Cell Journal (Yakhteh)
Year:2023 | Volume:25 | Issue:3|Papers Count:8

Vitiligo is an auto-immune disease, causing depigmentation of skin in 0. 2-1. 8% of global population. Topical corticosteroids and calcineurin inhibitors are the only treatments with firm evidence of optimal effectiveness with considerable side effects. Phototherapies might not induce serious side effects, although the effectiveness of the method is limited. Advanced therapy medicinal products (ATMPs) are emerging treatment modalities based on correction and replacement of affected genes, damaged tissues or cells in treatment of difficult-to-treat diseases. Due to optimal effectiveness and minimal side effects, ATMPs have recently gained much attention in order to develop new treatments. In this review, the ATMPs for treating vitiligo were along with its clinical success, affordability and cost-effectiveness. Currently, the main ATMP based products using in treatment of vitiligo are non-cultured epidermal cell, melanocytes, and hair follicle melanocytes. These products have shown promising results in the non-responding vitiligo patients. Furthermore, mesenchymal stem cells and multi-lineage differentiating stress enduring cells are other new potential modalities. Recently, Iranian Food and Drug Administration (IR-FDA) authorized the first cell-based product for vitiligo. This product is autologous suspension of keratinocytes and melanocytes. Although ATMPs are efficient and could be cost-effective in long term, the most important obstacle is affordability of them. This could be facilitated by insurance companies and instalments payment programs from manufacturers.

The goal of tissue engineering is to repair and regenerate diseased and damaged tissues and organs with functional and biocompatible materials that mimic native and original tissues which leads to maintaining and improvement of tissue function. Lignin and cellulose are the most abundant polymers in nature and have many applications in industry. Moreover, recently the physicochemical behaviors of lignin and cellulose, including biocompatibility, biodegradability, and mechanical properties, have been used in diverse biological applications ranging from drug delivery to tissue engineering. To assess these aims, this review gives an overview and comprehensive knowledge and highlights the origin and applications of lignin and cellulose-derived scaffolds in different tissue engineering and other biological applications. Finally, the challenges for future development using lignin and cellulose are also included. Plant-based tissue engineering is a promising technology for progressing areas in biomedicine, regenerative medicine, and nanomedicine, with much research focused on the development of newer material scaffolds with individual specific features to make functional and biocompatible tissues and organs for medical applications.

Objective: Stress may have an important role in the origin and progress of depression and can impair metabolic homeostasis. The one-carbon cycle (1-CC) metabolism and amino acid (AA) profile are some of the consequences related to stress. In this study, we investigated the Paroxetine treatment effect on the plasma metabolite alterations induced by forced swim stress-induced depression in mice. Materials and Methods: In this experimental study that was carried out in 2021, thirty male NMRI mice (6-8 weeks age, 30 ±,5 g) were divided into five groups: control, sham, paroxetine treatment only (7 mg/kg BW/day), depression induction, and Paroxetine+depression. Mice were subjected to a forced swim test (FST) to induce depression and then were treated with Paroxetine, for 35 consecutive days. The swimming and immobility times were recorded during the interventions. Then, animals were sacrificed, plasma was prepared and the concentration of 1-CC factors and twenty AAs was measured by spectrophotometry and high-performance liquid chromatography system (HPLC) techniques. Data were analyzed by SPSS, using One-Way ANOVA and Pearson Correlation, and P<0. 05 was considered significant. Results: Plasma concentrations of phenylalanine, glutamate, aspartate, arginine, ornithine, citrulline, threonine, histidine, and alanine were significantly reduced in the depression group in comparison with the control group. The Homocysteine (Hcy) plasma level was increased in the Paroxetine group which can be associated with hyperhomocysteinemia. Moreover, vitamin B12, phenylalanine, glutamate, ornithine, citrulline, and glycine plasma levels were significantly reduced in the depression group after Paroxetine treatment. Conclusion: This study has demonstrated an impairment in the plasma metabolites’,homeostasis in depression and normal conditions after Paroxetine treatment, although, further studies are required.

Objective: Beta-thalassemia is a group of inherited hematologic. The most HBB gene variant among Iranian beta-thalassemia patients is related to two mutations of IVSII-1 (G>A) and IVSI-5 (G>C). Therefore, our aim of this study is to use the knock in capability of CRISPR Cas9 system to investigate the correction of IVSII-1 (G>A) variant in Iran. Materials and Methods: In this experimental study, following bioinformatics studies, the vector containing Puromycin resistant gene (PX459) was cloned individually by designed RNA-guided nucleases (gRNAs), and cloning was confirmed by sequencing. Proliferation of TLS-12 was done. Then, the transfect was set up by the vector with GFP marker (PX458). The PX459 vectors carrying the designed gRNAs together with Single-stranded oligodeoxynucleotides (ssODNs) as healthy DNA pattern were transfected into TLS-12 cells. After taking the single cell clones, molecular evaluations were performed on single clones. Sanger sequencing was then performed to investigate homology directed repair (HDR). Results: The sequencing results confirmed that all three gRNAs were successfully cloned into PX459 vector. In the transfection phase, The TLS-12 containing PX459-gRNA/ssODN was selected. Molecular evaluations showed that the HBB gene was cleaved by the CRISPR/Cas9 system, that indicates that the performance of non-homologous end joining (NHEJ) repair system. Sequencing in some clones cleaved by the T7E1 enzyme showed that HDR was not confirmed in these clones. Conclusion: IVS-II-1 (G> A) mutation, which is the most common thalassemia mutation especially in Iran, the CRISPR/ Cas9 system was able to specifically target the HBB gene sequence. This could even lead to a correction in the mutation and efficiency of the HDR repair system in future research.

Objective: Umbilical cord blood (UCB) is an accessible and effective alternative source for hematopoietic stem cell (HSC) transplantation. Although the clinical application of UCB transplantation has been increased recently, quantitative limitation of HSCs within a single cord blood unit still remains a major hurdle for UCB transplantation. In this study we used microcarrier beads to evaluate the ex vivo expansion of UCB-derived HSCs in co-cultured with UCB-derived mesenchymal stem cells (MSC). Materials and Methods: In this experimental study, we used microcarrier beads to expand UCB-derived MSCs. We investigated the simultaneous co-culture of UCB-derived CD34 + cells and MSCs with microcarrier beads to expand CD34 + cells. The colony forming capacity and stemness-related gene expression on the expanded CD34 + cells were assessed to determine the multipotency and self-renewal of expanded cells. Results: Our results indicated that the microcarrier-based culture significantly increased the total number and viability of UCB-derived MSCs in comparison with the monolayer cultures during seven days. There was a significant increase in the UCB-derived CD34 + cells expanded in the presence of microcarrier beads in this co-culture system. The expanded UCB-derived CD34 + cells had improved clonogenic capacity, as evidenced by higher numbers of total colony counts, granulocyte, erythrocyte, monocyte, megakaryocyte colony forming units (CFU-GEMM), and granulocyte–, monocyte colony forming units (CFU-GM). There were significantly increased expression levels of key regulatory genes (CXCR4, HOXB4, BMI1) during CD34 + cells self-renewal and quiescence in the microcarrier-based co-culture. Conclusion: Our results showed that the increase in the expansion and multipotency of CD34 + cells in the microcarrierbased co-culture can be attributed to the enhanced hematopoietic support of UCB-derived MSCs and improved cell-cell interactions. It seems that this co-culture system could have the potential to expand primitive CD34 + cells.

Objective: We investigated whether co-incubation of 5-FU and gum-based cerium oxide nanoparticles (CeO2 NPs) would improve half-maximal inhibitory concentration (IC50 ) and apoptosis in the Caco-2 cancer cell line Materials and Methods: In this experimental study, we synthesized Ceo-2-XG by the nano perception method. X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), dynamic light scattering (DLS) and vibrating sample magnetometer (VSM) techniques were employed to characterize the synthesized nanoparticles. The Caco-2 cancer cells were cultured and treated with Ceo-2XG and 5-FU. Cytotoxicity analysis was carried out using MTT assay on Caco-2 cancer cells. CXCR1, CXCR2, CXCL8, BAX, BCL-2, P53, CASPASE-3, CASPASE-8 and CASPASE-9 gene expression changes were assessed by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR). The Caco-2 cancer cell mortality mechanism was analyzed using Annexin V-FITC/PI flow cytometry. Using the inverted microscope morphology changes of the Caco-2 cancer cells was observed. Results: With a sample size of roughly 11 nm, TEM analysis revealed spherical structures. Interestingly, after 72 hours, 400 μ, g/ml nanoparticles significantly lowered the IC50 of 5-FU from 101 to 71 μ, g/ml (P<000. 1). Furthermore, qRT-PCR analysis showed that BCL-2, CXCR1, CXCR2 and CXCR8 expressions were significantly decreased in the 5-FU and Ceo-2-XG nanoparticles co-incubated group, compared to the 5-FU alone (P<0. 001). Notably, gene expressions of BAX, P53, CASPASE-3, CASPASE-8 and CASPASE-9 were significantly higher in the 5-FU and Ceo2-XG nanoparticles co-incubated group, compared to the 5-FU alone (P<0. 001). The findings revealed that dead cells owing to apoptosis were more than two times higher in 5-FU and Ceo-2-XG nanoparticles cancer cells than in 5-FU alone treated cancer cells. Conclusion: Co-incubation of 5-FU and Ceo-2-XG nanoparticles significantly increased apoptosis in the Caco-2 cancer cells. The antiproliferative activity of co-incubated 5-FU and Ceo-2-XG nanoparticles on Caco-2 cancer cells was substantially higher than that of 5-FU alone.

Objective: This study aimed to introduce novel techniques for identifying the genes associated with developing chronic obstructive pulmonary disease (COPD) and to prioritize COPD candidate genes using regression methods. Materials and Methods: This is a secondary analysis of the data from an experimental study. We used penalized logistic regressions with three different types of penalties included least absolute shrinkage and selection operator (LASSO), minimax concave penalty (MCP), and smoothly clipped absolute deviation (SCAD). The models were trained using genome-wide expression profiling to define gene networks relevant to the COPD stages. A 10-fold cross-validation scheme was used to evaluate the performance of the methods. In addition, we validate our results by the external validity approach. We reported the sensitivity, specificity, and area under curve (AUC) of the models. Results: There were 21, 22, and 18 significantly associated genes for LASSO, SCAD, and MCP models, respectively. The most statistically conservative method (detecting less significant features) was MCP detected 18 genes that were all detected by the other two approaches. The most appropriate approach was a SCAD penalized logistic regression (AUC= 96. 26, sensitivity= 94. 2, specificity= 86. 96). In this study, we have a common panel of 18 genes in all three models that show a significant positive and negative correlation with COPD, in which RNF130, STX6, PLCB1, CACNA1G, LARP4B, LOC100507634, SLC38A2, and STIM2 showed the odds ratio (OR) more than 1. However, there was a slight difference between penalized methods. Conclusion: Regularization solves the serious dimensionality problem in using this kind of regression. More exploration of how these genes affect the outcome and mechanism is possible more quickly in this manner. The regression-based approaches we present could apply to overcoming this issue.

Vitiligo is an autoimmune skin disorder that can significantly affect the quality of life in patients. However, current treatment strategies such as phototherapy, topical glucocorticosteroids, or systemic immunosuppressants can be helpful in vitiligo management, and treatments for disease stability which is the main challenge in vitiligo management. Novel therapeutic modalities have made promising advances in this regard. Molecular targeted therapy to target Janus-activated kinase (JAK) such as Tofacitinib and Ruxolitinib in addition to the cell-based treatments are innovative therapeutic options, which are recently used for vitiligo treatment. Transplantation of non-cultured melanocytes-keratinocytes have been studied in Iran and phase one data published in 2010 on 10 patients that continued on 300 patients as phase three in 2018. Cell Tech Pharmed™,Co. registered and applied this cell-based product, ReColorCell ®, , as a GMP certified by Iranian Food and Drug Administration (IR-FDA). On 11 th December 2021, ReColorCell ®, officially registered as the first cell-based product in the Iranian drug list (IDL). Currently, the post-market study of this product is ongoing.