Cell Journal (Yakhteh)

158-164

Authors: Reza Chavooshi; Mohammad Reza Ranjkesh; Behnam Hashemi; Leila Roshangar

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.

176-183

Authors: Nazli Servatian; Saeid Abroun; Seyed Abolhassan Shahzadeh Fazeli; Masoud Soleimani

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.

194-202

Authors: Zahra Keramati; Gholamreza Motalleb; Abbas Rahdar; Mohammad Amin Kerachian

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-2- XG 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 μg/ml nanoparticles significantly lowered the IC50 of 5-FU from 101 to 71 μg/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 Ceo- 2-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.

212-214

Authors: Mohammad Amin Shahrbaf; Leila Ataei Fashtami; Massoud Vosough

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 11th 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.

85-91

Authors: Sepideh Shahkarami; Samareh Younesian; Shahrbano Rostami; Farzad Kompani; Davood Bashash; Seyed Asadollah Mousavi; Seyed H. Ghaffari

Objective: Minimal residual disease (MRD) is considered the greatest prognostic factor in acute lymphoblastic leukemia (ALL). MRD is a valuable tool for anticipating impending relapse and treatment response assessment. The objective of the present study was to investigate whether the detection of IgH gene rearrangement using polymerase chain reaction (PCR)-based GeneScan analysis could be a complementary method to monitor MRD along with the quantitative realtime PCR (qPCR). Materials and Methods: In this cross-sectional study, we valued the MRD levels, based on the GeneScanning analysis (GSA), and then compared the data with quantitative real-time polymerase chain reaction at different time points in peripheral blood (PB) samples of adult B-lineage ALL patients (n=35). The specific polymerase chain reaction (PCR) primers for IGH gene FR-1 and fluorescence-labeled J-primer were used and analyzed by capillary gel electrophoresis on a sequencer. The results of this study were compared with the previously reported MRD results obtained by the IGH rearrangements allele-specific oligonucleotide (ASO) -qPCR methods. Results: The total concordance rate was 86.7%, with a P<0.001. MRD results obtained by GSA and ASO-qPCR methods were concordant in all diagnostic samples and samples on the 14th and 28th days of induction therapy. The results of these 2.5 years’ follow-ups demonstrated a significant correlation between the two techniques (r=0.892, P<0.001). Conclusion: It seems that the PCR-based GeneScan analysis of IGH gene rearrangement detection may be a valuable molecular technique to distinguish monoclonality from polyclonality. And also, it may be a precise tool to detect the residual leukemic DNA in the PB follow-up samples of patients.

102-109

Authors: Behdokht Jamali; Malihe Entezari; Nahid Babaei; Mehrdad Hashemi

Objective: Parkinson’s disease (PD) is a severely debilitating disease for which no suitable treatment has been found so far. In recent years, nanoparticles (NPs) have shown therapeutic potential in PD. Thus, in the current research, for the first time, we investigated the effects of vitamin E and TiO2 nanoparticles (TiO2-NPs) on a rat model of PD. Materials and Methods: In this experimental study, after preparation and induction of PD, rats were administrated with vitamin E and TiO2-NPs. One day after receiving the last dose of treatments, rats were killed and substantia nigra was extracted from the brain and its cell suspension was prepared. In each group, female rats were mated, and after confirmation that the female rats were pregnant by vaginal smear test, the fetus was removed. Cell viability was studied by the MTT method and apoptosis, and necrosis were studied by the flow cytometry technique. Also, after RNA extraction and cDNA synthesis, the expression of Bcl-2 and circRNA 0001518 genes were studied using the real time polymerase chain reaction (RT-PCR) technique. For analyzing the data, two-way ANOVA was used. Results: The results showed a sharp decrease in cell viability in female PD rats and fetuses resulting from PD female rats. Vitamin E treatment showed the greatest effect on increasing cell viability. Decreased expression of the Bcl-2 gene and increased expression of circRNA 0001518 were observed in PD conditions. Conclusion: Administration of vitamin E may be a good option for reducing PD-induced cell death.

118-125

Authors: Junhao Fu; Min Yu; Wenxia Xu; Shian Yu

Objective: Chemotherapeutic drug resistance is the main obstacle that affects the efficacy of current therapies of hepatocellular carcinoma (HCC), which needs to be addressed urgently. High expression of histone methyltransferase G9a was reported to play a pivotal role in the progression of HCC. Regulatory mechanism of aberrant activation of G9a in HCC and the association with subsequent cisplatin (DDP) resistance still remains ambiguous. This study strived to investigate mechanism of G9a overexpression and its impact on cisplatin resistance in HCC cells. Materials and Methods: In this experimental study, we investigated effects of different concentrations of cisplatin in combination with BIX-01294 or PR-619 on viability and apoptosis of HuH7 and SNU387 cells via CCK-8 kit and flow cytometric analysis, respectively. Colony formation capacity was applied to evaluate effect of cisplatin with or without BIX-01294 on cell proliferation, and western blotting was used to verify expression level of the related proteins. Global mRNA expression profile analysis was adopted to identify differentially expressed genes associated with overexpression of G9a. Results: We observed that overexpression of G9a admittedly promoted cisplatin resistance in HCC cells. Global mRNA expression profile analysis after G9a inhibition showed that DNA repair and cell cycle progression were downregulated. Moreover, we identified that deubiquitination enzymes (DUBs) stabilized high expression of G9a in HCC through deubiquitination. Additionally, cisplatin could significantly inhibit proliferation of DUBs-deficient HCC cells, while promoting their apoptosis. Conclusion: Collectively, our data indicated that DUBs stabilize G9a through deubiquitination, thereby participating in the cisplatin resistance of HCC cells. The elucidation of this mechanism contributes to propose a potential alternative intervention strategy for the treatment of HCC patients harboring high G9a levels.

135-142

Authors: Somayeh Karimi Majd; Mandana Gholami; Behzad Bazgir

Objective: Satellite cells play an important role in muscle regeneration, which this process can be affected by different genes including PAX7 and MyoD. Exercise training known as an important strategy for mediating the satellite cell’s function. Therefore, the main purpose of the present study is to investigate the changes in PAX7 and MyoD protein expression in response to eccentric and concentric resistance exercise in healthy young men. Materials and Methods: In this semi-experimental and cross-sectional study, 10 healthy men (age range 18-30 years old) participated. They were randomly divided into two equal groups (n=5) to perform one of two high-intensity eccentric or concentric knee extensions muscle contraction protocols. The contractions included a maximum of 12 sets of 10 repetitions, with a 30 second rest time interval between sets. PAX7 and MyoD protein expression was assessed using Immunohistochemistry analysis from the Vastus Lateralis muscle needle biopsy samples that have been taken 24 hours before and 3 to 4 hours after the end of the exercise protocol. Results: We observed that the PAX7 protein expression level increased significantly after eccentric (47.75%) and concentric (39.21%) (P=0.01) intervention. While, the MyoD protein expression level reduced (38.14%) significantly following acute eccentric resistance exercise (P=0.01). Conclusion: It seems that eccentric or concentric muscular contraction modulates the expression of PAX7 and MyoD protein expression in the skeletal muscle, with further effects observed in eccentric resistance exercise.

822-828

Authors: Mansoureh Sabzalizadeh; Mohammad Reza Afarinesh; Ali Derakhshani; Vahid Sheibani

Objective: Stem cells (SCs) can improve the functional defects of brain injury. Rodents use their whiskers to get tactile information from their surroundings. The aim of this study was to investigate whether the transplantation of SCs into the lesioned barrel cortex can help neuronal function in the contralateral cortex. Materials and Methods: Sixteen male Wistar rats (200-230 g) were used in this experimental study. We induced a mechanical lesion in the right barrel cortex area of rats by removing this area by a 3 mm skin punch. Four groups containing one intact group of rats: group 1: control, and three lesion groups, group 2: lesion+un-differentiated dental pulp SCs (U-DPSCs), group 3: lesion+differentiated dental pulp SCs (D-DPSCs), and group 4: cell medium (vehicle) that were injected in the lesion area. Three weeks after transplantation of SCs or cell medium, the rats’ responses of left barrel cortical neurons to controlled deflections of right whiskers were recorded by using the extracellular single-unit recordings technique. Results: The results showed that the neural spontaneous activity and response magnitude of intact barrel cortex neurons in the lesion group decreased significantly (P<0.05) compared to the control group while ON and OFF responses were improved in the D-DPSCs (P<0.001) group compared to the vehicle group three weeks after transplantation. Conclusion: Transplantation of dental pulp mesenchymal SCs significantly improved the neural responses of the left barrel cortex that was depressed in the vehicle group.

839-846

Authors: Lei Yang; Liyi Hu

Objective: Non-small cell lung cancer (NSCLC) stands as a prominent contributor to cancer-related fatalities on a global scale, necessitating the search for novel therapeutic agents. SP-8356, a derivative of (1S)-(–)-verbenone, has shown promise as an anticancer agent in preclinical studies. However, specific mechanisms underlying its effects in NSCLC remain to be elucidated. The aim of this research was to explore the in vitro anti-NSCLC effects of SP-8356, elucidate its mechanisms of action, and assess its efficacy in inhibiting tumor formation in a murine model. Materials and Methods: In this experimental study, NSCLC cell lines were treated with various concentrations of SP- 8356. Cell viability and proliferation were assessed using MTT and colony formation assays, respectively. Cell cycle distribution was analyzed by flow cytometry, and apoptosis was evaluated by determining apoptotic protein expression. Western blot analysis was conducted to assess protein expression levels of the both p53 and MDM2. Additionally, we evaluated efficacy of the SP-8356 in inhibiting tumor formation of the nude mouse model. Results: SP-8356 demonstrated a concentration-dependent inhibition of cell proliferation in the NSCLC cell lines. Flow cytometric analysis showed that SP-8356 led to cell cycle arrest at the G2/M phase, indicating its potential influence on regulating the cell cycle. SP-8356 treatment was associated with the downregulation of CDK1 and Cyclin B1. Additionally, SP-8356 significantly enhanced apoptosis in NSCLC cells. SP-8356 treatment was associated with the downregulation of Bcl-2, while Bax expression was upregulated. Mechanistically, SP-8356 led to accumulation of the p53 protein levels within the NSCLC cells. This accumulation was mediated through inhibition of its negative regulator, MDM2. Using a nude mouse model demonstrated that SP-8356 effectively inhibited tumor formation in vivo. Conclusion: Our findings shed light on the molecular mechanisms underlying anticancer activity of SP-8356 and highlight its potential as a promising therapeutic candidate for NSCLC treatment.

854-862

Authors: Mahnaz Babaahmadi; Nima Makvand Gholipour; Behnoosh Tayebi; Jed Pheneger; Ensiyeh Hajizadeh-Saffar; Mohamadreza Baghaban Eslaminejad; Seyedeh-Nafiseh Hassani

Objective: The collagen-induced arthritis (CIA) model is the most commonly studied autoimmune model of rheumatoid arthritis (RA). In this study, we investigated the usefulness of collagen type II emulsified in Freund's incomplete adjuvant (CII/IFA) as a suitable method for establishing RA in Lewis rats. The aim of the present study was to present a straightforward and effective method for inducing CIA in rats. Materials and Methods: In this experimental study, animals were divided into two equal groups (n=5); control and CIA. Five rats were injected intradermally at the base of the tail with a 0.2 ml CII/IFA emulsion. On the seventh day, a 0.1 ml CII/IFA emulsion booster was injected. Arthritis symptoms that arose were evaluated at clinical, histological, radiological, and at protein expression levels to find out if the disease had been induced successfully. Results: Our finding showed a decreasing trend in the body weight during the RA induction period, while the arthritis score and paw thickness were increased during this period. The results of the enzyme-linked immunosorbent assay (ELISA) for serum samples revealed that the levels of proinflammatory cytokines, interleukin (IL)-1β, IL-6, IL-17, and tumor necrosis factor (TNF)-α and anti-CII IgG were significantly increased in CIA rats compared to the control group. After CIA induction, the level of anti-inflammatory protein IL-10 was decreased significantly. Radiographic examination of the hind paws showed soft tissue swelling, bone erosion, and osteophyte formation in CIA rats. Additionally, based on histological evaluations, the hind paws of the CIA group showed pannus formation, synovial hyperplasia, and bone and cartilage destruction. Conclusion: It seems that CII/IFA treatment can be an appropriate and effective method to induce RA disease in Lewis rats. This well-established and well-characterized CIA model in female Lewis rats could be considered to study aspects of RA and develop novel anti-arthritic agents.

874-882

Authors: Mansoureh Farhangniya; Farzaneh Mohamadi Farsani; Najmeh Salehi; Ali Samadikuchaksaraei

Objective: Wound healing is a complex process involving the coordinated interaction of various genes and molecular pathways. The study aimed to uncover novel therapeutic targets, biomarkers and candidate genes for drug development to improve successful wound repair interventions. Materials and Methods: This study is a network-meta analysis study. Nine wound healing microarray datasets obtained from the Gene Expression Omnibus (GEO) database were used for this study. Differentially expressed genes (DEGs) were described using the Limma package and shared genes were used as input for weighted gene co-expression network analysis. The Gene Ontology analysis was performed using the EnrichR web server, and construction of a protein-protein interaction (PPI) network was achieved by the STRING and Cytoscape. Results: A total of 424 DEGs were determined. A co-expression network was constructed using 7692 shared genes between nine data sets, resulting in the identification of seven modules. Among these modules, those with the top 20 genes of up and down-regulation were selected. The top down-regulated genes, including TJP1, SEC61A1, PLEK, ATP5B, PDIA6, PIK3R1, SRGN, SDC2, and RBBP7, and the top up-regulated genes including RPS27A, EEF1A1, HNRNPA1, CTNNB1, POLR2A, CFL1, CSNk1E, HSPD1, FN1, and AURKB, which can potentially serve as therapeutic targets were identified. The KEGG pathway analysis found that the majority of the genes are enriched in the "Wnt signaling pathway". Conclusion: In our study of nine wound healing microarray datasets, we identified DEGs and co-expressed modules using WGCNA. These genes are involved in important cellular processes such as transcription, translation, and posttranslational modifications. We found nine down-regulated genes and ten up-regulated genes, which could serve as potential therapeutic targets for further experimental validation. Targeting pathways related to protein synthesis and cell adhesion and migration may enhance wound healing, but additional experimental validation is needed to confirm the effectiveness and safety of targeted interventions.

753-763

Authors: Sabere Nouri; Rasoul Roghanian; Giti Emtiazi; Oguzhan Gunduz; Rasoul Shafiei

Objective: Multipotent cells derived from human exfoliated deciduous teeth (SHED) possess the ability to differentiate into various cell types, including osteoblasts. This study aims to simulate the growth induction and osteogenic differentiation of SHED cells using probiotics and their resultant biomaterials. Materials and Methods: This experimental study proceeded in two stages. Initially, we evaluated the effect of autoclaved nutrient agar (NA) grown probiotic Bacillus coagulans (B. coagulans) on the SHED and MG-63 cell lines. Subsequently, probiotics grown on the Pikovskaya plus urea (PVKU) medium and their synthesised hydroxyapatite (HA) were identified using X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray (EDX), and Fourier transform infrared spectroscopy (FTIR), and then used to stimulate growth and osteogenic differentiation of the SHED cell line. Osteoblast cell differentiation was assessed by morphological changes, the alkaline phosphatase (ALP) assay, and alizarin red staining. Results: There was a substantial increase in SHED cell growth of about 14 and 33% due to probiotics grown on NA and PVKU medium, respectively. The PVKU grown probiotics enhanced growth and induced stem cell differentiation due to HA content. Evidence of this differentiation was seen in the morphological shift from spindle to osteocyte-shaped cells after five days of incubation, an increase in ALP level over 21 days, and detection of intracellular calcium deposits through alizarin red staining-all indicative of osteoblast cell development. Conclusion: The osteogenic differentiation process in stem cells, improved by the nano-HA-containing byproducts of probiotic bacteria in the PVKU medium, represents a promising pathway for leveraging beneficial bacteria and their synthesised biomaterials in tissue engineering.

772-782

Authors: Yasmin Mirzaalikhan; Nasim Eslami; Amin Izadi; Faezeh Shekari; Sahar Kiani

Objective: Spinal cord injury (SCI) can disrupt membrane transmission by affecting transmembrane channels or neurotransmitter release. This study aimed to explore gene expression changes of transmembrane proteins underlying SCI through bioinformatics approaches and confirming in SCI model in rats. Materials and Methods: In this experimental study, the differentially expressed genes (DEGs) in acute and subacute SCI were obtained based on microarray data downloaded from the gene expression omnibus (GEO). Transmembrane proteins of DEGs were recognized by using the UniProt annotation and transmembrane helices prediction (TMHMM) methods. The model of SCI was established through a weight-dropping procedure in rats. To confirm the SCI model, hematoxylin and eosin (H&E) staining was performed. Total mRNA was extracted from spinal cord tissues, and the RNA expression profile of some of the significantly changed genes in the previous part that has been confirmed by real-time polymerase chain reaction (PCR). Blood was collected from rats before sacrificing. Extracellular vesicles (EVs) were isolated by high-speed centrifugation from plasma. For the assessment of protein expression, western blotting was used. Results: Based on bioinformatics analysis, we candidated a set of membrane proteins in SCI’s acute and sub-acute phases, and confirmed significant upregulation in Grm1, Nrg1, CD63, Enpp3, and Cxcr4 between the acute and control groups and downregulation in Enpp3 between acute and subacute groups at the RNA level. Considering CD63 as an EV marker, we examined the protein expression of CD9 and CD63 in the plasma-derived EVs, and CD9 has significant expression between acute and control groups. We also demonstrate no significant CD63 and Cxcr4 expressions between groups. Conclusion: Our results provide new insight into the relationship between candidate transmembrane protein expression and different stages of SCI using in-silico approaches. Also, results show the release of EVs in blood in each group after SCI helping enlarge strategies to enhance recovery following SCI.

790-800

Authors: Majid Kamali-Dolat Abadi; Gholamhossein Yousefi; Farzaneh Dehghani; Ali Akbar Alizadeh; Abolfazl Jangholi; Mohammad Amin Moadab; Maryam Naseh; Shima Parsa; Golara Nasiri; Negar Azarpira; Mehdi Dianatpour

Objective: Androgenetic alopecia (AGA) is a prevalent form of hair loss, mainly caused by follicular sensitivity to androgens. Despite developing different anti-androgen treatment options, the success rate of these treatments has been limited. Using animal models, this study evaluated the therapeutic effects of umbilical cord (UC) stem cell conditioned media (CM) combined with oral anti-androgens for hair regeneration. Materials and Methods: In this experimental study, Poloxamer 407 (P407) was used as a drug carrier for subcutaneous testosterone injection. AGA models were treated with oral finasteride, oral flutamide, and CM injections. Samples were thoroughly evaluated and compared using histological, stereological, and molecular analyses. Results: Injecting CM-loaded hydrogel alone or combined with oral intake of anti-androgens improved hair regeneration. These treatments could promote hair growth by inducing hair follicles in the anagen stage and shortening the telogen and catagen phases. Furthermore, the combination treatment led to an upregulation of hair induction gene expression with a downregulation of inflammation genes. Conclusion: Through a reduction in inflammation, injection of CM-loaded hydrogel alone or combined with oral intake of anti-androgens induces the hair cell cycle with regeneration in damaged follicles. Hence, this could be a promising therapeutic method for AGA patients.

809-812

Authors: Naeimeh Sadat Abtahi; Bita Ebrahimi; Firouzeh Ghaffari; Rohollah Fathi; Mojtaba Rezazadeh Valojerdi; Abolfazl Mehdizadeh Kashi; Sepideh Khodaverdi; Azar Yahyaei; Maziar Faridi

Today, timely diagnosis and therapeutic progress open a road of hope for survival in cancerous patients. Increased knowledge about the various cytotoxic treatment's impacts on ovarian function and fertility has resulted in a surge in the number of patients seeking to preserve their fertility before starting the anti-cancer treatment process. In this regard, embryo cryopreservation can be recommended for fertility preservation when the woman is married and has adequate time for ovarian stimulation. If patients are prepubertal girls or not married women, oocytes or ovarian tissue can be frozen instead to be used in the future. In this regard, the first attempts for ovarian tissue transplantations were conducted in 2016 and in 2019 for two cancerous patients whose ovarian tissue was cryopreserved in the Royan Human Ovarian Tissue Bank (Tehran, Iran). Unfortunately, the transplantations did not result in a live birth.

674-687

Authors: Ali Sayadmanesh; Mohamad Azadbakht; Kheirollah Yari; Ali Abedelahi; Hajar Shafaei; Dariush Shanehbandi; Behzad Baradaran; Mohsen Basiri

Objective: Chimeric antigen receptor (CAR) T cell therapy has recently emerged as a promising approach for the treatment of different types of cancer. Improving CAR T cell manufacturing in terms of costs and product quality is an important concern for expanding the accessibility of this therapy. One proposed strategy for improving T cell expansion is to use genetically engineered artificial antigen presenting cells (aAPC) expressing a membrane-bound anti-CD3 for T cell activation. The aim of this study was to characterize CAR T cells generated using this aAPC-mediated approach in terms of expansion efficiency, immunophenotype, and cytotoxicity. Materials and Methods: In this experimental study, we generated an aAPC line by engineering K562 cells to express a membrane-bound anti-CD3 (mOKT3). T cell activation was performed by co-culturing PBMCs with either mitomycin C-treated aAPCs or surface-immobilized anti-CD3 and anti-CD28 antibodies. Untransduced and CD19-CARtransduced T cells were characterized in terms of expansion, activation markers, interferon gamma (IFN-γ) secretion, CD4/CD8 ratio, memory phenotype, and exhaustion markers. Cytotoxicity of CD19-CAR T cells generated by aAPCs and antibodies were also investigated using a bioluminescence-based co-culture assay. Results: Our findings showed that the engineered aAPC line has the potential to expand CAR T cells similar to that using the antibody-based method. Although activation with aAPCs leads to a higher ratio of CD8+ and effector memory T cells in the final product, we did not observe a significant difference in IFN-γ secretion, cytotoxic activity or exhaustion between CAR T cells generated with aAPC or antibodies. Conclusion: Our results show that despite the differences in the immunophenotypes of aAPC and antibody-based CAR T cells, both methods can be used to manufacture potent CAR T cells. These findings are instrumental for the improvement of the CAR T cell manufacturing process and future applications of aAPC-mediated expansion of CAR T cells.

696-705

Authors: Hamid Reza Bidkhori; Moein Farshchian; Halimeh Hasanzadeh; Reza Jafarzadeh Esfehani; Reihaneh Alsadat Mahmoudian; Mahdi Moradi Marjaneh; Houshang Rafatpanah

Objective: The immunoregulatory properties of mesenchymal stromal/stem cells (MSCs) bring a promise for the treatment of inflammatory diseases. However, their ability to suppress the immune system is unstable. To enhance their effectiveness against immune responses, it may be necessary to manipulate MSCs. Although some dsRNA transcripts come from invading viruses, the majority of dsRNA has an endogenous origin and is known as endo-siRNA. DICER1 is a ribonuclease protein that can generate small RNAs to modulate gene expression at the post-transcriptional level. We aimed to evaluate the expression of several immune-related genes at mRNA and protein levels in MSCs overexpressing DICER1 exogenously. Materials and Methods: In this comparative transcriptomic experimental study, the adipose-derived MSCs (Ad-MSCs) were transfected using the pCAGGS-Flag-hsDicer vector for the DICER1 overexpression. Following the RNA extraction, mRNA expression level of DICER1 and several inflammatory cytokines were examined. We performed a relative real-time polymerase chain reaction (PCR) assay and transcriptome analysis between two groups including DICER1- transfected MSCs and control MSCs. Moreover, media from the transfected MSCs were evaluated for various interferon response factors by ELISA. Results: The overexpression of DICER1 is associated with a significant increase in the mRNA expression level of COX-2, DDX-58, IFIH1, MYD88, RNase L, TLR3/4, and TDO2 genes and a downregulation of the TSG-6 gene in MSCs. Moreover, the expression levels of IL-1, 6, 8, 17, 18, CCL2, INF-γ, TGF-β, and TNF-α were higher in the DICER1-transfected MSCs group. Conclusion: It seems that the ectopic expression of DICER1 in Ad-MSCs is linked to alterations in the expression level of immune-related genes. It is suggested that the manipulation of immune-related pathways in MSCs via the Dicer1 overexpression could facilitate the development of MSCs with distinct immunoregulatory phenotypes.

717-726

Authors: Negar Saliani; Shideh Montasser Kouhsari; Maryam Izad

Objective: Vaccinium arctostaphylos has traditionally been employed in Iranian folk medicine to treat diabetes. However, the precise molecular mechanisms underlying its antidiabetic properties remain incompletely understood. The current experiment intended to explore the modulatory effects of V. arctostaphylos fruit ethanolic extract (VAE) on biochemical and molecular events in the livers of diabetic rats. Materials and Methods: In this experimental study, male Wistar rats were randomly assigned to four groups: normal control, normal rats with VAE treatment, diabetic control, and diabetic rats with VAE treatment. Following 42 days of treatment, the impact of VAE on diabetes-induced rats was assessed by measuring various serum biochemical parameters, including insulin, free fatty acids (FFA), tumor necrosis factor-α (TNF-α), reactive oxygen species (ROS), and adiponectin levels. The activities of hepatic carbohydrate metabolic enzymes and glycogen content were determined. Additionally, expression levels of selected genes implicated in carbohydrate/lipid metabolism and miR-27b expression were evaluated. H&E-stained liver sections were prepared for light microscopy examination. Results: Treatment with VAE elevated levels of insulin and adiponectin that reduced levels of FFA, ROS, and TNF-α in the serum of diabetic rats. VAE-treated rats exhibited increased activities of hepatic glucokinase (GK), glucose-6-phosphate dehydrogenase (G6PD), and glycogen concentrations, in conjunction with decreased activities of glucose-6-phosphatase (G6Pase) and fructose-1,6-bisphosphatase (FBPase). Furthermore, VAE significantly upregulated the transcription levels of hepatic insulin receptor substrate 1 (Irs1) and glucose transporter 2 (Glut2), while considerably downregulated the expression of peroxisome proliferator-activated receptor gamma (Pparg) and sterol regulatory element-binding protein 1c (Srebp1c). VAE remarkably enhanced the expression of miR27-b in the hepatic tissues of diabetic rats. Abnormal histological signs were dramatically normalized in diabetic rats receiving VAE compared to those in the diabetic control group. Conclusion: Our findings underscore the hypoglycemic and hypolipidemic activities of V. arctostaphylos and assist in better comprehension of its antidiabetic properties.

738-740

Authors: Sarina omidshafiee; Moustapha Hassan; Andreas K. Nussler; Najimi M; Massoud Vosough

Theory of Forms" implies that a genuine version of creatures exists beyond the shapes in this world. Stem cell technology has adopted developmental cues to mimic real life. However, the functionality of the lab-made cells is far from primary ones. Perhaps it is time to switch from analytical to systematic perspective in stem cell science. This may be the way to define new horizons based on the systematic perspective and convergence of science in stem cell biology, bridging the current gap between the shadows of real knowledge in current research and reality in future.