The signaling mechanisms active within mesenchymal stromal cells (MSCs) influence the composition of microvesicles (MVs) and exosomes (Exos) secreted by them. Previously, we showed that priming MSCs with a p38 pharmacological inhibitor (pMSCs) rejuvenates them and improves their ability to promote ex vivo hematopoietic stem cell (HSC) expansion. This study examined whether pMSCs exerted HSC-supportive ability via MVs (pMVs) and Exos (pExos). Our findings demonstrate distinct regulation of HSC fate by pMVs and pExos. pMVs promoted the expansion of long-term HSCs (LT-HSCs), distinguished by their robust self-renewal capacity and superior engraftment ability. In contrast, pExos facilitated expansion of short-term HSCs (ST-HSCs) with high proliferative and differentiation potential. Infusing a combination of pMVs- and pExos-expanded HSCs as a composite graft resulted in significantly higher HSC engraftment, emphasizing the synergistic interaction between LT- and ST-HSC populations. Gene expression studies, functional and phenotypic experiments showed that pMVs regulate HSC quiescence via the Egr1/Cdkn1a axis, while pExos control HSC proliferation via the Nfya/Cdkn1a axis. These findings provide insights into the molecular mechanisms underlying the differential regulation of HSC function by pMVs and pExos. It also proposes a composite graft strategy of using pMVs and pExos as "MSC-derived biologics" for improving the HSC transplantation success.

Bovine mammosphere-derived epithelial cell (MDEC) cultures are heterogeneous and enriched for stem and progenitor cells. We previously reported that the bovine MDEC secretome, comprised of all bioactive factors secreted by the cells, displays regenerative properties, exerts antimicrobial effects, and modulates neutrophil activity, positioning it as a promising non-antibiotic biologic therapy for infectious diseases important to the dairy industry, like mastitis. Mastitis is defined as inflammation of the udder, and it is typically caused by bacterial infection. The effect of the MDEC secretome on macrophages, a first line of defense against bacterial infections in the udder, is unknown and could impact the utility of the secretome as a therapy for mastitis. To address this, we isolated bovine monocytes from peripheral blood and maintained them as an unpolarized (M0) population or polarized them into M1 or M2 phenotypes. Macrophages cultured with the secretome of bovine MDECs were assessed for their ability to phagocytose labeled bacterial particles and accumulate reactive oxygen species (ROS). We used single-cell RNA sequencing (scRNA-seq) and fluorescence-activated cell sorting (FACS) to isolate a subpopulation of MDECs that exert enhanced effects on macrophages. We found that the secretome of MDECs that do not express cluster of differentiation (CD) 73, a cell surface enzyme used as a marker for mesenchymal stromal cells, most strongly increased macrophage phagocytosis and ROS accumulation. These findings will help optimize the generation of the bovine MDEC secretome as a suitable treatment option for mastitis.

Mesenchymal Stromal/Stem Cells (MSCs) are among the most frequently studied cell types in clinical trials, and their small extracellular vesicles (sEVs) are now being extensively investigated for therapeutic applications. The RNA cargo of MSC-sEVs, particularly miRNAs and mRNAs, is widely believed to be a key therapeutic component of these vesicles. In this review, we critically examine using first principles and peer-reviewed literature, whether MSC- extracellular vesicles (MSC-EVs) can deliver sufficient quantity of functional miRNA or mRNA to target compartments within recipient cells to elicit a pharmacological response. Several RNA sequencing studies reveal that miRNAs are underrepresented in the small RNA population of MSC-sEVs compared to the parent MSCs. Additionally, the majority of miRNAs are mature forms that are not associated with Argonaute (AGO) proteins, essential for their function in RNA-induced silencing complexes (RISCs). Compounding this, cellular uptake of EVs is generally inefficient, with less than 1% being internalized, and only a fraction of these reaching the cytosol. This suggests that EVs may not deliver miRNAs in sufficient quantities to meaningfully interact with AGO proteins, either through canonical or non-canonical pathways, or with other proteins like Toll-like receptors (TLRs). Further, MSC-sEV RNAs are generally small, with sizes less than 500 nucleotides indicating that any mRNA present is likely fragmented as the average mammalian mRNA is approximately 2000 nucleotides, a fact confirmed by RNA sequencing data. Together, these findings challenge the notion that RNA, particularly miRNAs and mRNAs, are primary therapeutic attributes of MSC-sEVs.

The clinical application of human umbilical cord mesenchymal stem cells (hUCMSCs) in the treatment of liver failure faces challenges due to cell quality, short engraftment time, and limited efficacy. Here, gelatin microcryogel (GM) microcarriers with pore sizes ranging from 15 to 36 μm were tuned from mixed gelatin and glutaraldehyde to develop a 3D culture system of hUCMSCs with improved therapeutic effects. Bulk RNA sequencing and in vitro assays showed that 3D-hUCMSCs exhibited significant improvement in signaling pathways related to paracrine secretion and anti-inflammation. These 3D-hUCMSCs superior compared to 2D-hUCMSCs not only in terms of paracrine secretion, protection from oxidation, and resistance to mechanical force damage, but also had better liver function improvement effect than 2D-hUCMSCs when they were transplanted as single cells into liver injury mice. Furthermore, a gelatin sponge patch grafting (GSPG) strategy was developed to enable the direct engraftment of 3D-hUCMSCs within the GM microcarriers. The results showed that overall engraftment in the host liver was significantly improved, and the life span of transplanted hosts was extended. Our study provided a practical strategy to achieve high engraftment and long retraining time of 3D-hUCMSCs in rescuing acute liver failure with gelatin matrixes.

Despite the significant potential of mesenchymal stem cells (MSC) therapy in clinical settings, challenges persist regarding the efficient detection of consistency and uniformity of MSC populations. Raman spectroscopy is a fast, convenient, and nondestructive technique to acquire molecular properties of biomolecules across laboratory and mass-production settings. Here we utilized Raman spectroscopy to evaluate the heterogeneity of primary MSC from varying donors, passages, and distinct culture conditions, and compared its effectiveness with conventional techniques such as flow cytometry. Although these MSC exhibited insignificant differences in morphology and surface markers in flow cytometry analysis, they could be distinctly clustered into different populations by Raman spectroscopy and the subsequent machine learning using linear discriminant analysis. Principal component analysis demonstrated limited efficiency in clustering Raman data from diverse sources, which could be enhanced through combination with support vector machine or deterministic finite automation. These findings highlight the sensitivity of Raman spectroscopy in detecting subtle differences. Moreover, the analysis of characteristic Raman peaks attributed to cellular biomolecules in MSC from passages 2 (P2) to P10 revealed a gradual decrease in the levels of nucleic acids, lipids, and proteins with increasing passages, and a significant increase in carotenoids from P8. These results suggest the potential use of Raman spectroscopy to assess cellular biochemical characteristics such as aging, with carotenoids emerging as a potential marker of cell aging. In conclusion, Raman spectroscopy demonstrates the ability to rapidly and non-invasively detect cellular heterogeneity and biochemical status, offering significant potential for quality control in stem cell therapy.

Mesenchymal stromal cells (MSCs) are promising candidates for cell therapy. Most of the therapeutic applications have used adult bone marrow MSCs, adipose MSCs and perinatal tissue-derived MSCs. Recent evidence suggests that MSCs from mid-gestational fetal tissues are more primitive, grow faster and are biologically more closely related to embryonic stem cells than other sources of MSCs. However, the expression of pluripotency genes raises the question of whether these genes are safe for clinical application. In this study, we demonstrated that second-trimester fetal liver-derived MSCs lack the expression of pluripotent markers and maintain their proliferative and osteogenic differentiation potential beyond passage 12. Compared to other sources, FL-MSCs exhibit characteristics that are promising for use in skeletal regeneration.

MSCs were isolated from the second-trimester fetal liver and characterized for surface antigen expression, pluripotency marker expression and multilineage differentiation. The growth kinetics, population doubling, and number of colony-forming units were analyzed at the 3rd, 5th, 8th and 10th passages of FLMSCs and compared with those of BMMSCs. The immunomodulatory properties of FLMSCs were analyzed by a T-cell proliferation assay. The osteogenic differentiation potential of FL-MSCs was assessed at passages 3, 5, 8 and 12 and compared with that of BMMSCs.

We demonstrated that second-trimester fetal liver-derived MSCs exhibited a distinct fibroblast-like spindle-shaped morphology and expressed typical MSC surface antigens. Unlike first-trimester fetal MSCs, second-trimester FL-MSCs did not express pluripotent markers and showed significantly greater self-renewal and proliferative potential at higher passages and a lower apoptotic rate than BM-MSCs. Additionally, the osteogenic differentiation potential of FL-MSCs was 4-6 times greater than that of BM-MSCs at both early and late passages CONCLUSION: Our findings underscore the robust self-renewal and proliferative potential of second-trimester fetal liver-derived MSCs, which notably lack pluripotent markers. The ability of FL-MSCs to sustain osteogenic potential through multiple passages makes them promising candidates for bone tissue engineering and regenerative medicine.

The ultrastructure of the tendon-bone interface (TBI) is inherently complex. After arthroscopic reconstruction, it is often replaced by disorganized scar tissue, which increases the risk of re-tearing.Stem cell therapies offer a promising approach to regenerate the original tissue structure and enhance the healing environment. The effectiveness of these therapies depends on understanding the localization, proliferation, and overall behavior of the implanted stem cells. This study aimed to track the distribution of stem cells in a rat model of rotator cuff injury using Magnetic Resonance Imaging (MRI) and superparamagnetic iron oxide nanoparticles (SPIO) and to evaluate the mechanisms and therapeutic effects of stem cell therapy.

Adipose-derived mesenchymal stem cells (ADSCs) were isolated and expanded, then labeled with SPIO at an optimized concentration. The visibility of these labeled cells was assessed via MRI, along with evaluations of their viability, potential toxicity, and migration capacity in vitro.For the in vivo study, rats with rotator cuff tears were divided into two groups: a control group that received a PBS injection, and a treatment group that received SPIO-labeled ADSCs (designated as S-A). MRI scans were conducted at 1, 2, and 4 weeks post-surgery, followed by histological analysis after the rats were euthanized. At 8 weeks post-surgery, rats were sacrificed, and their shoulder joints were analyzed biomechanically and histologically to assess the overall treatment efficacy.

SPIO nanoparticles were successfully incorporated into ADSCs, and MRI imaging demonstrated that these SPIO-labeled cells significantly enhanced MRI contrast without affecting cell viability, proliferation, or migration ability. Both MRI and histological analyses confirmed that the implanted stem cells survived and remained localized for at least two weeks. Further histological and biomechanical evaluations indicated that the stem cells facilitated the repair of the TBI. This repair process appeared to be mediated by an increase in M2 macrophage activity within the injured tissue, promoting improved local healing conditions.

This study confirms that labeling ADSCs with SPIO nanoparticles is an effective method for tracking these cells in vivo using MRI, providing a non-invasive approach to monitor the repair of injured TBI. Moreover, the localized survival of transplanted stem cells supports their role in enhancing TBI repair by modulating the local inflammatory response.

Dopaminergic neuron loss caused by microglia activation is an important pathological factor of Parkinson's disease (PD). Previously, we reported that small extracellular vesicle from adipose derived stem cells (ADSC-sEVs) could inhibit the activation of microglia and protect neuron apoptosis from microglia activation. However, whether ADSC-sEVs have protective effect on the motor deficit of PD mouse and the exact mechanism remains unknown. In this study, ADSC-sEVs were delivered to experimental model of Parkinson's disease by tail vein injection to explore the in vivo effect of ADSC-sEVs on PD. Next, the potential key microRNA in ADSC-sEVs was screened by RNA sequencing (RNA-seq), and the exact mechanism was further explored. We found that ADSC-sEVs greatly alleviated the activation of microglia and reduced the loss of dopaminergic neurons in the substantia nigra of PD mice, the motor deficit was also significantly improved. By RNA-seq analysis, miR-100-5p was verified as a potential microRNA in this process, because knockdown of miR-100-5p in ADSC-sEVs weakened the protective effect of ADSC-sEVs on PD mouse as well as the anti-inflammatory effect on microglia activation. Finally, we found that miR-100-5p could target Deltex E3 ubiquitin ligase 3 L (DTX3L) and suppress its expression, which then decreased the expression and phosphorylation of Signal Transducers and Activators of Transcription 1 (STAT1), as well as alleviating the activation of microglia. Our findings illustrate that ADSC-sEVs are an effective therapy for PD, and it could be a promising therapy for the treatment of PD.

The human placenta is a transient yet crucial organ that plays a key role in sustaining the relationship between the maternal and fetal organisms. Despite its historical classification as "biowaste," placental tissues have garnered increasing attention since the early 1900s for their significant medical potential, particularly in wound repair and surgical application. As ethical considerations regarding human placental derivatives have largely been assuaged in many countries, they have gained significant attention due to their versatile applications in various biomedical fields, such as biomedical engineering, regenerative medicine, and pharmacology. Moreover, there is a substantial trend toward various animal product substitutions in laboratory research with human placental derivatives, reflecting a broader commitment to advancing ethical and sustainable research methodologies. This review provides a comprehensive examination of the current applications of human placental derivatives, explores the mechanisms behind their therapeutic effects, and outlines the future potential and directions of this rapidly advancing field.

Calcium oxalate (CaOx) is the predominant form of kidney stones, associated with significant morbidity and recurrence rates. Mesenchymal stem cells (MSCs) have shown promise in treating renal injury, but their impact on CaOx stone formation remains unclear.

We established a hyperoxaluria-induced AKI model in mice through intraperitoneal injection of glyoxylate. Two types of MSCs, bone marrow-derived MSCs (BMSCs) and umbilical cord-derived mesenchymal stem cells (UMSCs), were injected through tail vein injection. Histological evaluations and blood biochemical tests were performed to assess crystal deposition and kidney function. The inflammatory response and NLRP3 inflammasome activation were assessed using immunofluorescence, immunohistochemistry, TUNEL staining, and qPCR. In vitro, macrophages were cocultured in the presence of MSCs. ELISA was used to measure IL-1β and IL-18 release. MTS assays assessed renal epithelial cell protection. Western blotting evaluated NLRP3 inflammasome activation in macrophages.

Both BMSCs and UMSCs significantly inhibited CaOx crystal deposition and kidney injury by inhibiting NLRP3 inflammasome activation. In vitro, both MSC types suppressed NLRP3 inflammasome activation in macrophages through the NF-κB signaling pathway, leading to decreased release of IL-1β and IL-18 and enhanced protection of renal epithelial cells. This attenuation of renal tubular cell injury is a critical factor in preventing CaOx stone formation.

Our findings reveal that Both BMSCs and UMSCs effectively attenuate hyperoxaluria-induced kidney injury and crystal deposition by inhibiting NLRP3 inflammasome activation. This discovery is helpful for developing new effective therapeutic means for nephrolithiasis.

Despite the tremendous success in preclinical models, the translation of human mesenchymal stromal cells (hMSCs) as a therapy in the clinic is not up to the expectation. Intrinsic factors (age, sex, health status, life style of the donor, source, cellular senescence, and oxidative stress in hMSCs), extrinsic factors (culture system, batch-to-batch variations, choice of biomaterials, cell processing and preservation protocols), and host microenvironment (inflammatory milieu, oxidative stress, and hypoxia in the recipient) compromise the overall therapeutic efficacy of the transplanted hMSCs. In recent times, the approach of 'Disease Microenvironment Preconditioning (DMP)' has garnered attention to overcome the host-associated attributes involved in compromised hMSCs therapeutic potential. In this review, we discuss various approaches of DMP of hMSCs by employing serum and other body fluids obtained from diseased patients/animals and small molecules, including cytokines such as IFN-γ, IL-6, IL-10, IL- β, TGF-β1, IL-1α, IL-1β, TNF-α, HMGB1, IL-17 A, and IL-8 which are associated with disease conditions. DMP strengthens hMSCs ability to adapt/acclimatize and respond more efficiently to the hostile microenvironment they encounter upon transplantation. DMP modulate hMSCs to withstand inflammation, survive under hypoxic and nutrient-deprived conditions, and resist oxidative stress. Evidence from various disease models ranging from cardiovascular and neurodegenerative disorders to autoimmune diseases and tissue injuries supports the role of DMP in improving hMSC survival, integration, and functional efficacy. While the potential of DMP to revolutionize MSC-based therapies is evident, challenges such as standardizing/optimizing protocols for preconditioning is essential. This review synthesizes current advancements in the approach of DMP aiming to propel the area of regenerative medicine.

Menopausal syndrome is characterized by a wide range of physical and psychological symptoms in women aged 40s-50s as a result of hormonal fluctuations and age-related decline. Various treatments have been used to manage the symptoms, including hormone replacement therapy, but no effective causal therapies have yet been identified. Regenerative medicine has gained considerable attention as a promising approach to age-related problems, and mesenchymal stem cell therapies have been extensively studied. Recently, menstrual blood has emerged as a novel cell source of stem cells, called menstrual blood-derived stem cells (MenSCs), due to its non-invasive, regular and consistent collection from women. In this study, we have investigated the therapeutic potential of intravenous administration of autologous MenSCs on female menopausal syndromes.

Menstrual blood was collected from 15 patients aged 30s-60s with ovarian dysfunction using a menstrual cup, and MenSCs were isolated, cultured and expanded. Patients received either 3 × 107 cells or 1 × 108 cells intravenously 1 to 5 times at intervals of more than 1 month. Patient-reported symptoms were assessed using the Simplified Menopausal Index at pre-treatment and after 1, 3, 6, and 12 months, and safety assessments were performed. Serum estradiol and follicle-stimulating hormone levels were also measured by immunoassay.

Almost all patients who received MenSCs experienced a sharp reduction in menopausal symptoms, including vasomotor, neuropsychiatric, and motor symptoms, one month after the first administration, and these symptoms remained low for 6 months. The Simplified Menopausal Index score was significantly reduced after treatment. The reducing potency of 1 × 108 MenSCs was greater than that of 3 × 107 MenSCs. Patients who received a higher number of MenSCs showed an increasing trend in estradiol levels and a decreasing trend in follicle-stimulating hormone levels. When MenSCs were administered to postmenopausal patients, this trend was more pronounced. Overall, no apparent serious adverse events were observed during these treatments.

The present results suggest that the administration of MenSCs improved menopausal symptoms and regulated hormonal balance without any serious adverse events. This is the first report on the promising therapeutic potential of cell-based therapy using autologous MenSCs for female menopausal syndrome.

Osteoarthritis (OA) is a prevalent degenerative joint disease characterized by progressive cartilage loss, inflammation, and joint dysfunction. With profound effects on joint function and quality of life, OA imposes a substantial socio-economic burden. As of now, OA remains incurable, lacking approved medications, regenerative therapies, or procedures that can halt the progressive destruction of the joint. Intraarticular (IA) injections have emerged as a cornerstone in the management of knee OA, offering localized minimally invasive therapeutic options. Traditional IA therapies, including corticosteroids and hyaluronic acid (HA), primarily aim to reduce pain but lack regenerative capacity. Biologic IA therapies for knee OA including autologous blood-derived products like platelet-rich plasma (PRP), bone marrow aspirate concentrate (BMAC) and mesenchymal stromal cells (MSCs) have become more commonly used. Finally, newer IA therapies such as fibroblast growth factor 18 and gene therapy are being investigated. In this review, we highlight the current evidence around IA injections for the treatment of knee OA.

ObjectiveThis study aimed to isolate and compare the mesenchymal stem cell characteristics of CD90+ cells from different fibrocartilage tissues in the temporomandibular joint (TMJ), the knee joint, and the intervertebral joint to further understand the similarities and differences of these 4 fibrocartilage tissues.MethodsCD90+ cells were isolated from TMJ disc, condylar cartilage, meniscus, and intervertebral disc by using magnetic-activated cell sorting. Cellular assays including 4.5-ethynyl-2'-deoxyuridine labeling, multilineage differentiation, colony formation, and cell migration were conducted to compare their mesenchymal stem cell characteristics. Immunofluorescent staining was performed for observing the expression of actively proliferating CD90+ cells within the tissues. H&E staining and Safranine O staining were used to compare the histological features.ResultsThe CD90+ cells derived from these 4 fibrocartilage tissues exhibited comparable cell proliferation abilities. However, the cells from the TMJ disc displayed limited multilineage differentiation potential, colony formation, and cell migration abilities in comparison with the cells from the other fibrocartilage tissues. In vivo, there was relatively more abundant expression of CD90+ cells in the TMJ disc during the early postnatal stage. The limited EDU+ cell numbers signified a low proliferation capacity of CD90+ cells in the TMJ disc. In addition, we observed a significant decrease in cell density and a restriction in the synthesis of extracellular proteoglycans in the TMJ disc.ConclusionOur study highlights the spatial heterogeneity of CD90+ cells in the fibrocartilages of different joint tissues, which may contribute to the limited cartilage repair capacity in the TMJ disc.

In recent years, various clinical trials have been designed and implemented using mesenchymal stem cells (MSCs) for the treatment of heart diseases. Clinical trials exploring MSC-based treatments have proliferated, yet the lack of standardized protocols for MSC administration remains a significant challenge. Despite the growing popularity of MSC trials, questions persist regarding optimal dosing, administration routes, and frequency to achieve safety and efficacy, particularly in the context of cardiac regeneration. The current study has reviewed the clinical trials that have used MSCs for the treatment of heart diseases since 2009. The findings reveal diverse transplantation methods and varying MSCs quantities, highlighting the absence of a universal guideline for MSCs utilization in heart disease clinical trials.

Mesenchymal stem cell (MSC) therapy, with its unique properties, has garnered interest in cancer treatment. Exosomes (EXOs)-derived from MSC retain the paracrine components of MSCs and demonstrate increased stability, minimal immunogenicity, and low risk of unintended tumorigenesis. Enhanced endocytosis methods make them versatile delivery vehicles for therapeutic cargo. MSC-EXOs can either promote or inhibit carcinogenesis, mediated by paracrine factors and various RNA molecules, particularly microRNAs (miRNAs). The prospect of using MSC-EXOs as a delivery tool for antitumor miRNAs in solid tumor therapy is promising. Exosomes' intrinsic tumor-targeting abilities and low immunogenicity make them ideal for delivering miRNAs, which have shown potential as cancer therapeutics. miRNAs within MSC-EXOs molecules can stimulate tumor growth or induce non-apoptotic cell death pathways, such as ferroptosis, depending on context. Ferroptosis is a kind of controlled cell death that is associated with the pathophysiology of several illnesses and includes iron metabolism. There is growing evidence that miRNAs carried by exosomes derived from MSCs may control ferroptosis in tumor cells by altering key genes related to antioxidant defense, lipid peroxidation, and iron metabolism. Understanding their complex mechanisms in the tumor microenvironment and optimizing their cargo are critical steps toward harnessing their full therapeutic potential. This review provides a comprehensive overview of MSC-EXOs and their role in cancer treatment. We also discuss the potential of MSC-EXOs as delivery vehicles for miRNAs to enhance therapeutic efficacy, as well as the role of exosomal miRNAs in the induction of ferroptosis.

Post-mastectomy radiotherapy plays a crucial role in breast cancer treatment but can lead to an inflammatory response causing soft tissue damage, particularly radiation-induced capsular contracture (RICC), impacting breast reconstruction outcomes. Adipose-derived stem cells (ADSCs), known for their regenerative potential via paracrine capacity, exhibit inherent radiotolerance. The influence of tumor necrosis factor-alpha (TNF-α) on ADSCs has been reported to enhance the paracrine effect of ADSCs, promoting wound healing by modulating inflammatory responses.

This study investigates the potential of TNF-α-treated human ADSCs (T-hASCs) on silicone implants to alleviate RICC, hypothesizing to enhance suppressive effects on RICC by modulating inflammatory responses in a radiation-exposed environment.

In vitro, T-hASCs were cultured on various surfaces to assess viability after exposure to radiation up to 20 Gy. In vivo, T-hASC and non-TNF-α-treated hASC (C-hASCs)-coated membranes were implanted in mice before radiation exposure, and an evaluation of the RICC mitigation took place 4 and 8 weeks after implantation. In addition, the growth factors released from T-hASCs were assessed.

In vitro, hASCs displayed significant radiotolerance, maintaining consistent viability after exposure to 10 Gy. TNF-α treatment further enhanced radiation tolerance, as evidenced by significantly higher viability than C-hASCs at 20 Gy. In vivo, T-hASC-coated implants effectively suppressed RICC, reducing capsule thickness. T-hASCs exhibited remarkable modulation of the inflammatory response, suppressing M1 macrophage polarization while enhancing M2 polarization. The elevated secretion of vascular endothelial growth factor from T-hASCs is believed to induce macrophage polarization, potentially reducing RICC.

This study establishes T-hASCs as a promising strategy for ameliorating the adverse effects experienced by breast reconstruction patients after mastectomy and radiation therapy. The observed radiotolerance, anti-fibrotic effects, and immune modulation suggest the possibility of enhancing patient outcomes and quality of life. Further research and clinical trials are warranted for broader clinical uses.

A substantial proportion of patients diagnosed with rheumatologic and musculoskeletal diseases (RMDs) exhibit resistance to conventional therapies or experience recurrent symptoms. These diseases, which include autoimmune disorders such as multiple sclerosis, rheumatoid arthritis, and systemic lupus erythematosus, are marked by the presence of autoreactive B cells that play a critical role in their pathogenesis. The persistence of these autoreactive B cells within lymphatic organs and inflamed tissues impairs the effectiveness of B-cell-depleting monoclonal antibodies like rituximab. A promising therapeutic approach involves using T cells genetically engineered to express chimeric antigen receptors (CARs) that target specific antigens. This strategy has demonstrated efficacy in treating B-cell malignancies by achieving long-term depletion of malignant and normal B cells. Preliminary data from patients with RMDs, particularly those with lupus erythematosus and dermatomyositis, suggest that CAR T-cells targeting CD19 can induce rapid and sustained depletion of circulating B cells, leading to complete clinical and serological responses in cases that were previously unresponsive to conventional therapies. This review will provide an overview of the current state of preclinical and clinical studies on the use of CAR T-cells and other cellular therapies for RMDs. Additionally, it will explore potential future applications of these innovative treatment modalities for managing patients with refractory and recurrent manifestations of these diseases.

Stem cell-based therapies have emerged as a promising frontier in the treatment of gastrointestinal disorders, offering potential solutions for challenges posed by conventional treatments. This review comprehensively examines recent advancements in cell-based therapeutic strategies, particularly focusing on stem cell applications, immunotherapy, and cellular therapies for digestive diseases. It highlights the successful differentiation of enteric neural progenitors from pluripotent stem cells and their application in animal models, such as Hirschsprung disease. Furthermore, the review evaluates clinical trials and experimental studies demonstrating the potential of stem cells in regenerating damaged tissues, modulating immune responses, and promoting healing in conditions like Crohn's disease and liver failure. By addressing challenges, such as scalability, immunogenicity, and ethical considerations, the review underscores the translational opportunities and obstacles in realizing the clinical potential of these therapies. Concluding with an emphasis on future directions, the study provides insights into optimizing therapeutic efficacy and fostering innovations in personalized medicine for digestive disorders.

While adipose-derived stem cells (ADSCs) transplantation represents an appealing therapeutic strategy for bone defect repair, the osteogenic capacity of ADSCs is largely limited. Melatonin has been demonstrated to contribute to the bone marrow stem cell (BMSC) osteogenesis. However, its effect on the osteogenic differentiation of ADSCs has not yet been determined. This study aims to identify whether melatonin exerts influences on the osteogenic differentiation in rat ADSCs. Rat ADSCs were isolated and identified. Subsequently, the impact of melatonin on the proliferation of rat ADSCs was examined. The effects of melatonin on the phenotypic features as well as marker genes and proteins of osteogenic differentiation were determined through the use of alkaline phosphatase (ALP) staining, ALP activity assay, alizarin red staining (ARS), RT-qPCR, western blot assay, and cellular immunofluorescence assay. To investigate the potential molecular mechanism through which melatonin promotes osteogenic differentiation of rat ADSCs, RNA sequencing, MAPK signaling pathway blocking assay and p38 mRNA interference assay were carried out. The results showed that melatonin at concentrations of 0-100 μM was safe and nontoxic for the proliferation of rat ADSCs, with the concentration at 100 μM exhibiting the most pronounced osteogenesis. Additionally, melatonin was observed to activate the p38/MAPK signaling pathway in rat ADSCs. Moreover, the p38/MAPK pathway inhibitor (SB203580) and siRNA targeting p38 mRNA (p38 siRNA) were found to inhibit the melatonin-promoted osteogenic differentiation of rat ADSCs. In conclusion, the results of this study indicate that melatonin promotes osteogenic differentiation of rat ADSCs through the activation of the p38/MAPK signaling pathway. In light of these findings, melatonin treatment represents an effective strategy for promoting osteogenic differentiation of ADSCs.

Osteoarthritis (OA) is a degenerative joint condition leading to disability. The lack of effective treatment for OA creates a need for the development of new therapeutic approaches that may rely on stem cells including mesenchymal stem/stromal cells (MSCs) and their derivatives such as extracellular vesicles (EVs). The objective of this study was to evaluate the impact of MSCs derived from adipose tissue (AT-MSCs) and umbilical cord (UC-MSCs) and their EVs on cartilage-bone injury in vivo, to identify the specimen with the highest regenerative potential for further clinical applications in patients with OA. Humanized pigs underwent cartilage-bone injuries followed by intraarticular administration of products containing AT-MSCs, UC-MSCs, AT-MSC-EVs or UC-MSC-EVs mixed with hyaluronic acid (HA) or HA alone (for comparison). After 6-m follow-up, almost-fully-healed cartilage-bone defects were observed in the AT-MSC- and UC-MSC-treated pigs, and the defects were filled primarily with hyaline cartilage. In AT-MSC-EV- and UC-MSC-EV-treated pigs, a partial cartilage-bone tissue repair was observed, and the defects were filled primarily with fibrocartilage. The control pigs demonstrated limited regeneration capacity. The microcomputed tomography parameters of the subchondral bone indicated the ongoing progression of OA in controls, whereas in the MSC- and MSC-EV-treated pigs, the parameters indicated the cessation of OA progression. Moreover, no serious side effects were observed after the administration of products containing MSCs or MSC-EVs. The results indicate the safety and regenerative activity of MSCs on injured tissues, which favors not only the healing and improvement of bone structure but also the formation of hyaline cartilage. Superior tissue repair was observed after the administration of products containing AT-MSCs. The treatment of OA with MSC-EVs needs further standardization.

Mesenchymal stem cells have shown promise in many areas of regenerative medicine due to the anti-inflammatory and pro-regenerative effects of the secreted factors. However, successful delivery remains problematic, particularly for delivery to areas such as the brain. Spray delivery is a method investigated in wound care and lung injury, which may be applicable for brain delivery to patients already requiring surgery. To retain therapeutic mesenchymal stem cells at the delivery site, biomaterials can be employed; pectin is a biocompatible, sprayable, and mucoadhesive material, which could prove suitable for spray delivery of cells for therapeutic uses.

The biocompatibility of four grades of low-methyl pectin gelled by addition of calcium was assessed using SH-SY5Y cells. After, mesenchymal stem cells were suspended within the four different grades of low-methyl pectin solutions and sprayed using a syringe-driven spray device. The suitability was then assessed by cell viability testing, flow cytometry to test for surface markers, and differential gene expression studies to understand the effects of both the pectin and the spraying process on the gene expression of the cells.

All four grades of low-methyl pectin were biocompatible with SH-SY5Y cells. The syringe-driven spray device delivered human mesenchymal stem cells to well plates with high viability, and suspending these cells in pectin solutions for spraying did not negatively affect the viability. The grade of pectin named CU-701 was the best grade based on results of the flow cytometry, whereby the surface marker expression was not altered from the control cells. The RNA sequencing showing the differential expression showed that the process of spraying the cells did not alter gene expression compared to the control, however the pectin, and the presence of calcium used to induce gelation of the pectin, did lead to altered gene expression in cells.

Spraying is a suitable delivery method for the mesenchymal stem cells, showing no detrimental effect on the cells. Pectin shows little effect on the viability of the cells, however the use of calcium to gel the pectin appears to affect the expression of several genes.

Aging is the greatest risk factor for breast cancer, and although epithelial cells are the source of carcinomas, epithelial changes alone do not fully explain cancer susceptibility. Fibroblasts and macrophages are key stromal constituents around the cells of origin for cancer in breast tissue. With age, macrophages surrounding terminal ductal lobular units (TDLUs) become increasingly immunosuppressive. CD105+ fibroblasts intercalate within TDLUs, drive luminal differentiation, and give rise to immunosuppressive cancer-associated fibroblasts in other tissues. We propose that differences in fibroblasts are a crucial component of the stroma that shapes cancer susceptibility.

Primary peri-epithelial fibroblast cultures were established from prophylactic and reduction mammoplasties from 30 women ranging in age from 16 to 70 years and from BRCA1 mutation carriers. Growth characteristics, transcriptional profiles, differentiation potential, and secreted proteins were profiled for fibroblast subtypes from diverse donors. Co-cultures with fibroblasts, macrophages, and T cells were used to ascertain the functional role played by CD105+ fibroblasts in immune cell modulation.

We found that peri-epithelial CD105+ fibroblasts are enriched in older women as well as women who carry BRCA1 mutations. These CD105+ fibroblasts exhibit robust adipogenesis and secrete factors related to macrophage polarization. Macrophages cocultured with fibroblasts better maintain or enhance polarization states than media alone. CD105+ fibroblasts increased expression of immunosuppressive macrophage genes. CD105+ fibroblasts supported anti-inflammatory macrophage-mediated suppression of T cell proliferation, whereas CD105- fibroblasts significantly reduced the suppressive effect of anti-inflammatory macrophages on T cell proliferation.

Establishment of a coculture system to dissect the molecular circuits between CD105+ fibroblasts and macrophages that drive immunosuppressive macrophage polarization has broad utility in understanding mammary gland development and events that precede cancer initiation. CD105+ fibroblasts and macrophages may coordinate to suppress immunosurveillance and increase breast cancer susceptibility.

Recently, cell therapies, including chimeric antigen receptor (CAR) modified T cell therapy and mesenchymal stem cell (MSC) therapy, have demonstrated considerable potential for systemic lupus erythematosus (SLE). In this study, a CAR-MSC model was constructed, combining two cell therapies. The structural domains of the CAR were designed by using the anti-CD19 scFv, targeting the CD19 antigen on the surface of B cells and the intracellular region of the interferon-gamma receptor, activating the JAK-STAT1 signaling pathway. Then we screened and identified the most effective structural domain of CAR as CAR1, as it facilitates MSCs to maintain significantly higher levels of JAK2 phosphorylation and IDO expression, as shown by western blot analysis. We also demonstrated CAR1 could be consistently and stably expressed at high levels in MSCs, and CAR1 transduction did not significantly affect the surface antigenic phenotypic criteria of MSCs via flow analysis. Furthermore, immunofluorescence results showed CAR1-MSCs could stably bind CD19 antigen, and they were activated by human CD19 antigen resulting in significantly high JAK2 phosphorylation and IDO expression via western blot analysis following co-culture. Besides, when activated peripheral blood mononuclear cells (PBMCs) were co-cultured with untransduced MSCs (UTD-MSCs) and CAR1-MSCs in vitro, respectively, the results showed that the percentage of activated CD3+ T cells and CD19+ B cells was both significantly lower after co-culturing. The percentage of activated CD19+ B cells was lower in the CAR1-MSCs co-culture group than in the UTD-MSCs co-culture group, whereas the percentage of activated CD3+ T cells was similar in the two co-culture groups. This suggests that CAR1 increased the inhibitory ability of MSCs on activated CD19+ B cells and had no significant effect on the ability of MSCs to inhibit activated CD3+ T cells. In conclusion, CAR1-MSCs were successfully constructed and demonstrated the ability to enhance the inhibitory effect of MSCs on activated human CD19+ B cells, facilitating SLE therapy.

This study was conducted to evaluate the impact of sodium hydrogen sulfide (NaHS) on the therapeutic efficacy of bone marrow mesenchymal stem cells (BM-MSCs) in the treatment of collagen-induced arthritis (CIA) rats. MSCs were isolated and cultured from rat bone marrow, and their characteristics were determined. The CIA model was induced in rats by intradermal injections of type II collagen on days 0 and 21. A variety of treatments were administered, including naproxen, BM-MSCs, BM-MSC-conditioned media, NaHS, BM-MSCs preconditioned with NaHS, and BM-MSCs preconditioned with NaHS-conditioned media. The infused BM-MSCs homed to the bone trabeculae and cartilage of the knee joint, leading to significant improvements in gait scores and a reduction in paw withdrawal frequency (PWF). Treatment with BM-MSCs and NaHS also significantly suppressed serum levels of CRP, RF, and 14-3-3η, while downregulating TNF-α gene expression and MMP-1 protein levels in the synovial membrane. Histopathological analysis confirmed these biochemical and molecular genetic findings. Notably, CIA rats treated with BM-MSCs preconditioned with NaHS showed the most significant improvements, with outcomes closely resembling those of healthy controls. This study concludes that NaHS enhances the therapeutic efficacy of BM-MSC therapy for rheumatoid arthritis (RA) by augmenting their anti-inflammatory, immunomodulatory, and regenerative properties.

Ginsenoside Rg1 is a key bioactive compound in ginseng, a traditional herbal medicine known for tonifying qi and nourishing blood, traditionally used to replenish "qi" by regulating hematopoietic function. But its underlying mechanism remains to be elucidated.

This study aims to delve into the role of Rg1 on senescent hematopoiesis and its mechanisms.

A model of D-galactose-induced hematopoietic injury was established with ginsenoside Rg1. The hematopoietic supportive effect of Rg1 was assessed by quantifying the levels of hematopoietic supportive factors VCAM1, CXCL12 and SCF, CFU-Mix formation and cellular senescence; and the levels of inflammatory factors and oxidative stress were measured by ELISA in the serum and cellular supernatant of mice. Co-culture technique was used to examine the ability of Rg1 to restore impaired hematopoiesis by improving the inflammatory hematopoietic microenvironment. For mechanism exploration, RNA-Seq was used to detect differential genes in Rg1-treated MSCs, GO- and KEGG-based enrichment analyses were used to screen the key pathways in which Rg1 exerts its effects, and molecular docking was used to demonstrate the feasibility of molecular interconnections between Rg1 and TLR2. To further explore the mechanism, pathway activators were further used and the expression levels of target proteins downstream of the TLR2 pathway were quantified using Western blotting.

Rg1 decreased the levels of inflammatory factors IL-1β, IL-6 and TNFα, while enhancing the expression of hematopoietic support factors in senescent MSCs, thereby improving the self-renewal and differentiation of aged HSPCs. Additionally, Rg1 also delayed HSPC senescence and reduced the level of oxidative stress. KEGG and GO were enriched for the Toll/NF-κB signaling pathway, based on differential genes obtained by transcriptional sequencing. Rg1 could inhibit the elevated levels of MyD88, NF-κB-p65 and IκBα proteins, and their phosphorylation levels by binding to TLR2 protein and inhibiting them. In conclusion, Rg1 ameliorates the inflammatory hematopoietic microenvironment induced by MSCs senescence via the TLR2/NF-κB-p65 signaling pathway, alleviating HSPCs senescence.

Our results reveal the mechanism by which Rg1 regulates HSPCs function and represent a potential therapeutic strategy for hematopoietic dysfunction, highlighting the potential value of traditional Chinese medicine extracts in clinical applications.

The acquisition of suitable stem cell sources is a significant issue in regenerative medicine. There has been considerable interest in utilizing mesenchymal stem cells (MSCs) derived from endometrial and menstrual blood as a promising resource of MSCs, owing to their unique biochemical properties and prospective use in clinical therapies. This population of stem cells has distinct characteristics in terms of immunophenotype, proliferation rate, and differentiation capacity. A notable characteristic of these stem cells is their capacity to develop into mesodermal lineages, highlighting their regenerative capability. Moreover, the presence of certain surface markers facilitates the augmentation of clonogenic endometrial MSCs. Their distinctive characteristics, along with their swift multiplication ability, underscore their significant promise for therapeutic applicability in regenerative medicine and cell-based treatments. Current investigations are examining possible usage of diverse stem cell resources in the treatment of inflammatory diseases and perhaps intractable illnesses like Parkinson's disease, utilizing their immunomodulatory properties. This review aims to analyze stem cell-related research that has utilized endometrial and menstrual blood-derived MSCs (enMSCs and MenSCs) with a special focus on their clinical application. We will explore the existing evidence about the therapeutic potential for these stem cells across many medical diseases and address the obstacles and prospective trajectories in this domain. Additionally, we will study the unique properties of enMSCs and MenSCs that make them promising candidates for regenerative medicine.

In regenerative medicine, mesenchymal stem cells (MSCs) constitute a promising therapeutic route for many diseases. The current quality-by-design guidelines do not clearly define a framework for MSC production. Here, we suggest and experimentally validate a model-based method to determine design spaces (DSs) for MSC cultivation. A kinetic model used in previous work was employed; part of the experimental data was used to re-estimate the maximum specific growth rate in the kinetic model and then calculate the prediction intervals of this parameter. Subsequently, regions of seeding density and harvesting time where both the upper and lower limits of growth predictions met the acceptable number of cells and confluency with given risk levels were defined as DSs. Finally, the established DS was validated with the remaining data; it allowed better predictions of the cell numbers and confluency under specific cultivation conditions and improved the overall robustness of MSC cultivation processes.

This study aims to investigate the mechanisms of MSC therapy for acute kidney injury, focusing on the regulation of mitochondrial function and pyroptosis in renal tubular epithelial cells (RTECs).

An in vivo ischemia/reperfusion (I/R) model was used to assess the effects of MSC treatment on mitochondrial membrane potential, mitochondrial function, cell pyroptosis, and PGC-1α expression in RTECs.

MSCs significantly improved mitochondrial function in RTECs by upregulating PGC-1α expression, regulating mitochondrial fusion and fission proteins, reducing mitochondrial ROS production, and suppressing NLRP3 inflammasome activation. Furthermore, MSC treatment reduced the levels of pyroptotic markers, such as IL-18, and exhibited a marked anti-fibrotic effect in the long-term. These findings suggest that MSCs not only repair acute kidney injury but also offer long-term protection against fibrosis.

MSCs improve the repair of acute kidney injury by modulating mitochondrial function and inhibiting pyroptosis, providing new theoretical support for MSC-based therapies in AKI treatment.

Periodontitis is a chronic inflammatory disease and macrophages play a pivotal role in the progression of periodontitis. Mesenchymal stem cells (MSCs) have emerged as potential therapeutic agents for the treatment of periodontitis due to their immunomodulatory properties and capacity for tissue regeneration. Compared to conventionally derived MSCs, induced pluripotent stem cell-derived MSCs (iMSCs) offer distinct advantages as promising candidates for MSC-based therapies, owing to their non-invasive acquisition methods and virtually unlimited availability. This study aims to investigate the effects and mechanisms of iMSCs in modulating macrophage polarization and alleviating periodontitis-related alveolar bone loss.

iMSCs were generated from iPSCs and characterized for differentiation potential. The effects of iMSCs on macrophage polarization were evaluated using THP-1-derived macrophages under inflammatory conditions (LPS and IFN-γ stimulation). Co-culture assays, cytokine analysis, reactive oxygen species (ROS) detection, transcriptomic analysis, flow cytometry, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and western blot analysis were performed to elucidate the underlying mechanisms. The therapeutic potential of iMSCs was assessed in a ligature-induced periodontitis mouse model using micro-CT, histological analysis, and immunofluorescence staining.

iMSCs inhibit M1 macrophage polarization through the suppression of the NF-κB signaling pathway. Additionally, iMSCs reduce the production of pro-inflammatory cytokines (IL-1β, IL-17) and reactive oxygen species (ROS), while enhancing the secretion of anti-inflammatory cytokines (IL-10) and growth factors (VEGF), thereby improving the inflammatory microenvironment. Under inflammatory conditions, iMSCs preserve the osteogenic potential of periodontal ligament stem cells (PDLSCs) and alleviate alveolar bone loss in mice with periodontitis. In vivo, iMSCs reduce the number of M1 macrophages and inhibit the activation of NF-κB in periodontal tissues, supporting their anti-inflammatory and immunomodulatory effects.

iMSCs demonstrate significant therapeutic potential in periodontitis by modulating macrophage polarization, reducing oxidative stress, and mitigating alveolar bone loss associated with the disease. These findings provide new insights into the mechanisms of iMSCs and their application as cell-based therapies for periodontal diseases.

Aging affects the reparative potency of mesenchymal stem/stromal cells (MSCs) by diminishing their proliferation and differentiation capability; making them unsuitable for regenerative purposes. Earlier we showed that MSCs acquire the expression of CD45 as a consequence of aging, and this increased expression is associated with downregulated expression of osteogenic markers and upregulated expression of adipogenic and osteoclastogenic markers. However, whether CD45 is actively involved in the aging-mediated deregulated differentiation in the MSCs was not elucidated.

In the present study, we showed that pharmacological inhibition of CD45-specific phosphatase activity in the aged MSCs restores their differentiation potential to young-like. Investigation of the molecular mechanism involved in the process showed that several regulatory kinases like p38, p44/42, Src, and GSK3β are in their dephosphorylated form in the aged MSCs, and importantly, this status gets reversed by the application of a CD45-specific PTP inhibitor. Conversely, pharmacological inhibition of these kinases in young MSCs imposes an aged-like gene expression profile on them. Additionally, we also showed that the secretome of aged MSCs affects the viability and differentiation of primary chondrocytes, and this detrimental effect is reversed by treating aged MSCs with the PTP inhibitor. Our data demonstrate that the aging-mediated expression of CD45 in MSCs alters their differentiation profile by dephosphorylating several kinases and treating the aged MSCs with a CD45 PTP activity inhibitor rejuvenates them.

CD45 can be used as an aging marker for mesenchymal stem cells. Alteration of CD45 phosphatase activity could have significant implications for the use of MSCs in regenerative medicine.

The complex of biofactors secreted by mesenchymal stem cells, termed the secretome, can promote wound healing. Studies using this secretome often utilise material collected from short term and sub-confluent lab-scale cultures. Secretome was derived from prolonged culture of high-density industrial scale human Wharton's jelly stem cells and its effects on wound healing was assessed. In vitro cell proliferation and scratch closure assays showed that secretome treatment dose-dependently increased cell proliferation and promoted scratch closure. Subsequently, using biopsy punch, circular wounds were created on three-dimensional de-epidermised dermis human skin equivalent (in vitro) where secretome-treated wounds showed accelerated wound closure, and enhanced epithelial proliferation and differentiation were observed and quantified. In an in vivo rat acute wound model, secretome applied to the back of test animals greatly enhanced wound healing by promoting re-epithelialisation, vascularisation and granulation maturation. In conclusion, secretome derived from prolonged culture of high-density industrial scale two-dimensional human Wharton's jelly stem cells possesses potent wound healing properties. This could greatly lower the cost of production and facilitate development of highly efficacious secretome-based wound healing products.

Histone lysine demethylases (KDMs), as important epigenetic regulators, are involved in various biological processes such as energy metabolism, apoptosis, and autophagy. Recent research shows that KDMs activate or silence downstream target genes by removing lysine residues from histone tails, and participate in the regulation of bone marrow mesenchymal stem cells (BM-MSCs), osteoblasts (OB), osteoclasts (OC), chondrocytes and other skeletal cell development, differentiation and formation. Moreover, several members of the KDM family affect the occurrence and development of bone diseases such as osteoporosis (OP), osteoarthritis (OA), osteosarcoma (OS), by regulating target genes. Specific regulation mechanisms of KDMs suggest new strategies for bone disease treatment and prevention. Despite the unique function and importance of KDMs in the skeletal system, previous studies have never systematically summarized their specific role, molecular mechanism, and clinical treatment in bone physiology and pathology. Therefore, this review summarises the expression pattern, intracellular signal transduction, and mechanism of action of the KDM family in several bone physiological and pathological conditions, aiming to highlight the important role of KDMs in bone diseases and provide a reference for the future treatment of bone diseases.

Tracheal cartilage provides structural support to the airways to enable breathing. However, it can become damaged or impaired, sometimes requiring surgical resection and reconstruction. Previously, we clinically applied an artificial trachea composed of a polypropylene mesh and collagen sponge, with a favorable postoperative course. However, the artificial trachea presents a limitation, as the mesh is not biodegradable and cannot be used in pediatric patients. Compared to a polypropylene mesh, regenerated cartilage represents an ideal material for reconstruction of the damaged trachea. The use of mesenchymal stem cells (MSCs) as a source for cartilage regeneration has gained widespread acceptance, but challenges such as the invasiveness of harvesting and limited cell supply persist. Therefore, we focused on the potential of human-induced pluripotent stem cell (hiPSC)-derived mesenchymal stem cells (iMSCs) for tracheal cartilage regeneration. In this study, we aimed to regenerate tracheal cartilage on an artificial trachea as a preliminary step to replace the polypropylene mesh. iMSCs were induced from hiPSCs through neural crest cells and transplanted with a polypropylene mesh covered with a collagen sponge into the damaged tracheal cartilage in immunodeficient rats. Human nuclear antigen (HNA)-positive cells were observed in all six rats at 4 weeks and in six out of seven rats at 12 weeks after transplantation, indicating that transplanted iMSCs survived within the tracheal cartilage defects of rats. The HNA-positive cells coexpressed SOX9, and type II collagen was detected around HNA-positive cells in four of six rats at 4 weeks and in three of seven rats at 12 weeks after transplantation, reflecting cartilage-like tissue regeneration. These results indicate that the transplanted iMSCs could differentiate into chondrogenic cells and promote tracheal cartilage regeneration. iMSC transplantation thus represents a promising approach for human tracheal reconstruction.

The focus on breast cancer treatment has shifted from the cytotoxic effects of single drugs on tumor cells to multidimensional multi‑pathway synergistic intervention strategies targeting the tumor microenvironment (TME). The activation of the Wnt signaling pathway in the TME of breast cancer cells serves a key regulatory role in tissue homeostasis and is a key driver of the carcinogenic process. Modulating the crosstalk between the Wnt pathway and TME of breast cancer is key for understanding the biological behavior of breast cancer and advancing the development of novel antitumor drugs. The present review aimed to summarize the complex mechanisms of the Wnt signaling pathway in the breast cancer TME, interactions between the Wnt signaling pathway and components of the breast cancer TME and breast cancer‑associated genes, as well as the interactions between the Wnt signaling pathway and other signaling cascades at the molecular level. Furthermore, the present review aimed to highlight the unique advantages of the Wnt signaling pathway in the macro‑regulation of the TME and the current therapeutic strategies targeting the Wnt signaling pathway, their potential clinical value and future research directions in breast cancer treatment.

The stroma of healthy pancreases contains various non-hematopoietic, non-endothelial mesenchymal cells. It is altered by chronic inflammation which in turn is a major contributor to the development of pancreatic adenocarcinoma (PDAC). In PDAC, the stroma plays a decisive and well-investigated role for tumor progression and therapy response. This review addresses the central role of stromal cells in the early inflammation-driven development of PDAC. It focuses on major subpopulations of pancreatic mesenchymal cells, i.e., fibroblasts, pancreatic stellate cells, and multipotent stroma cells, particularly their activation and functional alterations upon chronic inflammation including the development of different types of carcinoma-associated fibroblasts. In the second part, the current knowledge on the impact of activated stroma cells on acinar-to-ductal metaplasia and the transition to pancreatic intraepithelial neoplasia is summarized. Finally, putative strategies to target stroma cells and their signaling in early pancreatic carcinogenesis are reflected. In summary, the current data show that the activation of pancreatic stroma cells and the resulting fibrotic changes has pro- and anti-carcinogenetic effects but, overall, creates a carcinogenesis-promoting microenvironment. However, this is a dynamic process and the therapeutic targeting of specific pathways and cells requires in-depth knowledge of the molecular interplay of various cell types.

We explore the interaction between corneal immunity and mesenchymal stem/stromal cells (MSCs) and their potential in treating corneal and ocular surface disorders. We outline the cornea's immune privilege mechanisms and the immunomodulatory substances involved. In this realm, MSCs are characterized by their immunomodulatory properties and regenerative potential, making them promising for therapeutic application. Therefore, we focus on the role of MSCs in immune-mediated corneal diseases such as dry eye disease, corneal transplantation rejection, limbal stem cell deficiency, and ocular graft-versus-host disease. Preclinical and clinical studies demonstrate MSCs' efficacy in promoting corneal healing and reducing inflammation in these conditions. Overall, we emphasize the potential of MSCs as innovative therapies in ophthalmology, offering promising solutions for managing various ocular surface pathologies.

Arthroscopic repair is currently the gold standard for the surgical treatment of rotator cuff tears, but the retear rates remain unacceptably high. Mesenchymal stem cells (MSCs) may play a role in the local biology and enhance tendon-to-bone healing during rotator cuff repair. However, the scientific literature is still not well systematized on the effects of injection of MSCs as an augmentation tool for rotator cuff repair. Our goal was to investigate the effect of injections of MSCs to augment rotator cuff repair in patients with rotator cuff tear.

PubMed and EMBASE were searched up to June 2022 for clinical studies that applied MSCs injections to augment rotator cuff repair. Imaging, patient-reported outcomes measures, shoulder range of motion and strength were collected. Quantitative synthesis included within- and between-group mean differences with the within-group percentage of minimal clinically important difference for each study and continuous outcomes, and relative risks (RR) for retears and adverse events. Quantitative synthesis was computed with 95% confidence intervals (CIs).

We included 5 studies comprising a total of 228 individuals with a weighted mean age of 59.3 ± 1.2 years. Three studies used bone marrow MSCs and two studies applied adipose-derived MSCs. Patient-reported outcomes measures, shoulder range of motion, and strength improved significantly in all MSCs groups, with minimal clinically important differences ranging from 120% to 679% of established cut-off. When compared to rotator cuff repair alone, the MSCs groups did not result in improved outcomes. The MSCs group showed significant protective effect at the mid-term (RR = 0.52, 95% CI 0.27-0.98) and long-term (RR = 0.24, 95% CI 0.11-0.53).

There are no differences in clinical and functional outcomes between rotator cuff repair with or without augmentation with MSCs. However, there may be a protective effect against retear at the mid-term and long-term follow-up when augmenting the repair with MSCs. The literature on this topic is still preliminary and the quality and certainty of evidence is limited.

The study investigated in vitro the influences of shear stress preconditioning on human periodontal ligament stem cells (hPDLSCs) under serum deprivation.

hPDLSCs were subjected to shear stress at 0.5 and 5 dyn/cm², both with and without serum starvation. Cell viability and apoptosis were assessed using the Resazurin assay and flow cytometry analysis, respectively. Gene and protein expressions were analysed by real-time polymerase chain reaction, immunofluorescent staining, and Western blotting.

Our results revealed that shear stress potentially mitigated serum derivation-induced cell death by inducing cell viability, enhancing colony formation, and inhibiting cell apoptosis. The addition of an ERK inhibitor inhibited the shear stress-induced cell apoptosis resistance. Shear stress treatment upregulated cell viability-related gene expression, including SOX2, SOD1 and BIRC5. In particular, shear stress promoted the nuclear translocation of SOX2. Meanwhile, the expression of BIRC5 was not inhibited by cycloheximide. Shear stress-induced SOX2 and BIRC5 expression was attenuated by PI3K and ERK inhibitors, respectively.

Shear stress contributes to promoting SOX2 and BIRC5 expression by hPDLSCs, implicating the promotion of stemness and cell survival under serum starvation.