Tingyu Yang
Jiapan An1
Bin Li
Zhimin Dou- 1The First School of Clinical Medicine, Lanzhou University, Lanzhou, China
- 2Department of Critical Care Medicine, The First Hospital of Lanzhou University, Lanzhou, China
Pyroptosis is a newly discovered form of inflammatory programmed cell death, which is frequently involved in the occurrence and development of various diseases. The primary mechanism underlying pyroptosis is the formation of membrane pores mediated by activated pyroptosis-related proteins. The expression levels of these pyroptosis-related proteins serve as crucial biomarkers for assessing the degree of pyroptosis. Modulating pyroptosis can alleviate tissue and organ damage in diseases and promote tissue and organ repair. Therefore, regulating pyroptosis is considered a potential therapeutic strategy. In recent years, mesenchymal stem cell-derived exosomes (MSC-Exos) have emerged as a novel therapeutic tool for pyroptosis due to their carrier properties. MSC-Exos can mitigate tissue damage in various diseases by regulating pyroptosis, thus emerging as strong candidates for disease treatment. Owing to their multifunctionality, MSC-Exos exert different effects by mediating different pathways in the treatment of various diseases. This review summarizes the mechanisms of pyroptosis and the research progress on MSC-Exos-regulated pyroptosis and outlines the existing challenges for the clinical translation of MSC-Exos. Collectively, MSC-Exos can not only precisely regulate the pyroptosis process but also provide new perspectives and approaches for future disease treatment. Therefore, MSC-Exos possess substantial potential for clinical translation.
1 Introduction
Pyroptosis is a form of inflammatory programmed cell death regulated by genes that can be activated in response to external stimuli and plays a significant role in the pathogenesis of various diseases (1, 2). The hallmarks of pyroptosis include pore formation in the plasma membrane, cell swelling, membrane rupture, and the release of pro-inflammatory cytokines, ultimately triggering an inflammatory response (3). Under physiological conditions, pyroptosis serves as an important defense mechanism against pathogen invasion by inhibiting intracellular pathogen replication, activating immune cells to phagocytose and kill pathogens, and releasing cellular contents as danger signals to modulate the innate immune response (4, 5). However, once pyroptosis becomes dysregulated, it can activate the inflammatory response in neighboring cells and tissues, thereby exacerbating inflammatory damage. In severe cases, this can lead to tissue damage and disease progression, adversely affecting patient prognosis (6). As a result, it is crucial to identify effective means to precisely regulate pyroptosis.
In recent years, stem cells with self-renewal and multidirectional differentiation potential have been proven to be applicable for the treatment of various diseases (7–9). Among them, mesenchymal stem cells (MSCs) have become the most commonly studied type due to their minimal ethical controversy, wide availability, and ease of acquisition (10, 11). Studies have shown that MSCs possess a variety of biological functions, including the regulation of immune responses, modulation of cell death, and promotion of tissue repair. MSCs can be used to treat a range of diseases, such as osteoarthritis, pulmonary fibrosis, bone marrow injury, myocardial injury, and knee cartilage damage (12, 13). Meanwhile, an increasing amount of evidence indicates that exosomes derived from MSCs (MSC-Exos) not only retain the therapeutic effects of the parental MSCs but also avoid the risks associated with live cell therapy (14, 15). Thus, applying MSC-Exos as a substitute for MSCs in cell-free therapy may be a focus of future research and clinical treatment.
This review aimed to elucidate the pathological mechanisms of pyroptosis, summarize the mechanisms by which MSC-Exos modulate pyroptosis and the associated signaling pathways, and review the recent research progress in this area. This effort is intended to explore the clinical application potential of MSC-Exos and provide theoretical support and practical references for future therapies targeting pyroptosis in various diseases.
2 Pyroptosis
Pyroptosis is a form of programmed cell death mediated by the gasdermin (GSDM) family protein. The gasdermin protein family functions as the effector molecules of pyroptosis. These proteins can be cleaved by active caspases, releasing an N-terminal domain (GSDM-NT) with pore-forming potential (16). The GSDM-NT can insert into the cell membrane and oligomerize, leading to the formation of pores with an inner diameter of approximately 15 nm on the membrane surface, thereby triggering pyroptosis (17, 18). Depending on the type of inflammatory caspase, pyroptosis can be categorized into the canonical pathway and the non-canonical pathway (19, 20).
2.1 Classical pyroptosis pathway
The classical pathway is regulated through the activation of caspase-1 by the inflammasome complex (Figure 1). To be specific, pattern recognition receptors (PRRs) on the cell surface can recognize a variety of pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs) (21). Activated PRRs can activate the nuclear factor-κB gene (NF-κB), which in turn promotes the transcription of pyroptosis-related molecules such as nucleotide-binding oligomerization domain-like receptor 3 (NLRP3) and pro-interleukin (IL)-1β. Subsequently, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and pro-caspase-1 are recruited to assemble into the inflammasome (22, 23). After the formation of the inflammasome, pro-caspase-1 undergoes self-cleavage to form the active caspase-1. Active caspase-1 can cleave and remove the C-terminal domain of the GSDMD to form active GSDMD-NT, thereby triggering pyroptosis (24). Meanwhile, active caspase-1 can also cleave pro-IL-1β and pro-IL-18 to form mature IL-1β and IL-18. These inflammatory mediators can be released into the extracellular space through GSDMD pores to activate local immune cells and trigger an inflammatory storm (25, 26).
Figure 1. Classical pyroptosis pathway. DAMPs, pathogen-associated molecular patterns; DAMPs, damage-associated molecular patterns; PRR, pattern recognition receptors; NF-κB, nuclear factor-κB gene; ASC, apoptosis-associated speck-like protein containing CARD; PYD, PYRIN Domain; GSDM, gasdermin; ESCRT, endosomal sorting complex required for transport; IL-1β, interleukin-1β; IL-18, interleukin-18.
2.2 Non-classical pyroptosis pathway
The non-canonical pyroptosis pathway is directly triggered by lipopolysaccharide (LPS) and mediated by caspase-4/5/11 (Figure 2). Initially, LPS can enter cells through damage to the host cell membrane, bacterial outer membrane vesicles, or binding to high-mobility group box 1 (HMGB1) (27–29). Pro-caspase-4/5/11 can directly bind to intracellular LPS, inducing its own activation to produce caspase - 4/5/11 (30, 31). Active caspase-4/5/11 can cleave GSDMD into GSDMD-NT, causing pyroptosis (32–36). However, caspase-4/5/11 cannot cleave pro-IL-1β/pro-IL-18, but they can mediate the maturation and secretion of IL-1β/IL-18 by activating the NLRP3/caspase-1 pathway (37). In addition, recent studies have found that apoptotic caspases can also mediate pyroptosis (38–40). For instance, caspase-4/11 can cleave and activate pro-caspase-3, cleaving GSDME and triggering pyroptosis (41, 42). Caspase-6/8 can drive pyroptosis by modulating the activity of caspase-11 (43–46). Notably, several studies have revealed that granzyme A (GzmA) and GzmB can cleave GSDMB and GSDME, respectively, to produce active GSDMB/E-NT fragments (38, 47, 48). Neutrophil elastase is also capable of cleaving GSDMD to induce pyroptosis (39, 49).
Figure 2. Non-classical pyroptosis pathways. LPS, lipopolysaccharide; IL-1β, interleukin-1 β; IL-18, interleukin-18; GSDM, gasdermin; ELANE, Neutrophil elastase.
These findings enrich our understanding of pyroptosis. It is noteworthy that recent studies have shown that the organism can remove GSDMD pores through the endosomal sorting complex required for transport (ESCRT) to inhibit pyroptosis and thus repair the plasma membrane (50). In addition, GSDMD pores can regulate the secretion of IL-1β in living cells without causing cell death (51, 52). These results suggest that pyroptosis is modulatable, providing a theoretical basis for targeting pyroptosis in disease treatment.
3 Overview of MSC-exos
In recent years, an increasing number of studies have demonstrated that MSCs mainly exert therapeutic effects such as anti-inflammation, immune regulation, antioxidation, and promotion of tissue injury repair through their paracrine factors, demonstrating great potential in treating various diseases (53–55). Exos, as key substances secreted by MSCs, are gradually being regarded as an alternative to stem cell therapy due to their multiple advantages, such as biocompatibility, modifiability, and non-cytotoxicity (56–59).
3.1 Biogenesis of exos
Exosomes originate from lipid raft microdomains of the endoplasmic reticulum’s plasma membrane and are nanoscale particles with a diameter of approximately 40–160 nm (60, 61). The biogenesis of exosomes is a continuous and strictly regulated process, mainly involving two invaginations of the plasma membrane and the formation of multivesicular bodies (MVBs) (Figure 3A). Initially, the plasma membrane invaginates from the cell surface and the extracellular environment to encapsulate proteins and bud inward, forming early sorting endosomes (ESEs) [77, 85]. Subsequently, ESEs fuse with each other to form late sorting endosomes (LSEs). The limiting membrane of LSEs invaginates again to form MVBs [84, 86]. Ultimately, the interaction between the generated MVBs and the plasma membrane releases vesicular components, which are referred to as exosomes (62, 63). This process can be mediated by either the ESCRT pathway or non-ESCRT pathways (64–67). After secretion, exosomes facilitate intercellular communication by interacting with and fusing with recipient cells (Figure 3C). Additionally, various molecules, including lipids like ceramide, heat shock proteins, lactadherin, GTPases, annexins, platelet-derived growth factor receptors, and tetraspanins, play roles in the formation of exosomes (68–70).
Figure 3. Schematic illustration of the biogenesis, compositions, and also release of the exosome. (A) Biogenesis of exosomes. (B) Compositions of exosomes. (C) Uptake mechanisms of exosomes by target cells. Following MVBs incorporation with the cellular membrane, the release of exosome into the extracellular space is accomplished, and finally the released molecules are conveyed to recipient cells through endocytosis, or direct membrane fusion, or receptor‐ligand interfaces. GA, Golgi apparatus; ER, endoplasmic reticulum; ESEs, early sorting endosomes; LSEs, late-stage sorting endosomes; ILVs, intraluminal vesicles; MVB, multivesicular bodies; MHC, major histocompatibility complex; HSPs, heat shock proteins.
Although we have gained some understanding of the molecules involved in the biogenesis of exosomes, it is crucial to further investigate the underlying mechanisms by which these molecules precisely regulate exosome biogenesis. This will not only enhance our ability to produce exosomes in a targeted manner but also hold significant implications for the development of cell-free therapies.
3.2 Isolation of exosomes
To date, there is no consensus on the “gold standard” method for exosome isolation (71). Although differential ultracentrifugation, density gradient ultracentrifugation, and tangential flow filtration have all been successfully employed to isolate exosomes, each method has its own advantages and disadvantages (72). Among them, differential high-speed ultracentrifugation is the most widely utilized and traditional method due to its simple protocol, efficiency, and high purity (73). Density gradient ultracentrifugation can isolate exosomes with higher purity than traditional ultracentrifugation by separating particles through layers of biocompatible media with different densities (74). However, none of these methods can distinguish exosomes from microvesicles and other vesicles with overlapping size ranges (75). Although capture methods based on immune affinity can differentiate various exosomes via surface markers of extracellular vesicles, their yield is too low (75, 76). In addition, in recent years, a variety of new methods have emerged, such as low-speed centrifugation based on polyethylene glycol, enrichment methods based on antibodies and filters, methods combining acoustics and microfluidics, and commercial kits (77–79). However, whether these new methods can effectively isolate exosomes has not been fully evaluated.
3.3 Characteristics of MSC-exos
MSC-Exos selectively package a variety of biomolecules, including proteins, mRNA, long noncoding RNA, miRNA, metabolic enzymes, and lipids (80–83) (Figure 3B). By facilitating the transfer of these molecules to target cells to mediate intercellular communication, exosomes exert biological effects similar to those of MSCs (84–86). Additionally, other molecules, including tetraspanins, heat shock proteins, and RNA-binding proteins, can also be packaged into exosomes, where they assist in the assembly and intracellular transport of exosomes (80, 87, 88). In addition to their inherent qualities, MSC-Exos also serve as an ideal delivery system for therapeutic compounds such as genes, drugs, enzymes, and RNA to reach specific cells (89–92). MSC-Exos have been shown to protect their cargo from degradation and promote intracellular uptake via endocytosis (93). Compared with MSCs, MSC-Exos possess a series of advantages, including low immunogenicity, high stability, preferential targeting of damaged tissues, and simple storage (94–96). Exosomes can initiate various pathophysiological responses in recipient cells, such as cell proliferation, differentiation, and development, immune regulation, homeostasis, and neurocommunication, by interacting with receptors and mediating signaling pathways (97). These advantages make them a promising cell-free alternative to existing stem cell therapies and hold great potential for disease treatment.
4 Effects of MSC-Exos on pyroptosis
Exosomes, as the principal mediators of the therapeutic effects of MSCs, possess inherent nanocarrier properties and contain a variety of effector molecules, including miRNAs, proteins, and long non-coding RNAs (98). A substantial body of research indicates that MSC-Exos can modulate the progression of various diseases through various mechanisms, such as influencing inflammasome activation, pyroptosis-related molecules, or the activation of specific signaling pathways, thereby exerting therapeutic effects (Table 1).
Table 1. Effects of application of mesenchymal stem cell-derived exosomes regulate pyroptosis in various disease models.
4.1 Pyroptosis regulation by MSC-exos-derived miRNAs
MiRNAs are a type of endogenous small non-coding RNA molecules that regulate various cellular activities, including gene expression, cell differentiation, proliferation, and apoptosis. They function by binding to complementary sequences in the 3’ untranslated region (3’ UTR) of target mRNAs, thereby promoting the translational inhibition and degradation of these mRNAs and thus regulating the expression and translation efficiency of mRNAs in target cells (99, 100). MiRNAs carried by MSC-exos can modulate pyroptosis by affecting the expression of inflammasomes and pyroptosis-related signaling pathways, thereby influencing the occurrence and development of diseases and providing new strategies and targets for the treatment of diseases.
4.1.1 Regulation of inflammasome activation
MSC-Exos can deliver miRNAs that directly target inflammasome mRNAs, leading to mRNA degradation and reduced protein expression, thereby modulating gene expression and influencing the occurrence and development of diseases, providing new ideas for the treatment of pyroptosis-related diseases.
Numerous studies have shown that miR-223 in MSC-Exos can exert protective effects in various disease models by targeting and inhibiting NLRP3 (101–104). Specifically, in an ovalbumen-induced asthma rat model, MSC-Exos can deliver miR-223-3p to inhibit the formation of the NLRP3 inflammasome in tracheal epithelial cells, thereby blocking the ASC/caspase-1/GSDMD signaling pathway and reducing airway inflammation and remodeling (104). In a rheumatoid arthritis model, miR-223 in MSC-Exos can target NLRP3 mRNA in macrophages, thereby reducing the release of pro-inflammatory factors such as IL-1β, TNF-α, and IL-18 and alleviating joint damage in rats (102). In an LPS-induced acute uterine injury model, miR-223-3p from MSC-Exos can promote the degradation of NLRP3 in endothelial progenitor cells, reverse the toxic effects of LPS on endothelial progenitor cells, and improve LPS-induced acute uterine injury in mice (103). In the autoimmune hepatitis cellular model, MSC-Exos can deliver miR-223b to the target and inhibit the expression of NLRP3 and caspase-1, thereby alleviating hepatocyte damage (101). In a silicosis mouse model, miR-223-3p derived from MSC-exos can bind to circPWWP2A in macrophages, thereby targeting and inhibiting the activation of NLRP3, which leads to the alleviation of silica-induced pulmonary inflammation and fibrosis (105).
Besides, other miRNAs also possess unique regulatory characteristics. For instance, studies have shown that miR-539-5p from MSC-Exos can target NLRP3 in intestinal epithelial cells, modulating the process of pyroptosis in a mouse model of inflammatory bowel disease (IBD), thereby alleviating the progression of IBD (106). MSC-Exos carrying miR-22 can improve memory in APP/PS1 double-transgenic Alzheimer’s disease model mice by inhibiting the expression of GSDMD, NLRP3, and caspase-1 in neurons (107). MSC-Exos overexpressing miR-188-3p can inhibit autophagy and pyroptosis in Parkinson’s disease mouse and cell models by targeting CDK5 and NLRP3 (108). The miR-26a-5p derived from MSC-Exos can reduce NLRP3 inflammasome-mediated pyroptosis and alleviate high-glucose induced pyroptosis in retinal ganglion cells by suppressing the protein expression levels of NLRP3, caspase-1, and GSDMD (109). MiR-302c and miR-410 in MSC-Exos can reduce the expression of caspase-1, IL-1β, and GSDMD by inhibiting the activation of the NLRP3 inflammasome, thereby inhibiting pyroptosis of nucleus pulposus cells (NPCs) in a mouse model of intervertebral disc degeneration (110, 111). MiR-320b and miR-182-5p in MSC-Exos can negatively regulate the expression of NLRP3, reducing cell pyroptosis of cardiomyocytes caused by myocardial ischemia/reperfusion, thereby alleviating myocardial injury in animal models (112, 113).
These findings indicate that a variety of miRNAs in MSC-Exos can regulate the pathological process of cells by inhibiting inflammasome activation and blocking downstream inflammatory cascades. However, the specific mechanisms by which MSC-Exos regulate NLRP3 to inhibit pyroptosis are still not fully understood.
4.1.2 Regulation of caspase family activity
The caspase family plays a crucial role as the key protease in pyroptosis execution and is responsible for cleaving the GSDM family and activating proinflammatory factors. Studies have shown that specific miRNAs can inhibit the progression of pyroptosis-related diseases by modulating the activity of caspase family proteins.
It has been found that miR-203a-3p.2 from MSC-Exos can reduce macrophage pyroptosis and thus alleviate colitis by inhibiting the activation of caspase - 11/4 in mice (114). In addition, it has been suggested that MSC-Exos can deliver miR-128-3p, miR-138-5p, and miR-221-3p to inhibit the expression of caspase-3, thereby reducing pyroptosis and inflammation in hypoxia-reperfusion myocardial cells (115). In a high-glucose-induced renal tubular epithelial cell pyroptosis model, miR-30e-5p from MSC-derived exosomes can inhibit caspase-1 activation and mediate GSDMD cleavage by targeting the RNA-binding protein embryonic lethal abnormal vision-like 1, thereby suppressing cell pyroptosis (116).
These observations demonstrate that miRNAs from MSC-Exos can also effectively inhibit cell pyroptosis by regulating the activity of the caspase family, providing new strategies and targets for the treatment of related diseases. However, the detailed mechanisms underlying miRNA regulation of caspase family activity are still not well understood.
4.1.3 Regulation of pyroptosis-related signaling pathways
MSC-Exos can precisely regulate key signaling pathways such as NF-κB, PI3K/AKT, YAP/β-catenin, and STAT1 by delivering miRNAs, thereby forming a signaling network that inhibits pyroptosis (117–120).
For example, miR-326 from MSC-Exos can target HDAC3 and the STAT1/NF-κB p65 signaling pathway, inhibiting the expression of HDAC3 and NF-κB p65 while promoting the expression of STAT1, acetylated STAT1, and acetylated NF-κB p65 in chondrocytes. This process reduces the expression of pyroptosis-related proteins such as NLRP3, ASC, GSDMD, and caspase-1, thereby inhibiting chondrocyte pyroptosis and improving osteoarthritis in rats (120). Additionally, it has been found that the bioactive substances in MSC-Exos can target and regulate key upstream targets such as TRAF6, TXNIP, FOXO3, ELAVL1, and CMPK2 (116, 121–123). For example, both in vitro and in vivo studies have revealed that miR-202-5p from MSC-Exos can target and inhibit CMPK2 in alveolar epithelial cells, thereby suppressing the expression of NLRP3 and inhibiting the progression of lung ischemia-reperfusion injury (123). MiR-17 in MSC-Exos can inhibit the activation of the inflammasome in hepatic macrophages by targeting TXNIP, thereby reducing liver damage in a mouse model of acute liver failure (124). MiR-324-5p in MSC-Exos can inhibit the expression of pyroptosis-related proteins by suppressing the NF-κB signaling pathway, alleviating hypoxia-reperfusion injury to cardiomyocytes (115). MiR-100-5p in MSC-Exos can inhibit pyroptosis-related proteins by targeting the miR-100-5p/FOXO3/NLRP3 pathway, thereby reducing pyroptosis in hypoxia-reperfusion cardiomyocytes (125). MiR-146a-5p derived from MSC-Exos targets and inhibits TRAF6, thereby reducing pyroptosis of microglia in mouse models of chronic inflammatory pain. This intervention effectively diminishes neuroinflammation and alleviates inflammatory pain (122). MiR-140-3p delivered by MSC-Exos targets HMGB1 to modulate S-lactoylglutathione metabolism, thereby inhibiting pyroptosis and inflammatory responses in microglia induced by LPS. This mechanism alleviates sepsis-associated brain injury in mice (126). MiR-155-5p from MSC-Exos can promote autophagy and inhibit pyroptosis by targeting TGFβR2 in NPCs, thereby alleviating intervertebral disc degeneration in mice (127). MSC-Exos can modulate the nuclear factor erythroid 2-related factor 2 signaling pathway by delivering miR-23 b and targeting PTEN in microglia to inhibit the activation of NLRP3, ultimately alleviating pyroptosis and promoting neurological recovery in rats with cerebral hemorrhage (128). Exosomes carrying miR-367-3p derived from MSCs can inhibit muscle pyroptosis in a mouse model of hindlimb ischemia-reperfusion (I/R) injury by targeting the enhancer of zeste homolog 2 (129).
The above studies have elaborated in detail the specific pathways by which MSC-Exos regulate pyroptosis by delivering internal miRNAs, not only providing new ideas for the treatment of related diseases but also offering directions for subsequent therapeutic research.
4.2 Other molecular components in MSC-exos regulate pyroptosis
Apart from the extensively studied miRNAs, other types of RNAs present in MSC-Exos also have the ability to inhibit pyroptosis.
CircRNA, a newly discovered non-coding RNA, forms a circular structure through backsplicing and has remained conserved during evolution. Current research has linked it to various disease processes (130, 131). For example, a recent study reported that circ-003564 from MSC-Exos can alleviate pyroptosis in spinal cord neurons and mitigate spinal cord injury in rats by suppressing the activation of the pyroptosome and inflammatory factors (132). CircHIPK3 from MSC-Exos targeted the miR-421/FOXO3a pathway in myoblasts. By downregulating miR-421 and increasing the expression of FOXO3a, it reduces pyroptosis and repairs ischemic muscle injury in mice (133).
Moreover, lncRNAs, a non-coding RNA molecule longer than 200 nucleotides, can regulate gene expression by interacting with miRNA, mRNA, or proteins. Research indicates that lncRNA KLF3-AS1 derived from MSC-Exos can modulate the miR-138-5p/Sirt1 axis, inhibiting pyroptosis in cardiomyocytes, thereby slowing the progression of myocardial infarction in rats (134). Derived from MSC-Exos, the lncRNA XIST is capable of interacting with miR-214-3p to relieve the inhibitory effect of miR-214-3p on Arl2, thereby reducing pyroptosis of cardiomyocytes induced by atrial fibrillation and alleviating cardiac injury in a mouse model (135).
Although research on the regulation of pyroptosis by MSC-Exos is limited, existing studies have sufficiently demonstrated that MSC-exos can alleviate diseases by modulating pyroptosis-related pathways. A deeper investigation into these regulatory mechanisms will enhance our understanding of the roles of MSC-Exos and the mechanisms of diseases, while providing a theoretical basis and practical references for targeting pyroptosis in disease treatment.
5 Critical gaps and challenges
Despite the promising therapeutic potential of MSC-Exos in modulating pyroptosis, several critical gaps and challenges must be addressed before their clinical application.
5.1 Identification of pyroptosis
Although pyroptosis can be distinguished from other modes of programmed cell death, such as apoptosis and necrosis, based on its unique morphological characteristics, key molecular events, and associated inflammatory responses, the complex interplay among programmed cell death pathways means that no single identification method is absolutely specific. For instance, studies have revealed that caspase-1/8 exhibits functional pleiotropy, capable of inducing not only pyroptosis but also apoptosis. Moreover, the morphological similarities at the terminal stages of different death modalities further complicate the identification process (136). Therefore, it is currently not feasible to rely solely on any single indicator to confirm pyroptosis.
In basic research, a multifaceted approach is typically employed to assess the occurrence of pyroptosis. This includes microscopic observation of cellular morphological changes, molecular biological detection of caspase-1 activation, and the activation of the pyroptosis execution protein GSDMD, as well as the detection of mature IL-1β and IL-18 release in the cell supernatant (136). Only through this multi-angular, corroborative method can a rigorous and reliable identification of pyroptosis be achieved.
However, this multi-step, multi-technological identification method is not only technically complex but also costly. Additionally, although the cleavage of GSDMD, the activation of the NLRP3 inflammasome, and the release of IL-1β/IL-18 are important markers of pyroptosis, the specificity and sensitivity of these markers still pose certain challenges. The heterogeneity of diseases further increases the complexity of marker identification. Thus, the precise and efficient identification of pyroptosis remains an urgent challenge that needs to be addressed.
5.2 MSC-exos heterogeneity
The heterogeneity of MSCs-Exos may be conceptualized based on their size, content, and particularly cellular origin (80). Based on the refined classification of extracellular vesicles, exosomes contain subpopulations defined by a distinct size range (137). Size heterogeneity could be due to uneven invagination of the limiting membrane of MVBs, leading to different total amounts of material within the vesicles, or because the isolation process includes other types of vesicles (138–140). The microenvironment of cells and their inherent biological properties may also influence the types and amounts of exosome contents and potentially affect the surface biological markers of exosomes. Therefore, as observed in the analysis of miRNA content within exosomes, not all exosomes contain a similar abundance of a given molecule (141).
Moreover, the effects of exosomes on target cells can vary due to the different expression of cell surface receptors. In different target cell types, this functional heterogeneity can result in the coexistence of exosome-induced cell survival, apoptosis, and immunomodulation. Heterogeneity can also be based on the organ and tissue of origin of the exosomes. Exosomes secreted by MSCs from different tissues carry distinct biological components, endowing exosomes with unique biological functions (142). For example, exosomes secreted by MSCs derived from adipose tissue have weaker immunomodulatory properties (143), while exosomes from MSCs derived from umbilical cord tissue have stronger renewal capabilities and more effective gene transfection (144). The combination of all these features has the potential to increase the complexity of exosome-based therapies.
5.3 Insufficient in vivo validation
Over the past few years, the therapeutic potential of MSC-Exos has been validated through animal experiments in the majority of studies (Table 1), and their safety and potential efficacy have been demonstrated in a few reported clinical studies (Table 2) (145). However, given the complexity of human disease mechanisms, which may influence the activity and function of exosomes, the safety and efficacy of exosomes in disease treatment still require further in vivo experiments, particularly in large animal models and clinical studies, to confirm the application effects of MSC-Exos in various diseases.
Table 2. Ongoing clinical studies of mesenchymal stem cell-derived exosomes.
5.4 Standardization of therapeutic applications
Although MSCs can be isolated from various tissues, in current basic experiments and registered clinical trials, MSCs derived from adipose tissue, bone marrow, or umbilical cord are commonly used as sources of exosomes. Among these, bone marrow is the most frequently used source of MSC-Exos in basic research (62), while adipose tissue is the most commonly used source in clinical research (Tables 2, 3). Given the non-immunogenic nature of MSC-Exos, allogeneic sources have been used in all but two studies. In addition, the routes of administration for MSC-Exos in existing studies encompass intravenous infusion, inhalation, or local delivery (Tables 1–3). The majority of these studies employ intravenous infusion or inhalation as the administration routes. Consistent with the basic research (Table 1), the dosing regimens in these clinical studies vary according to the route of administration and the specific disease being treated. Moreover, the units used to quantify MSC-Exos also differ: some studies measure the quantity of MSC-Exos by weight (in micrograms), others by particle number, while some merely specify the number of MSCs used to generate the MSC-Exos. These inconsistencies highlight the lack of consensus on the application of MSC-Exos, rendering it impossible to conduct a robust assessment of their therapeutic efficacy. Therefore, standardizing the application of exosomes across different diseases remains a significant challenge at present.
Table 3. Published clinical studies of mesenchymal stem cell-derived exosomes.
6 Conclusions and prospects
Pyroptosis is a form of programmed cell death whose mechanism involves the activation of caspases and the GSDM family, as well as the release of inflammatory factors. These key steps will trigger cell death and severe inflammatory responses, ultimately leading to the progression of various diseases (146). Therefore, it is essential to identify key biomarkers for pyroptosis and to seek therapeutic strategies to modulate this process (147).
MSCs have shown great potential in the treatment of various diseases due to their multidifferentiation, self-renewal, and immunomodulatory properties. Exosomes derived from MSCs not only retain the therapeutic functions of the parental cells but also possess characteristics such as non-proliferative capacity, low immunogenicity, and the capacity for cargo loading and targeted delivery (148). These characteristics make exosomes a strong candidate for replacing MSCs in disease diagnosis and treatment (Figure 4).
Figure 4. Applications of mesenchymal stem cells (MSCs) derived exosomes. GSDM, gasdermin.
In terms of diagnostic potential, MSC-Exos may contain differentially expressed miRNAs, ncRNAs, and proteins under various disease conditions. By extracting exosomes from diseased individuals and identifying the differential expression of specific substances within them, a highly sensitive method for the identification and monitoring of pyroptosis in diseases may be provided. These substances may help accurately reflect the state and extent of pyroptosis in diseases, thereby improving the precision of pyroptosis identification. This will render exosomes a promising candidate for early pyroptosis detection, monitoring, and the development of personalized therapeutic strategies.
From a therapeutic perspective, MSC-Exos hold distinct advantages over traditional drugs. Traditional drugs exert broad-spectrum effects on the entire body, whereas MSC-Exos can selectively target specific cells and tissues, offering more precise and manageable therapeutic options. Moreover, exosomes, characterized by their low toxicity, weak immunogenicity, and high permeability, can serve as efficient delivery vehicles for substances such as miRNAs, enzymes, and proteins. The lipid bilayer membrane of exosomes protects the encapsulated substances, ensuring their stability and reducing their susceptibility to enzymatic degradation. These attributes render MSC-Exos a potentially more practical therapeutic strategy. Emerging evidence indicates that MSC-Exos can target and modulate key nodes in the pyroptosis pathways to inhibit pyroptosis, thus becoming a research focus in regenerative medicine.
Over the past few years, with the progress of genetic engineering and nanotechnology, MSC-Exos have gradually emerged as an alternative strategy to stem cell-based regenerative therapies. Exosomes can be genetically engineered to deliver various therapeutic components to the desired targets (149). Through genetic engineering and chemical modification, the targeting ability of exosomes can be enhanced, and their circulation half-life in the body can be extended, thereby improving their therapeutic efficacy (150). Compared with traditional exogenous nanocarriers, engineered exosomes possess high bioavailability, low toxicity, drug protection, and precise targeting capabilities, making them one of the most promising drug delivery vehicles currently available. Moreover, the adoption of combination therapy strategies may also aid in disease treatment. Studies have shown that the therapeutic efficacy of single MSC-Exos can be enhanced through co-culture, combined drugs, combined physical factors, and genetic modification (151, 152).
In summary, MSC-Exos have the potential to alleviate disease progression by modulating pyroptosis, thus offering a promising therapeutic avenue. However, the clinical translation of MSC-exos therapy faces several challenges. Nevertheless, with the advancement of research, the clinical potential of MSC-exos is expected to be enhanced through several approaches: elucidating the specific biomarkers, key targets, and signaling pathways of pyroptosis in various diseases; optimizing the tissue sources, production techniques, and administration strategies of MSC-exos; and developing engineered exosomes with specificity and combination therapies. In the future, MSC-exos are anticipated to serve as a tool for early detection, precise monitoring, and targeted modulation of pyroptosis, thereby treating diseases and preventing disease progression, offering new therapeutic options for patients.
Author contributions
TY: Conceptualization, Writing – original draft, Writing – review & editing. JA: Writing – review & editing. XX: Writing – review & editing. BL: Conceptualization, Writing – review & editing. ZD: Conceptualization, Funding acquisition, Project administration, Writing – review & editing.
Funding
The author(s) declare that financial support was received for the research and/or publication of this article. This work was supported by the Gansu Provincial Health Industry Research Program (GSWSKY2023-24), the Gansu Provincial Youth Science and Technology Foundation (20JR10RA710), and the Foundation of the First Hospital of Lanzhou University(ldyyyn2019-13).
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Generative AI statement
The author(s) declare that no Generative AI was used in the creation of this manuscript.
Any alternative text (alt text) provided alongside figures in this article has been generated by Frontiers with the support of artificial intelligence and reasonable efforts have been made to ensure accuracy, including review by the authors wherever possible. If you identify any issues, please contact us.
Publisher’s note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.
Glossary
MSCs: mesenchymal stem cells
MSC-Exos: exosomes derived from MSCs
GSDM: gasdermin
GSDM-NT: gasdermin N-terminal fragment
PRRs: pattern recognition receptors
PAMPs: pathogen-associated molecular patterns
DAMPs: damage-associated molecular patterns
NLRP3: NOD-like receptor pyrin domain containing 3
IL-1β: interleukin-1β
NF-κB: nuclear factor-κB gene
ASC: apoptosis-associated speck-like protein containing a caspase recruitment domain
LPS: lipopolysaccharide
HMGB1: high-mobility group box 1
ZBP1: Z-DNA Binding Protein 1
TNF: tumor necrosis factor
Gzm: granzyme
ESCRT: endosomal sorting complex required for transport
ESEs: early sorting endosomes
LSEs: late sorting endosomes
MVBs: multivesicular bodies
NPCs: nucleus pulposus cells.
References
1. Broz P. Pyroptosis: molecular mechanisms and roles in disease. Cell Res. (2025) 35:334–44. doi: 10.1038/s41422-025-01107-6
2. Liu Y, Pan R, Ouyang Y, Gu W, Xiao T, Yang H, et al. Pyroptosis in health and disease: mechanisms, regulation and clinical perspective. Signal Transduct Target Ther. (2024) 9:245. doi: 10.1038/s41392-024-01958-2
3. Wright SS, Kumari P, Fraile-Ágreda V, Wang C, Shivcharan S, Kappelhoff S, et al. Transplantation of gasdermin pores by extracellular vesicles propagates pyroptosis to bystander cells. Cell. (2025) 188:280–91.e17. doi: 10.1016/j.cell.2024.11.018
4. Miao EA, Leaf IA, Treuting PM, Mao DP, Dors M, Sarkar A, et al. Caspase-1-induced pyroptosis is an innate immune effector mechanism against intracellular bacteria. Nat Immunol. (2010) 11:1136–42. doi: 10.1038/ni.1960
5. Aachoui Y, Leaf IA, Hagar JA, Fontana MF, Campos CG, Zak DE, et al. Caspase-11 protects against bacteria that escape the vacuole. Science. (2013) 339:975–8. doi: 10.1126/science.1230751
6. Aglietti RA and Dueber EC. Recent insights into the molecular mechanisms underlying pyroptosis and gasdermin family functions. Trends Immunol. (2017) 38:261–71. doi: 10.1016/j.it.2017.01.003
7. Tao YC, Wang ML, Chen EQ, and Tang H. Stem cells transplantation in the treatment of patients with liver failure. Curr Stem Cell Res Ther. (2018) 13:193–201. doi: 10.2174/1574888X13666180105123915
8. Schacher FC, Martins Pezzi da Silva A, Silla L, and Álvares-da-Silva MR. Bone marrow mesenchymal stem cells in acute-on-chronic liver failure grades 2 and 3: A phase I-II randomized clinical trial. Can J Gastroenterol Hepatol. (2021) 2021:3662776. doi: 10.1155/2021/3662776
9. Choi J, Kang S, Kim B, So S, Han J, Kim GN, et al. Efficient hepatic differentiation and regeneration potential under xeno-free conditions using mass-producible amnion-derived mesenchymal stem cells. Stem Cell Res Ther. (2021) 12:569. doi: 10.1186/s13287-021-02470-y
10. Wang JL, Ding HR, Pan CY, Shi XL, and Ren HZ. Mesenchymal stem cells ameliorate lipid metabolism through reducing mitochondrial damage of hepatocytes in the treatment of post-hepatectomy liver failure. Cell Death Dis. (2021) 12:111. doi: 10.1038/s41419-020-03374-0
11. Marofi F, Vahedi G, Hasanzadeh A, Salarinasab S, Arzhanga P, Khademi B, et al. Mesenchymal stem cells as the game-changing tools in the treatment of various organs disorders: Mirage or reality? J Cell Physiol. (2019) 234:1268–88. doi: 10.1002/jcp.27152
12. Park YB, Ha CW, Lee CH, Yoon YC, and Park YG. Cartilage regeneration in osteoarthritic patients by a composite of allogeneic umbilical cord blood-derived mesenchymal stem cells and hyaluronate hydrogel: results from a clinical trial for safety and proof-of-concept with 7 years of extended follow-up. Stem Cells Transl Med. (2017) 6:613–21. doi: 10.5966/sctm.2016-0157
13. Han Y, Li X, Zhang Y, Han Y, Chang F, and Ding J. Mesenchymal stem cells for regenerative medicine. Cells. (2019) 8. doi: 10.3390/cells8080886
14. Liang X, Ding Y, Zhang Y, Tse HF, and Lian Q. Paracrine mechanisms of mesenchymal stem cell-based therapy: current status and perspectives. Cell Transplant. (2014) 23:1045–59. doi: 10.3727/096368913X667709
15. Jafarinia M, Alsahebfosoul F, Salehi H, Eskandari N, and Ganjalikhani-Hakemi M. Mesenchymal stem cell-derived extracellular vesicles: A novel cell-free therapy. Immunol Invest. (2020) 49:758–80. doi: 10.1080/08820139.2020.1712416
16. Zhu C, Xu S, Jiang R, Yu Y, Bian J, and Zou Z. The gasdermin family: emerging therapeutic targets in diseases. Signal Transduct Target Ther. (2024) 9:87. doi: 10.1038/s41392-024-01801-8
17. Broz P, Pelegrín P, and Shao F. The gasdermins, a protein family executing cell death and inflammation. Nat Rev Immunol. (2020) 20:143–57. doi: 10.1038/s41577-019-0228-2
18. Liu X, Zhang Z, Ruan J, Pan Y, Magupalli VG, Wu H, et al. Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature. (2016) 535:153–8. doi: 10.1038/nature18629
19. Yu P, Zhang X, Liu N, Tang L, Peng C, and Chen X. Pyroptosis: mechanisms and diseases. Signal Transduct Target Ther. (2021) 6:128. doi: 10.1038/s41392-021-00507-5
20. Man SM and Kanneganti TD. Regulation of inflammasome activation. Immunol Rev. (2015) 265:6–21. doi: 10.1111/imr.12296
21. Zhao C and Zhao W. NLRP3 inflammasome-A key player in antiviral responses. Front Immunol. (2020) 11:211. doi: 10.3389/fimmu.2020.00211
22. Bergsbaken T, Fink SL, and Cookson BT. Pyroptosis: host cell death and inflammation. Nat Rev Microbiol. (2009) 7:99–109. doi: 10.1038/nrmicro2070
23. Hornung V, Ablasser A, Charrel-Dennis M, Bauernfeind F, Horvath G, Caffrey DR, et al. AIM2 recognizes cytosolic dsDNA and forms a caspase-1-activating inflammasome with ASC. Nature. (2009) 458:514–8. doi: 10.1038/nature07725
24. Sborgi L, Rühl S, Mulvihill E, Pipercevic J, Heilig R, Stahlberg H, et al. GSDMD membrane pore formation constitutes the mechanism of pyroptotic cell death. EMBO J. (2016) 35:1766–78. doi: 10.15252/embj.201694696
25. Xia S, Zhang Z, Magupalli VG, Pablo JL, Dong Y, Vora SM, et al. Gasdermin D pore structure reveals preferential release of mature interleukin-1. Nature. (2021) 593:607–11. doi: 10.1038/s41586-021-03478-3
26. Ruan J, Xia S, Liu X, Lieberman J, and Wu H. Cryo-EM structure of the gasdermin A3 membrane pore. Nature. (2018) 557:62–7. doi: 10.1038/s41586-018-0058-6
27. Vanaja SK, Russo AJ, Behl B, Banerjee I, Yankova M, Deshmukh SD, et al. Bacterial outer membrane vesicles mediate cytosolic localization of LPS and caspase-11 activation. Cell. (2016) 165:1106–19. doi: 10.1016/j.cell.2016.04.015
28. Santos JC, Dick MS, Lagrange B, Degrandi D, Pfeffer K, Yamamoto M, et al. LPS targets host guanylate-binding proteins to the bacterial outer membrane for non-canonical inflammasome activation. EMBO J. (2018) 37. doi: 10.15252/embj.201798089
29. Deng M, Tang Y, Li W, Wang X, Zhang R, Zhang X, et al. The endotoxin delivery protein HMGB1 mediates caspase-11-dependent lethality in sepsis. Immunity. (2018) 49:740–53.e7. doi: 10.1016/j.immuni.2018.08.016
30. Fernández-Duran I, Quintanilla A, Tarrats N, Birch J, Hari P, Millar FR, et al. Cytoplasmic innate immune sensing by the caspase-4 non-canonical inflammasome promotes cellular senescence. Cell Death Differ. (2022) 29:1267–82. doi: 10.1038/s41418-021-00917-6
31. Chu LH, Indramohan M, Ratsimandresy RA, Gangopadhyay A, Morris EP, Monack DM, et al. The oxidized phospholipid oxPAPC protects from septic shock by targeting the non-canonical inflammasome in macrophages. Nat Commun. (2018) 9:996. doi: 10.1038/s41467-018-03409-3
32. Hagar JA, Powell DA, Aachoui Y, Ernst RK, and Miao EA. Cytoplasmic LPS activates caspase-11: implications in TLR4-independent endotoxic shock. Science. (2013) 341:1250–3. doi: 10.1126/science.1240988
33. Shi J, Zhao Y, Wang Y, Gao W, Ding J, Li P, et al. Inflammatory caspases are innate immune receptors for intracellular LPS. Nature. (2014) 514:187–92. doi: 10.1038/nature13683
34. Aglietti RA, Estevez A, Gupta A, Ramirez MG, Liu PS, Kayagaki N, et al. GsdmD p30 elicited by caspase-11 during pyroptosis forms pores in membranes. Proc Natl Acad Sci U S A. (2016) 113:7858–63. doi: 10.1073/pnas.1607769113
35. Yi YS. Caspase-11 non-canonical inflammasome: a critical sensor of intracellular lipopolysaccharide in macrophage-mediated inflammatory responses. Immunology. (2017) 152:207–17. doi: 10.1111/imm.12787
36. Zamyatina A and Heine H. Lipopolysaccharide recognition in the crossroads of TLR4 and caspase-4/11 mediated inflammatory pathways. Front Immunol. (2020) 11:585146. doi: 10.3389/fimmu.2020.585146
37. Ding J and Shao F. SnapShot: the noncanonical inflammasome. Cell. (2017) 168:544–.e1. doi: 10.1016/j.cell.2017.01.008
38. Zhou Z, He H, Wang K, Shi X, Wang Y, Su Y, et al. Granzyme A from cytotoxic lymphocytes cleaves GSDMB to trigger pyroptosis in target cells. Science. (2020) 368. doi: 10.1126/science.aaz7548
39. Sollberger G, Choidas A, Burn GL, Habenberger P, Di Lucrezia R, Kordes S, et al. Gasdermin D plays a vital role in the generation of neutrophil extracellular traps. Sci Immunol. (2018) 3. doi: 10.1126/sciimmunol.aar6689
40. Wang Y, Gao W, Shi X, Ding J, Liu W, He H, et al. Chemotherapy drugs induce pyroptosis through caspase-3 cleavage of a gasdermin. Nature. (2017) 547:99–103. doi: 10.1038/nature22393
41. Wu Y, Pan B, Zhang Z, Li X, Leng Y, Ji Y, et al. Caspase-4/11-mediated pulmonary artery endothelial cell pyroptosis contributes to pulmonary arterial hypertension. Hypertension. (2022) 79:536–48. doi: 10.1161/HYPERTENSIONAHA.121.17868
42. Shi Y, He T, Liu H, Li X, Li Z, Wen Q, et al. Ganglioside GA2-mediated caspase-11 activation drives macrophage pyroptosis aggravating intimal hyperplasia after arterial injury. Int J Biol Sci. (2025) 21:433–53. doi: 10.7150/ijbs.97106
43. Zheng M, Karki R, Kancharana B, Berns H, Pruett-Miller SM, and Kanneganti TD. Caspase-6 promotes activation of the caspase-11-NLRP3 inflammasome during gram-negative bacterial infections. J Biol Chem. (2021) 297:101379. doi: 10.1016/j.jbc.2021.101379
44. Mandal P, Feng Y, Lyons JD, Berger SB, Otani S, DeLaney A, et al. Caspase-8 collaborates with caspase-11 to drive tissue damage and execution of endotoxic shock. Immunity. (2018) 49:42–55.e6. doi: 10.1016/j.immuni.2018.06.011
45. Santos RA, Cerqueira DM, Zamboni DS, and Oliveira SC. Caspase-8 but not caspase-7 influences inflammasome activation to act in control of Brucella abortus infection. Front Microbiol. (2022) 13:1086925. doi: 10.3389/fmicb.2022.1086925
46. Sheng M, Weng Y, Cao Y, Zhang C, Lin Y, and Yu W. Caspase 6/NR4A1/SOX9 signaling axis regulates hepatic inflammation and pyroptosis in ischemia-stressed fatty liver. Cell Death Discov. (2023) 9:106. doi: 10.1038/s41420-023-01396-z
47. Liu Y, Fang Y, Chen X, Wang Z, Liang X, Zhang T, et al. Gasdermin E-mediated target cell pyroptosis by CAR T cells triggers cytokine release syndrome. Sci Immunol. (2020) 5. doi: 10.1126/sciimmunol.aax7969
48. Zhang Z, Zhang Y, Xia S, Kong Q, Li S, Liu X, et al. Gasdermin E suppresses tumour growth by activating anti-tumour immunity. Nature. (2020) 579:415–20. doi: 10.1038/s41586-020-2071-9
49. Kambara H, Liu F, Zhang X, Liu P, Bajrami B, Teng Y, et al. Gasdermin D exerts anti-inflammatory effects by promoting neutrophil death. Cell Rep. (2018) 22:2924–36. doi: 10.1016/j.celrep.2018.02.067
50. Rühl S, Shkarina K, Demarco B, Heilig R, Santos JC, and Broz P. ESCRT-dependent membrane repair negatively regulates pyroptosis downstream of GSDMD activation. Science. (2018) 362:956–60. doi: 10.1126/science.aar7607
51. Semino C, Carta S, Gattorno M, Sitia R, and Rubartelli A. Progressive waves of IL-1β release by primary human monocytes via sequential activation of vesicular and gasdermin D-mediated secretory pathways. Cell Death Dis. (2018) 9:1088. doi: 10.1038/s41419-018-1121-9
52. Evavold CL, Ruan J, Tan Y, Xia S, Wu H, and Kagan JC. The pore-forming protein gasdermin D regulates interleukin-1 secretion from living macrophages. Immunity. (2018) 48:35–44.e6. doi: 10.1016/j.immuni.2017.11.013
53. Xu HK, Chen LJ, Zhou SN, Li YF, and Xiang C. Multifunctional role of microRNAs in mesenchymal stem cell-derived exosomes in treatment of diseases. World J Stem Cells. (2020) 12:1276–94. doi: 10.4252/wjsc.v12.i11.1276
54. Fayazi N, Sheykhhasan M, Soleimani Asl S, and Najafi R. Stem cell-derived exosomes: a new strategy of neurodegenerative disease treatment. Mol Neurobiol. (2021) 58:3494–514. doi: 10.1007/s12035-021-02324-x
55. Yin K, Wang S, and Zhao RC. Exosomes from mesenchymal stem/stromal cells: a new therapeutic paradigm. biomark Res. (2019) 7:8. doi: 10.1186/s40364-019-0159-x
56. Zhang X, Han C, Du B, Nan D, Zhang W, and He G. Isolation and identification of adipose stem cell exosomes and the study of its potential as drug delivery carrier in vitro. Appl Biochem Biotechnol. (2022) 194:2594–603. doi: 10.1007/s12010-022-03835-6
57. Yadav S, Maity P, and Kapat K. The opportunities and challenges of mesenchymal stem cells-derived exosomes in theranostics and regenerative medicine. Cells. (2024) 13. doi: 10.3390/cells13231956
58. Hu P, Yang Q, Wang Q, Shi C, Wang D, Armato U, et al. Mesenchymal stromal cells-exosomes: a promising cell-free therapeutic tool for wound healing and cutaneous regeneration. Burns Trauma. (2019) 7:38. doi: 10.1186/s41038-019-0178-8
59. Ma ZJ, Yang JJ, Lu YB, Liu ZY, and Wang XX. Mesenchymal stem cell-derived exosomes: Toward cell-free therapeutic strategies in regenerative medicine. World J Stem Cells. (2020) 12:814–40. doi: 10.4252/wjsc.v12.i8.814
60. Tan SS, Yin Y, Lee T, Lai RC, Yeo RW, Zhang B, et al. Therapeutic MSC exosomes are derived from lipid raft microdomains in the plasma membrane. J Extracell Vesicles. (2013) 2. doi: 10.3402/jev.v2i0.22614
61. Théry C, Amigorena S, Raposo G, and Clayton A. Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol. (2006) Chapter 3:Unit 3.22. doi: 10.1002/0471143030.cb0322s30
62. Hassanzadeh A, Rahman HS, Markov A, Endjun JJ, Zekiy AO, Chartrand MS, et al. Mesenchymal stem/stromal cell-derived exosomes in regenerative medicine and cancer; overview of development, challenges, and opportunities. Stem Cell Res Ther. (2021) 12:297. doi: 10.1186/s13287-021-02378-7
63. Zhang Y, Liu Y, Liu H, and Tang WH. Exosomes: biogenesis, biologic function and clinical potential. Cell Biosci. (2019) 9:19. doi: 10.1186/s13578-019-0282-2
64. Hessvik NP and Llorente A. Current knowledge on exosome biogenesis and release. Cell Mol Life Sci. (2018) 75:193–208. doi: 10.1007/s00018-017-2595-9
65. Jadli AS, Ballasy N, Edalat P, and Patel VB. Inside(sight) of tiny communicator: exosome biogenesis, secretion, and uptake. Mol Cell Biochem. (2020) 467:77–94. doi: 10.1007/s11010-020-03703-z
67. Zhen Y and Stenmark H. Cellular functions of Rab GTPases at a glance. J Cell Sci. (2015) 128:3171–6. doi: 10.1242/jcs.166074
68. McMahon HT and Boucrot E. Membrane curvature at a glance. J Cell Sci. (2015) 128:1065–70. doi: 10.1242/jcs.114454
69. Henne WM, Stenmark H, and Emr SD. Molecular mechanisms of the membrane sculpting ESCRT pathway. Cold Spring Harb Perspect Biol. (2013) 5. doi: 10.1101/cshperspect.a016766
70. Gurunathan S, Kang MH, Jeyaraj M, Qasim M, and Kim JH. Review of the isolation, characterization, biological function, and multifarious therapeutic approaches of exosomes. Cells. (2019) 8. doi: 10.3390/cells8040307
71. Théry C, Witwer KW, Aikawa E, Alcaraz MJ, Anderson JD, Andriantsitohaina R, et al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles. (2018) 7:1535750. doi: 10.1080/20013078.2018.1535750
72. Wang J, Chen D, and Ho EA. Challenges in the development and establishment of exosome-based drug delivery systems. J Control Release. (2021) 329:894–906. doi: 10.1016/j.jconrel.2020.10.020
73. Gardiner C, Di Vizio D, Sahoo S, Théry C, Witwer KW, Wauben M, et al. Techniques used for the isolation and characterization of extracellular vesicles: results of a worldwide survey. J Extracell Vesicles. (2016) 5:32945. doi: 10.3402/jev.v5.32945
74. Li P, Kaslan M, Lee SH, Yao J, and Gao Z. Progress in exosome isolation techniques. Theranostics. (2017) 7:789–804. doi: 10.7150/thno.18133
75. Weng Z, Zhang B, Wu C, Yu F, Han B, Li B, et al. Therapeutic roles of mesenchymal stem cell-derived extracellular vesicles in cancer. J Hematol Oncol. (2021) 14:136. doi: 10.1186/s13045-021-01141-y
76. Zhu L, Sun HT, Wang S, Huang SL, Zheng Y, Wang CQ, et al. Isolation and characterization of exosomes for cancer research. J Hematol Oncol. (2020) 13:152. doi: 10.1186/s13045-020-00987-y
77. Rider MA, Hurwitz SN, and Meckes DG Jr. ExtraPEG: A polyethylene glycol-based method for enrichment of extracellular vesicles. Sci Rep. (2016) 6:23978. doi: 10.1038/srep23978
78. Wu M, Ouyang Y, Wang Z, Zhang R, Huang PH, Chen C, et al. Isolation of exosomes from whole blood by integrating acoustics and microfluidics. Proc Natl Acad Sci U S A. (2017) 114:10584–9. doi: 10.1073/pnas.1709210114
79. Liu F, Vermesh O, Mani V, Ge TJ, Madsen SJ, Sabour A, et al. The exosome total isolation chip. ACS Nano. (2017) 11:10712–23. doi: 10.1021/acsnano.7b04878
80. Kalluri R and LeBleu VS. The biology, function, and biomedical applications of exosomes. Science. (2020) 367. doi: 10.1126/science.aau6977
81. Hade MD, Suire CN, and Suo Z. Mesenchymal stem cell-derived exosomes: applications in regenerative medicine. Cells. (2021) 10. doi: 10.3390/cells10081959
82. Liu T, Zhang Q, Zhang J, Li C, Miao YR, Lei Q, et al. EVmiRNA: a database of miRNA profiling in extracellular vesicles. Nucleic Acids Res. (2019) 47:D89–d93. doi: 10.1093/nar/gky985
83. Kowal J, Arras G, Colombo M, Jouve M, Morath JP, Primdal-Bengtson B, et al. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proc Natl Acad Sci U S A. (2016) 113:E968–77. doi: 10.1073/pnas.1521230113
84. Naji A, Favier B, Deschaseaux F, Rouas-Freiss N, Eitoku M, and Suganuma N. Mesenchymal stem/stromal cell function in modulating cell death. Stem Cell Res Ther. (2019) 10:56. doi: 10.1186/s13287-019-1158-4
85. Pote MS and Gacche RN. Exosomal signaling in cancer metastasis: Molecular insights and therapeutic opportunities. Arch Biochem Biophys. (2025) 764:110277. doi: 10.1016/j.abb.2024.110277
86. Yudintceva N, Mikhailova N, Fedorov V, Samochernych K, Vinogradova T, Muraviov A, et al. Mesenchymal stem cells and MSCs-derived extracellular vesicles in infectious diseases: from basic research to clinical practice. Bioengineering (Basel). (2022) 9. doi: 10.3390/bioengineering9110662
87. Squadrito ML, Baer C, Burdet F, Maderna C, Gilfillan GD, Lyle R, et al. Endogenous RNAs modulate microRNA sorting to exosomes and transfer to acceptor cells. Cell Rep. (2014) 8:1432–46. doi: 10.1016/j.celrep.2014.07.035
88. Statello L, Maugeri M, Garre E, Nawaz M, Wahlgren J, Papadimitriou A, et al. Identification of RNA-binding proteins in exosomes capable of interacting with different types of RNA: RBP-facilitated transport of RNAs into exosomes. PloS One. (2018) 13:e0195969. doi: 10.1371/journal.pone.0195969
89. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science. (1999) 284:143–7. doi: 10.1126/science.284.5411.143
90. Lu X, Guo H, Wei X, Lu D, Shu W, Song Y, et al. Current status and prospect of delivery vehicle based on mesenchymal stem cell-derived exosomes in liver diseases. Int J Nanomedicine. (2023) 18:2873–90. doi: 10.2147/IJN.S404925
91. Feng Y, Guo K, Jiang J, and Lin S. Mesenchymal stem cell-derived exosomes as delivery vehicles for non-coding RNAs in lung diseases. BioMed Pharmacother. (2024) 170:116008. doi: 10.1016/j.biopha.2023.116008
92. Lavi Arab F, Hoseinzadeh A, Hafezi F, Sadat Mohammadi F, Zeynali F, Hadad Tehran M, et al. Mesenchymal stem cell-derived exosomes for management of prostate cancer: An updated view. Int Immunopharmacol. (2024) 134:112171. doi: 10.1016/j.intimp.2024.112171
93. Bagno L, Hatzistergos KE, Balkan W, and Hare JM. Mesenchymal stem cell-based therapy for cardiovascular disease: progress and challenges. Mol Ther. (2018) 26:1610–23. doi: 10.1016/j.ymthe.2018.05.009
94. Oveili E, Vafaei S, Bazavar H, Eslami Y, Mamaghanizadeh E, Yasamineh S, et al. The potential use of mesenchymal stem cells-derived exosomes as microRNAs delivery systems in different diseases. Cell Commun Signal. (2023) 21:20. doi: 10.1186/s12964-022-01017-9
95. Cui M, Chen F, Shao L, Wei C, Zhang W, Sun W, et al. Mesenchymal stem cells and ferroptosis: Clinical opportunities and challenges. Heliyon. (2024) 10:e25251. doi: 10.1016/j.heliyon.2024.e25251
96. Zhang ZG, Buller B, and Chopp M. Exosomes - beyond stem cells for restorative therapy in stroke and neurological injury. Nat Rev Neurol. (2019) 15:193–203. doi: 10.1038/s41582-018-0126-4
97. Hu S, Hu Y, and Yan W. Extracellular vesicle-mediated interorgan communication in metabolic diseases. Trends Endocrinol Metab. (2023) 34:571–82. doi: 10.1016/j.tem.2023.06.002
98. Roszkowski S. Therapeutic potential of mesenchymal stem cell-derived exosomes for regenerative medicine applications. Clin Exp Med. (2024) 24:46. doi: 10.1007/s10238-023-01282-z
99. Kandettu A, Kuthethur R, and Chakrabarty S. A detailed review on the role of miRNAs in mitochondrial-nuclear cross talk during cancer progression. Biochim Biophys Acta Mol Basis Dis. (2025) 1871:167731. doi: 10.1016/j.bbadis.2025.167731
100. Ying W, Riopel M, Bandyopadhyay G, Dong Y, Birmingham A, Seo JB, et al. Adipose tissue macrophage-derived exosomal miRNAs can modulate in vivo and in vitro insulin sensitivity. Cell. (2017) 171:372–84.e12. doi: 10.1016/j.cell.2017.08.035
101. Chen L, Lu FB, Chen DZ, Wu JL, Hu ED, Xu LM, et al. BMSCs-derived miR-223-containing exosomes contribute to liver protection in experimental autoimmune hepatitis. Mol Immunol. (2018) 93:38–46. doi: 10.1016/j.molimm.2017.11.008
102. Huang Y, Lu D, Ma W, Liu J, Ning Q, Tang F, et al. miR-223 in exosomes from bone marrow mesenchymal stem cells ameliorates rheumatoid arthritis via downregulation of NLRP3 expression in macrophages. Mol Immunol. (2022) 143:68–76. doi: 10.1016/j.molimm.2022.01.002
103. Liu Y, Zhang S, Xue Z, Zhou X, Tong L, Liao J, et al. Bone mesenchymal stem cells-derived miR-223-3p-containing exosomes ameliorate lipopolysaccharide-induced acute uterine injury via interacting with endothelial progenitor cells. Bioengineered. (2021) 12:10654–65. doi: 10.1080/21655979.2021.2001185
104. Li X and Yang N. Exosome miR-223-3p in the bone marrow-derived mesenchymal stem cells alleviates the inflammation and airway remodeling through NLRP3-induced ASC/Caspase-1/GSDMD signaling pathway. Int Immunopharmacol. (2023) 123:110746. doi: 10.1016/j.intimp.2023.110746
105. Hou L, Zhu Z, Jiang F, Zhao J, Jia Q, Jiang Q, et al. Human umbilical cord mesenchymal stem cell-derived extracellular vesicles alleviated silica induced lung inflammation and fibrosis in mice via circPWWP2A/miR-223-3p/NLRP3 axis. Ecotoxicol Environ Saf. (2023) 251:114537. doi: 10.1016/j.ecoenv.2023.114537
106. Wang D, Xue H, Tan J, Liu P, Qiao C, Pang C, et al. Bone marrow mesenchymal stem cells-derived exosomes containing miR-539-5p inhibit pyroptosis through NLRP3/caspase-1 signalling to alleviate inflammatory bowel disease. Inflammation Res. (2022) 71:833–46. doi: 10.1007/s00011-022-01577-z
107. Zhai L, Shen H, Sheng Y, and Guan Q. ADMSC Exo-MicroRNA-22 improve neurological function and neuroinflammation in mice with Alzheimer’s disease. J Cell Mol Med. (2021) 25:7513–23. doi: 10.1111/jcmm.16787
108. Li Q, Wang Z, Xing H, Wang Y, and Guo Y. Exosomes derived from miR-188-3p-modified adipose-derived mesenchymal stem cells protect Parkinson’s disease. Mol Ther Nucleic Acids. (2021) 23:1334–44. doi: 10.1016/j.omtn.2021.01.022
109. Tang L, Zhang J, and Gao J. Extracellular vesicles from adipose-derived mesenchymal stem cells prevent high glucose-induced retinal ganglion cell pyroptosis through a microRNA-26a-5p-dependent mechanism. J Diabetes Investig. (2025) 16:1597–609. doi: 10.1111/jdi.70100
110. Yu Y, Li W, Xian T, Tu M, Wu H, and Zhang J. Human embryonic stem-cell-derived exosomes repress NLRP3 inflammasome to alleviate pyroptosis in nucleus pulposus cells by transmitting miR-302c. Int J Mol Sci. (2023) 24. doi: 10.3390/ijms24087664
111. Zhang J, Zhang J, Zhang Y, Liu W, Ni W, Huang X, et al. Mesenchymal stem cells-derived exosomes ameliorate intervertebral disc degeneration through inhibiting pyroptosis. J Cell Mol Med. (2020) 24:11742–54. doi: 10.1111/jcmm.15784
112. Yue R, Lu S, Luo Y, Zeng J, Liang H, Qin D, et al. Mesenchymal stem cell-derived exosomal microRNA-182-5p alleviates myocardial ischemia/reperfusion injury by targeting GSDMD in mice. Cell Death Discov. (2022) 8:202. doi: 10.1038/s41420-022-00909-6
113. Tang J, Jin L, Liu Y, Li L, Ma Y, Lu L, et al. Exosomes derived from mesenchymal stem cells protect the myocardium against ischemia/reperfusion injury through inhibiting pyroptosis. Drug Des Devel Ther. (2020) 14:3765–75. doi: 10.2147/DDDT.S239546
114. Xu Y, Tang X, Fang A, Yan J, Kofi Wiredu Ocansey D, Zhang X, et al. HucMSC-Ex carrying miR-203a-3p.2 ameliorates colitis through the suppression of caspase11/4-induced macrophage pyroptosis. Int Immunopharmacol. (2022) 110:108925. doi: 10.1016/j.intimp.2022.108925
115. Della Rocca Y, Diomede F, Konstantinidou F, Trubiani O, Soundara Rajan T, Pierdomenico SD, et al. Protective effect of oral stem cells extracellular vesicles on cardiomyocytes in hypoxia-reperfusion. Front Cell Dev Biol. (2023) 11:1260019. doi: 10.3389/fcell.2023.1260019
116. Lv J, Hao YN, Wang XP, Lu WH, Xie LY, and Niu D. Bone marrow mesenchymal stem cell-derived exosomal miR-30e-5p ameliorates high-glucose induced renal proximal tubular cell pyroptosis by inhibiting ELAVL1. Ren Fail. (2023) 45:2177082. doi: 10.1080/0886022X.2023.2177082
117. Zhang T, Lu L, Li M, Zhang D, Yu P, Zhang X, et al. Exosome from BMMSC attenuates cardiopulmonary bypass-induced acute lung injury via YAP/β-catenin pathway: downregulation of pyroptosis. Stem Cells. (2022) 40:1122–33. doi: 10.1093/stmcls/sxac063
118. Zeng H, Yang Y, Tou F, Zhan Y, Liu S, Zou P, et al. Bone marrow stromal cell-derived exosomes improve oxidative stress and pyroptosis in doxorubicin-induced myocardial injury in vitro by regulating the transcription of GSDMD through the PI3K-AKT-Foxo1 pathway. Immun Inflammation Dis. (2023) 11:e810. doi: 10.1002/iid3.810
119. Tao Y, Xu X, Yang B, Zhao H, and Li Y. Mitigation of sepsis-induced acute lung injury by BMSC-derived exosomal miR-125b-5p through STAT3-mediated suppression of macrophage pyroptosis. Int J Nanomedicine. (2023) 18:7095–113. doi: 10.2147/IJN.S441133
120. Xu H and Xu B. BMSC-Derived Exosomes Ameliorate Osteoarthritis by Inhibiting Pyroptosis of Cartilage via Delivering miR-326 Targeting HDAC3 and STAT1//NF-κB p65 to Chondrocytes. Mediators Inflamm. (2021) 2021:9972805. doi: 10.1155/2021/9972805
121. Hu J, Jiang Y, Wu X, Wu Z, Qin J, Zhao Z, et al. Exosomal miR-17-5p from adipose-derived mesenchymal stem cells inhibits abdominal aortic aneurysm by suppressing TXNIP-NLRP3 inflammasome. Stem Cell Res Ther. (2022) 13:349. doi: 10.1186/s13287-022-03037-1
122. Hua T, Yang M, Song H, Kong E, Deng M, Li Y, et al. Huc-MSCs-derived exosomes attenuate inflammatory pain by regulating microglia pyroptosis and autophagy via the miR-146a-5p/TRAF6 axis. J Nanobiotechnology. (2022) 20:324. doi: 10.1186/s12951-022-01522-6
123. Sun ZL, You T, Zhang BH, Liu Y, and Liu J. Bone marrow mesenchymal stem cell-derived exosomes miR-202-5p inhibited pyroptosis to alleviate lung ischemic-reperfusion injury by targeting CMPK2. Kaohsiung J Med Sci. (2023) 39:688–98. doi: 10.1002/kjm2.12688
124. Liu Y, Lou G, Li A, Zhang T, Qi J, Ye D, et al. AMSC-derived exosomes alleviate lipopolysaccharide/d-galactosamine-induced acute liver failure by miR-17-mediated reduction of TXNIP/NLRP3 inflammasome activation in macrophages. EBioMedicine. (2018) 36:140–50. doi: 10.1016/j.ebiom.2018.08.054
125. Liang C, Liu Y, Xu H, Huang J, Shen Y, Chen F, et al. Exosomes of Human Umbilical Cord MSCs Protect Against Hypoxia/Reoxygenation-Induced Pyroptosis of Cardiomyocytes via the miRNA-100-5p/FOXO3/NLRP3 Pathway. Front Bioeng Biotechnol. (2020) 8:615850. doi: 10.3389/fbioe.2020.615850
126. Ma Y, She X, Liu Y, and Qin X. MSC-derived exosomal miR-140-3p improves cognitive dysfunction in sepsis-associated encephalopathy by HMGB1 and S-lactoylglutathione metabolism. Commun Biol. (2024) 7:562. doi: 10.1038/s42003-024-06236-z
127. Chen D, Jiang X, and Zou H. hASCs-derived exosomal miR-155-5p targeting TGFβR2 promotes autophagy and reduces pyroptosis to alleviate intervertebral disc degeneration. J Orthop Translat. (2023) 39:163–76. doi: 10.1016/j.jot.2023.02.004
128. Hu LT, Wang BY, Fan YH, He ZY, and Zheng WX. Exosomal miR-23b from bone marrow mesenchymal stem cells alleviates oxidative stress and pyroptosis after intracerebral hemorrhage. Neural Regener Res. (2023) 18:560–7. doi: 10.4103/1673-5374.346551
129. Sun H, Wang J, Bi W, Zhang F, Zhang K, Tian X, et al. Mesenchymal stem cell-derived exosomal microRNA-367-3p mitigates lower limb ischemia/reperfusion injury in mouse skeletal muscle via EZH2 targeting. J Pharm Pharmacol. (2024) 76:1634–46. doi: 10.1093/jpp/rgae086
130. Caba L, Florea L, Gug C, Dimitriu DC, and Gorduza EV. Circular RNA-is the circle perfect? Biomolecules. (2021) 11. doi: 10.3390/biom11121755
131. Chen X, Zhou M, Yant L, and Huang C. Circular RNA in disease: Basic properties and biomedical relevance. Wiley Interdiscip Rev RNA. (2022) 13:e1723. doi: 10.1002/wrna.1723
132. Zhao Y, Chen Y, Wang Z, Xu C, Qiao S, Liu T, et al. Bone Marrow Mesenchymal Stem Cell Exosome Attenuates Inflammasome-Related Pyroptosis via Delivering circ_003564 to Improve the Recovery of Spinal Cord Injury. Mol Neurobiol. (2022) 59:6771–89. doi: 10.1007/s12035-022-03006-y
133. Yan B, Zhang Y, Liang C, Liu B, Ding F, Wang Y, et al. Stem cell-derived exosomes prevent pyroptosis and repair ischemic muscle injury through a novel exosome/circHIPK3/FOXO3a pathway. Theranostics. (2020) 10:6728–42. doi: 10.7150/thno.42259
134. Mao Q, Liang XL, Zhang CL, Pang YH, and Lu YX. LncRNA KLF3-AS1 in human mesenchymal stem cell-derived exosomes ameliorates pyroptosis of cardiomyocytes and myocardial infarction through miR-138-5p/Sirt1 axis. Stem Cell Res Ther. (2019) 10:393. doi: 10.1186/s13287-019-1522-4
135. Yan B, Liu T, Yao C, Liu X, Du Q, and Pan L. LncRNA XIST shuttled by adipose tissue-derived mesenchymal stem cell-derived extracellular vesicles suppresses myocardial pyroptosis in atrial fibrillation by disrupting miR-214-3p-mediated Arl2 inhibition. Lab Invest. (2021) 101:1427–38. doi: 10.1038/s41374-021-00635-0
136. Hu XM, Li ZX, Lin RH, Shan JQ, Yu QW, Wang RX, et al. Guidelines for regulated cell death assays: A systematic summary, A categorical comparison, A prospective. Front Cell Dev Biol. (2021) 9:634690. doi: 10.3389/fcell.2021.634690
137. Zhang H, Freitas D, Kim HS, Fabijanic K, Li Z, Chen H, et al. Identification of distinct nanoparticles and subsets of extracellular vesicles by asymmetric flow field-flow fractionation. Nat Cell Biol. (2018) 20:332–43. doi: 10.1038/s41556-018-0040-4
138. Mathieu M, Martin-Jaular L, Lavieu G, and Théry C. Specificities of secretion and uptake of exosomes and other extracellular vesicles for cell-to-cell communication. Nat Cell Biol. (2019) 21:9–17. doi: 10.1038/s41556-018-0250-9
139. Bebelman MP, Smit MJ, Pegtel DM, and Baglio SR. Biogenesis and function of extracellular vesicles in cancer. Pharmacol Ther. (2018) 188:1–11. doi: 10.1016/j.pharmthera.2018.02.013
140. Ciardiello C, Cavallini L, Spinelli C, Yang J, Reis-Sobreiro M, de Candia P, et al. Focus on extracellular vesicles: new frontiers of cell-to-cell communication in cancer. Int J Mol Sci. (2016) 17:175. doi: 10.3390/ijms17020175
141. Chevillet JR, Kang Q, Ruf IK, Briggs HA, Vojtech LN, Hughes SM, et al. Quantitative and stoichiometric analysis of the microRNA content of exosomes. Proc Natl Acad Sci U S A. (2014) 111:14888–93. doi: 10.1073/pnas.1408301111
142. Álvarez-Viejo M. Mesenchymal stem cells from different sources and their derived exosomes: A pre-clinical perspective. World J Stem Cells. (2020) 12:100–9. doi: 10.4252/wjsc.v12.i2.100
143. Wu D, Tao S, Zhu L, Zhao C, and Xu N. Chitosan hydrogel dressing loaded with adipose mesenchymal stem cell-derived exosomes promotes skin full-thickness wound repair. ACS Appl Bio Mater. (2024) 7:1125–34. doi: 10.1021/acsabm.3c01039
144. Sharma R, Kumari M, Mishra S, Chaudhary DK, Kumar A, Avni B, et al. Exosomes secreted by umbilical cord blood-derived mesenchymal stem cell attenuate diabetes in mice. J Diabetes Res. (2021) 2021:9534574. doi: 10.1155/2021/9534574
145. Lotfy A, AboQuella NM, and Wang H. Mesenchymal stromal/stem cell (MSC)-derived exosomes in clinical trials. Stem Cell Res Ther. (2023) 14:66. doi: 10.1186/s13287-023-03287-7
146. Wu J, Lin S, Wan B, Velani B, and Zhu Y. Pyroptosis in liver disease: new insights into disease mechanisms. Aging Dis. (2019) 10:1094–108. doi: 10.14336/AD.2019.0116
147. Gao L, Jiang Z, Han Y, Li Y, and Yang X. Regulation of pyroptosis by ncRNA: A novel research direction. Front Cell Dev Biol. (2022) 10:840576. doi: 10.3389/fcell.2022.840576
148. Wang Y, Zhang Y, Li T, Shen K, Wang KJ, Tian C, et al. Adipose mesenchymal stem cell derived exosomes promote keratinocytes and fibroblasts embedded in collagen/platelet-rich plasma scaffold and accelerate wound healing. Adv Mater. (2023) 35:e2303642. doi: 10.1002/adma.202303642
149. Batrakova EV and Kim MS. Using exosomes, naturally-equipped nanocarriers, for drug delivery. J Control Release. (2015) 219:396–405. doi: 10.1016/j.jconrel.2015.07.030
150. Mohammadi AH, Ghazvinian Z, Bagheri F, Harada M, and Baghaei K. Modification of extracellular vesicle surfaces: an approach for targeted drug delivery. BioDrugs. (2023) 37:353–74. doi: 10.1007/s40259-023-00595-5
151. Xu Y, Wang X, Zhou X, Zeng W, Yuan J, and Ye J. Multiple strategies enhance the efficacy of MSC-Exos transplantation for spinal cord injury. Exp Neurol. (2025) 383:115038. doi: 10.1016/j.expneurol.2024.115038
152. Xu Y, Zhou X, Wang X, Jin Y, Zhou L, and Ye J. Progress of mesenchymal stem cells (MSCs) & MSC-Exosomes combined with drugs intervention in liver fibrosis. BioMed Pharmacother. (2024) 176:116848. doi: 10.1016/j.biopha.2024.116848
153. Allogeneic mesenchymal stem cell-derived exosome therapy for progressive multiple sclerosis (2025). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT07146087 (Accessed November 11, 2025).
154. Effect of microvesicles and exosomes therapy on β-cell mass in type I diabetes mellitus (T1DM) (2014). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT02138331 (Accessed November 11, 2025).
155. MSC-exos promote healing of MHs (MSCs). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT03437759 (Accessed November 11, 2025).
156. Allogenic mesenchymal stem cell derived exosome in patients with acute ischemic stroke (2021). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT03384433 (Accessed November 11, 2025).
157. A pilot clinical study on inhalation of mesenchymal stem cells exosomes treating severe novel coronavirus pneumonia (2020). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT04276987 (Accessed November 11, 2025).
158. Evaluation of safety and efficiency of method of exosome inhalation in SARS-coV-2 associated pneumonia. (COVID-19EXO) (2020). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT04491240 (Accessed November 11, 2025).
159. Exosome of mesenchymal stem cells for multiple organ dysfuntion syndrome after surgical repaire of acute type A aortic dissection (2020). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT04356300 (Accessed November 11, 2025).
160. Effect of UMSCs Derived Exosomes on Dry Eye in Patients With cGVHD (2022). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT04213248 (Accessed November 11, 2025).
161. The safety and the efficacy evaluation of allogenic adipose MSC-exos in patients with alzheimer’s disease (2021). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT04388982.
162. MSC EVs in dystrophic epidermolysis bullosa (2025). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT04173650 (Accessed November 11, 2025).
163. A clinical study of mesenchymal stem cell exosomes nebulizer for the treatment of ARDS (2024). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT04602104 (Accessed November 11, 2025).
164. A clinical study of mesenchymal progenitor cell exosomes nebulizer for the treatment of pulmonary infection (2024). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT04544215 (Accessed November 11, 2025).
165. Evaluation of adipose derived stem cells exo.in treatment of periodontitis (2020). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT04270006 (Accessed November 11, 2025).
166. iExosomes in treating participants with metastatic pancreas cancer with krasG12D mutation (2025). USNational Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT03608631 (Accessed November 11, 2025).
167. Intra-articular injection of MSC-derived exosomes in knee osteoarthritis (ExoOA-1) (2021). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT05060107 (Accessed November 11, 2025).
168. Expanded Access for Use of bmMSC-Derived Extracellular Vesicles in Patients With COVID-19 Associated ARDSs (2024). US: National Library of Medicine. Available online at: https://clinicaltrials.gov/study/NCT04657458 (Accessed November 11, 2025).
169. Nassar W, El-Ansary M, Sabry D, Mostafa MA, Fayad T, Kotb E, et al. Umbilical cord mesenchymal stem cells derived extracellular vesicles can safely ameliorate the progression of chronic kidney diseases. Biomater Res. (2016) 20:21. doi: 10.1186/s40824-016-0068-0
170. Cho BS, Lee J, Won Y, Duncan DI, Jin RC, Lee J, et al. Skin brightening efficacy of exosomes derived from human adipose tissue-derived stem/stromal cells: A prospective, split-face, randomized placebo-controlled study. Cosmetics. (2020) 7:90. doi: 10.3390/cosmetics7040090
171. Kwon HH, Yang SH, Lee J, Park BC, Park KY, Jung JY, et al. Combination treatment with human adipose tissue stem cell-derived exosomes and fractional CO2 laser for acne scars: A 12-week prospective, double-blind, randomized, split-face study. Acta Derm Venereol. (2020) 100:adv00310. doi: 10.2340/00015555-3666
172. IRB approved pilot safety study of an extracellular vesicle isolate product evaluating the treatment of osteoarthritis in combat-related injuries. (2020). doi: 10.52793/JSCR.2020.1(2)-09
Keywords: mesenchymal stem cells, exosomes, pyroptosis, pathway, gasdermin
Citation: Yang T, An J, Xu X, Li B and Dou Z (2025) Research progress on pyroptosis regulated by mesenchymal stem cell-derived exosomes. Front. Immunol. 16:1717465. doi: 10.3389/fimmu.2025.1717465
Received: 02 October 2025; Accepted: 19 November 2025; Revised: 11 November 2025;
Published: 04 December 2025.
Edited by:
Dhanu Gupta, Karolinska University Laboratory, SwedenReviewed by:
Mohammad Asad, Albert Einstein College of Medicine, United StatesYanjuan Song, Hubei University of Chinese Medicine, China
Copyright © 2025 Yang, An, Xu, Li and Dou. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
*Correspondence: Zhimin Dou, bGR5eV9kb3V6aGltaW5AbHp1LmVkdS5jbg==