JOURNAL OF CELL & TISSUE
Year:2024 | Volume:15 | Issue:4|Papers Count:6

Aim: Potato (Solanum tuberosum L.) is a crop belongs to the Solanaceae family. It is the fourth strategic crops following wheat, rice, and corn. Salicylic acid or orthohydroxybenzoic acid, act as a regulator that can regulate physiological, biochemical and signaling functions in plant metabolic processes. Aim: Salicylic acid as a growth regulator can interact with auxin and gibberellin hormones and effectively induce growth and metabolic processes in potato plants. Potato is one of the most important agricultural plants and in vitro culture can be used as a model in laboratory conditions. The aim of this study is to investigate the effect of salicylic acid on growth indicators and its interaction with other hormones.Material and method: In this study, potato seedlings with uniform growth were grown on MS culture medium with concentrations of zero, 0.1, 10 and 100 μM salicylic acid. The seedlings were placed in the growth room under light conditions of 16 hours of light and 8 hours of darkness at room temperature. After four weeks, the grown seedlings were used for wet weight, dry weight, shoot and root length, photosynthetic pigments, auxin and gibberellin hormones content were measured. The statistical analysis was performed using SPSS software.Results: According to the data, growth parameters in the samples treated with salicylic acid showed similar patter. Our results showed that relative water capacity, root and shoot length and pigments under 0.1, 1 and 10 μM significantly increased compared to control samples. While at the concentration of 100 μM, a decreasing trend was observed in the growth parameters and showed a significant decrease compared to the control samples. In this study, the maximum values ​​in all growth parameters were obtained under the concentration of 10 μM salicylic acid. Furthermore, based on the results, the concentration of 0.1 μM salicylic acid had no significant difference on gibberellin hormone content. While the gibberellin hormone content under salicylic acid treatment at concentrations of 1 and 10 μM significantly increased compared to the control sample, and the maximum level was observed at 10 μM. The concentration of 100 μM, unlike the other concentrations, showed a significant difference compared to the control sample with a decreasing trend. Also the results in auxin levels showed a similar pattern with growth parameters and gibberellin hormone. The auxin levels showed a significant increase under the concentrations of 1, 10 and 100 μM compared to the control sample, and the maximum levels were observed at the concentration of 10 μM. However, concentrations of 1 and 10 μM did not show significant differences. Similar to the results of gibberellin hormone, auxin hormone showed a significant difference under the concentration of 100 μM with a decreasing trend compared to the control sample. Conclusion: It seems that salicylic acid increased in low concentrations due to the direct effects on the metabolic and enzymatic processes of growth parameters, photosynthetic pigments, and auxin and gibberellin content. On the other hand, at high concentrations, the growth parameters were inhibited due to the high accumulation of reactive oxygen species produced by salicylic acid and the resulting oxidative stress.

Aim: Turmeric contains a high amount of curcumin (Cur), which exhibits strong antioxidant activity. Cur binds to antioxidant response elements for gene, thus suppressing reactive oxygen species (ROS). Reactive oxygen species are short-lived, highly electrophilic molecules. When intracellular antioxidants are reduced or ROS accumulate excessively, it brings about the imbalance in the redox state which leading to oxidative stress. DEHP consists of a pair of eight-carbon esters linked to a benzene-dicarboxylic acid ring with chemical formula of C24H38O4. DEHP is used as a plasticizer in many polyvinyl chloride products, especially in medical devices such as intravenous bags and tubing, umbilical artery catheters, blood bags, infusion tubing, enteral nutrition feeding bags, nasogastric tubes and dialysis bags. Due to non-covalent binding to the plastic products, DEHP leaches into biological fluids during medical treatment causing contamination. There is ample evidence that DEHP induces oxidative stress and causes apoptosis in cells. Therefore, the study aimed to investigate the effects of Cur on DEHP-induced oxidative stress in bone marrow mesenchymal stem cells (BMSCs), as DEHP has been found to induce oxidative stress in BMSCs. Materialand Methods: Rat BMSCs was extracted and after the 3rd passage, their cell viability and proliferation were examined in the presence of various concentrations (0.05,0.1, 0.25, 1, 2.5 and 5 μM) of Cur for a period of 4 days. Subsequently, a concentration of 0.1 μM Cur was chosen for further assessment of viability and proliferation abilities (measured by population doubling number (PDN)), total protein, malondialdehyde (MDA), total antioxidant capacity (TAC), catalase and superoxide dismutase (SOD) activity as well as the expression of NFkB and Nrf2 both individually and in conjunction with 100 or 500 μM of DEHP (chosen based on previous study) for durations of 4 and8 days. The data underwent statistical analysis with P<.05 considered as the minimum level of significance. Results: Concentrations of 0.05, 0.1, and 0.25 μM of Cur were found to have no effect on the viability (P<0.05) of the BMSCs but significantly increased proliferation ability (P<0.0001). Conversely, concentrations of 1, 2.5, and 5 μM of Cur significantly reduced viability (P<0.0001). The concentration of 0.1 μM was chosen for further analysis based on viability tests and PDN analysis. This selected concentration notably increased BMSCs proliferation after 4 days, while higher concentrations (1, 2.5, and 5 μM) reduced both viability and proliferation. Furthermore, this concentration counteracted the effects of DEHP at both day 4 and day 8 by reducing MDA levels and increasing TAC levels as well as catalase and SOD activity - particularly at day 8 compared to the DEHP-treated groups. Additionally, Cur was found to reduce NFkB expression while increasing Nrf2 expression in BMSCs.Conclusion: Curcumin mitigated oxidative stress induced by DEHP through the Nrf/NFkB pathway. Moreover, it enhanced the cell viability and proliferation capacity of BMSCs in the presence of DEHP. Therefore, patients undergoing hemodialysis and blood transfusion, who may be exposed to DEHP from medical devices like tubes and other plastic products, should consider consuming turmeric to counteract the oxidative impact of this chemical.

Aim: Breast cancer remains a significant health challenge, being one of the most common cancers among women and a leading cause of cancer-related deaths globally. The need for effective treatment options is paramount, and recent research has spotlighted sulfonamide derivatives for their notable antitumor properties. These compounds have shown considerable cytotoxic effects against various cancer cell lines, including breast cancer, through mechanisms such as apoptosis induction and inhibition of heat shock protein 70 (HSP70), which is crucial for cancer cell survival. This study focuses on synthesizing a solid lipid nanocarrier that incorporates a sulfonamide derivative, aiming to enhance its anticancer efficacy against the MCF-7 breast cancer cell line. The encapsulation of sulfonamide within these nanoparticles is expected to improve its bioavailability by increasing solubility and stability, thereby allowing higher concentrations to effectively reach tumor sites. Additionally, these nanoparticles are designed to minimize systemic toxicity and protect healthy tissues from adverse effects, resulting in enhanced therapeutic efficacy.methods: The research involved synthesizing solid lipid nanoparticles (SLNs) containing the sulfonamide derivative, followed by a comprehensive analysis of their physicochemical properties. Key parameters assessed included drug loading capacity, morphology, size distribution, and release profile using techniques such as Fourier Transform Infrared Spectroscopy (FTIR), Dynamic Light Scattering (DLS), Atomic Force Microscopy (AFM), and dialysis methods. The cytotoxicity of the nanocarrier was evaluated on MCF-7 cells through the MTT assay, alongside assessments of cell proliferation and migration using scratch assays.Results: The synthesized nanocarrier had an average size of 315 nanometers with a Polydispersity Index (PDI) of 0.3, indicating good uniformity. AFM analysis confirmed its spherical morphology. The nanocarrier demonstrated a high drug loading capacity of approximately 82% for the sulfonamide derivative and facilitated a controlled release over time. Cytotoxicity assays revealed that while the blank nanocarrier was safe, the formulation containing the sulfonamide derivative exhibited significantly enhanced cytotoxic effects on MCF-7 cells at lower concentrations compared to the free sulfonamide alone. This improvement in efficacy is attributed to the controlled release mechanism that allows for sustained exposure of cancer cells to the active compound. Moreover, the nanocarrier was more effective in inhibiting both growth and migration of cancer cells than treatments with free sulfonamide, as evidenced by scratch assays.Conclusion: This research highlights the potential of solid lipid nanoparticles in improving the therapeutic effectiveness of sulfonamide derivatives against breast cancer. The synthesized nanocarrier demonstrates favorable physicochemical properties, including high drug loading capacity and controlled release, which are essential for effective treatment outcomes. Notably, encapsulating sulfonamide derivatives significantly enhances cytotoxicity against MCF-7 cells at lower doses compared to their unencapsulated counterparts, emphasizing its promise for improved therapeutic strategies. Furthermore, these nanoparticles effectively inhibit cell proliferation and migration, contributing to tumor growth suppression. This innovative approach aims not only to enhance treatment efficacy but also to minimize systemic toxicity and protect healthy tissues from adverse effects. Future research should focus on innovative clinical applications and further optimization of these nanocarriers to maximize their potential in oncology, paving the way for more effective treatments for breast cancer.

Aim: The stinging nettle (Urtica dioica L.) is a very important medicinal plant that is used in the treatment of many diseases. Considering the important medicinal properties (flavonoid compounds and antioxidant enzymes) of nettle and the numerous advantages of using elicitors in plant tissue culture to enhance the production of medicinal compounds, this study investigated the effect of different concentrations of the chitosan elicitor on the production of some secondary metabolites and antioxidant enzyme activities in callus suspension culture of nettle.Material and methods: Nettle seeds were cultured in 1/2MS medium after surface sterilization (with 1% sodium hypochlorite and 70% ethanol). leaf explants were isolated from sterile seedling (at the four-leaf stage). The explants were cultured in MS hormone-containing medium (NAA and BA 2.5 and 1 mg L-1, respectively) and maintained under light conditions of 16 : 8 (light : darkness) hours  at 25 °C. After several stages of subculturing, uniform callus was produced. Eight days after transfer of calli to MS liquid culture medium, calli were treated with chitosan elicitor at concentrations of 0, 50, and 100 mg L-1. Calli were sampled at 24, 48, and 120 hours after treatment. Finally, the production of secondary metabolites (quercetin, kaempferol, and rutin) was determined using HPLC and the standard line equation. Also, the activities of antioxidant enzymes (phenylalanine ammonia-lyase, polyphenol oxidase, and peroxidase) and the total content of phenol, flavonoid and protein were measured and analyzed by spectrophotometry method. The experiment was conducted in completely randomized design with three replications and was analyzed as a split-time statistical design.Results: According to the results of the analysis of variance, the effect of chitosan elicitor concentration, sampling time after elicitor application and the interaction effect of elicitor concentration and sampling time on all measured parameters except for the total flavonoid content (the effect of elicitor concentration was not significant) were significant. The highest phenylalanine ammonia-lyase and polyphenol oxidase enzyme activities were at sampling times of 120 and 24 hours after elicitor application at 100 mg L-1 chitosan concentration, respectively. The peroxidase enzyme activity at the sampling time of 120 hours and a concentration of 50 mg L-1 of chitosan was 2.57 times that of the control sample. The total protein content decreased at concentrations of 50 and 100 mg L-1 chitosan compared with the control. The highest amount of quercetin production was in the treatment of 100 mg L-1 chitosan and sampling time of 48 hours after elicitor application, which increased 25 times compared with the control. Kaempferol and rutin had the highest production at sampling times of 48 and 24 hours and a concentration of 50 mg L-1, respectively, which increased by 72.6 and 51.2 percent compared with the control.Conclusion: Chitosan elicitor is known as a biotic elicitor with a positive effect on medicinal and antioxidant properties in various plants. In this study, chitosan elicitor increased the production of quercetin, kaempferol, and rutin metabolites as well as the activity of antioxidant enzymes in nettle callus culture. So, chitosan elicitor can be introduced as an enhancer of medicinal properties of nettle plant under in vitro culture conditions. Obviously, for the commercial production of medicinal compounds, the conditions for large-scale callus production in bioreactors must be optimized.

Aim: Oregano (Mentha longifolia) is an important medicinal plant from the mint family, which is widely used in food and pharmaceutical industries due to its active compounds. As an efficient approach, the use of biological elicitors (yeast, bacterial, and fungal agents) as well as non-biological elicitors (inactive enzymes, ultraviolet rays, heavy metal salts, polysaccharides, salicylic acid, jasmonic acid, and melatonin) can provide a way to increase the production of secondary metabolites in plant cell cultures. Given the importance of Oregano in the pharmaceutical and therapeutic industries, this study was carried out to assess melatonin's effect on the percentage of cell viability and the level of important bioactive compounds of oregano.  Materials and methods: This study was performed in a factorial experiment with a completely randomized design and three replications in the lab. Melatonin (0, 100, and 200 μM based on the previous studies) was added to the MS culture medium containing 2,4-D (1 mg.L-1). Then, 24, 48, and 72 h after treatment, the percentage of cell survival and the level of important oregano compounds were measured by tetrazolium and HPLC tests, respectively. In SAS software version 9.4, analysis of variance (ANOVA) was used to analyze the data and the LSD method to compare the means.Results: Based on the results of the tetrazolium test, there was no significant difference in the percentage of cell viability in various concentrations of melatonin. However, the survival percentage of cells decreased with the passage of time, so that the lowest survival percentage was observed 72 h after melatonin treatment. Adding melatonin to the culture medium increased the production of secondary metabolites. At the concentration of 100 µM, the amount of menthol, menthone, pulegone, and 1,8-cineole increased by 77, 108, 46, and 80%, respectively, after 48 hours in contrast to the control. The increase in the level of menthol, menthone, pulegone, and 1,8-cineole compounds in the concentration of 200 µM of melatonin was respectively 125, 130, 140, and 120%, after 48 hours when compared to the control. Therefore, increasing melatonin concentration from 100 to 200 µM in cell culture significantly increased the level of compounds compared to the control. The levels of menthol, menthone, pulegone, and 1,8-cineole increased up to 48 hours after treatment, but after that time, they showed a significant decrease. Since the ultimate goal is to increase the production of biologically active compounds and not the highest cell viability, therefore, 48 hours after applying the treatment of 200 μM melatonin provides the best conditions for obtaining a high level of valuable oregano metabolites. The above treatment increased menthol, menthone, pulegone, and 1,8-cineole by 125, 130, 140, and 120%, respectively.Conclusion: In this study, for the first time, the effect of melatonin elicitor on important secondary metabolites of the Oregano plant was investigated to determine the potential of using this metabolic stimulant in research and commercial areas. Overall, The use of melatonin at a concentration of 200 μM along with the extraction of metabolites 48 hours after the treatment can enhance menthol, menthone, pulegone, and 1,8-cineol, while maintaining the viability of oregano cells

Aims: Salvia L. with more than 1000 taxa is the largest genus in Lamiaceae family, that naturally grows in diverse regions of the world. The genus is represented in Iran by about 60 species, which 17 of them are endemic for the country. Salvia macrosiphon Boiss., is an annual aromatic herb of this genus, which is widely distributed in diverse parts of Iran. Due to the presence of a wide range of secondary metabolites (especially essential oil), this plant has been widely applied in the traditional medicine to cure different diseases. Various types of glandular and non-glandular trichomes have been detected in Lamiaceae taxa.  Essential oils are composed of several compounds, and are biosynthesized  and stored in the glandular trichomes. Additionally, the non-glandular trichomes play the prominent roles in growth and development of plants.  This study was aimed to detect different types of trichomes on the leaf epidermal surface of S. macrosiphon, and their variations among different Iranian populations. Material and methods: Eight natural populations of this plant were harvested from diverse habitats in Iran. Plant samples were identified, according to the morphological descriptions are available in the  valuable references. Three flowering plants were selected from each population and one mature and intact leaf was obtained per individual. The leaves were fixed in fixative (F.A.A) solution for 48 h. The hand-made cuttings of leaf's blade were double-stained with methylene blue and carmine colors. Then, the thin slices of each population were examined using a light microscopy (Olympus CH2, Japan) at different magnifications. Results: the leaf epidermal surfaces were covered by a dense indumentum, which were composed of the glandular and non-glandular trichomes. The non-glandular trichomes had the simple and unbranched structures with one to five linear-arranged cells. The non-glandular trichomes had two cell types: long and short. However, the more frequent non-glandular trichomes were the long two to four-celled types. Meanwhile, in  Amir kabir population,  the five-celled non-glandular trichomes also had a high density. The glandular trichomes were detected as capitate, digitate, and peltate types. Two types of capiate trichomes were observed on the leaves surfaces: short-stalked and long-stalked capitate. The main difference between these types relates to the cell number of trichomes stalk. However, the short-stalked capitate trichomes were the dominant glandular type in all the populations, except for Arak and Mashhad populations, which had the peltate trichome as a more frequent form.  Conclusion: the capitate and peltate trichomes do not have the same ability to maintain the biosynthesized essential oil in their cellular structures. The stored essential oil in the capitate trichomes seeps out through the micropores in their apical cells. Since, the short-stalked capitate was the dominant form in most populations, the secretion of essential oil makes this species very fragrant. But, these plants are less fragrant in populations that have a higher number of peltate trichomes. The non-glandular hairs play a key role in protecting the plant from herbivore insects and the ultraviolet rays of the sun light. Moreover, they protect the leaf epidermal surface from extreme heat and cold. The type and density of other glandular and non-glandular trichomes widely differed among the populations which explored their adaptive importance in this species.