ObjectiveTo comprehensively analyze the recent advancements in the field of mesenchymal stem cells (MSCs) derived exosomes (MSCs-exosomes) in tissue repair. MethodsThe literature about MSCs-exosomes in tissue repair was reviewed and analyzed. ResultsExosomes are biologically active microvesicles released from MSCs which are loaded with functional proteins, RNA, and microRNA. Exosomes can inhibit apoptosis, stimulate proliferation, alter cell phenotype in tissue repair of several diseases through cell-to-cell communication. ConclusionMSCs-exosomes is a novel source for the treatment of tissue repair. Further research of MSCs-exosomes biofunction, paracellular transport, and treatment mechanism will help the transform to clinical application.
Epilepsy is a common neurological disease with complex etiology and various seizure forms. It can affect people of all ages. Although a variety of antiseizure medications are available, one-third of patients still have poor drug treatment. Therefore, better methods for the diagnosis and treatment of epilepsy are particularly important. Exosomes are extracellular vesicles with a diameter of 30 ~ 150 nm that have powerful intercellular information transmission functions and also play an important role in the central nervous system. Exosomes released by nerve cells in the local microenvironment can participate in nerve development and plasticity, regulate neuroinflammation, and reduce neuronal loss. Moreover, some proteins or micro ribonucleic acid (miRNA) in exosomes are highly correlated with epilepsy and are changed in epileptogenesis, so they play an important role in the prevention and early diagnosis of epilepsy. In addition, exosomes have better biocompatibility and lower immunogenicity. Its small size can effectively avoid the phagocytosis of mononuclear macrophages. Moreover, the proteins carried on its surface have a strong homing ability to target tissues or cells and can penetrate the blood-brain barrier to the intracranial, so exosomes have the advantage of natural drug delivery. Therefore, this study reviews the application of exosomes in epilepsy to improve the understanding of exosomes in scientific research and clinical workers.
Objective To observe the effects of co-transfection of Nogo extracellular peptide residues 1-40 (NEP1-40) and neurotrophin 3 (NT-3) genes with Schwann cell-derived exosomes (SCDEs) on the survival and differentiation of neural stem cells (NSCs), and lay the foundation for the in vivo experiments of SCDE and NSC co-transplantation. Methods The NEP1-40 and NT-3 genes were transfected into Schwann cells by lentiviral vector, and SCDEs were collected for identification. The NSCs that have been passaged for 3 times were selected and inoculated into the inoculation plate, and they were divided into conventional culture group, simple exosome culture group (adding empty vector plasmid to modify SCDE for culture) and two genes exosome culture group (adding two genes modified SCDE for culture). The activity of cells in each group was detected. The survival and differentiation of NSCs were evaluated by immunofluorescence detection of neuronal nuclei (NeuN), glial fibrillary acidic protein (GFAP) and galactosylceramidase (GALC) positive cells. Results After transfection of these two genes, the fluorescence intensity was higher and the cell state was better. The relative expression levels of messenger RNA and protein of NEP1-40 and NT-3 in the two gene groups were higher than those in the empty plasmid group (P<0.05). The relative expression levels of NEP1-40 and NT-3 proteins in SCDE of the two gene groups were higher than those of the empty vector group (P<0.05). There was no significant difference in the relative expression level of CD63 protein in SCDE between the two groups (P>0.05). In terms of cell activity, the cell activity of the two genes exosome culture group was the strongest, followed by the simple exosome culture group, and the conventional culture group was the weakest. The differences between any two groups were statistically significant (1.28±0.04 vs. 0.72±0.09 vs. 0.41±0.04, P<0.05). In terms of cell survival, NeuN-positive cells (5.23±0.22 vs. 2.36±0.09 vs. 1.00±0.01) and GALC-positive cells (2.29±0.06 vs. 1.75±0.02 vs. 1.00±0.04) of the two genes exosome culture group were the best, followed by the simple exosome culture group, and the conventional culture group were the weakest. The differences between any two groups were statistically significant (P<0.05). In terms of cell differentiation, NeuN-positive cells (0.44±0.02 vs. 0.29±0.01 vs. 0.16±0.01) and GALC-positive cells (0.38±0.07 vs. 0.23±0.02 vs. 0.12±0.01) of the two genes exosome culture group were the best, followed by the simple exosome culture group, and the conventional culture group were the weakest. The differences between any two groups were statistically significant (P<0.05). The differentiation of GFAP-positive cells in the conventional culture group was the best, followed by the simple exosome culture group, and the two genes exosome culture group was the worst (0.52±0.05 vs. 0.42±0.03 vs. 0.30±0.09). The differences between any two groups were statistically significant (P<0.05). Conclusion NEP1-40 and NT-3 genes can be successfully transfected into Schwann cells by lentiviral vector, which can effectively increase the content of related proteins in SCDE, and the exosomes can effectively promote the survival and differentiation of NSCs in vitro.
Exosomes are a type of tiny vesicles released by cells, which contain bioactive molecules such as proteins, nucleic acids, and lipids secreted by cells. Exosomes released by different cells play an important role in tumor development and metastasis. These exosomes can regulate the tumor microenvironment, promote the tumor growth and invasion, and participate in the process of distant metastasis by carrying specific proteins and nucleic acids. In addition, some biomarkers in exosomes can serve as potential biomarkers for early diagnosis and prognosis evaluation of osteosarcoma. This article reviews the research progress of exosomes in osteosarcoma, aiming to gain a deeper understanding of their mechanisms of action in this disease and provide a reference for the development of new treatment strategies and prognostic evaluation indicators.
This research aims to investigate the encapsulation and controlled release effect of the newly developed self-assembling peptide R-LIFE-1 on exosomes. The gelling ability and morphological structure of the chiral self-assembling peptide (CSAP) hydrogel were examined using advanced imaging techniques, including atomic force microscopy, transmission electron microscopy, and cryo-scanning electron microscopy. The biocompatibility of the CSAP hydrogel was assessed through optical microscopy and fluorescent staining. Exosomes were isolated via ultrafiltration, and their quality was evaluated using Western blot analysis, nanoparticle tracking analysis, and transmission electron microscopy. The controlled release effect of the CSAP hydrogel on exosomes was quantitatively analyzed using laser confocal microscopy and a BCA assay kit. The results revealed that the self-assembling peptide R-LIFE-1 exhibited spontaneous assembly in the presence of various ions, leading to the formation of nanofibers. These nanofibers were cross-linked, giving rise to a robust nanofiber network structure, which further underwent cross-linking to generate a laminated membrane structure. The nanofibers possessed a large surface area, allowing them to encapsulate a substantial number of water molecules, thereby forming a hydrogel material with high water content. This hydrogel served as a stable spatial scaffold and loading matrix for the three-dimensional culture of cells, as well as the encapsulation and controlled release of exosomes. Importantly, R-LIFE-1 demonstrated excellent biocompatibility, preserving the growth of cells and the biological activity of exosomes. It rapidly formed a three-dimensional network scaffold, enabling the stable loading of cells and exosomes, while exhibiting favorable biocompatibility and reduced cytotoxicity. In conclusion, the findings of this study support the notion that R-LIFE-1 holds significant promise as an ideal tissue engineering material for tissue repair applications.
ObjectiveTo investigate whether exosomes derived from atorvastatin (ATV)-pretreated human umbilical cord mesenchymal stem cells (ATV-MSC-EXO) alleviate high glucose-induced injury in human retinal vascular endothelial cells (HREC) via the protein kinase B (AKT)/endothelial nitric oxide synthase (eNOS) signaling pathway. MethodsThe optimal pretreatment concentration of ATV was determined using the cell counting Kit-8 (CCK-8) assay. Exosomes derived from mesenchymal stem cells (MSC-EXO) and ATV-pretreated MSC (ATV-MSC-EXO) were isolated and extracted, and their morphology and surface markers were characterized by transmission electron microscopy, nanoparticle tracking analysis, and Western blotting (WB). The uptake capacity of exosomes by human retinal vascular endothelial cells (HREC) was evaluated using a fluorescence labeling assay. In vitro cultured HREC were divided into the following groups: normal control group (NC group), high glucose group (HG group), high glucose+MSC-EXO group (MSC-EXO group), high glucose+ATV-MSC-EXO group (ATV-MSC-EXO group), high glucose+ATV-MSC-EXO+AKT inhibitor group (ATV-MSC-EXO-MK-2206-2HCL group), and high glucose+ATV-MSC-EXO+eNOS inhibitor group (ATV-MSC-EXO-L-NAME group). Cell proliferation and apoptosis were detected using CCK-8 and flow cytometry, respectively. The protein expression levels of B-cell lymphoma/leukemia-2 (Bcl-2), Bcl-2-associated protein (Bax), and Caspase-3 were measured by WB. In addition, the regulatory effects of ATV-MSC-EXO on the AKT/eNOS signaling pathway and its downstream functional molecules were analyzed by detecting the phosphorylation levels of AKT (P-AKT/AKT) and eNOS (P-eNOS/eNOS) via WB, the mRNA expression levels of AKT and eNOS by quantitative real-time polymerase chain reaction, and the concentrations of nitric oxide (NO) and endothelin-1 (ET-1) using commercial NO and ET-1 assay kits. ResultsThe optimal pretreatment concentration of ATV was 1 μmol/L. ATV-MSC-EXO exhibited similar morphology and particle size to MSC-EXO and were efficiently taken up by HREC. Under high glucose conditions, ATV-MSC-EXO significantly enhanced the viability of HREC (F=83.24, P＜0.000 1) and inhibited apoptosis (F=77.39, P＜0.000 1). WB analysis further confirmed that ATV-MSC-EXO upregulated the expression of the anti-apoptotic protein Bcl-2 (F=53.17), while downregulating the pro-apoptotic proteins Bax (F=36.49) and Caspase-3 (F=60.75) (P＜0.001). In addition, ATV-MSC-EXO markedly increased the protein levels of P-AKT/AKT (F=107.60) and P-eNOS/eNOS (F=38.59), as well as the relative mRNA expression of AKT, eNOS (F=203.60, 315.00; P＜0.000 1). Furthermore, ATV-MSC-EXO promoted NO production (F=407.40) and suppressed the relative expression of ET-1 (F=49.76) (P＜0.000 1). ConclusionATV-MSC-EXO enhances the viability and inhibits apoptosis of HREC under high glucose conditions by activating the AKT/eNOS signaling pathway.
Bone malignancies exhibit the characteristics of high incidence, poor prognosis, and strong chemoresistance. Exosomal microRNAs can regulate the proliferation of bone malignant cells, improve chemoresistance, influence cell communication and the microenvironment, and have significant potential in the diagnosis and treatment of bone malignancies. Due to their stability, exosomal microRNAs can serve as non-invasive biomarkers for diagnosis and prognosis. However, their widespread application in clinical settings requires standardized research. This review summarizes the progress of exosomal microRNA research in various bone malignancies including osteosarcoma, chondrosarcoma, Ewing sarcoma, and fibrosarcoma, to provide new theoretical foundations and perspectives for the field.
Mesenchymal stem cells (MSCs) are considered as an ideal treatment for multiple diseases including ocular disease. Recent studies have demonstrated that MSCs-derived exosomes have similar functions with MSCs. Exosomes are nanovesicles surrounded by a phospholipid layer that shuttle active cargo between different cells. They are capable of passing the biological barrier and have potentials to be utilized as natural carrier for the ocular drug delivery.
ObjectiveTo observe the effect of exosomes derived from human umbilical cord blood mesenchymal stem cells (hUCMSC) on the expression of vascular endothelial growth factor (VEGF) A in blue light injured human retinal pigment epithelial (RPE) cells. MethodshUCMSC were cultured with exo-free fetal bovine serum for 48 hours, and then the supernatants were collected to isolate and purify exosomes by gradient ultracentrifugation method. Transmission electron microscopy was used to identify the morphology of exosomes. Surface specific maker protein CD63 and CD90 were detected via Western blot. Cultured ARPE-19 cells were divided into normal control group, blue light injured group and hUCMSC exosomes treated group. Cells were exposed to the blue light at the intensity of (2000±500) Lux for 12 hours to establish the light injured models. The cells of hUCMSC exosomes treated group were treated by different concentrations of exosomes for 8, 16, 24 hours. The mRNA and protein of VEGF-A were determined by real time-polymerase chain reaction and Western blot. Immunofluorescence assay were used to detect the expression levels of VEGF-A. ResultshUCMSC exosomes were successfully isolated, they exhibited round or oval shape and their diameter ranged from 50 to 100 nm with membrane structure through electron microscope. hUCMSC exosomes expressed the common surface marker protein CD63 and the surface marker protein CD90 of hUCMSC. The protein and mRNA level of VEGF A in the blue light injured group increased significantly compared to that in normal control group (t=-16.553, -19.456; P < 0.05). After treating with low, middle and high concentration of hUCMSC exosomes for 8, 16 and 24 hours, the protein and mRNA level of VEGF A of injured RPE were significantly decreased (P < 0.05). With the treated time and concentration of hUCMSC exosomes improved, the protein and mRNA level of VEGF A of injured RPE gradually decreased (P < 0.05). Immunofluorescence assay showed the protein level of VEGF-A of injured RPE gradually decreased with the same concentration of hUCMSC exosomes treated over time. ConclusionhUCMSC exosomes can effectively down-regulate the mRNA and protein level of VEGF-A in blue light injured RPE, the effect depends on the concentration and treated time of hUCMSC exosomes.
ObjectiveTo observe the effects of exosomes derived from rat mesenchymal stem cells (MSC-exosomes) on the rat experimental autoimmune uveitis (EAU) model.MethodsTwelve Lewis rats were randomly divided into experimental group and control group by random number table, with 6 rats in each group. Rats in the experimental group were established with EAU model, 100 μl of MSC-exosomes (50 μg) were periocular injected on the 9th day after modeling while the control rats were injected with the same volume of phosphate buffer. At different time points after modeling, the retinal structure was observed by hematoxylin and eosin (HE) staining, and the clinical and pathological manifestations were evaluated. T cells from the two groups were analyzed by flow cytometry. Immunohistochemical staining was used to observe the expression of macrophage surface marker CD68. The effect of MSC-exosomes on T cells was measured by lymphocyte proliferation assays. And flow cytometry was used to detect Th1, Th17 and regulatory T cells Variety. Electroretinogram (ERG) was used to evaluate the retinal function. Data were compared between the two groups using the t test.ResultsHE staining showed that the retina structure of the experimental group was more complete than that of the control group on the 15th day after modeling. Immunohistochemical staining showed that the positive expression of CD68 in the experimental group was significantly less than that in the control group. On the 15th day after modeling, the retinal pathological score of the experimental group was lower than that of the control group. On the 9th to 13th day after modeling, compared to the control group, the average clinical scores of the retina in the experimental group were lower, and the difference was statistically significant (t=3.665, 3.21, 3.181, 4.121, 3.227; P＜0.01). The results of T cell proliferation assay showed that exosomes (1.0, 10.0 μg/ml) inhibited the proliferation of T cells under different concentrations of R16 (1, 10, 30 μg/ml), and the difference was statistically significant (F=11.630, 4.188, 6.011; P＜0.05). The results of flow cytometry showed that the number of Th1, Th17 and Treg cell subsets in the experimental group was decreased compared with the control group, and the difference was statistically significant (t=7.374, 4.525, 6.910; P＜0.01). There was no difference in the proportion of cells in the T cells and lymph nodes (t=1.126, 0.493, 0.178; P=0.286, 0.632, 0.862). The results of ERG showed that, compared with the control group, the amplitudes of 0.01, 3.0 cd/m2 a wave and b wave of the experiment group were all increased on the 15th day after modeling, and the differences were statistically significant (t=3.604, 4.178, 4.551, 2.566, P＜0.05).ConclusionsMSC-exosomes can reduce the clinical and pathological manifestations of EAU, protect retinal function, reduce ocular macrophage infiltration, down-regulate the proportion of inflammatory cells in the eye, and inhibit T cell proliferation.