cytotoxicity. Enalapril, captopril, valsartan, and losartan protected breast cancer cells from doxorubicin cytotoxicity in a concentration- and time-dependent manner. Captopril and losartan treatment increased caspase-7/3 activity in MDA-MB-231 cells. The increase in caspase-7/3 activity in MDA-MB-231 cells suggests that captopril and losartan-induced cytotoxicity may be accompanied by apoptosis. Apoptosis is a desirable outcome in cancer treatment as it allows for the elimination of cancer cells while preserving normal tissue function.
Conclusion: RAS inhibitors can interfere with the cytotoxic effects of doxorubicin on breast cancer cells, depending on the concentration and duration of exposure. Further research is required to understand the optimal use and dosage of RAS system inhibitors in this context to ensure a balance between protecting the heart from doxorubicin-induced cardiotoxicity and maintaining the efficacy of cancer treatment.
 

Abstract

Abstract Aim: The aim of this study was to compare the anti-proliferative and apoptotic effects of hydroalcoholic extracts of different herbal medicines (Achillea wilhelmsii, Silybum marianumseed, Echinacea purpurea, Adiantum capillus-venerisand apricot kernel) against breast cancer cells (Mcf-7).
Material and method: For this purpose, the plants were dried and milled, then, soaked in 70% ethanol for 72 hours and their extracts were extracted using a rotary evaporator.Different concentrations of herbal extracts (12.5, 25, 50 and 100 μg/ml) were added to the cancer cell culture medium and their cytotoxicity and apoptotic effects were investigated after 24h by MTT assay and acridine orange - ethidium bromide staining, respectively.Data was analyzed by SPSS software at the significant level of 5%.
Results: Addition of the highestconcentration of all extracts to the culture mediumshowed the most significant anti-proliferative and apoptotic effects (p < 0.05) compared to other concentrations of the same extracts.Also, among the high concentrations (100μg/ml), the highest cell cytotoxicity effects were related to the extracts of Echinacea purpureaand Adiantum capillus-veneris (p < 0.05).
Conclusion: The results of this study indicated that the addition of Echinacea purpurea and Adiantum capillus-venerisextractsto the cell culturesin high concentrationshad the most significant anti proliferative and apoptotic effects on breast cancer cells in comparison with other plant extractsand concentrations.