Cell and Tissue Journal

Abstract Introduction: Plant-derived exosome-like nanoparticles (PDENs) are nano-sized vesicles released by plant cells. They contain a lipid bilayer membrane and carry bioactive molecules such as proteins, lipids, nucleic acids, and secondary metabolites. Due to their low toxicity and natural drug delivery potential, they have gained attention for therapeutic applications. Plant extracts are also rich in phenolic compounds and flavonoids with known antioxidant and antimicrobial effects. The rising problem of antibiotic resistance in pathogens like Escherichia coli, Staphylococcus aureus, and Streptococcus pyogenes has created an urgent need for new natural agents.
Aim: This study aimed to isolate and characterize exosomes from ginger, lavender, onion, and lemon, and to compare their antioxidant and antibacterial activities with those of aqueous extracts from the same plants.
Materials and methods: Fresh ginger, lavender, onion, and lemon were used for the preparation of plant extracts and exosome isolation. For aqueous extraction, 2 g of ginger and lavender samples were homogenized with 20 mL of distilled water and incubated in a water bath at 70°C for 90 minutes. The mixtures were centrifuged at 3000 × g for 10 minutes, and the supernatants were filtered and stored at 4°C until analysis. Exosomes were isolated using the Exosun Exosome Isolation Kit (EXOSUN Company). Plant materials were homogenized, filtered, and subjected to differential centrifugation. The resulting supernatants were processed according to the manufacturer's instructions using buffers A and B. Due to the acidic nature of lemon juice, modification of the protocol was required by increasing the concentration of buffer A to facilitate exosome precipitation. Protein concentration of isolated exosomes was determined using the Bradford assay with bovine serum albumin (BSA) as the standard. Exosome morphology was characterized by transmission electron microscopy (TEM). The antioxidant activity of extracts and exosomes was evaluated using the Ferric Reducing Antioxidant Power (FRAP) assay. Samples were analyzed in 96-well microplates, and absorbance was measured at 570 nm. Antibacterial activity was assessed against Escherichia coli, Staphylococcus aureus, and Streptococcus pyogenes using the disk diffusion method. Sterile blank disks were impregnated with plant extracts or exosome preparations at concentrations of 25, 50, and 100 mg/mL and placed on Mueller-Hinton agar plates inoculated with bacterial suspensions adjusted to a 0.5 McFarland standard. Gentamicin and vancomycin served as positive controls. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were determined using the broth microdilution method in 96-well microplates. All experiments were performed in triplicate. Statistical analyses were conducted using SPSS version 27, applying one-way ANOVA followed by Tukey’s post hoc test, with p <0.05 considered statistically significant.
Results: TEM confirmed successful isolation of exosome-like vesicles from all plants. Bradford assay indicated protein-containing nanoparticles. FRAP analysis showed that lavender had the highest antioxidant capacity. Plant extracts generally showed greater antioxidant activity than their corresponding exosomes. Antibacterial testing revealed that exosomes from all four plants had no detectable antibacterial effect. Similarly, aqueous extracts of ginger, lavender, and onion showed no inhibition. However, lemon extract exhibited significant antibacterial activity against all three bacteria. MIC values of lemon extract were 3.12 mg/mL for S. aureus, 6.25 mg/mL for S. pyogenes, and 50 mg/mL for E. coli. Inhibition zones for S. aureus reached 22 mm at 100 mg/mL. Gram-positive bacteria were more susceptible than Gram-negative E. coli.
Discussion:The spread of antibiotic-resistant pathogens has increased the need for new treatments. In this study, exosomes isolated from ginger, lavender, onion, and lemon showed no antibacterial activity, while among aqueous extracts, only lemon extract displayed good antibacterial properties. The lack of activity in aqueous lavender extract compared to alcoholic extracts in previous reports may be due to solvent differences affecting phenolic compound extraction. Aqueous ginger extract also showed no effect, which contrasts with some alcoholic extracts. However, lavender exosomes demonstrated the highest antioxidant capacity among all samples, highlighting their potential for antioxidant applications.
Conclusion: Although plant-derived exosomes from these four species did not show antibacterial effects under the tested conditions, lemon aqueous extract exhibited promising antimicrobial activity, especially against Gram-positive bacteria. Lavender exosomes showed the strongest antioxidant potential. These findings suggest that lemon extract could serve as a natural antibacterial agent, while lavender-derived exosomes and extracts may be valuable sources of natural antioxidants. Further in vivo studies are needed to explore their efficacy and safety.

Abstract Introduction: Cartilage, a tissue without blood vessels and nerves, possesses inherently limited regenerative capacity following injury, often leading to progressive joint degeneration and conditions like osteoarthritis (OA) if left untreated. Current clinical interventions, such as surgical microfracture or autologous chondrocyte implantation (ACI), face significant challenges, including donor site morbidity, immune rejection, and the formation of fibrocartilage with inferior biomechanical properties. These limitations underscore the urgent need for novel therapeutic strategies that can effectively stimulate hyaline cartilage regeneration. In this context, mesenchymal stem cell-derived exosomes (MSC-Exos) have garnered attention as a cell-free regenerative tool, leveraging their cargo of bioactive molecules (e.g., miRNAs, cytokines, and growth factors) to modulate chondrogenesis, suppress inflammation, and enhance extracellular matrix (ECM) synthesis. Concurrently, glucosamine, a natural amino sugar and precursor for glycosaminoglycan (GAG) biosynthesis, has demonstrated dual functionality in joint health: not only does it serve as a building block for proteoglycans critical to cartilage integrity, but it also exhibits chondroprotective effects by mitigating ECM degradation and promoting stem cell chondrogenic differentiation. The potential synergy between MSC-Exos and glucosamine could thus address multiple facets of cartilage repair, combining anabolic stimulation (via exosomal signaling) with metabolic support (via glucosamine supplementation), offering a promising combinatorial approach to halt OA progression and restore functional cartilage.
Aims: This study aimed to investigate the combined effect of mouse bone marrow stem cell-derived exosomes and glucosamine on the expression of cartilage-specific genes, including Sox9, Acan, Col2a1, and Col10a1.
Materials and Methods: Bone marrow mesenchymal stem cells were prepared from NMRI mice. The mice were euthanized by cervical dislocation, the femoral heads were removed, and the bone marrow contents were transferred into a cell culture flask using a syringe containing culture medium. The bone marrow cells were cultured and were ready for use after 3 to 5 passages. The cell supernatant was separated, and exosomes were extracted from it by successive rounds of centrifugation followed by ultracentrifugation. Mesenchymal stem cell viability and determining the appropriate concentration of exosomes and glucosamine were performed using the MTT assay. The experiments were performed on mesenchymal stem cells in 4 groups: control, exosome, glucosamine, and exosome + glucosamine. The effects of exosomes and glucosamine on the expression of Sox9, Acan, Col2a1, and Col10a1 genes in mesenchymal stem cells were investigated in the presence of chondrogenic medium.
Results: According to the MTT assay results demonstrating the synergistic effect of exosomes and glucosamine, the combined concentrations of 15 μg/mL exosomes and 25 μg/mL glucosamine were chosen for subsequent applications. Real-time PCR results showed that the expression of Sox9, Acan, and Col2a1 genes in stem cells treated with exosomes and glucosamine significantly increased compared to the other groups after 14 days, while the expression of the Col10a1 gene significantly decreased compared to the other groups.
Discussion: The combined treatment of bone marrow–derived mesenchymal stem cell (BMSC) exosomes and glucosamine significantly upregulated the expression of key chondrogenic markers, including Sox9, Acan, and Col2a1, while downregulating the hypertrophic marker Col10a1. This gene expression profile suggests a dual beneficial effect: (1) promotion of chondrogenic differentiation and extracellular matrix (ECM) synthesis, and (2) suppression of hypertrophic differentiation, a critical factor in preventing cartilage calcification and osteoarthritis progression. These findings highlight the synergistic potential of BMSC exosomes and glucosamine as a combinatorial therapy for cartilage regeneration. By enhancing anabolic processes (Sox9-mediated chondrogenesis and aggrecan/collagen II deposition) and concurrently inhibiting catabolic pathways (Col10a1-associated hypertrophy), this strategy may offer a promising approach to delay or reverse early-stage cartilage degeneration in degenerative joint diseases
Conclusion: Our study reveals that combining bone marrow stem cell-derived exosomes with glucosamine synergistically enhances chondrogenesis by upregulating key cartilage markers (Sox9, Acan, Col2a1) while suppressing hypertrophy-related Col10a1. This dual action suggests that exosomes promote cartilage matrix synthesis through their bioactive cargo (e.g., miRNAs/growth factors), while glucosamine likely inhibits hypertrophic differentiation, potentially via modulation of the Wnt/β-catenin pathway. These findings support this combination as a promising strategy for improving cartilage repair and preventing OA progression, though further in vivo validation is needed.

Abstract Aim: Infertility is a life crisis that affects patients worldwide. Infertility is defined as failure to conceive after 12 months of sexual activity, affecting 15-17% of couples worldwide. and about 50% of them are related to the factors of female infertility. Activating the process of meiosis in the oocyte and its maturation has been one of the important therapeutic goals of infertility researchers. Today, inducing oocyte growth and development outside the body is one of the methods used in assisted reproduction technology. Bone marrow is a complex organ in which different lineages of hematopoietic and stromal cells support hematopoiesis. Extracellular vesicles (EVs) with a size of 20-100 nm are released by different types of cells in culture media. In addition to proteins, exosomes are enriched with an array of cytokines, specific lipid rafts such as phosphoglyceride, cholesterol, ceramide, fatty-acyl chains, as well as mRNAs, miRNAs, non-coding RNAs, tRNAs, rRNAs, and rarely DNA.
  The purpose of this experimental research is to investigate the effect of exosomes derived from the bone marrow stem cells of small laboratory mice on the maturation of preantral follicles. Material and methods: Exosomes were isolated and cultured from the mesenchymal stem cells of the bone marrow of small laboratory mice by the flushing method. Identification of stem cells was done by flow cytometry method and separation and purification of exosomes was done by ultracentrifuge, identification of exosome was also checked by atomic force microscope (AFM). The effects of exosomes on the viability of follicles were measured by the MTT method, and developmental parameters such as the diameter and the formation of the antrum cavity in the follicles were examined and the follicles were examined on days 0, 2, 3 and 4 of culture with 20 magnification and inverted microscope. Photographs were taken and the diameter of the follicles was measured in micrometers by Image J software. The expression of GDF-9, BMP-15 and BMP-7 genes as genes involved in the growth and maturation of follicles was investigated using Real Time-PCR method. GraphPad Prism 8 software and one way Anova statistical test were used to analyze the data of this research.
Results: The evaluation of the viability of the follicles showed that compared to the control group, the follicles treated with exosome 25, 50 and 100 micrograms/ml showed an increase in viability, as well as the rate of antrum formation increased significantly in the group with the concentration 100 μg/ml showed a significant level (p<0.01**) compared to the control group. The diameter of the follicles increased with increasing the concentration of exosomes compared to the control group. GDF-9, BPM-15 and BMP-7 genes also increased in the treatment groups. Conclusion: According to the findings of this experimental research, it can be stated that exosomes derived from bone marrow stem cells Small laboratory mice have a positive effect on survival and maturity, as well as the growth of ovarian follicles.

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