Figure 3 - available via license: Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Content may be subject to copyright.
![A, Western blot analysis of mouse mesenchymal stem cells (MSCs) and exosomes by TSG101, CD63, and CD9. B, Transmission electron microscope images of control (blank-exosomes [Exos]) and ischemic myocardium-targeting peptide CSTSMLKAC (IMTP)-positive exosomes (IMTP-Exos). Bar=200 nm. C, Size distribution of blank-Exos and IMTP-Exos on the basis of nanoparticle tracking analysis measurements peaking at 134 and 135 nm, respectively.](publication/327175479/figure/fig3/AS:688267674546177@1541107209335/A-Western-blot-analysis-of-mouse-mesenchymal-stem-cells-MSCs-and-exosomes-by-TSG101.png)
A, Western blot analysis of mouse mesenchymal stem cells (MSCs) and exosomes by TSG101, CD63, and CD9. B, Transmission electron microscope images of control (blank-exosomes [Exos]) and ischemic myocardium-targeting peptide CSTSMLKAC (IMTP)-positive exosomes (IMTP-Exos). Bar=200 nm. C, Size distribution of blank-Exos and IMTP-Exos on the basis of nanoparticle tracking analysis measurements peaking at 134 and 135 nm, respectively.
Source publication
Background
Exosomes are membranous vesicles generated by almost all cells. Recent studies demonstrated that mesenchymal stem cell–derived exosomes possessed many effects, including antiapoptosis, anti‐inflammatory effects, stimulation of angiogenesis, anticardiac remodeling, and recovery of cardiac function on cardiovascular diseases. However, targ...
Contexts in source publication
Context 1
... obtained 400 to 600 lg (3.2- 4.8910 10 particles) of exosomes per 10 7 cells. Western blot analysis confirmed that mouse MSC exosomes expressed typical exosomal protein TSG101, CD63, and CD9 without contamination of cellular protein ( Figure 3A). The exosomes were morphologically homogeneous, with size ranging from 30 to 150 nm and peaking at 134 nm, as examined by NTA. ...
Context 2
... did not affect the physical properties of the IMTP-exosomes, according to the transmission electron microscope images ( Figure 3B) and the NTA results ( Figure 3C). ...
Citations
... Therefore, it is possible to modify the surface membrane of EVs through genetic engineering or chemical modification of their parent cells. To achieve this, fusion gene vectors can be constructed to encode membrane proteins associated with EVs, such as LAMP2B [60,61], CD63 [62,63], CD9 [64,65], or a mutant form of Nef (Nef mut ) [66]. The targeting peptides or ligands, such as ischemic myocardium targeting peptide (IMTP) [60], RVG peptide [61], the TNF-α binding domain of human TNF receptor-1 (hTNFR1) [62], or the RNA binding protein HuR [64], can be coexpressed on the membrane surface of specific EVs. ...
... To achieve this, fusion gene vectors can be constructed to encode membrane proteins associated with EVs, such as LAMP2B [60,61], CD63 [62,63], CD9 [64,65], or a mutant form of Nef (Nef mut ) [66]. The targeting peptides or ligands, such as ischemic myocardium targeting peptide (IMTP) [60], RVG peptide [61], the TNF-α binding domain of human TNF receptor-1 (hTNFR1) [62], or the RNA binding protein HuR [64], can be coexpressed on the membrane surface of specific EVs. This can be accomplished by creating fusion gene vectors and then transferring them into parent cells. ...
Background
Extracellular vesicles (EVs) derived from human adipose-derived mesenchymal stem cells (hADSCs) have shown great therapeutic potential in plastic and reconstructive surgery. However, the limited production and functional molecule loading of EVs hinder their clinical translation. Traditional two-dimensional culture of hADSCs results in stemness loss and cellular senescence, which is unfavorable for the production and functional molecule loading of EVs. Recent advances in regenerative medicine advocate for the use of three-dimensional culture of hADSCs to produce EVs, as it more accurately simulates their physiological state. Moreover, the successful application of EVs in tissue engineering relies on the targeted delivery of EVs to cells within biomaterial scaffolds.
Methods and Results
The hADSCs spheroids and hADSCs gelatin methacrylate (GelMA) microspheres are utilized to produce three-dimensional cultured EVs, corresponding to hADSCs spheroids-EVs and hADSCs microspheres-EVs respectively. hADSCs spheroids-EVs demonstrate excellent production and functional molecule loading compared with hADSCs microspheres-EVs. The upregulation of eight miRNAs (i.e. hsa-miR-486-5p, hsa-miR-423-5p, hsa-miR-92a-3p, hsa-miR-122-5p, hsa-miR-223-3p, hsa-miR-320a, hsa-miR-126-3p, and hsa-miR-25-3p) and the downregulation of hsa-miR-146b-5p within hADSCs spheroids-EVs show the potential of improving the fate of remaining ear chondrocytes and promoting cartilage formation probably through integrated regulatory mechanisms. Additionally, a quick and innovative pipeline is developed for isolating chondrocyte homing peptide-modified EVs (CHP-EVs) from three-dimensional dynamic cultures of hADSCs spheroids. CHP-EVs are produced by genetically fusing a CHP at the N-terminus of the exosomal surface protein LAMP2B. The CHP + LAMP2B-transfected hADSCs spheroids were cultured with wave motion to promote the secretion of CHP-EVs. A harvesting method is used to enable the time-dependent collection of CHP-EVs. The pipeline is easy to set up and quick to use for the isolation of CHP-EVs. Compared with nontagged EVs, CHP-EVs penetrate the biomaterial scaffolds and specifically deliver the therapeutic miRNAs to the remaining ear chondrocytes. Functionally, CHP-EVs show a major effect on promoting cell proliferation, reducing cell apoptosis and enhancing cartilage formation in remaining ear chondrocytes in the M1 macrophage-infiltrated microenvironment.
Conclusions
In summary, an innovative pipeline is developed to obtain CHP-EVs from three-dimensional dynamic culture of hADSCs spheroids. This pipeline can be customized to increase EVs production and functional molecule loading, which meets the requirements for regulating remaining ear chondrocyte fate in the M1 macrophage-infiltrated microenvironment.
Graphical Abstract
... CTPs can potentially be utilized in various applications. One prominent application is that these peptides can serve as vehicles for delivering therapeutic agents specifically to the damaged myocardium by conjugating with these agents [26][27][28] . This approach could minimize off-target effects of therapeutic agents and enhancing their therapeutic efficacy. ...
Heart failure remains a leading cause of mortality. Therapeutic intervention for heart failure would benefit from targeted delivery to the damaged heart tissue. Here, we applied in vivo peptide phage display coupled with high-throughput Next-Generation Sequencing (NGS) and identified peptides specifically targeting damaged cardiac tissue. We established a bioinformatics pipeline for the identification of cardiac targeting peptides. Hit peptides demonstrated preferential uptake by human induced pluripotent stem cell (iPSC)-derived cardiomyocytes and immortalized mouse HL1 cardiomyocytes, without substantial uptake in human liver HepG2 cells. These novel peptides hold promise for use in targeted drug delivery and regenerative strategies and open new avenues in cardiovascular research and clinical practice.
... Surface engineering of EVs involves modifying their membrane with targeted ligands, such as antibodies or peptides, to achieve specific interactions with receptors in target cells [80,87]. In the context of CVDs, this approach can be tailored to direct EVs to ECs, CMs, or immune cells. ...
Extracellular vesicles (EVs) are tiny vesicles released by various cells that contain a variety of proteins, lipids, and nucleic acids, which can have a wide range of effects on other cells. The dynamic composition and contents of EVs can serve as sensitive biomarkers for diagnosing and monitoring various cardiovascular diseases (CVDs). In addition to their diagnostic potential, EVs are therapeutic agents capable of precise modulation and amelioration of CVDs, because of their innate ability to encapsulate and deliver bioactive molecules. This growing field reveals the intricate interplay between EVs and cardiovascular pathophysiology, showing that EVs can act as messengers of intercellular communication for CVD regenerative therapy. Extracellular vesicles serve as dual agents in the field of theranostics, both as diagnostic biomarkers able to decode nuanced molecular signatures of CVDs and as potent vehicles for targeted therapeutic interventions. This review delves into the evolving landscape of EVs, uncovering their diagnostic and therapeutic prospects and emphasizing their growing importance in shaping the future of cardiovascular theranostics.
... Second, the wide application of stem cell-derived exosomes is hampered by problems in high-yield and scalable manufacturing processes for both stem cell culture and isolation [114]. Third, systemic delivery of exosomes required surface modification for targeted delivery and prevention of non-specific trapping in organs like the liver, lung, and spleen [115]. ...
Atherosclerotic cardiovascular disease (ASCVD) is an advanced chronic inflammatory disease and the leading cause of death worldwide. The pathological development of ASCVD begins with atherosclerosis, characterised by a pathological remodelling of the arterial wall, lipid accumulation and build-up of atheromatous plaque. As the disease advances, it narrows the vascular lumen and limits the blood, leading to ischaemic necrosis in coronary arteries. Exosomes are nano-sized lipid vesicles of different origins that can carry many bioactive molecules from their parental cells, thus playing an important role in intercellular communication. The roles of exosomes in atherosclerosis have recently been intensively studied, advancing our understanding of the underlying molecular mechanisms. In this review, we briefly introduce exosome biology and then focus on the roles of exosomes of different cellular origins in atherosclerosis development and progression, functional significance of their cargoes and physiological impact on recipient cells. Studies have demonstrated that exosomes originating from endothelial cells, vascular smooth muscle cells, macrophages, dendritic cells, platelets, stem cells, adipose tissue and other sources play an important role in the atherosclerosis development and progression by affecting cholesterol transport, inflammatory, apoptotic and other aspects of the recipient cells' metabolism. MicroRNAs are considered the most significant type of bioactive molecules transported by exosomes and involved in ASCVD development. Finally, we review the current achievements and limitations associated with the use of exosomes for the diagnosis and treatment of ASCVD.
... For example, expression of a fusion of the cardiomyocyte binding peptide with Lamp2b on EVs results in increased efficiency of EV uptake into cardiomyocytes and decreased cardiomyocyte apoptosis associated with higher cell retention (63). A fusion of the Lamp2b protein with the ischemic myocardium targeting peptide (CSTSMLKAC) expressed on MSC-derived EVs reduced fibrosis and enhanced vascular neogenesis, and cardiac function could be monitored in ischemic heart tissue (64). The therapeutic potential of EVs has been observed in skin repair (65) as well as in kidney (66). ...
... In addition to the use of phage display to target cardiomyocytes, this approach can be used for various other cell types to screen for peptides that have cell-type-specific affinity. In the example of targeting ischemic cardiac tissue, phage display was performed with a fusion with Lamp2b protein on the surface of EVs (64). EVs engineered with silk fibroin patch (SFP)-biding peptides that were identified by phage display showed dose-dependent accumulation of the SFP and enhanced wound closured efficacy in the diabetic wound model (89). ...
Extracellular vesicles (EVs) have important roles as mediators of cell-to-cell communication, with physiological functions demonstrated in various in vivo models. Despite advances in our understanding of the biological function of EVs and their potential for use as therapeutics, there are limitations to the clinical approaches for which EVs would be effective. A primary determinant of the biodistribution of EVs is the profile of proteins and other factors on the surface of EVs that define the tropism of EVs in vivo. For example, proteins displayed on the surface of EVs can vary in composition by cell source of the EVs and the microenvironment into which EVs are delivered. In addition, interactions between EVs and recipient cells that determine uptake and endosomal escape in recipient cells affect overall systemic biodistribution. In this review, we discuss the contribution of the EV donor cell and the role of the microenvironment in determining EV tropism and thereby determining the uptake and biological activity of EVs.
... It has been postulated that EV miRNAs with an ability to improve regenerative activities of recipient cells might attenuate the progression of HF [127]. In this study, EVs internalized by hypoxia-injured H9C2 cardiac cell attenuated inflammation, apoptosis, fibrosis, enhanced angiogenesis and cardiac function. ...
Physiologically, extracellular vesicles (EVs) have been implicated as crucial mediators of immune response, cell homeostasis, angiogenesis, cell differentiation and growth, and tissue repair. In heart failure (HF) they may act as regulators of cardiac remodeling, microvascular inflammation, micro environmental changes, tissue fibrosis, atherosclerosis, neovascularization of plaques, endothelial dysfunction, thrombosis, and reciprocal heart-remote organ interaction. The chapter summaries the nomenclature, isolation, detection of EVs, their biologic role and function physiologically as well as in the pathogenesis of HF. Current challenges to the utilization of EVs as diagnostic and predictive biomarkers in HF are also discussed.
... Their ability to serve as nanocarriers facilitates cell-mediated drug delivery, thereby maximizing therapeutic efficacy. Notably, certain exosomal proteins exhibit selective homing abilities, enhancing the efficiency of delivery [127]. The yield of exosomes is influenced by the type of cells involved, with immune cells often producing consistent and therapeutically potent yields. ...
Introduction
This study evaluates the effectiveness of a combined regimen involving injectable hydrogels for the treatment of experimental myocardial infarction.
Patient concerns
Myocardial infarction is an acute illness that negatively affects quality of life and increases mortality rates. Experimental models of myocardial infarction can aid in disease research by allowing for the development of therapies that effectively manage disease progression and promote tissue repair.
Diagnosis
Experimental animal models of myocardial infarction were established using the ligation method on the anterior descending branch of the left coronary artery (LAD).
Interventions
The efficacy of intracardiac injection of hydrogels, combined with cells, drugs, cytokines, extracellular vesicles, or nucleic acid therapies, was evaluated to assess the functional and morphological improvements in the post-infarction heart achieved through the combined hydrogel regimen.
Outcomes
A literature review was conducted using PubMed, Web of Science, Scopus, and Cochrane databases. A total of 83 papers, including studies on 1332 experimental animals (rats, mice, rabbits, sheep, and pigs), were included in the meta-analysis based on the inclusion and exclusion criteria.
The overall effect size observed in the group receiving combined hydrogel therapy, compared to the group receiving hydrogel treatment alone, resulted in an ejection fraction (EF) improvement of 8.87% [95% confidence interval (CI): 7.53, 10.21] and a fractional shortening (FS) improvement of 6.31% [95% CI: 5.94, 6.67] in rat models, while in mice models, the improvements were 16.45% [95% CI: 11.29, 21.61] for EF and 5.68% [95% CI: 5.15, 6.22] for FS.
The most significant improvements in EF (rats: MD = 9.63% [95% CI: 4.02, 15.23]; mice: MD = 23.93% [95% CI: 17.52, 30.84]) and FS (rats: MD = 8.55% [95% CI: 2.54, 14.56]; mice: MD = 5.68% [95% CI: 5.15, 6.22]) were observed when extracellular vesicle therapy was used. Although there have been significant results in large animal experiments, the number of studies conducted in this area is limited.
Conclusion
The present study demonstrates that combining hydrogel with other therapies effectively improves heart function and morphology. Further preclinical research using large animal models is necessary for additional study and validation.
Graphical abstract
... Previous studies revealed different ways of MSC-exos modification to improve their therapeutic potential [100] through loading exosomal lumen with endogenous or exogenous biomolecules such as nucleic acids, peptides, or drugs or through modifying MSC-exos' surface for targeting of a particular type of cells or tissues [101]. ...
Diabetic and chemotherapy-induced peripheral neuropathies are known for long-term complications that are associated with uncontrolled hyperglycemia and cancer treatment, respectively. Peripheral neuropathy often requires long-term therapy and could persist after treatment provoking detrimental effects on the patient’s quality of life. Despite continuous drug discoveries, development of efficient therapies is still needed for the significant management of diabetic and chemotherapy-induced peripheral neuropathy. Exosomes are nanosized extracellular vesicles that show great promise recently in tissue regeneration and injury repair compared to their parent stem cells. Herein, we provided a summary for the use of mesenchymal stem cell–derived exosomes in diabetic and chemotherapy-induced peripheral neuropathy in addition to recent advancements and ways proposed for the enhancement of their efficacy in these diseases.
Graphical abstract
... The lysosome-associated membrane glycoprotein 2b (Lamp2b) was a protein specifically expressed on exosome membrane, which can be genetically modified to induce engineering exosomes to specifically target tumor cells or myocardial cells (Tian et al., 2014;Wang et al., 2018a). A study fused an MSC-binding peptide E7 with the lamp2b yields to prepare engineering exosomes, which had SMSC targeting capability. ...
The treatment of bone or cartilage damage and inflammation-related diseases has been a long-standing research hotspot. Traditional treatments such as surgery and cell therapy have only displayed limited efficacy because they can’t avoid potential deterioration and ensure cell activity. Recently, exosomes have become a favorable tool for various tissue reconstruction due to their abundant content of proteins, lipids, DNA, RNA and other substances, which can promote bone regeneration through osteogenesis, angiogenesis and inflammation modulation. Besides, exosomes are also promising delivery systems because of stability in the bloodstream, immune stealth capacity, intrinsic cell-targeting property and outstanding intracellular communication. Despite having great potential in therapeutic delivery, exosomes still show some limitations in clinical studies, such as inefficient targeting ability, low yield and unsatisfactory therapeutic effects. In order to overcome the shortcomings, increasing studies have prepared genetically or chemically engineered exosomes to improve their properties. This review focuses on different methods of preparing genetically or chemically engineered exosomes and the therapeutic effects of engineering exosomes in bone regeneration and anti-inflammation, thereby providing some references for future applications of engineering exosomes.
... EVs carrying cardiac tissuespecific peptides have shown improved cardiac targeting [175]. In another study, the ischemic myocardial targeting peptide was incorporated into the EV membrane protein LAMP-2B derived from MSCs through genetic engineering [176]. After being intravenously injected into mouse models, these modified EVs were shown to aggregate higher in the ischemic heart region than unmodified EVs. ...
... After being intravenously injected into mouse models, these modified EVs were shown to aggregate higher in the ischemic heart region than unmodified EVs. Additionally, in a mouse model of myocardial infarction, these modified EVs significantly reduced inflammation and cardiomyocyte apoptosis, promoted angiogenesis, and enhanced cardiac function ( Figure 6) [176]. ...
Targeted delivery of anti‐tumor drugs and overcoming drug resistance in malignant tumor cells remain significant clinical challenges. However, there are only few effective methods to address these issues. Extracellular vesicles (EVs), actively secreted by cells, play a crucial role in intercellular information transmission and cargo transportation. Recent studies have demonstrated that engineered EVs can serve as drug delivery carriers and showed promising application prospects. Nevertheless, there is an urgent need for further improvements in the isolation and purification of EVs, surface modification techniques, drug assembly processes, and precise recognition of tumor cells for targeted drug delivery purposes. In this review, we summarize the applications of engineered EVs in cancer treatment and overcoming drug resistance, and current challenges associated with engineered EVs are also discussed. This review aims to provide new insights and potential directions for utilizing engineered EVs as targeted delivery systems for anti‐tumor drugs and overcoming drug resistance in the near future.
