Mohammad Nabiuni

Background: Male infertility affects a substantial portion of the population, and extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) have been proposed as a cell‐free therapeutic approach. This study evaluated the effects of adipose tissue‐derived MSC EVs (ADSC‐EVs) on spermatogonial stem cell (SSC) function in vitro and on spermatogenesis in vivo. Methods and Results: Rat ADSCs were cultured to passage 3, and EVs were isolated from serum‐free conditioned medium by ultracentrifugation (100,000 × g , 70 min). In vitro, SSCs co‐cultured with sertoli cells were treated with 10, 50, and 100 µg/mL ADSC‐EVs; cell viability was measured by MTT after 4 days and gene expression (SYCP3, DAZL) assessed by qRT‐PCR after 10 days. In vivo, immature male Wistar rats (21 days old, n = 6 per group) were randomly assigned to control or intratesticular injection of ADSC‐EVs at 0.25, 1.25, or 2.5 mg/kg; testes were collected 10 days postinjection for histological (hematoxylin and eosin [H&E]) assessment and qRT‐PCR analysis. In vitro, 10 µg/mL ADSC‐EVs produced the highest SSC viability (78.06%) and significantly upregulated SYCP3 (~5.3‐fold, p < 0.001) while DAZL showed a nonsignificant change. In vivo, low‐dose ADSC‐EVs (0.25 mg/kg) increased the presence of spermatozoa within the lumen of seminiferous tubules and moderately upregulated SYCP3 (fold change ~1.95), whereas ADSC‐EVs at 2.5 mg/kg induced seminiferous epithelium degeneration and absence of mature spermatozoa. DAZL expression was downregulated in vivo at the tested doses. Conclusions: ADSC‐EVs exert dose‐dependent, dual effects on spermatogenesis: at low concentrations, they promote SSC differentiation (SYCP3‐associated), while higher concentrations can damage seminiferous architecture and impair sperm release. These findings emphasize the importance of dose optimization for potential therapeutic applications.

Citalopram (CT) has indeed been demonstrated to induce sexual dysfunction, although its capacity to be harmful to the male reproductive system has not been fully explored. The current investigation analyzed the mechanism underlying the protective impact of date palm pollen (DPP) on CT‐induced Sertoli cell (TM4 cells) apoptosis and clarified the harmful effects of CT on the mouse males’ reproductive system. Using flow cytometry, the apoptotic rate in each group was determined. The impact of DPP on the expression of Bcl-2 , Nrf2 , and SOD1 molecular targets was identified. Also, male NMRI mice were orally treated simultaneously with CT and DPP for 30 days, and the testes underwent histological analysis. Our findings demonstrated that adding different doses of DPP to Sertoli cells raised the mRNA expression levels of Nrf2 , SOD1 , and Bcl-2 to varying degrees ( p < 0.01). Our findings also demonstrated that DPP decreased the rate of TM4 cell death brought on by CT ( p < 0.01). In addition, many abnormalities were observed in testicular tissue in mice treated with CT. Our results showed that DPP can reduce the harmful effects of CT on testicular tissue and restore spermatogenesis. These results lend credence to the idea that CT can result in testicular injury and infertility. In contrast, treatment with DPP can lessen the negative effects of CT, most likely through the regeneration of testicular tissue and antioxidant systems.

Polycaprolactone (PCL) is a suitable material for bone repair due to good biocompatibility and mechanical properties. However, low bioactivity and hydrophobicity pose major challenges for its biomedical applications. To overcome these limitations, PCL-based scaffolds loaded with bioactive agents have been developed. Salicin (Sal) is an anti-inflammatory and analgesic herbal glycoside with osteogenic potential. In the present study, we aimed to produce a Sal-laden PCL (PCL–Sal) scaffold for bone healing applications. Three-dimensional scaffolds were produced and their biocompatibility, and physical-chemical characteristics were determined. The osteogenic potential of the PCL (PCL) and PCL–Sal scaffolds was evaluated using bone marrow mesenchymal stem cells (BMSCs). Scaffolds were implanted into a 5 mm bone defect created in the femur of adult rats, and the new bone fraction was determined using micro-computed tomography scanning at one-month follow-up. PCL–Sal scaffold had a structure, porosity, and fiber diameter suitable for bone construction. It also possessed a higher rate of hydrophilicity and bioactivity compared to the PCL, providing a suitable surface for the proliferation and bone differentiation of BMSCs. Furthermore, PCL–Sal scaffolds showed a higher capacity to scavenge free radicals compared to PCL. The improved bone healing potential of the PCL–Sal scaffold was also confirmed according to in vivo implantation results. Our findings revealed that the Sal-laden implant could be considered for bone repair due to desirable characteristics of Sal such as hydrophilicity, surface modification for cell attachment, and antioxidant properties.

Ovarian hyperstimulation syndrome (OHSS) mostly occurs in undergoing gonadotropin therapy women sensitive to endogenous gonadotropins. Honey bee venom (HBV) contains biologically active components that has pharmaceutical properties. This study was designed to assess the possibility of HBV application as an anti-inflammatory and anti-angiopathy therapeutic agent to suppress main inflammatory mediators in OHSS. OHSS induced by Pregnant Mare’s Serum Gonadotropin) PMSG) and hCG intraperitoneal in immature female Wistar rats. OHSS induction followed by intraperitoneal injection of HBV and metformin for 10 consecutive days;. At the end of the treatment period, metabolic parameters as well as ovarian expression of IL-6, COX2, and VEGF proteins were evaluated. This data showed that the OHSS+HBV group had a significant reduction in peritoneal and ovarian vascular permeability compared to the OHSS group. The ovarian expression of COX2, IL-6 and VEGF proteins in group OHSS+HBV.. The results of this study showed that HBV can affect the inflammatory feature of OHSS by reducing the angiogenesis factors.

Objective: Melittin is one of the natural components of bee venom (Apis mellifera), and its anticancer and anti metastatic properties have been well established, but the underlying mechanism remains elusive. The MDA-MB-231 is a triple-negative cell line that is highly aggressive and invasive. Besides, many critical proteins are involved in tumor invasion and metastasis. In this study, we investigated whether melittin inhibits the migration and metastasis of epidermal growth factor (EGF)-induced MDA-MB-231 cells via the suppression of SDF-1 alpha/CXCR4 and Rac1-mediated signaling pathways. Materials and Methods: In this experimental study, cells were treated with melittin (0.5-4 mu g/ml), and the toxicity of melittin was assessed by the MTT assay. Afterward, the migration assay was conducted to measure the degree of the migration of EGF-induced cells. The western blot technique was performed to analyze the rate of Rac1, p-Rac1, SDF-1 alpha, and CXCR4 expression in different groups. Results: The results demonstrated that melittin markedly suppressed the migration of EGF-induced cells and decreased the expression of p-Rac1, CXCR4, and SDF-1 alpha proteins. Conclusion: The results of the present study suggested that the anti-tumor properties of melittin could be through the blocking of the SDF-1 alpha/CXCR4 signaling pathway, which is beneficial for the reduction of tumor migration and invasion.

Steroid-associated osteonecrosis (SAON) is a chronic disease that leads to the destruction and collapse of bone near the joint that is subjected to weight bearing, ultimately resulting in a loss of hip and knee function. Zn²⁺ ions, as an essential trace element, have functional roles in improving the immunophysiological cellular environment, accelerating bone regeneration, and inhibiting biofilm formation. In this study, we reconstruct SAON lesions with a three-dimensional (3D)-a printed composite made of poly (epsilon-caprolactone) (PCL) and nanoparticulate Willemite (npW). Rabbit bone marrow stem cells were used to evaluate the cytocompatibility and osteogenic differentiation capability of the PCL/npW composite scaffolds. The 2-month bone regeneration was assessed by a Micro-computed tomography (micro-CT) scan and the expression of bone regeneration proteins by Western blot. Compared with the neat PCL group, PCL/npW scaffolds exhibited significantly increased cytocompatibility and osteogenic activity. This finding reveals a new concept for the design of a 3D-printed PCL/npW composite-based bone substitute for the early treatment of osteonecrosis defects.

HIF-1α has critical roles in the formation of tumor microenvironment by regulating genes involved in angiogenesis and anaerobic respiration. TME fuels tumors' growth and metastasis and presents therapy with several challenges. Therefore, we aimed to investigate if Melittin disrupts HIF-1α signaling pathway in breast adenocarcinoma cell line MDA-MB-231. Breast adenocarcinoma cell line MDA-MB-231 was cultured in the presence of different doses of Melittin, and MTT assay was carried out to measure Melittin’s cytotoxic effects. Cells were exposed to 5% O₂ to mimic hypoxic conditions and Melittin. Western blot was used to measure HIF-1α protein levels. Gene expression analysis was performed using real-time PCR to measure relative mRNA abundance of genes involved in tumor microenvironment formation. Our results revealed that Melittin effectively inhibits HIF-1α at transcriptional and translational/post-translational level. HIF-1α protein and mRNA level were significantly decreased in Melittin-treated groups. It is found that inhibition of HIF-1α by Melittin is through downregulation of NFκB gene expression. Furthermore, gene expression analysis showed a downregulation in VEGFA and LDHA expression due to inhibition of HIF-1α protein by Melittin. In addition, cell toxicity assay showed that Melittin inhibits the growth of MDA-MB-231 cell line through activation of extrinsic and intrinsic apoptotic pathways by upregulating TNFA and BAX expression. Melittin suppresses the expression of genes responsible for formation of TME physiological hallmarks by suppressing HIF-1α signaling pathway. Our results suggest that Melittin can modulate tumor microenvironment by inhibition of VEGFA and LDHA.