Cell and Tissue Journal

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: Low back pain (LBP) is one of the most common musculoskeletal diseases in the world. Apoptosis of nucleus pulposus (NP) cells and structural changes in the intervertebral disc (IVD) matrix cause its degeneration and are directly related to LBP. Because of exosomes derived from mesenchymal stem cells (MSC-Exo) are high therapeutic potential and may be valuable options for decreasing intervertebral disc degeneration (IDD). 
The present study evaluates changes in the expression level of genes responsible for repairing NP cells (collagen and aggrecan) and tumor necrosis factor-alpha (TNF-α) gene and the effectiveness of exosomes in inhibiting excessive apoptosis of NP cells under inflammation.
Material and Methods: Exosomes were separated by ultracentrifuge. They were identified with the help of transmission electron microscopy (TEM), atomic force microscope (AFM), and dynamic light scattering (DLS). Cell viability was evaluated by MTT assay and DAPI staining. Real-time PCR measured the expression of collagen, aggrecan, and TNF-α genes.
Results: Exosomes are vesicles with a diameter of about 35.5 to 100 nm. Based on the findings of the MTT assay, the survival rate of the inflammatory cells treated with exosomes was significantly different from the inflammation group without treatment (*P<0.05 at the dose of 100 μg/ml, **P<0.01 at the doses of 25 and 50 μg/ml). DAPI results showed a decrease in cell death in the exosome-treated group. Real-time PCR results showed increased expression of collagen (*P<0.05) and aggrecan (***P<0.001) genes and decreased the TNF-α (**P<0.01) gene in inflammatory cells treated with exosomes.
Conclusion: Exosomes derived from mesenchymal stem cells can increase collagen and aggrecan gene expression and decrease TNF-α gene expression in treated cells.

Abstract

Abstract Aim: The purpose of this study is to investigate the pro-apoptotic and anti-metastatic potentials of Lippia citriodora alcoholic leaf extract on A2780 ovarian cancer cells and to evaluate the expression of E-cadherin as one of the most important marker in metastasis.
Material and Methods: A2780 ovarian cancer cell line was prepared from Pasteur cell bank and cultured in RPMI1640 medium containing 10% FBS and 1% antibiotic. After cell seeding and treatment with different concentrations of extract (10-400 µg/ml), the cells viability and pro-apoptotic potential were examined by MTT assay and acridine orange/ propodium iodide, respectively. Caspase-3 assay and anti-invasive effect were analyzed by migration assay and assessment of E-cadherin expression, respectively. Then, one way ANOVA test was employed for analysis of quantitative data.
Results: Data indicated that L. citriodora alcoholic extract attenuated ovarian cancer cell viability. Acridine orange/ propodium iodide and caspase-3 assays showed that this extract in IC50 concentration (100 µg/ml) induced apoptosis mainly through caspase dependent pathway in the ovarian cancer cells. Migration assay and RT-PCR exhibited that this extract has anti-invasive capability and by up- regulation of E-cadherin prevents the loss of attachment between cancer cells.
Conclusion: The results revealed that L. citriodora alcoholic extract has apoptosis inducing capacity and ability of restoration E-cadherin expression in A2780 cancer cells, which it able to suppress invasive potential of ovarian cancer cells.
 

Abstract Aim: This study was conducted to determine the neuroprotective effects of mint extract on alpha motor neuron degeneration at the anterior horn of the spinal cord after sciatic nerve compression in rats.
Material and Methods: In this study, 30 male Wistar rats weighing 200-250 g were randomly divided into 5 groups including control, compression and treatment groups of 50,75 and 100 mg.  In order to induce the compression, sciatic nerve was undergone to compress by using locking- scissors for 60 seconds. Hydroalcoholic extract of mint was injected intraperitoneally during the first and second weeks after the compression. After 28 days, rats were undergone by the perfusion method and after sampling the lumbar spinal cord, neuronal density was calculated by using dissector and stereological methods and the findings were compared together.
Results: a significant decrease was observed in compression group compared to the control for the neuronal density and also a significant increase was seen in treatment groups rather than the compression group (p

Abstract Aim: In this study, the effects of curcumin- coated silver nanoparticles were examined on induction of apoptosis in ovarian cancer A2780 cell.
Material and Methods: Silver nanoparticles coated with curcumin were biosynthesized, then A2780 cells were treated with different concentration of silver nanoparticles. Cytotoxicity effects of silver nanoparticles in A2780 cells were assessed by MTT assay and apoptotic effects of these nanoparticles were examined using DAPI and acridine orange/ propidium iodide staining and caspase 3/9 activation assay. Changes in Bax and Bcl-2 gene expression were analyzed by Real Time PCR.
Results: Findings showed that silver nanoparticles inhibit A2780 cells proliferation in a dose dependent manner. 8 µg/ml 50% decreased cell viability at 24 hours, DAPI and, acridine orange propidium iodide staining represent that percentage of apoptotic cells in the treated groups was increased. The results of Real Time PCR showed that Bax gene expression in cells treated with silver nanoparticles increased, while Bcl-2 gene expression in the treated cells was decreased.
Conclusion: Silver nanoparticles coated with curcumin induce apoptosis in A2780 cancer cells. The use of the nanoparticles should be considered a promising strategy for the treatment of ovarian cancer.

Abstract Aim: In this study the synergic effect of Chiton lamyi foot alcoholic extract and decellulation rat brain’s tissue on angiogenesis in hen (chick???) chorioalantoic membrane (CAM) was investigated. Material and Methods: In this study, 70 fertilized eggs of Ross strain were divided into 7 treatments groups in ten repetitions: 1. control, 2. DMSO, 3. decellulation rat brain’s tissue, 4. 10mgμl^-1 Chiton lamyi foot alcoholic extract, 5. 20mgμl^-1 Chiton lamyi foot alcoholic extract, 6. decellulation rat brain’s tissue & 10mgμl^-1 Chiton lamyi foot alcoholic extract and 7. decellulation rat brain’s tissue & 20mgμl^-1 Chiton lamyi foot alcoholic extract. All of eggs were placed in the incubator for eight days and treatments induction was done in 8^th day. Then they photographed using photo steriomicroscope in 12^th day. Blood vessels branches length and number were evaluated by Image J software in treatment sites on chorioalantoic membrane. Data were analyzed using ANOVA, SPSS Test (p < /em><0.05). Results: Results showed that Chiton lamyi foot alcoholic extract, 10 and 20mgμl^-1 Chiton lamyi foot alcoholic extracts with decellulation rat brain’s tissue treatments significantly decreased blood vessels branches length and number in comprising with control and DMSO treatments (p < /em>< 0.05). Conclusion: The Synergic usage of Chiton lamyi foot alcoholic extract and decellulation (acellular) rat brain’s tissue has dose inhibitory effect on hen chorioalantoic membrane (CAM) angiogenesis.  

Abstract Aim: Sea cucumber has been paid attention in treatment of bone defects due to possessing bioactive compound such as chondroitin sulfate. There were done successful researches in the field of application of mesenchymal stem cells for treatment of bone injury recently. Therefore, this study performed to define the osteogenic and adipogenic potency of Persian Gulf sea cucumber alcoholic extract on differentiation rat bone marrow derived stem cells into osteogenic and adipogenic. Material and Methods: In this experimental study, bone marrow stromal cells were isolated by flushing from male Wistar rat and demonstrated stemness by immunocytochemistry. The cells in 9 experimental group were exposed to various concentration (3.5, 6.25, 12.5, 25, 50, 100, 200, 400, 800 µg/ml) of sea cucumber alcoholic extract and cytotoxicity of sea cucumber alcoholic extract determined by MTT assay. Eventually, after 21 day, osteogenic and adipogenic differentiation were evaluated with alizarin red, oil red staining and alkalin phosphatase kit. Data were analyzed by SPSS software, ANOVA and the level of p ≤ 0.05 was considered significant. Result: An appropriate dosage for treatment of mesenchymal stem cells was determined less than 50 µg/ml. Also, oil red staining showed that sea cucumber alcoholic extract does not have capacity of adipogenic induction while alizarin red staining and alkaline phosphatase revealed that concentration of 25 µg/ml sea cucumber alcoholic extract was most efficient concentration into osteogenic differentiation.  Conclusion: Sea cucumber extract was able to differentiate rat bone marrow mesenchymal stem cells into osteogenic lineage.

Abstract Angiogenesis is a necessary process in many physiological and pathological events such as tumor growth. Therefore angiogenesis therapy can be a strategy to cancer treatment. It is important to identify different aspects of tumor’s angiogenesis.  Formation of functional vessels within tumors is essential for its growth and progression. The induction of new blood vessel growth by tumors is mediated through different molecules. A balance between pro-angiogenic and anti-angiogenic growth factors strictly control the process. This fact has led to design of therapeutic agents against tumor angiogenesis. Recently, endogenous (such as interleukin -8) and exogenous angiogenesis inhibitors (such as: Avastin) have been described. Furthermore because of the low mutagenic potential of endothelial cells, tumor’s vessel do not show resistance to the effects of many of these compounds. In addition, it is recognized that anti-angiogenesis and anti-cancer tumors can be more effective by combination of factors than the use of either alone. This article would review different aspects of angiogenesis in tumors. Since tumor growth is dependent on the development of blood vessels in the tumor, inhibition of angiogenesis maybe considered an appropriate treatment for cancer.