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Gene expression analysis. (a) Gene expression analysis (mean of independent experiments ± SEM) for hTERT-MSCs following treatment with IFN-γ (250 U/ml, 48 h), poly(I:C) (20 µg/ml, 48 h) and BiP (10 µg/ml, 72 h) in comparison to untreated cells using qRT-PCR analysis. (b) Gene expression analysis (mean of independent experiments ± SEM) of the hTERT-MSCs bearing different gene constructs in comparison to GFP transduced cells via qRT-PCR analysis. Among different candidate immune encountered genes, genes depicting statistically significant differences (significance level p < 0.05) after modification in comparison to untreated cells/GFP transduced cells are demonstrated. Data analysis was performed by CFX Manager Software (version 1.6) and GraphPad Prism, version 6.00. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001.
Source publication
The therapeutic potential of mesenchymal stem cells (MSCs) is out of the question. Yet, recent drawbacks have resulted in a strategic shift towards the application of MSC-derived cell-free products such as extracellular vesicles (EVs). Recent reports revealed that functional properties of MSCs, including EV secretion patterns, correlate with microe...
Citations
... It is also worth pointing out that this rationale also holds true for EVs isolated from primary MSCs, as some researchers have observed pro-tumorigenic effects associated with their use [75][76][77][78]. While others have begun to assess the immunomodulatory properties of hTERT-immortalized MSCs [79,80], to the best of our knowledge, this is the first study to investigate the reparative effects of hTERT-immortalized MSC EVs in the context of eye repair. ...
As the economic burden associated with vision loss and ocular damage continues to rise, there is a need to explore novel treatment strategies. Extracellular vesicles (EVs) are enriched with various biological cargo, and there is abundant literature supporting the reparative and immunomodulatory properties of stem cell EVs across a broad range of pathologies. However, one area that requires further attention is the reparative effects of stem cell EVs in the context of ocular damage. Additionally, most of the literature focuses on EVs isolated from primary stem cells; the use of EVs isolated from human telomerase reverse transcriptase (hTERT)-immortalized stem cells has not been thoroughly examined. Using our large-scale EV-manufacturing platform, we reproducibly manufactured EVs from hTERT-immortalized mesenchymal stem cells (MSCs) and employed various methods to characterize and profile their associated cargo. We also utilized well-established cell-based assays to compare the effects of these EVs on both healthy and damaged retinal pigment epithelial cells. To the best of our knowledge, this is the first study to establish proof of concept for reproducible, large-scale manufacturing of hTERT-immortalized MSC EVs and to investigate their potential reparative properties against damaged retinal cells. The results from our studies confirm that hTERT-immortalized MSC EVs exert reparative effects in vitro that are similar to those observed in primary MSC EVs. Therefore, hTERT-immortalized MSCs may represent a more consistent and reproducible platform than primary MSCs for generating EVs with therapeutic potential.
... Among them, MSC and neural stem cells have demonstrated significant effectiveness in promoting neuroprotection, immune regulation, astrogenesis, oligodendrogenesis, synaptogenesis, and improvements in motor and cognitive functions. They also play a role in secreting neurotrophic factors and growth factors and in remodeling the extracellular matrix (Zhang et al., 2020;Napoli et al., 2016;Napoli et al., 2018;Nguyen et al., 2019;Haghighitalab et al., 2021;Dabrowska et al., 2019;Regenhardt et al., 2020;Tuazon et al., 2019). ...
Strokes are among the diseases with the highest mortality and morbidity worldwide. Despite this, current stroke treatments have several limitations, prompting the exploration of new therapeutic options to address the post-acute phase. A key innovation involves the transplantation of stem cells, which exert their effects through various mechanisms. Notably, they release extracellular vesicles that contain anti-inflammatory factors, thereby reducing cellular damage and promoting neurogenesis. While the use of stem cells and extracellular vesicles is still under investigation, this review highlights that the utilization of extracellular vesicles presents several advantages. These advantages suggest that extracellular vesicles could potentially be a superior treatment option for recovering cognitive and motor function.
... The drake aneurysm clip (Codman, Sheffield, UK) was placed on the spinal cord for one min. 33 Injection of 60.00 µL hydrogel or 60.00 µL hydrogel containing 100 µg sEVs was performed over the injured site immediately following the induction of SCI. The sham group only received laminectomy without SCI and did not receive any drugs (Fig. 1). ...
Spinal cord injury has challenging and unpredictable neurological recovery. During inflammatory conditions, the amount of serum albumin and nutrition consumption decrease, nowadays, it is proposed to measure serum albumin and glucose content in human or animal subjects to predict the recovery rate and the efficiency of treatments following SCI. In this study, the effect of extracellular vesicles (EVs) from immortalized human adipose tissue-derived mesenchymal stem cells (Ad-MSCs) equipped with the ectopic expression of human IDO1 gene was investigated on the serum albumin and glucose levels. After pre-clearing steps of 72-hour conditioned media, small EVs (sEVs) were isolated based on the ultrafiltration method. They were encapsulated with a chitosan-based hydrogel. Five experimental groups (female rats, N=30, ~230 g) were considered, including SCI, sham, hydrogel, hTERT-MSCs-GFP-EVs, and hTERT-MSCs-IDO1-EVs. 60 µl of hydrogel or hydrogels containing 100 µg sEVs from control GFP-EVs or IDO1-EVs were locally injected immediately after SCI (laminectomy of the T10 and clip compression). Eight weeks later, non-fasting serum glucose and albumin levels were measured. Results indicated that the level of serum albumin in the animals received IDO1-EVs (3.52±0.04) was increased in comparison to the SCI group (3.00±0.94).Also, these animals indicated higher glucose levels in their serum (250.17±69.61) in comparison to SCI rats (214.00±45.34). Although, these changes were not statistically significant, this could be considered as an evidence for the beneficial effects of IDO1-EVs administration in the context of SCI to reduce hypoalbuminemia and improve energy consumption. However, more detailed experiments are required to confirm these results.
... Among them, MSC and neural stem cells have demonstrated significant effectiveness in promoting neuroprotection, immune regulation, astrogenesis, oligodendrogenesis, synaptogenesis, and improvements in motor and cognitive functions. They also play a role in secreting neurotrophic factors and growth factors and in remodeling the extracellular matrix (Zhang et al., 2020;Napoli et al., 2016;Napoli et al., 2018;Nguyen et al., 2019;Haghighitalab et al., 2021;Dabrowska et al., 2019;Regenhardt et al., 2020;Tuazon et al., 2019). ...
Strokes are among the diseases with the highest mortality and morbidity worldwide. Despite this, current stroke treatments have several limitations, prompting the exploration of new therapeutic options to address the post-acute phase. A key innovation involves the transplantation of stem cells, which exert their effects through various mechanisms. Notably, they release extracellular vesicles that contain anti-inflammatory factors, thereby reducing cellular damage and promoting neurogenesis. While the use of stem cells and extracellular vesicles is still under investigation, this review highlights that the utilization of extracellular vesicles presents several advantages. These advantages suggest that extracellular vesicles could potentially be a superior treatment option for recovering cognitive and motor function.
... Thus, the decreased Tregs infiltration may result from synergistic PTX-mediated ICD and IDO inhibition by NLG919. However, it should not be ignored that IDO overexpression in multidrug resistant carcinoma may predict poor responses with single PTX treatment [40]. Besides, IDO can also suppress systemic and tumor local immune responses through TAMs-Tregs axis and Tregs dependent MDSCs recruitment [41]. ...
Hepatocellular carcinoma (HCC) remains a global health challenge and accounts for ~ 90% of liver cancers. Immunotherapy is demonstrating potent potential for HCC treatment. However, the therapeutic efficacy has been impeded by poor immunogenicity and immunosuppressive tumor microenvironment. Herein, we developed a dual-stimulus responsive nanocarrier (PN@GPB-PEG NPs) to co-deliver chemotherapeutic agent paclitaxel (PTX) and indoleamine 2,3-dioxygenase (IDO) inhibitor NLG919 for HCC chemoimmunotherapy. The resulting nanoparticles demonstrated tumor-specific accumulation and efficient cellular uptake in HCC cells. Furthermore, rapid cargo release could be observed as reactive oxygen species (ROS) triggered disassembly within tumor cells. PTX triggered immunogenic cell death (ICD) to provoke antitumor immune responses meanwhile NLG919 mediated IDO inhibition relieved immunosuppression in tumor microenvironment. As a result, the dual-stimulus responsive nanoparticles efficiently boosted the synergistic antitumor effects and inhibited postoperative tumor recurrence in a mouse HCC model. Hence, the nanoparticles provide a promising strategy for HCC chemoimmunotherapy.
... The metabolic activity of IDO, which participated in the activation of the kynurenine pathway, has been widely investigated as a potential mechanism of the MSC immunosuppressive effect on the immune cells. Meisel showed increased IDO activity and enhanced immunosuppressive capacities, especially inhibition of T cell proliferation [107][108][109], which was widely investigated in immune and inflammatory diseases such as GVHD [110], EAE [111] and CCl4-induced liver injury [112]. Moreover, the application of competitive inhibitors of IDO, such as 1-methyl-tryptophan (1-MT) rescued the inhibition effects of MSCs on immune cells, including T cells [108]. ...
Background
Mesenchymal stromal/stem cells (MSCs) are the most promising stem cells for the treatment of multiple inflammatory and immune diseases due to their easy acquisition and potent immuno-regulatory capacities. These immune functions mainly depend on the MSC secretion of soluble factors. Recent studies have shown that the metabolism of MSCs plays critical roles in immunomodulation, which not only provides energy and building blocks for macromolecule synthesis but is also involved in the signaling pathway regulation.
Aim of Review
A thorough understanding of metabolic regulation in MSC immunomodulatory properties can provide new sights to the enhancement of MSC-based therapy.
Key scientific Concepts of Review
MSC immune regulation can be affected by cellular metabolism (glucose, fatty acid and amino acid metabolism), which further mediates MSC therapy efficiency in inflammatory and immune diseases. The enhancement of glycolysis of MSCs, such as signaling molecule activation, inflammatory cytokines priming, or environmental control can promote MSC immune functions and therapeutic potential. Besides glucose metabolism, inflammatory stimuli also alter the lipid molecular profile of MSCs, but the direct link with immunomodulatory properties remains to be further explored. Arginine metabolism, glutamine-glutamate metabolism and tryptophan-kynurenine via indoleamine 2,3-dioxygenase (IDO) metabolism all contribute to the immune regulation of MSCs. In addition to the metabolism dictating the MSC immune functions, MSCs also influence the metabolism of immune cells and thus determine their behaviors. However, more direct evidence of the metabolism in MSC immune abilities as well as the underlying mechanism requires to be uncovered.
... In fact, most if not all parental cell constituents were found in their derived EVs when EV signatures were compared with their parental cell with -omics methods (proteomics, lipidomics, RNAseq) [2]. EVs raised a particular interest in the community when they were reported to have therapeutic activities in a myriad of preclinical models ranging from cardiovascular, neurodegenerative, skin, COVID-19 pneumopathy, inflammatory diseases and regenerative medicine via the induction of various pleiotropic effects: angiogenesis [3], immunomodulation [4], cell proliferation [5], inhibiting fibrosis [6], resolving inflammation [7], etc. Hence, a growing number of clinical trials are launched based on the therapeutic properties of EVs [8]. ...
... It is therefore very likely that MSC-EVs would also benefit from this pleiotropic effect as MSC-EVs are now reported to be the mediator of MSC therapeutic effect [118]. Some teams either tried to inhibit [3,6,7,[119][120][121][122] or activate particular pathways [4] that were expected to be involved in the MSC-EV mechanism of action and have shown that in both cases the results were not unequivocal, each pathway having been only responsible for a fraction of the effect. ...
... Then, challenges faced for the clinical development of EV mediated RNA delivery concern both EV and RNA massive bio-production, as well as efficient encapsulation of RNA in EVs with controlled and scal-able processes. Furthermore, EV based delivery may benefit from intrinsic EV properties depending on its parental cell type (angiogenesis [3], immunomodulation [4], increased cell proliferation [5], fibrosis inhibition [6], inflammation resolution [7], etc.) compared to synthetic vectors. Of note, quantitative results on which are based our estimations are usually very limited (if any) and subject to caution. ...
Extracellular vesicles (EVs) are 50–1000 nm vesicles secreted by virtually any cell type in the body. They are expected to transfer information from one cell or tissue to another in a short- or long-distance way. RNA-based transfer of information via EVs at long distances is an interesting well-worn hypothesis which is ~15 years old. We review from a quantitative point of view the different facets of this hypothesis, ranging from natural RNA loading in EVs, EV pharmacokinetic modeling, EV targeting, endosomal escape and RNA delivery efficiency. Despite the unique intracellular delivery properties endowed by EVs, we show that the transfer of RNA naturally present in EVs might be limited in a physiological context and discuss the lessons we can learn from this example to design efficient RNA-loaded engineered EVs for biotherapies. We also discuss other potential EV mediated information transfer mechanisms, among which are ligand–receptor mechanisms.
... Stem cell types examined in preclinical stroke models include embryonic stem cells, hematopoietic stem cells (HSCs), neural stem cells (NSCs), mesenchymal stem cells (MSCs), adult tissue-derived stem cells, and induced pluripotent stem cells (IPSCs) [28,29]. Preclinical studies for stem cell transplantation post-stroke, particularly MSCs and NSCs, have shown improved cognitive and motor function through replacement, neuroprotection, growth factor secretion (bystander effect), neurotrophic factor secretion, extracellular matrix (ECM) remodeling, recruitment of endogenous NSCs (biobridge), immune regulation, neurogenesis, angiogenesis, astrogenesis, synaptogenesis, and oligodendrogenesis [12,13,[30][31][32][33][34][35][36]. To determine the optimal cell type to use for exogenous stem cell transplantation, issues surrounding efficacy, availability, and ethics are to be considered [29]. ...
Stem cell transplantation with rehabilitation therapy presents an effective stroke treatment. Here, we discuss current breakthroughs in stem cell research along with rehabilitation strategies that may have a synergistic outcome when combined together after stroke. Indeed, stem cell transplantation offers a promising new approach and may add to current rehabilitation therapies. By reviewing the pathophysiology of stroke and the mechanisms by which stem cells and rehabilitation attenuate this inflammatory process, we hypothesize that a combined therapy will provide better functional outcomes for patients. Using current preclinical data, we explore the prominent types of stem cells, the existing theories for stem cell repair, rehabilitation treatments inside the brain, rehabilitation modalities outside the brain, and evidence pertaining to the benefits of combined therapy. In this review article, we assess the advantages and disadvantages of using stem cell transplantation with rehabilitation to mitigate the devastating effects of stroke.
... Chin et al. claimed that the anti-inflammatory cytokine levels remained elevated from baseline up until six months post-MSC transfusion [46]. MSCs are known to secrete immunosuppressive factors such as transforming growth factor-beta (TGF-β), vascular endothelial growth factor (VEGF), IL-10, prostaglandin E2 (PGE2), indoleamine 2,3-dioxygenase (IDO), and galectin-1 into the circulatory system [47][48][49][50][51]. These molecules interact with the immune cells such as T and B cells to suppress their proliferation and differentiation, causing the polarization of macrophage to an anti-inflammatory phenotype and reduction of the pro-inflammatory milieu consists of cytokines such as tumor necrosis factor-alpha (TNF-α), interferongamma (IFN-γ), and IL-6 [46,47,49,[52][53][54][55]. ...
Cell therapy involves the transplantation of human cells to replace or repair the damaged tissues and modulate the mechanisms underlying disease initiation and progression in the body. Nowadays, many different types of cell-based therapy are developed and used to treat a variety of diseases. In the past decade, cell-free therapy has emerged as a novel approach in regenerative medicine after the discovery that the transplanted cells exerted their therapeutic effect mainly through the secretion of paracrine factors. More and more evidence showed that stem cell-derived secretome, i.e., growth factors, cytokines, and extracellular vesicles, can repair the injured tissues as effectively as the cells. This finding has spurred a new idea to employ secretome in regenerative medicine. Despite that, will cell-free therapy slowly replace cell therapy in the future? Or are these two modes of treatment still needed to address different diseases and conditions? This review provides an indepth discussion about the values of stem cells and secretome in regenerative medicine. In addition, the safety, efficacy, advantages, and disadvantages of using these two modes of treatment in regenerative medicine are also critically reviewed.
Zero‐dimensional boron nitride quantum dots (BNQDs) are arousing interest for their versatile optical, chemical, and biochemical properties. Introducing carbon contents in BNQDs nanostructures is a great challenge to modulate their physicochemical properties. Among the carbon moieties, phenolic groups have attracted attention for their biochemical properties and phenol‐containing nanomaterials are showing great promise for biomedical applications. Herein, the first example of direct synthesis of water dispersible BNQDs exposing phenolic and carboxylic groups is presented. The carbon‐BNQDs are prepared in a single‐step by solvent‐assisted reaction of urea with boronic reagents and are characterized by optical absorption, luminescence, Raman, Fourier transform infrared and NMR spectroscopy, X‐ray photoelectron spectroscopy, dynamic light scattering, and atomic force microscopy. The carbon‐BNQDs exhibit nanodimension, stability, high photothermal conversion efficiency, pH‐responsive luminescence and Z‐potential. The potential of the carbon‐BNQDs to provide photothermal materials in solid by embedding in agarose substrate is successfully investigated. The carbon‐BNQDs exhibit biocompatibility on colorectal adenocarcinoma cells (Caco‐2) and protective effects from chemical and oxidative stress on Caco‐2, osteosarcoma (MG‐63), and microglial (HMC‐3) cells. Amplicon mRNA‐seq analyses for the expression of 56 genes involve in oxidative‐stress and inflammation are performed to evaluate the molecular events responsible for the cell protective effects of the carbon‐BNQDs.
