Mesenchymal Stem Cell Exosomes Research Articles

Exosomes of mesenchymal stem cells delivered from methacrylated hyaluronic acid patch improve the regenerative properties of endothelial and dermal cells.

Wound care management urgently needs the development of innovative smart wound dressings. The complexity of the wound often requires the use of personalized medication and the advent of three-dimensional (3D) bioprinting fits strongly with this need. In this view, in the present work a methacrylated hyaluronic acid (MeHA) bioink was tested for the fabrication of advanced smart patches as a delivery system of exosomes derived from human mesenchymal stem cells (hMSC-EXOs) suitable for wound healing purposes. MeHA patches were realized by 3D bioprinting technique and they were loaded with hMSC-EXOs. The 3D printed MeHA patches revealed improved mechanical performance, appropriate swelling ratio, extended degradation time, and suitable biocompatibility. Furthermore, MeHA patches loaded with hMSC-EXOs improved the proliferation, migration, angiogenic ability, and expression of specific markers related to wound healing process in human fibroblasts and human endothelial cells.

Infarct-preconditioning exosomes of umbilical cord mesenchymal stem cells promoted vascular remodeling and neurological recovery after stroke in rats

BackgroundStroke is the leading cause of disability worldwide, resulting in severe damage to the central nervous system and disrupting neurological functions. There is no effective therapy for promoting neurological recovery. Growing evidence suggests that the composition of exosomes from different microenvironments may benefit stroke. Therefore, it is reasonable to assume that exosomes secreted in response to infarction microenvironment could have further therapeutic effects.MethodsIn our study, cerebral infarct tissue extracts were used to pretreat umbilical cord mesenchymal stem cells (UCMSC). Infarct-preconditioned exosomes were injected into rats via tail vein after middle cerebral artery occlusion (MCAO). The effect of infarct-preconditioned exosomes on the neurological recovery of rats was examined using Tunel assay, 2,3,5-triphenyltetrazolium chloride (TTC) assay, magnetic resonance imaging (MRI) analyses, modified Neurological Severity Score (mNSS), Morris water maze (MWM), and vascular remodeling analysis. Mi-RNA sequencing and functional enrichment analysis were used to validate the signal pathway involved in the effect of infarct-preconditioned exosomes. Human umbilical vein endothelial cells (HUVECs) were co-cultured with the isolated exosomes. Cell Counting Kit-8 (CCK-8) assay, scratch healing, and Western blot analysis were used to detect the biological behavior of HUVECs.ResultsThe results showed that compared with normal exosomes, infarct-preconditioned exosomes further promoted vascular remodeling and recovery of neurological function after stroke. The function of upregulated miRNAs and their target genes which is beneficial to vascular smooth muscle cells verified the importance of vascular remodeling in improving stroke. Better resistance to oxygen–glucose deprivation/reoxygenation (OGD/R), reduced apoptosis, and enhanced migration were observed in infarct-preconditioned exosomes-treated umbilical vein endothelial cells.ConclusionsOur results demonstrated that infarct-preconditioned exosomes promoted neurological recovery after stroke by enhancing vascular endothelial remodeling, suggested that infarct-preconditioned exosomes could be a novel way to alleviate brain damage following a stroke.

Three-Dimensional-Cultured MSC-Derived Exosome-Hydrogel Hybrid Microneedle Array Patch for Spinal Cord Repair.

Exosomes derived from mesenchymal stem cells (MSCs) have been proven to exhibit great potentials in spinal cord injury (SCI) therapy. However, conventional two-dimensional (2D) culture will inevitably lead to the loss of stemness of MSCs, which substantially limits the therapeutic potency of MSCs exosomes (2D-Exo). Exosomes derived from three-dimensional culture (3D-Exo) possess higher therapeutic efficiency which have wide applications in spinal cord therapy. Typically, conventional exosome therapy that relies on local repeated injection results in secondary injury and low efficiency. It is urgent to develop a more reliable, convenient, and effective exosome delivery method to achieve constant in situ exosomes release. Herein, we proposed a controlled 3D-exohydrogel hybrid microneedle array patch to achieve SCI repair in situ. Our studies suggested that MSCs with 3D-culturing could maintain their stemness, and consequently, 3D-Exo effectively reduced SCI-induced inflammation and glial scarring. Thus, it is a promising therapeutic strategy for the treatment of SCI.

TGFβ1-Pretreated Exosomes of Wharton Jelly Mesenchymal Stem Cell as a Therapeutic Strategy for Improving Liver Fibrosis

Background: Mesenchymal stem cells (MSCs) are the most promising tools for cell treatment and human tissue regeneration, e.g., in liver fibrosis. Mesenchymal stem cells repair tissue damage through paracrine mediators such as exosomes. Types and concentrations of inflammatory mediators, including transforming growth factor-beta (TGFβ1), in MSCs microenvironment can affect MSCs’ function and therapeutic potency. Objectives: This experimental study aimed to explore the effects of Wharton jelly MSCs (WJ-MSCs) exosomes on fibrotic gene expression and Smad2/3 phosphorylation (phospho-Smad2/3 (p-Smad2/3)). Moreover, we further investigated whether WJ-MSCs pretreatment with different concentrations of TGFβ1 changes the anti-fibrotic properties of their exosomes. Methods: After isolation from the umbilical cord, WJ-MSCs were characterized by observing differentiation and measuring surface biomarkers using flowcytometry. The WJ-MSC-derived exosomes were extracted and identified using transmission electron microscopy (TEM), dynamic light scattering (DLS), and western blotting. Real-time PCR and western blot for extracellular matrix (ECM) and p-Smad2/3 expression detection were used to investigate the effect of exosomes from untreated and TGFβ1-pretreated WJ-MSCs on activated hepatic stellate cells (HSCs). Results: Phospho-Smad2/3, α-smooth muscle actin (α-SMA), and collagen1α1 levels were enhanced following treatment with TGFβ1, whereas E-cadherin was decreased. However, the outcomes were reversed after treatment with WJ-MSC-derived exosomes. Exosomes from TGFβ1-pretreated WJ-MSCs induced a significant decrease in p-Smad2/3 levels in activated HSCs, accompanied by the upregulation of E-cadherin gene expression and downregulation of α-SMA and collagen1α1 when compared to untreated WJ-MSC-derived exosomes. The p-Smad2/3 proteins were significantly decreased (fold change: 0.23, P-value < 0.0001) after exposure to low-dose TGFβ1-pretreated WJ-MSC-derived exosomes (0.1 ng/mL), showing the best effect on activated HSCs. Conclusions: Exosomes derived from untreated WJ-MSCs could regress TGFβ-Smad2/3 signaling and the expression of fibrotic markers in activated LX-2 cells. However, these effects were significantly profound with applying exosomes derived from 0.1 ng/mL TGFβ-pretreated WJ-MSCs. We also observed the dose-response effects of TGFβ on WJ-MSCs-derived exosomes. Therefore, exosomes derived from TGFβ-pretreated WJ-MSCs may be critical in improving fibrosis and benefit liver fibrosis patients.

Mesenchymal Stem Cell Exosomes Encapsulated Oral Microcapsules for Acute Colitis Treatment.

Mesenchymal stem cells derived exosomes (MSC-exos) exhibit an intrinsic and directed efficiency for multiple diseases, while their versatile and effective delivery to the target site is still a challenge. Herein, inspired by the acids and enzymes resistant property of sealing gelatin capsules, novel MSC-exo-encapsulated oral microcapsules are presented for colitis treatment. Based on a microfluidic electrospray technique, MSC-exos are first encapsulated in sodium alginate (SA) hydrogel microspheres with sustainable bioactivity. The resultant SA microspheres are then coated with a middle gelatin layer to protect MSC-exos from degradation. Especially, with an enteric coating-Eudragit FS30D on the outer layer, the resistance of the microcapsules in gastric juice is further enhanced. The prepared microcapsules maintain the stability and bioactivity of the MSC-exos during storage, protect them from the harsh conditions in the gastrointestinal tract, and enable the release of actives in the suitable sites for exerting their biological functions. In addition, these MSC-exos encapsulated microcapsules reduce the proinflammatory cytokines levels of inflammatory macrophages and impaired colonic epithelial cells, which exhibit superior damage repair ability in injured colon sites. Thus, it is believed that the proposed oral MSC-exos encapsulated microcapsules are valuable for many practically clinical treatments.

Association between Mesenchymal Stem Cells and COVID-19 Therapy: Systematic Review and Current Trends.

Background The novel coronavirus first emerged in Wuhan, China, and quickly spread across the globe, spanning various countries and resulting in a worldwide pandemic by the end of December 2019. Given the current advances in treatments available for COVID-19, mesenchymal stem cell (MSC) therapy seems to be a prospective option for management of ARDS observed in COVID-19 patients. This present study is aimed at exploring the therapeutic potential and safety of using MSC obtained by isolation from health cord tissues in the treatment of patients with COVID-19. Methods A systematic search was done based on the guidelines of the PRISMA 2020 statement. A literature search was executed using controlled vocabulary and indexing of trials to evaluate all the relevant studies involving the use of medical subject headings (MeSH) in electronic databases like PubMed, Embase, Scopus, Cochrane Central Register of Controlled Trials (CENTRAL), and clinicaltrials.gov up to 31 December 2021. The protocol was registered in the PROSPERO register with ID CRD42022301666. Findings. After screening finally, 22 remaining articles were included in this systematic review. The studies revealed that MSC exosomes are found to be superior to MSC alone in terms of safety owing to being smaller with a lesser immunological response which leads to free movement in blood capillaries without clumping and also cannot further divide, thus reducing the oncogenic potential of MSC-derived exosomes as compared to MSC only. The studies demonstrated that the lungs healed with the use of exosomes compared to how they presented initially at the hospital. MSCs are found to increase the angiogenesis process and alveolar reepithelization, reducing markers like TNF alpha, TGF beta, and COL I and III, reducing the growth of myofibroblasts and increasing survivability of endothelium leading to attenuated pulmonary fibrosis and even reversing them. Interpretation. We can conclude that the use of mesenchymal stem cells or their derived exosomes is safe and well-tolerated in patients with COVID-19. It improves different parameters of oxygenation and helps in the healing of the lungs. The viral load along with different inflammatory cells and biomarkers of inflammation tend to decrease. Chest X-ray, CT scan, and different radiological tools are used to show improvement and reduced ongoing destructive processes.

Mesenchymal Stem Cell Exosomes as Nanotherapeutic Agents for Neurodegenerative Diseases

Neurodegenerative diseases are systemic diseases with high heterogeneity and complicated etiology dependent on proper interneuronal communication, resulting in severe syndromes including cognitive impairment and dementia. The blood-brain barrier (BBB) remains Central nervous system (CNS) therapeutic delivery, a significant challenge without effective vivo therapeutic methods in clinical practice. Mesenchymal stem cells (MSC) with multi-directional differentiation potential have the characteristics of low immunogenicity, strong proliferation ability, immune regulation, and multi-directional differentiation potential. The repair effects have been identified mediated by transplanted MSCs paracrine factors, including exosomes and nanometer-sized cell communication mediators, to reduce tissue injury and enhance repair, growth, and regeneration. MSC-derived exosomes have become an attractive vehicle by passing through the blood-brain barrier (BBB), delivering therapeutic agents targeting the brain for treating autoimmune and neurodegenerative diseases. Safeties, convenience, and the effectiveness of MSC-derived exosomes have been demonstrated mainly through mechanistic clinical and preclinical evidence of potential nanotherapeutic agents for further prevalent use. Thus, we want to investigate the clinical applications of MSC-derived exosomes to reveal their regenerative treatment capacity from direct and indirect neuron repairment effect, reduced neuroinflammation, and nanotherapeutic agent advantage. This paper discusses the potential and practicality of using this novel cell-free entity of mesenchymal stem cell derivatives such as exosomes in vivo administration as a therapeutic modality for treating degenerative disease and pathologies and innovation and emerging trends in the field.

Mesenchymal Stem Cell Exosomal miR-146a Mediates the Regulation of the TLR4/MyD88/NF-κB Signaling Pathway in Inflammation due to Diabetic Retinopathy.

Diabetic retinopathy (DR) is the main cause of vision loss in diabetic patients, which cannot be completely resolved by typical blood sugar control. Inflammation influences the development of DR, so reducing the inflammatory response in DR patients is crucial to the prevention of DR. Therefore, we explored the regulatory effect of bone marrow mesenchymal stem cell (BMSC) exosomes on inflammation in DR mice. In order to analyze the mechanism of action, we used BMSC exosomal miR-146a to treat microglias in DR mice to observe cellular changes and expression of inflammatory factors. It was found that BMSC exosomal miR-146a reduced the levels of proliferating cell antigen and B-cell lymphoma-2 in microglias of DR mice and increased Bcl-2-related X with cysteine aspartic protease-3. By analyzing the expression of inflammatory factors, we found that BMSC exosomal miR-146a reduced the levels of TNF-α, IL-1β, and IL-6, which suggested that miR-146a can alleviate inflammation in DR mice. Further exploration found that miR-146a reduced the activity of TLR4 and increased the activity of MyD88 and NF-κB. Furthermore, the overexpression of TLR4 reversed the effects of miR-146a on the proliferation, apoptosis, and inflammation of microglias. Our study demonstrated that BMSC exosomal miR-146a can regulate the inflammatory response of DR by mediating the TLR4/MyD88/NF-κB pathway, providing an experimental basis for the prevention and treatment of DR.

Expression profile analysis of lncRNA in bone marrow mesenchymal stem cells exosomes of postmenopausal osteoporosis patients through microarray and bioinformatics analyses

BackgroundPostmenopausal osteoporosis (PMOP) is the most common bone metabolic disease affecting women worldwide. In this study, we investigate the role of long non-coding RNA (lncRNA) expression in exosomes obtained from bone marrow mesenchymal stem cells (BMSCs) of patients with PMOP. MethodsBMSCs from patients diagnosed with PMOP and healthy post-menopausalfemales as controls were isolated and cultured before exosome extraction. RNA microarray technology was used to identify differentially expressed lncRNAs in exosomes from BMSCs. Bioinformatics technology was utilized to analyze the roles of differentially expressed lncRNAs. Further, RT-qPCR was used to validate differentially expressed lncRNAs in 20 pairs of clinical samples. ResultsA total of 286 differentially expressed lncRNAs were detected in the exosomes from BMSCs unlike in the control group, among which 148 were up-regulated, whereas 138 were down-regulated. RT-qPCR identified five critical lncRNAs, including ENST00000593078, NR_120593, ENST00000422343, MEG3 and NR_029192. This was consistent with the microarray results and with a significant difference (P < 0.01). Based on the differentially expressed lncRNAs, we constructed lncRNA-miRNA-mRNA interaction networks. Functional analysis revealed that differentially expressed lncRNAs in patients with PMOP potentially target Wnt/β-catenin, MAPK, and PI3K-Akt pathways. ConclusionIn summary, we detected several dysregulated lncRNAs regulating PMOP progression in exosomes extracted from BMSCs of affected patients acting as novel biomarkers. This in turn provides valuable data for targeted treatment of PMOP. SubjectsGenomics; Molecular biology; Orthopedics; Women’s Health

Abstract 5369: LOXL1-AS1 contributes to medulloblastoma metastasis: A potential target for mesenchymal stem cell exosome-delivered gene therapy

Abstract Introduction: Medulloblastoma (MB) is the most common malignant tumor in children. MB-related treatment failures and deaths mostly result from metastasis. One of the key genetic drivers in MB is MYCN, whose aberrant expressions are associated with tumor aggressiveness and poor prognosis. Long non-coding (lnc)RNAs have been implied as potential targets for cancer treatment. More insights into the role of lncRNAs in MYCN-mediated MB progression would have both prognostic and therapeutic significance in metastatic MB. Cell-derived vesicles such as exosome can encapsulate siRNAs to improve their bioavailability and targeted delivery to brain tumors. Mesenchymal stem cell (MSC) is a readily available and modifiable source to produce functional exosomes. Objective: To identify the downstream lncRNAs of MYCN that contributes to MB metastasis, and to develop a novel gene therapy for MB by MSC exosome-delivered siRNAs to target these prometastatic lncRNAs. Methods: An RNA sequencing analysis was used to identify LOXL1-AS1 as a MYCN targeting gene in MYCN-overexpressing MB cells. The association of LOXL1-AS1 and MYCN was validated in MB cell lines by RT-qPCR and Western blot assay, and in tumor tissues by in-situ hybridization and immunohistochemistry. The anti-tumor effect of LOXL1-AS1 knockdown in MYCN-overexpressing MB cells was assessed in vitro by migration and sphere formation assays, and in an orthotopic xenograft mouse model by intracranial injection. To generate therapeutic exosomes, MSCs were transfected with exogenous LOXL1-AS1 siRNAs and then subjected to exosome characterization of size distribution and marker expression. The effect of exosome treatment on MB cells was evaluated using RT-qPCR and wound healing assay. Results: Elevated LOXL1-AS1 was found in MYCN-overexpressed MB cells compared to parental cells, and this association was confirmed in clinical tissue samples. LOXL1-AS1 expression was correlated with poor prognosis in SHH-α and SHH-γ MB subtypes of patients. Knockdown of LOXL1-AS1 significantly reduced cell migration and cancer stemness properties of MB cells in vitro, and suppressed orthotopic tumor growth, metastasis, and enhanced survival in xenografted mice. Exogenous siRNAs were successfully packaged into MSC exosomes by cell transfection. Treatment of MB cells with LOXL1-AS1 siRNA-loaded exosomes effectively reduced LOXL1-AS1 expression and inhibited cell migration compared to those treated with unmodified exosomes. Conclusion: Our study has proven the contributive role of LOXL1-AS1 in promoting cancer progression and metastasis of MYCN-associated MB, and provided in vitro evidence to support the feasibility of LOXL1-AS1 gene therapy through siRNA-loaded MSC exosomes. In vivo biodistribution and therapeutic efficacy assessment of MSC exosome-delivered LOXL1-AS1 siRNAs are being investigated in animal models. Citation Format: Anh Duy Do, Chia-Ling Hsieh, Tai-Tong Wong, Shian-Ying Sung. LOXL1-AS1 contributes to medulloblastoma metastasis: A potential target for mesenchymal stem cell exosome-delivered gene therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5369.

Current regenerative medicine-based approaches for skin regeneration: A review of literature and a report on clinical applications in Japan.

Current trends indicate a growing interest among healthcare specialists and the public in the use of regenerative medicine-based approaches for skin regeneration. The approaches are categorised in either cell-based or cell-free therapies and are reportedly safe and effective. Cell-based therapies include mesenchymal stem cells (MSCs), tissue induced pluripotent stem cells (iPSCs), fibroblast-based products, and blood-derived therapies, such as those employing platelet-rich plasma (PRP) products. Cell-free therapies primarily involve the use of MSC-derived extracellular vesicles/exosomes. MSCs are isolated from various tissues, such as fat, bone marrow, umbilical cord, menstrual blood, and foetal skin, and expanded ex vivo before transplantation. In cell-free therapies, MSC exosomes, MSC-derived cultured media, and MSC-derived extracellular vesicles are collected from MSC-conditioned media or supernatant. In this review, a literature search of the Cochrane Library, MEDLINE (PubMed), EMBASE, and Scopus was conducted using several combinations of terms, such as ‘stem’, ‘cell’, ‘aging’, ‘wrinkles’, ‘nasolabial folds’, ‘therapy’, ‘mesenchymal stem cells’, and ‘skin’, to identify relevant articles providing a comprehensive update on the different regenerative medicine-based therapies and their application to skin regeneration. In addition, the regulatory perspectives on the clinical application of some of these therapies in Japan are highlighted.

MiR-199a-3p in mouse bone marrow mesenchymal stem cell exosomes increases epithelial sodium channel expression in lung injury.

Acute lung injury (ALI) causes significant morbidity and mortality in critically ill patients, which often presents with extensive accumulation of activated inflammatory cells and diffused alveolar damage accompanied by oxidative stress. Exosomes are nanovesicles, which have notable anti-inflammatory and repair properties, thus alleviating the symptoms of ALI. Epithelial sodium channel (ENaC) is essential for the transepithelial absorption of Na+ and fluid from alveolar spaces. We studied the effects of bone marrow mesenchymal stem cell exosomes (BMSC-exo) on the apoptosis and protein expression of ENaC in primary mouse alveolar epithelial type 2 cells (AT 2 cells). Moreover, the change of miR-199a-3p in AT 2 cells was detected by qRT-PCR, and we studied the regulation of miR-199a-3p on ENaC protein expression. Our results demonstrated that BMSC-exo could not only improve viability and reduce apoptosis in AT 2 cells, but also enhance the expression of ENaC protein and miR-199a-3p. Meanwhile, the upregulation of miR-199a-3p resulted in increased expression of ENaC protein. In summary, the BMSC-exo could participate in the regulation of ENaC through miR-199a-3p originated from BMSC-exo, thereby providing a new pharmacological tool for the treatment of ALI.

Nebulization Therapy with Umbilical Cord Mesenchymal Stem Cell-Derived Exosomes for COVID-19 Pneumonia

BackgroundScientists have been facing numerous challenges in the development of an effective therapeutic strategy for the treatment of COVID-19 pneumonia. Several studies have suggested that improving patient immunity and reducing lung injury induced by SARS-CoV-2 may be effective for treating patients with COVID-19.MethodsA pilot trial of nebulization therapy with exosomes of mesenchymal stem cells (MSCs) was performed on seven patients with COVID-19 pneumonia. Exosomes secreted from MSCs were collected and purified using multiple ultrafiltration steps. All patients were treated with nebulization of MSC-derived exosomes, and primary safety and efficacy outcomes were evaluated.ResultsOur clinical study demonstrated that nebulization of MSC-derived exosomes is a novel method that might be utilized in the treatment of COVID-19 pneumonia. Nebulization of MSC-derived exosomes did not induce acute allergic or secondary allergic reactions but did promote the absorption of pulmonary lesions and reduce the duration of hospitalization for mild cases of COVID-19 pneumonia.ConclusionsNebulization of MSC-derived exosomes is a safe, effective, and simple method, and their application at the beginning of treatment may be more beneficial.Trial RegistrationChinese Clinical Trial Registry, ChiCTR2000030261. Registered on 26 February 2020.Graphical

Bone Marrow Mesenchymal Stem Cell-Exosomes (BMSC-ExO) Promote Osteogenic Differentiation In Vitro and Osteogenesis In Vivo by Regulating miR-318/Runt-Related Transcription Factor 2 (RUNX2)

Primary osteoporosis (PMOP) is characterized by bone mass reduction and bone microstructure destruction, increased bone fragility and prone to fracture, which is partially caused by ovarian dysfunction and decreased estrogen content. Bone marrow mesenchymal stem cell exosomes (BMSC-ExO) can improve PMOP. In this study, BMSC-EXO was used to study the role and function of miR-318 and Runx2 in PMOP. Human osteogenitor cells were isolated from PMOP patients with primary osteoporosis. After BMSC-exo treatment, miR-318 and Runx 2 level was tested by RT-qPCR and Western blot. In addition, mice in OVX group were treated with BMSC-ExO (bilateral ovaries were removed) to observe the effect of BMSC-ExO on bone tissue. Our results showed that BMSC-exo treatment significantly decreased miR-318 level, upregulated RUNX2 expression and increased ALP activity. In addition, BMSC-exo administration ameliorated the declined bone mass and bone formation in osteoporotic femurs in OVX mice. In conclusion, BMSC-Exo enhances Runx2 levels by down-regulation of miR-318, thereby promoting osteogenic differentiation of osteogenitor cells, providing new potential therapeutic targets for treating PMOP.

The Effect of Bone Marrow Mesenchymal Stem Cells-Exosomes (BMSC-EXO) on Tumor Angiogenesis and Its Mechanism in Ovarian Cancer Microenvironment

In ovarian cancer microenvironment, BMSC cells can differentiate into a variety of stem cells, thereby reducing the damage to tissues, and this effect lies in the exosomal substances secreted by BMSC cells. Then, in ovarian cancer microenvironment, whether BMSC-exo exhibited an effect on angiogenesis at the tumor site, and its possible molecular mechanism remains unclear. BALA nude mice and ovarian cancer tumor tissues were collected to isolate vascular endothelial cells which were then assigned into Control group, 40 μg/ml BMSC-exo group, 80 μg/ml BMSC-exo group, 120 μg/ml BMSC-exo group in the presence of Wnt/β-catenin inhibitor (PNU-74654) followed by analysis of proliferation and migration of ovarian cancer vascular endothelial cells (OCVECs) and the angiogenesis. 40 μg/ml and 80 μg/mlBMSC-exo group showed significantly higher cell proliferation than control group with higher cell number in 80 μg/ml BMSC-exo group than 40 μg/ml BMSC-exo group (P &lt; 0.05). The number of cell migration after BMSC-exo treatment was increased (P &lt; 0.05) and the tumor tissue showed obvious angiogenesis with more CD31-positive cells (P &lt; 0.05). PNU-74654 group showed significantly downregulated Wnt and β-catenin proteins (P &lt; 0.05) and lower cell number and higher migration rate of vascular endothelial cells (P &lt; 0.05). In conclusion, exosomes secreted by BMSC can repair damaged tissues possibly through activation of Wnt/β-catenin signaling pathway.

Human umbilical cord mesenchymal stem cells-derived exosomes for treating traumatic pancreatitis in rats

BackgroundThe therapeutic and protective effects of human umbilical cord mesenchymal stem cells-exosomes (hucMSC-Exs) on traumatic pancreatitis (TP) remain unknown. Here, we established a rat model of TP and evaluated and compared the therapeutic effects of hUC-MSCs and hucMSC-Exs.MethodsHucMSC-Exs were obtained by ultracentrifugation and identified using transmission electron microscopy and western blot analysis. TP rats were treated by tail vein injection of hUC-MSCs and hucMSC-Exs. Their homing in rats was observed by performing fluorescence microscopy. The degree of pancreatic tissue damage was assessed by HE staining, the expression levels of amylase, lipase, and inflammatory cytokines were detected by ELISA, apoptosis was detected by TUNEL assay, and the expression levels of various apoptosis-related proteins were detected by western-blot. The expression levels of apoptosis-related molecular markers were detected by RT-qPCR.ResultsThe colonization of exosomes was observed in pancreatic tissue. Compared to TP group, the histopathological score of pancreas was significantly decreased in the TP + hUC-MSCs group and TP + hucMSC-Exs group (P < 0.05). Compared to TP group, the activity of serum amylase and lipase was significantly decreased (P < 0.05). The expression levels of IL-6 and TNF-α were significantly decreased, while those of IL-10 and TGF-β were significantly increased (P < 0.05). The apoptosis index of the TP group was significantly increased (P < 0.05), whereas that of the TP + hUC-MSCs and TP + hucMSC-Exs groups was significantly decreased (P < 0.05). Compared to TP group, the expression levels of Bax, Bcl-2, and Caspase-3 were significantly decreased in the TP + hUC-MSCs group and TP + hucMSC-Exs group (P < 0.05).ConclusionHucMSC-Exs can colonize injured pancreatic tissue, inhibit the apoptosis of acinar cells, and control the systemic inflammatory response to facilitate the repair of pancreatic tissue.

Assessment of ultra-structure, viability and function of lipopolysaccharides-stimulated human dermal fibroblasts treated with chrysin and exosomes (in vitro study)

BackgroundLipopolysaccharides (LPS) stimulate production of inflammatory cytokines. Chrysin is flavonoid beneficial for treatment of inflammatory conditions. Bone marrow mesenchymal stem cell (BM-MSC) exosomes have regenerative ability in different tissues. ObjectiveTo assess potential role of chrysin and BM-MSC exosomes on ultra-structure, viability and function of humandermalfibroblasts-adult (HDFa) stimulated by LPS. MethodsHDFa cells were divided into: Group I: Cells received no treatment. Group II: Cells were stimulated with LPS. Group III: LPS stimulated cells were treated with chrysin. Group IV: LPS stimulated cells were treated with exosomes. ResultsAfter 48 h, ultrastructural examination of HDFa cells in Group I revealed intact plasma membrane and numerous cytoplasmic organelles. Group II displayed destructed plasma membrane and apoptotic bodies. Group III showed intact plasma membrane with loss of its integrity at some areas. Group IV demonstrated intact plasma membrane that showed fusion with exosomes at some areas. Statistical analysis of MTT represented highest mean value of cell viability% in Group IV followed by Groups III, I and II respectively. Statistical analysis of enzyme-linked immunosorbent assay (ELISA) showed the highest mean value of interleukin-1β (IL-1β) was in Group II followed by Groups III, IV and I, while highest mean values of interleukin-10 (IL-10), nuclear factor-erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins were in Group I, followed by Groups IV, III and II respectively. ConclusionsLPS have harmful consequences on ultra-structure, viability and function of HDFa cells. BM-MSC exosomes have better regenerative action on inflamed fibroblasts in comparison to chrysin.

Low-Intensity Pulsed Ultrasound Enhances the Efficacy of Bone Marrow-Derived MSCs in Osteoarthritis Cartilage Repair by Regulating Autophagy-Mediated Exosome Release.

ObjectiveThe present study explored whether low-intensity pulsed ultrasound (LIPUS) enhances the therapeutic efficacy of mesenchymal stem cells (MSCs) in osteoarthritis (OA) cartilage repair by regulating autophagy-mediated exosome release.DesignMSCs were isolated from the rat bone marrow and treated with rapamycin, 3-methyladenine, or LIPUS. The mechanism of the LIPUS-stimulated exosome release by MSCs was analyzed by inhibiting autophagy. In addition, the MSCs were co-cultured with OA chondrocytes and stimulated by LIPUS, with or without exosome release inhibitor intervention. The exosome release was detected through transmission electron microscopy (TEM), nanoparticle tracking analysis, and biomarker expression analysis. Autophagy was analyzed through TEM, autophagy-related gene expression analysis, and immunofluorescence analysis in vitro. Furthermore, a rat knee OA model was constructed and treated with MSCs, GW4869, and LIPUS. The cartilage repair was assessed through histopathological analysis and extracellular matrix protein expression analysis.ResultsThe in vitro results indicated that LIPUS promoted MSC exosome release by activating autophagy. The in vivo results demonstrated that LIPUS significantly enhanced the positive effects of MSCs on OA cartilage. These effects were significantly blocked by GW4869, an inhibitor of exosome release.ConclusionsLIPUS can enhance the therapeutic efficacy of MSCs in OA cartilage repair, and the underlying mechanism is related to the increase in autophagy-mediated exosome release.

MiR-139-5p Was Identified as Biomarker of Different Molecular Subtypes of Breast Carcinoma.

Located on chromosome 11q13.4, miR-139-5p has been confirmed by several studies as a possible attractive biomarker for cancer, including breast cancer, but its mechanism of correlation in different molecular subtypes of breast cancer has not been reported. In this study, comprehensive bioinformatics analysis was used to evaluate the expression of miR-139-5p in different molecular subtypes of breast cancer (luminal A, luminal B, HER2-enriched, and basal-like). The target genes of miR-139-5p were predicted by using an online database TargetScan and miRDB, and three key genes, FBN2, MEX3A, and TPD52, were screened in combination with differentially expressed genes in different molecular subtypes of breast cancer. The expression of the three genes was verified separately, and the genes were analyzed for pathway and functional enrichment. Bone marrow mesenchymal stem cells (BMSC) are another kind of highly plastic cell population existing in bone marrow besides hematopoietic stem cells. BMSC can affect the proliferation and migration of cancer cells, promote the metastasis and development of cancer, and regulate the tumor microenvironment by secreting exosome mirnas, thus affecting the malignant biological behavior of tumor cells. Finally, human bone marrow mesenchymal stem cells exosomes were obtained by ultracentrifugation, and the morphology of exosomes was observed by transmission electron microscopy. The expression of miR-139-5p in normal breast cells MCF-10A, human breast cancer cell line MDA-MB-231 cells, and BMSCs-derived exosomes were compared; the exosomes and MDA-MB-231 cells were co-cultured to observe their effects on the proliferation of the MDA-MB-231 cells. Human bone marrow mesenchymal stem cell-derived exosomes inhibited the growth of breast cancer cells and promoted the expression of FBN2, MEX3A, and TPD52 by transporting miR-139-5p.

ADSC-Exosomes Alleviate MTX-induced Rat Neuronal Damage by Activating Nrf2-ARE Pathway

The aim of this study was to analyze the efficacy and underlying mechanism of adipose-derived mesenchymal stem cell exosome (ADSC-exosomes)–mediated protection on methotrexate (MTX)–induced neuronal damage. We established a H2O2­induced oxidative stress model in vitro, as well as an MTX-induced neuronal damage rat model in vivo. We analyzed the effects of ADSC-exosomes on neuronal damage and Nrf2-ARE signaling pathway in rats and related mechanisms. The morphological and functional recovery of rat hippocampal neurons by ADSC-exosomes was examined by Nissl staining and modified neurological severity score (mNSS) score. The activation of Nrf2-ARE pathway effectively inhibited H2O2-induced oxidative stress. ADSC-exosomes treatment restored the activity of hippocampal neuronal cells, reduced ROS production, and inhibited hippocampal neuronal cells apoptosis. In in vivo experiments, ADSC­exosomes ameliorates MTX-induced hippocampal neuron damage by triggering Nrf2­ARE pathway, decreasing IL-6, IFN-, and TNF-a levels and TUNEL positive cells in hippocampus, and repairing hippocampal neuronal cell damage. ADSC­exosomes ameliorated MTX-induced neuronal damage and suppressed oxidative stress induced by neuronal damage through the activation of Nrf2-ARE signaling pathway.