جستجوی مقالات مرتبط با کلیدواژه « t cells » در نشریات گروه « پزشکی »

Peripheral nerve injury (PNI) is a critical clinical issue primarily caused by trauma. Tissue engineering approaches using nanofiber scaffolds have been extensively explored to improve material quality and create an environment resembling the natural extracellular matrix (ECM). 

In this study, we employed electrospinning technique to fabricate a composite scaffold comprising poly(ɛ-caprolactone) (PCL) and collagen (Col) loaded with all-trans retinoic acid (RA), a neural patterning and signaling chemical known to promote nerve regeneration. 

The synthesized nanofiber scaffold exhibited a diameter of 391±79 nm and a tensile strength of 250±13 MPa, providing sufficient support for native peripheral nerve regeneration. The inclusion of Col enhanced the scaffold’s hydrophilic behavior (contact angle: 43±6°), ensuring stability in an aqueous solution. Moreover, the results demonstrated the proliferation and adhesion of nerve cells on the scaffold, aligning with the directions of the warp and weft of the nanofiber mat. Importantly, the scaffolds demonstrated non-toxicity, making them a promising substitute for the native ECM for enhanced cell attachment and proliferation. Finally, immune-histochemistry analyses further confirmed that the scaffolds supported the release and growth of neurites, promoting cell differentiation toward nerve repair. 

The RA-loaded scaffolds demonstrated the enhanced biocompatibility, supported neurite growth, and showed potential as a capable candidate for nerve regeneration.

Scientists have focused on the development of new drug delivery systems including pH-sensitive nanomaterials adaptive to tumor microenvironments. We aimed to fabricate a microfluidic system to synthesize and characterize curcumin (Cur)-containing PCL and Chitosan (CSN) polymeric nanoparticles against MCF7 breast cancer cells.

The microfluidic chip was fabricated by photolithography and polydimethylsiloxane (PDMS) molding procedure. The chip was Y-shaped and equipped with two inlets and one outlet. PCL and Chitosan (CSN) were dissolved in acetic acid overnight and mixed with Cur for three hours. The prepared solution was injected from one inlet and a solution of tween 80 in distilled water was injected from the other inlet. The nanoparticles were characterized in size, electrical charge, structure, drug loading, and drug release efficiency. Finally, the cytotoxicity was assessed using the MTT assay at specific concentrations after 24 and 48 hr. 

The mean diameter/zeta potentials of spherical-shaped nanoparticles with and without Cur were 209 ±2 nm / +15 and 219 ± 4 nm /+3 , respectively. FTIR results confirmed the presence of all components in the nanoparticles. The Cur loading rate was 1.5%, and Cur represented a sustained release manner. Also, the release profile showed faster release in a low-pH medium. MTT assay results showed that Cur-containing nanoparticles exerted a significant effect on cell viability. 

It can be concluded that microfluidic systems can pave the way for nanoparticle synthesis easily rapidly and cost-effectively for cancer agent delivery. Based on our observations, PCL-CSN-loaded Cur nanoparticles represent appropriate characteristics and suitable anti-cancer effects.

Levels of the human epidermal growth factor receptor 2 (HER2) gene are low in normal breast tissue, but half of the patients with breast cancer exhibit higher levels of this receptor. The differential expression of the HER2 gene in normal and malignant cells makes it an excellent biomarker for therapeutic purposes. In this study, we evaluated the degree of HER2 overexpression in patients and its relationship with age and the occurrence of metastases.

In this retrospective, registry-based, two-center cohort study, information on 1500 breast cancer patients recruited from Shahid Mostafa Khomeini Hospital in Ilam Province was collected from 2020 to 2023.

 The likelihood of metastasis in cancer patients with HER2 gene expression was three times higher (adjusted OR: 2.82; 95% CI: 1.79–3.29; P=0.001). Additionally, the involvement of lymph nodes (adjusted OR: 2.01; 95% CI: 0.87–3.79; P=0.03) was significantly associated with increased metastasis.

 This study demonstrates that HER2 gene expression and the number of involved lymph nodes are significant prognostic factors that increase the risk of metastasis. Therefore, implementing comprehensive breast cancer screenings can play an important role in the treatment and prevention of metastasis in breast cancer patients.

Dendritic cells (DCs) are remarkable professional antigen-presenting cells (APCs) that are pivotal in bridging the gap between innate and adaptive immunity (1). Given this unique ability, these cells are a promising target for immunotherapy of various diseases (2). Monocytes serve as a valuable resourcefor generating DCs in vitro. To generate monocyte-derived DCs and investigate morphological changes during the differentiation phenomenon, following the reception of written informed consent, peripheral blood samples were obtained from healthy donors using falcon tubes containing heparin. Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll solution with a density of 1.077 g/ml and density gradient centrifugation. Monocytes were subsequently extracted from PBMCs through the magnetic-activated cell sorting method (MACS) following the instructions provided with the kit. The culture of monocytes was carried out in a concentration of 1.5 × 106/mLin a 6-well plate using complete culture media (RPMI-1640 supplemented with 15% FBS, 100 μg/mL streptomycin, 100 IU/mL penicillin, and 2 mM L-glutamine), in conjunction with 50 μM 2-mercaptoethanol solution and recombinant human GM-CSF and IL-4 cytokines at concentrations of 40 and 25 ng/mL, respectively. The plate was incubated at 37°C with 5% CO2 and humidification. After three days, half of the culture media was substituted with a new mixture containing GM-CSF and IL-4 cytokines. Immature DC (iDCs) cells were collected on the fifth day. Following this, iDCs were treated with 100 ng/mL of lipopolysaccharide (LPS) and incubated for 24 hours to induce the maturation of iDCs. On the sixth day, mature DCs were harvested. Monocytes and DCs were examined for their morphology, with images captured utilizing an inverted light microscope. In this regard, microscopic analysis revealed morphological alterations between monocytes derived from PBMCs at the initiation of culture and differentiated mature DCs acquired on day 6. Monocytes appeared as round cells with no visible dendrites, in contrast to DCs whichwere distinguishable by their visible dendrites (Figure1).

Atrophy of the muscles following denervation can lead to the death of myofibers. This study evaluated the sciatic nerve and tibialis cranialis muscle (TCM) regeneration using scaffold and cells. 

Ninety adult male Wistar rats were divided into six main groups (n=15) and three subgroups (2, 4, and 8 weeks). Groups: control; without surgery, Tr; sciatic nerve transected in silicone tube, S; collagen gel put inside the silicone tube, MC; placed 3×104 mast cells mixed with scaffold, MSC; placed 3×104 mesenchymal stem cells mixed with scaffold, and MC+MSC; 3×104 of each of the mast cell and mesenchymal stem cells along with scaffold. Animals were euthanized and sampled for muscle and nerve histological and nerve immunohistochemical evaluations. 

Diameter of muscle fibers, ratio of the muscle fiber’s nuclei to the fibrocyte nuclei (mn/fn), ratio of the muscle fibers nuclei number to the muscle fiber’s number (mn/mf), and ratio of the blood vessels number to the number of muscle fibers (v/mf) in all treatment groups, especially the MC + MSC group, increased compared to the Tr group but the number of mast cells, the percentage of sarcoplasmolysis, and necrosis fibers decreased. Histomorphometric results of the nerve in its various parts and immunohistochemistry results also showed improved nerve conduction, especially in the MC + MSC group. 

In this study, nerve improvement happened mainly for two reasons: cells and time. So, the most obvious improvement occurred in the group with mast and mesenchymal cells in the 8th week.

Statement of the Problem:

 Squamous cell carcinomas (SCCs) and premalignant disorders such as leukoplakia are common oral cavity lesions. Although these lesions are epithelial in nature, they are also associated with juxta-epithelial chronic inflammation. Mast cells play a significant role in inflammation initiation and propagation.

Previous studies have yielded conflicting results in this field. Therefore, this research aimed to assess the number of mast cells in oral SCC and dysplastic leukoplakia and explore their possible role in these lesions.

In this retrospective cross-sectional study, sixty-three archival cases, including 22 OSCCs, 28 dysplastic leukoplakias as epithelial dysplasia (ED), and 13 normal oral mucosal tissues, were examined for mast cells, using toluidine blue staining. Hotspot areas were identified at 10× magnification and mast cells were counted in 5 fields at 40× magnification. The average cell numbers were calculated, and the severity of inflammation was scored. Statistical analysis was performed using SPSS statistical software 20, including One-way ANOVA, Two-way ANOVA, paired-t test, and independent t-test. p Value < 0.05 was considered significant.

Among the 51 pathologic lesions, 54.9% were males and 45.1% were females, with a mean age of 56.34±15.35 years. The most common locations were the tongue and buccal mucosa. The mast cell count was significantly lower in SCC compared to ED (p= 0.009). There was no correlation between mast cell count and inflammation score (p= 0.345).

In this study, the mast cell count was higher in ED compared to OSCC, sugg-esting an increase in these cells during the pre-malignant stages. However, the number of mast cells decreased after connective tissue invasion and microenvironmental changes occurred.

Statement of the Problem: 

Success of pulpotomy of primary teeth depends on biological and cytotoxic effects of pulp capping agents. Mineral trioxide aggregate (MTA), Biodentine, calcium enriched mixture (CEM) cement, and ferric sulfate (FS) are among the commonly used pulp capping agents (PCAs) for pulpotomy, and their successful application has been previously evaluated.

This study aimed to compare the cytotoxicity of PCAs against mesenchymal stem cells isolated from human exfoliated deciduous teeth (SHEDs).

In this in vitro study, SHEDs were exposed to MTA, Biodentine, CEM cement, and FS for 24 and 72 hours. The methyl thiazolyl tetrazolium (MTT) assay was performed for five different concentrations of PCAs after 24 and 72 hours of exposure. Data were analyzed by ANOVA.

Generally, the biocompatibility increased by reduction in concentration. All tested concentrations showed higher biocompatibility at 72 hours compared with 24 hours (p< 0.0001). Comparison of cytotoxicity of different biomaterials revealed no significant difference at any time point (p> 0.05).

In general, the cytotoxicity of MTA, Biodentine, CEM cement, and FS was comparable, with no significant difference. Cytotoxicity decreased over time and by a reduction in concentration of biomaterials. MTA and Biodentine showed maximum biocompatibility followed by FS, and CEM cement.

 Laser therapy employs a concentrated and slender light beam to eliminate or eradicate cancerous cells and pre-cancerous abnormalities. The specific wavelength of 808 nm light is preferentially absorbed by tumor cells compared to healthy cells. This study aimed to assess the combined therapeutic impact of laser and gold, given that gold exhibits photothermal properties when exposed to laser radiation.

 In this in vitro study, two cell lines, namely healthy HuGu cells (human gingival fibroblast cells) and head and neck cancer cells (HN-5), were obtained from the Pasteur Institute. The effect of the laser diode with a density of 3 J/cm2 and wavelength of 808 nm on the proliferation and the survival rate of oral squamous cell carcinoma (HN-5) and human gingival fibroblast (HUGU) was assessed in 60seconds. MTT assay, DAPI test, and trypan blue staining were used to determine the growth and proliferation of HN-5 and HUGU cell lines.

 Findings showed that the laser diode along with gold decreased the rate of proliferation and survival cells in HN-5 compared to healthy cells. The changes in the cell population treated with gold and laser diode 808 were insignificant.

 Findings reveal that using a low-power laser can effectively inhibit the growth of oral cancer cells. It seems that photothermal therapy is a novel approach to oral cancer therapy.

Although assisted reproductive technology has been improved, the success rate is only 30%. Since the interaction between oocytes and cumulus cells (CCs) is necessary for the formation of a fertile oocyte, increasing the survival rate of CCs can improve the function of oocytes in infertile women.

This study aimed to investigate the effects of magnetic graphene oxide (MGO) nanocomposite on the biocompatibility and antioxidant activity of human CCs.

In this lab-trial study, from July 2021-2023 human CCs were collected from 37 women aged 20-37 yr and cultured in a medium containing Dulbecco’s Modified Eagle’s/F12, fetal bovine serum (10%), and penicillin-streptomycin (1%). Then CCs were treated with increasing concentrations of nano-MGO for 24, 48, and 72 hr (3[4, 5-dimethylthiazole-2-yl]-2, 5-diphenyltetrazolium bromide) assay and flow cytometry technique were used to compare the survival rate and apoptosis of CCs before and after treatment. Western blot test was used for expressing nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant in 2 groups.

The results of the present study showed that treatment with MGO increased the viability of CCs at a concentration of 50 µg/ml after 48 hr (p > 0.01). At higher doses (100 µg/ml) MGO decreased the survival rate of CCs (p > 0.05). Also, treatment with MGO at a concentration of 50 µg/ml increased the expression level of antioxidant protein Nrf2 in human CCs.

Our results highlight the use of MGO in a new strategy that improves CCs viability and secretion of antioxidant protein Nrf2, thereby potentially increasing in vitro fertilization outcomes. This article has been extracted from Ph.D. Thesis. (Fahimeh Kabiri)

Cell therapy, especially with mesenchymal stem cells (MSCs), is a potent treatment for many diseases or disorders. Meanwhile, MSCs-based cell-free products, such as extracellular vesicles (EVs) have been suggested as an alternative to MSCs. These MSC-EVs have been used in different trials to treat various inflammatory-dependent disorders. MSCs, according to their isolated tissue source, could present different therapeutic features and their derived EVs. One of the new sources for MSC isolation is amniotic fluid (AF). In addition, other than MSCs, new studies have used AF as an acceptable source for EV isolation. These isolated EVs from AF (AF-EVs) or AF-derived MSCs EVs (AF-MSC-EVs) have been used in different in-vitro and animal studies to treat a wide variety of inflammatory-dependent pathological conditions due to their confirmed anti-inflammatory potentials (through suppressing different pro-inflammatory cytokines). Meanwhile, in other conditions requiring repairing properties (e.g. wound healing or myocardial infarction), considering their regenerative and angiogenic potentials, these EVs have shown proper therapeutic results. Furthermore, other than the in-vitro and animal studies, AF-EVs containing treatment have successfully been used in some clinical trials and showed no adverse events among the patients and expressed potent anti-inflammatory properties through suppression of two very important pro-inflammatory cytokines, namely interleukin 6 and tumor necrosis factor α. Accordingly, AF-EVs and AF-MSC-EVs could be suitable choices for treatment due to their anti-inflammatory and regenerative properties. However, further clinical studies are needed.

The therapeutic effect of mesenchymal stem cells (MSCs) in liver cirrhosis is limited by their entrapment in the pulmonary vessels. Thus, the use of MSC-derived exosomes has become a promising strategy. The current work aimed to compare the role of human umbilical cord blood-MSCs (hUCB-MSCs) and their derived exosomes in the alleviation of liver cirrhosis focusing on the role of miR-23b and miR-221 and their direct effectors in inflammatory and autophagic pathways.  Rats were divided into six groups: normal controls (negative control), liver cirrhosis group (positive control), liver cirrhotic rats that received conditioned media, liver cirrhotic rats that received hUCB-MSCs, cirrhotic rats that received exosomes, and cirrhotic rats that received both hUCB-MSCs and exosomes. The messenger RNA expression of transforming growth factor-β (TGF-β), Matrix metalloproteinase 9 (MMP 9), fibronectin, collagen type-1 (col1), alpha-smooth muscle actin (α-SMA), Suppressor of Mothers Against Decapentaplegic (SMAD) 2 and 7, Beclin, P62, and light chain 3 (LC3) were evaluated by quantitative real-time polymerase chain reaction. Immunohistochemical staining for Beclin, P62, and LC3 was performed.  The treatment of cirrhotic rats with hUCB-MSCs, exosomes, or the combination of them significantly downregulated miRNA-221, fibronectin, collagen I, α-SMA, Smad2 (P<0.001, for each), and P62 (P=0.032, P<0.001, P<0.001, respectively). Additionally, the treatment of cirrhotic rats with hUCB-MSCs, exosomes, or the combination of them significantly upregulated mTOR, Beclin, LC3, and Smad7 (P<0.001, for each) and miRNA-23 (P=0.021, P<0.001, P<0.001, respectively). hUCB-MSCs and their derived exosomes ameliorated liver cirrhosis by anti-inflammatory and anti-fibrotic effects besides modulation of autophagy. The exosomes had a better improvement effect either alone or combined with hUCB-MSCs, as proved by improvement in liver function tests, and molecular, histopathological, and immunohistochemical profiles.

This study aims to investigate the effects of carob (Ceratonia siliqua L.) pod extract (CPE) on theviability of human endometrial mesenchymal stromal/stem cells (EnMSCs) and its impact on mRNA and protein expressionsof DNA methyltransferases (DNMT1, DNMT3A, and DNMT3B), histone deacetylase 1 (HDAC1), matrixmetalloproteinase-2 (MMP2), and cyclooxygenase-2 (COX-2) in endometriotic patients. In this experimental study, EnMSCs were derived from endometrium of patients with ovarianendometrioma (OMA-EnMSCs group) and deep infiltrative endometriosis (DIE) samples of 10 endometriosisassociatedinfertility (EAI) women (E-EnMSCs group) and compared to EnMSCs derived from the endometrium ofan endometriosis-free, normal woman as the control group (C-EnMSCs). The metabolic activity of the control andcase groups were evaluated by treating them with different concentrations of CPE. Cell viability was analysed byMTT. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to evaluatethe expression of specific genes at the mRNA and protein levels, respectively. Treatment with 0.8 and 2 μg/mL of CPE downregulated COX-2 and HDAC1 in the E-EnMSC groupcompared to the C-EnMSCs group. Treatment with 0.8 μg/mL of CPE also decreased MMP2 and DNMT3B geneexpressions. The COX-2 and DNMT3A genes were significantly upregulated after treatment with 2 μg/mL of CPE.Expressions of the COX-2, HDAC1, DNMT1, DNMT3A, and DNMT3B peptides decreased in the all three groups after treatment with 0.8 and 2 μg/mL of CPE. Gas chromatography-mass spectroscopy (GC-MS) analysis of CPEidentified 14 bioactive compounds. Molecular docking showed the best position of each bioactive compound on thedifferent target proteins that are involved in the process of apoptosis in EnMSCs. In vitro and in silico analyses of CPE bioactive compounds show that they may downregulate the cellinflammatory pathway involved in the pathophysiology of endometriosis.

Mesenchymal stem cell (MSC) transplantation represents a promising approach for treating Alzheimer’s disease (AD). These stem cells, however, have a short lifespan following transplantation into recipient animals. Selenium nanoparticles, due to their size, aid in drug delivery for brain disorders. This study investigated the therapeutic effect of MSCs and polyvinyl alcohol (PVA)-coated selenium nanoparticles (SeNPs) in a rat model of AD.

An Alzheimer-like phenotype was induced through intracerebroventricular (ICV) administration of streptozotocin (STZ).  Rats were assigned to five groups: control, Alz (STZ; 3 mg/kg, 10 μl, ICV), Alz+stem cell (ICV transplantation), Alz+SeNP (0.4 mg/kg, orally), and Alz+stem cell+SeNPs. The ICV administration of STZ mimicked some aspects of AD in the Alz groups. SeNPs were administrated for 30 days following STZ administration. The novel object recognition (NOR) and passive avoidance learning (PAL) tests were used to evaluate cognition and memory. Oxidative stress biomarkers and brain-derived neurotrophic factor (BDNF) were assessed by biochemical analysis, ELISA kits, and Congo red staining, respectively. 

The combined therapy of PVA-coated SeNPs and MSC transplantation was more effective in enhancing memory reacquisition compared to either SeNPs or MSCs alone. The use of stem cells in conjunction with PVA-coated SeNPs significantly boosted anti-oxidant capacity.

The results suggest that the joint treatment with PVA-coated SeNPs and MSCs offers considerable neuroprotection against AD in animal models.

Rheumatoid arthritis (RA) and osteoarthritis (OA) are prevalent chronic joint disorders with immunological pathogenesis. However, the causal relationships between circulating immune cells and them remain largely unknown. Therefore, we conducted a bidirectional two-sample Mendelian randomization (MR) study to determine their causal relationship.

Genome-wide association study summary statistics were extracted from publicly available databases regarding immune cell phenotypes, RA, and OA. MR analysis was conducted using five MR methods, with inverse-variance-weighted (IVW) as the primary analysis method. False discovery rate correction (FDR) was used to reduce the likelihood of type 1 errors. We also conducted MR-Egger intercept tests to evaluate horizontal pleiotropy.

After FDR adjustment of the P values for the IVW method, the CD27 expression on memory B cells was negatively related to the risk of RA (P < 0.001), and the human leukocyte antigen (HLA)--DR expression on CD14+ monocytes was negatively related to the risk of OA (P < 0.001). We also found that RA was negatively associated with the expression of HLA-DR on myeloid dendritic cells (P < 0.001), but significant horizontal pleiotropy was observed.

Our study demonstrates a causal relationship between specific immune cell traits and RA as well as OA, providing further insight into the role of immune cells in the pathogenesis of these disorders.

The aim of this study is to review the role of renin-angiotensin in skin regeneration and wound healing with a focus on molecular mechanisms. Angiotensin receptor type 1 (AT1R) are abundant in the wounded area, and thus, lead to the activation of ERK, STAT1, and STAT3 which can lead to epidermal self-renewal. The expression of Renin Angiotensin System (RAS) components was significantly lower in wounds caused by burning, rather than intact skin, noting that RAS is involved in the re-epithelialization of skin. ERK, STAT and STAT3 are the targets of Ang II, indicating that RAS active components are involved in fibroblast, stem cells and keratinocyte migration. The effect of inhibiting the RAS on wound healing is context-dependent. On one hand, it is suggested that inhibiting RAS during this phase may slow down wound healing speed. On the other hand, studies have shown that RAS inhibition in this phase can lead to α-SMA activation, ultimately accelerating the wound healing process. Most of the investigations indicate that the inhibition of RAS with Angiotensin Receptor Blockers (ARBs) and Angiotensin Converting Enzyme (ACE) plays a significant role in tissue remodeling in the last phase of wound healing. It has been shown that the inhibition of RAS can inhibit scar formation and fibrosis through the downregulation of inflammatory and fibrogenic agents, such as TGF-β, SMAD2/3, and TAK1, PDGF-BB, and HSP47. To sum up, that local administration of RAS regulators might lead to less scar formation and inflammation in the last phase of wound closure.

Natural killer (NK) cells play a critical role in the pathogenesis of endometriosis. Moreover, a normal vitamin D level is remarkably associated with an optimal immune response. So, there may be a probable relationship between these factors and the endometriotic women.

This study aimed to evaluate the percentage of NK cells and their subsets and their relationship with serum levels of vitamin D and interferon gamma (IFN-γ) in women with endometriosis.

In this case-control study, 29 women with stage III-IV endometriosis and 30 healthy controls were enrolled. The study was conducted in the Immunology Department of Isfahan University of Medical Sciences, Isfahan, Iran between November 2021 and June 2022. The percentage of NK cells and their subsets, including CD56dim CD16+, CD56bright CD16- and CD56bright CD16bright were measured in the peripheral blood samples using flow cytometry. Serum levels of vitamin D and IFN-γ were also measured using the enzyme-linked immunosorbent assay.

The mean percentage of NK cells in women with endometriosis increased significantly compared to the control group (p = 0.03). The percentage of CD56dim CD16+ (p = 0.007) and CD56bright CD16 bright (p = 0.043) increased significantly in women with endometriosis in comparison with the control group, but the percentage of CD56bright CD16- subset was not significantly different. No relationship was observed between NK cells and their subsets with vitamin D and IFN-γ in the studied groups.

The study of NK cell subsets and their related factors can be useful in assessing and treating women suffering from endometriosis. However, more comprehensive studies are required to draw definitive conclusions about these observations.

In endemic malarial nations, repeated use of antimalarial drugs has increased due to resistance, misuse, and unrestricted availability, which could contribute to infertility rates. Therefore, we investigated the effects of long-term repeated treatment with two commonly prescribed artemisinin-based combination therapies, artemether/lumefantrine (A/L) and artesunate-amodiaquine (A/A), on reproductive potential in mice. 

Sixty male mice were divided into three groups: control, A/L, and A/A treatment.  Mice underwent treatment for three consecutive days per week, and this regimen was repeated every two weeks for a total of six cycles. Sperm parameters were evaluated after the 1st, 2nd, 3rd, and 6th exposure cycles, after which treated male mice were paired with female mice for mating.

Sperm viability was significantly reduced by 21% (P<0.001) following the 6th exposure to A/L, whereas the 2nd, 3rd, and 6th exposures to A/A resulted in significant decreases in sperm viability of 26% (P<0.001), 12% (P<0.01), and 31% (P<0.001), respectively, compared to the control group. Treatment with A/A during the 3rd and 6th periods led to a significant decline (P<0.001) in sperm mass activity by 20% and 28%, respectively, compared to the control group. However, long-term therapeutic exposure to A/L or A/A did not affect testosterone levels, epididymal content, or the ability to impregnate female mice.

Long-term treatment with A/L or A/A did not affect testosterone levels or epididymal content. However, a decrease in sperm viability was observed, even though the mice remained fertile.

Researchers are looking for a way to improve the myogenic differentiation of stem cells. Adipose-derived stem cells (ADSCs), known for their multipotency and regenerative capabilities, have been extensively studied for their therapeutic potential. Meanwhile, PC12 cells, derived from rat pheochromocytoma, have been found pivotal in neuroscience research, particularly as a neuronal model system. The current study investigated the effect of the PC12 adrenal pheochromocytoma cell line on the myogenic differentiation of ADSCs.  This experimental study was conducted during 2019-2022 (Ahvaz, Iran). Differentiation of ADSCs was induced by using 3 μg/mL 5-azacytidine for 24 hours. Then, the culture media was changed with Dulbecco’s Modified Eagle-High Glucose (DMEM-HG) containing 5% horse serum (HS) and kept for 7 days. Different percentages of differentiated ADSCs and PC12 (100:0, 70:30, 50:50, 30:70) were cocultured for 7 days in DMEM-HG containing 5% HS. PC12 was labeled with cell tracker C7000. The real-time polymerase chain reaction and Western blotting techniques were utilized to assess gene and protein expression. All experiments were repeated three times. Data were analyzed using GraphPad Prism 8.0.2 software with a one-way analysis of variance. P<0.05 was considered statistically significant. PC12 visualization confirmed the accuracy of the co-culture process. The differentiated cells showed an aligned, multinucleated shape. The differentiated ADSCs revealed significantly elevated levels of Myh1, Myh2, and Chrn-α1 gene expression compared with undifferentiated ADSCs (P<0.0001). The ADSCs cocultured with PC12 cells showed significantly higher Myh1, Myh2, and Chrn-α1 gene expression than differentiated ADSCs (P<0.001). ADSCs cocultured with 50% PC12 revealed significantly higher MYH and nAchR protein expression than the differentiated group (P<0.01 and P<0.001).  Coculturing PC12 cells and ADSCs improves the efficiency of myogenic differentiation. However, the effectiveness of myogenic differentiation depends on the proportions of administered PC12 cells.