Current Stem Cell Research & Therapy - Online First

Rheumatoid arthritis (RA) is an inflammatory disease that causes significant disability and persistent inflammation. Currently, there are no appropriate treatments for RA other than systemic immunosuppressants, which have a variety of undesirable effects after long-term use. Thus, this study aims to determine the anti-arthritis effect of chrysin (5,7-dihydroxyflavone) and/or bone marrow-derived mesenchymal stem cells (BM-MSCs), separately and combined, on CFA (complete Freund’s adjuvant)-induced arthritis in rats as an animal model of RA.

Male Wistar rats were subcutaneously injected with 100 μL of CFA/rat/day in the paw of the right hind limb for two consecutive days to induce RA. Arthritic rats received chrysin in an oral dose of 100 mg/kg bw each day, BM-MSCs at 1 × 10⁶ cells/rat once per week in DMEM (Dulbecco’s modified Eagle’s medium) into the lateral tail vein, and a combination for 21 days.

The oral administration of chrysin and intravenous injection of BM-MSCs significantly reduced the increased anteroposterior thickness, volume, and circumference of the right hind paw, as well as serum levels of RF, IL-1β, TNF-α, and IL-17, as well as serum MDA level, besides augmenting serum levels of GPx, GST, GSH, and SOD. The arthritic rats treated with chrysin and/or BM-MSCs exhibited a significant improvement in the elevated expression levels of IκBα, NF-κB p50, and NF-κB p65 proteins in ankle joint articular tissue. Similarly, the histopathological score and histological sections provided additional evidence of the improvement in arthritic lesions.

The treatment with chrysin and BM-MSCs has potential anti-arthritic effects, which may be attributed to their abilities to suppress the inflammation and oxidative stress and enhance the antioxidant defense system. The combinatory effect of chrysin and BM-MSCs was found to be the most effective. However, further clinical studies are required to assess their safety and efficacy in patients with arthritis.

Due to their strong antioxidant and anti-inflammatory properties, the combined administration of chrysin and BM-MSCs was found to be more effective in treating arthritis than either treatment alone in Wistar rats.

Diabetes mellitus (DM) is a condition that arises from the dysfunction or disruption of pancreatic islets, characterized by elevated blood glucose levels. The advent and development of islet organoids have facilitated insulin-independent treatments and the reproduction of complex tissue or organ development.

This review focuses on the potential and value of islet organoids in both basic research and clinical applications, particularly in addressing the limitations of current diabetes treatments. We further discuss the structural characteristics of islets and explore various methods for obtaining seed cells, constructing organoids, and identifying factors that influence the formation and development of islet organoids.

The online databases, including Pubmed, Google Scholar, Science Direct, Web of Science, Embase, and reference lists were searched using the keywords diabetes mellitus, islet organoids, beta cells, material, development, three-dimensional, extracellular matrix, biomechanical, to identify published articles relevant to pancreatic islet organoids.

We examine the structural characteristics of islets and investigate various methods for obtaining seed cells, constructing organoids, and identifying factors that influence the formation and maturation of islet organoids.

To achieve a cure for diabetes, researchers have made significant efforts in islet transplantation and cell-derived insulin-secreting devices. However, organoids still require substantial improvements in cell sources, assembly techniques, and vascularization.

Islet organoids derived from stem cells may enable them to achieve insulin-independent regulation of blood glucose levels, thereby offering new hope for the individuals with diabetes.

Stem cells have recently gained prominence in regenerative medicine, particularly in the treatment of neurological disorders. As a result, Human-induced Pluripotent Stem Cells (hiPSCs) have become a significant focus.

This study aimed to differentiate hiPSCs into neural lineages under in vitro conditions using forskolin and retinoic acid in an induction medium combined with chondroitin 4-sulfate and chondroitinase.

Optimal component concentrations were determined using the MTT assay and acridine orange/ethidium bromide (AO/EB) staining. Subsequently, neural-specific genes (NSE, MAP-2, β-tubulin III, Oligo-2, and GFAP) and proteins (gamma enolase, MAP-2, and β-tubulin III) were assessed using Real-time PCR analysis and immunofluorescence staining to provide a comprehensive evaluation of differentiated cells.

Our study demonstrated a significant enhancement in neural-specific gene and protein markers during the 7th and 14th days of differentiation in the presence of combined chondroitin 4-sulfate and chondroitinase, demonstrating a higher efficacy compared with the application of isolated enzymes or substrates.

These findings emphasize the potential importance of chondroitin 4-sulfate and chondroitinase as important factors in promoting the neural differentiation of hiPSCs. It seems that chondroitin 4-sulfate may activate cellular signaling pathways that are effective in inducing neural differentiation. Our findings in this research provide new opportunities to advance regenerative therapies for neurological disorders.

Parkinson's disease (PD) is a chronic progressive neurodegenerative disease with debilitating clinical presentations. Common therapeutic approaches for symptomatic improvement are often effective for a temporary period of time, after which patients often experience progressive disabilities. Cell replacement therapy is a potential therapeutic method that aims to replace depleted mesencephalic dopaminergic (DA) neurons, which may control symptoms and halt disease progression. Preclinical studies have investigated the efficacy of these PSC-derived DA cells in animal models of PD.

In this study, we comprehensively examine preclinical data on the therapeutic effect of primate PSC-derived DA progenitors on motor deficits in animal models of PD as a precursor for conducting human clinical trials. Relevant articles published before August 14^th, 2023, were obtained from PubMed, Scopus, and Web of Science.

Through several rounds of screening, 46 studies that met our inclusion criteria were included in this study. The quality of each study was assessed using CAMARADES and SYRCLE approaches. Although no included studies were judged to have an overall high risk of bias, several studies exhibited domain-specific methodological limitations. The analyzed studies demonstrate that cell therapy significantly improves motor dysfunction in rodent and non-human primate models of PD.

This systematic review and meta-analysis demonstrate that PSC-based cell therapy significantly improves motor dysfunction in rodent and NHP models of PD and could be a promising approach for halting disease progression, improving behavioral manifestations of the disease, and increasing the overall quality of life in PD.

Burn injuries pose a significant health challenge, leading to intense physiological stress compared to other types of trauma. Myrtle has been traditionally used for treating various skin ailments, while mesenchymal stem cells (MSCs) have introduced innovative approaches for burn treatment. This study aimed to evaluate the impact of myrtle compared to MSCs on the healing of deep second-degree burns.

Fifty adult male albino rats were randomly divided into five groups: Group A served as the control, Group B received an excision burn without treatment, Group C was treated with topical myrtle paste, Group D received an intradermal injection of mesenchymal stem cells (MSCs), and Group E received both topical myrtle paste and intradermal MSC injection. Burn healing was assessed based on visible characteristics over 21 days. At the end of the treatment, skin samples were collected for biochemical analysis, histological examination using hematoxylin and eosin (H&E) staining, and VEGF concentration measurement via ELISA. Additionally, PCR analysis was conducted to assess the expression levels of COL1a1, COL3a1, TNF-α, and IL-6, providing insights into collagen production and inflammatory response.

Gross evaluation and histopathological analysis indicated that Groups D and E exhibited complete skin regeneration compared to the burn group. VEGF analysis demonstrated enhanced angiogenesis in the treated groups. PCR analysis revealed upregulation of COL1a1 and COL3a1, along with downregulation of TNF-α and IL-6, suggesting reduced inflammation and improved skin healing.

The study demonstrated that both MSCs and myrtle contributed to significant burn healing. The combination of myrtle and MSCs (Group E) exhibited the most effective skin regeneration, likely due to enhanced collagen production, reduced inflammation, and improved angiogenesis. These findings suggest that combining traditional herbal treatments with stem cell therapy may offer a promising strategy for burn management.

Organoids are three-dimensional structures that faithfully mimic the intricate internal environment of the human body. Compared to conventional models, they demonstrated superior performance. Recently, they have emerged as valuable platforms for modeling obesity-related diseases and advancing therapeutic strategies.

This review not only aimed to simply discuss the limitations of 2D cellular and animal models for obesity-related diseases but also highlighted the importance of developing organoids to better understand the relationship between obesity, lipid metabolism, glucose homeostasis, and chronic inflammation. It also identifies the challenges and potential directions for organoid applications in these diseases.

We searched for keywords related to organoids, obesity, lipid metabolism, glucose homeostasis, chronic inflammation, disease models, and drug screening in scientific research databases.

Organoids have emerged as promising tools for investigating the pathophysiology of diseases and developing therapeutic interventions. They have effectively bridged the gap in research on obesity-related diseases between conventional experimental models and the human body. They could offer more efficient and physiologically relevant experimental models while also improving the treatment efficacy for individuals with obesity-related conditions.

Organoids are beneficial for investigating obesity-related diseases. However, it is imperative to implement standardised culture procedures to improve reproducibility and broaden their application. Combining medicine and science to create these processes and minimise variation can increase the reliability and consistency of organoid cultures and provide new opportunities for addressing obesity-related diseases.

Alzheimer's disease (AD) is a neurodegenerative disease that is still incurable. Therapy with stem cell or extracellular vesicles is a promising strategy for AD treatment. Therefore, we evaluated whether stem cell-derived extracellular vesicles could improve cognitive function and pathological features in AD model mice.

PubMed, Web of Science, Embase, and The Cochrane Library were searched for studies reporting stem cell-derived extracellular vesicles treatment of AD mice from the establishment of each database to 1^st August 2023. SYRCLE was used to assess the risk of bias. The extracted data were analyzed using RevMan 5.4 and Stata 15 software.

19 studies were included in the analysis. Meta-analysis showed that treatment with stem cell-derived extracellular vesicles significantly improved cognitive performance of AD mice in the Morris water maze test and the novel object recognition test, reduced β-amyloid deposition, alleviated neuroinflammation and decreased levels of the proinflammatory cytokines and glial fibrillary acidic protein (GFAP) in the brain of AD mice. However, stem cell-derived extracellular vesicle did not affect the level of brain phosphorylated tau (p-Tau).

stem cell-derived extracellular vesicles may promote the degradation of β-amyloid plaques in the brain, regulate immunity and protect nerves, which result in cognitive improvement in AD mice.

Mesenchymal stem cells derived from umbilical cord blood (UCB-MSCs) have a well-known role in fastening the wound healing process due to their less immune rejection, anti-inflammatory effects, and their role in cellular growth. Campesterol is a nutritional phytosterol with extensive health values and a competitor of cholesterol in the blood. Campesterol shares some anti-inflammatory effects via its regulation of inflammatory markers by inhibiting the pro-inflammatory cytokines (including TNF-α, TGF-β1, and IL-6) levels.

The purpose of this study was to assess the ameliorative role of combined therapy (campesterol and UCB-MSCs) in wound healing without immune rejection. The study comprised both in-vitro and in-vivo experiments. In-vitro analysis included assessments of the cell viability of campesterol on UCBMSCs using MTT, crystal blue, trypan blue, and cell scratch assays. For in-vivo trials, superficial burn wounds were created on Sprague Dawley rats to evaluate the effects of campesterol, UCB-MSCs, and their combination on healing outcomes. Tissue regeneration progress in the wound vicinity was assessed using H&E staining and ELISA (inflammatory and growth markers) analysis.

Results of in-vitro experiments indicated that campesterol at concentrations of 10µg, 20µg, and 30µg demonstrated the most efficient cell viability. Moreover, a 30ug dose of campesterol along with UCBMSCs was further applied, leading to smooth and uncomplicated healing in the animal models. H&E staining showed nearly normal skin tissue while hematological and biochemical markers were near to control. Serum levels of tissue growth promoter factors, including VEGF and collagen-3, were higher, and pro-inflammatory markers (such as TGF-β1, TNF-α, and IL-6) were lower at the same time.

The results of the combined (MSCs and campesterol) therapy showed enhanced wound healing abilities. However, further studies are recommended to explore new aspects of this promising therapeutic approach of UCB-MSCs along with steroid derivative campesterol.

Diabetic foot (DF) poses a great challenge to us due to its poor therapeutic effect. To seek a new cure, the human dental pulp mesenchymal stem cells (hDP-MSCs) were modified by vascular endothelial growth factor A (VEGFA) and basic fibroblast growth factor (bFGF) (hEDP-MSCs) to investigate their curative effect on DF wound in animal models.

Forty-eight rats with DF constructed with streptozotocin and ligation of femoral arteries, were randomly divided into six equal groups, which respectively received an intramuscular injection of normal saline (Control group), hDP-MSCs, VEGFA-modified hDP-MSCs, bFGF-modified hDP-MSCs, hEDP-MSCs, and Ad.VEGF.FGF (Ad.FV). The tissues around DF wound were collected to investigate the level of CD31, alpha-smooth muscle actin (α-SMA), and cytokines. The expression of Notch1, Hes1, and CD105 were assessed via Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) after administration.

The hEDP-MSCs increased capillaries and decreased wound area (%). QRT-PCR showed that hEDP-MSCs over-expressed the mRNA of Notch1, hairy and enhancer of split 1 and CD105 in peri-wound tissue post-treatment. Meanwhile, the hEDP-MSCs expressed more CD31 and α-SMA than other groups. The hEDP-MSCs expressed more VEGFA and bFGF than hDP-MSCs, and yet less than Ad.FV. Compared with hDP-MSCs, the hEDP-MSCs down-regulated the expressions of interleukin-1 beta (IL-1β), interleukin (IL-6), and tissue necrosis factor α (TNF-a) post-treatment.

This study highlights the curative effect of hEDP-MSCs in the wound healing process, and demonstrates the decisive function of hEDP-MSCs in promoting angiogenesis and reducing inflammation.

Osteoarthritis (OA) is a degenerative joint disease that can affect the many tissues of the joint. There are no officially recognized disease-modifying therapies for clinical use at this time probably due to a lack of complete comprehension of the pathogenesis of the disease. In recent years, emerging regenerative therapy and treatments with stem cells both undifferentiated and differentiated cells have gained much attention as they can efficiently promote tissue repair and regeneration.

To determine how bone marrow-derived mesenchymal stem cells (BM-MSCs) and chondrogenic differentiated MSCs (CD-MSCs) can treat OA in rats, OA was induced in Wistar rats by injecting three doses of 100 μL physiological saline containing 1 mg of MIA into rat ankle joint of the right hind leg for three consecutive days. Following the induction, the osteoarthritic rats were injected weekly with BM-MSCs or CD-MSCs at a dose of 1x10⁶ cells/rat/dose for three weeks. In addition to morphological and histological investigations of the ankle, spectrophotometric, ELISA, and Western blot analyses were applied to detect various immunological and molecular parameters in serum and ankle.

The results of the study showed that in osteoarthritic rats, BM-MSCs and CD-MSCs significantly reduced right hind paw circumference, total leucocyte count (TLC), differential leukocyte count (DLC) of neutrophils, monocytes, lymphocytes, and eosinophils, serum rheumatoid factor (RF), prostaglandin E2 (PGE2) and interleukin (IL-) 1β levels, while they elevated serum IL-10 level. Additionally, BM-MSCs and CD-MSCs markedly reduced lipid peroxides (LPO) levels while they elevated superoxide dismutase (SOD) and glutathione-S-transferase (GST) activities. The monocyte chemoattractant protein-1 (MCP-1) level was significantly downregulated in ankle joint articular tissues by treatment with BM-MSCs or CD-MSCs while nuclear factor erythroid 2-related factor 2 (Nrf2) was upregulated; CD-MSCs treatment was more effective.

According to these findings, it can be inferred that BM-MSCs and CD-MSCs have anti-arthritic potential in MIA-induced OA; CD-MSCs therapy is more effective than MSCs. The ameliorative anti-arthritic effects may be mediated by suppressing inflammation and oxidative stress through the downregulation of MCP-1 and upregulation of Nrf2. Based on the obtained results, BM-MSCs and CD-MSCs therapies are promising new options that can be associated with other clinical treatments to improve cartilage regeneration and joint healing. However, more preclinical and clinical research is required to assess the benefits and safety of treating osteoarthritic patients with BM-MSCs and CD-MSCs.

This study aims to explore the therapeutic potential of mesenchymal stem cells (MSC) in treating diabetic nephropathy (DN) by investigating their effect on IL-11 modulation in a mouse model.

The effects of MSC therapy on DN were examined both in vivo and in vitro. Sixty adult male C57BL/6 mice were divided into the streptozotocin (STZ) diabetes (T1D) and the high-fat diet diabetes (T2D) models, with both groups receiving MSC treatment or saline for 4 or 8 weeks. Blood glucose, serum urea, interleukin-11 (IL-11), and kidney fibrosis markers were measured. Additionally, western blotting was used to assess levels of Type I and III collagen, E-Cadherin, α-smooth muscle actin (α-SMA), Vimentin, and ferroptosis suppressor protein 1 (FSP-1).

MSC-treated T1D and T2D mice showed reduced blood glucose, serum urea, IL-11, TGF-β, and fibrosis markers (type I and III collagen, α-SMA, Vimentin, FSP-1), alongside increased E-Cadherin expression. Similar effects were observed in vitro using mouse glomerular epithelial cells, confirming MSC-mediated suppression of fibrosis pathways.

MSC therapy improves nephropathy, likely by inhibiting IL-11 and reducing fibrosis-related markers, making it a promising treatment for DN.

Human adipose-derived stem cells (hADSCs) are considered a promising source for cell replacement therapy in degenerative and traumatic conditions. This study explores the effects of phenylacetate and calcium on the neural differentiation of hADSCs for regenerative medicine. We assessed cell viability and cytotoxicity using the MTT assay, revealing that treatment with 1μM phenylacetate significantly enhanced cell viability compared to control groups over five days, while higher concentrations resulted in cytotoxic effects.

Additionally, qualitative analysis through Acridine orange/ethidium bromide (AO/EB) staining indicated normal cellular characteristics at lower phenylacetate concentrations, whereas higher doses led to observable cell death. A subsequent evaluation of intracellular calcium levels demonstrated a significant increase when hADSCs were treated with both phenylacetate and calcium.

The neural differentiation potential was further assessed through the relative quantification of neuronal-specific genes, showing marked upregulation of NSE, Oligo-2, β-tubulin III, and MAP-2 in all treatment groups compared to controls. Immunohistochemistry confirmed elevated protein expression of neural markers in cultures supplemented with phenylacetate and calcium.

These findings suggest that phenylacetate, particularly in conjunction with calcium, enhances the neural differentiation of hADSCs, highlighting its potential utility in regenerative medicine strategies targeting neurodegenerative conditions.

Trigeminal Neuralgia (TN) is an extremely painful condition without an established treatment other than symptom-suppressive medications or temporary relief from corticosteroid injections. Mesenchymal Stem Cells (MSCs) have demonstrated the ability to enhance healing and reduce inflammation and pain without side effects. Our objective was to evaluate the safety and efficacy of CT-guided foramen ovale MSC injection in the treatment of TN.

A 48-year-old woman presented with a 22-year history of severe TN. Previous treatments, including microvascular decompression, acupuncture, chiropractic adjustment, and hypnotism had failed. Medications decreased pain, but produced severe bothersome mental clouding. After proper informed consent, the patient elected trigeminal nerve injection in the foramen ovale with AlloRx (vitrobiopharma.com Golden Colorado) umbilical cord-derived Mesenchymal Stem Cells (MSCs). An experienced pain specialist with previous experience using CT guidance with sedation to inject the trigeminal nerve in the foramen ovale with corticosteroids performed the injection using 20 million MSCs. The patient reported no adverse events or complications related to the treatment.

At 1 month post-treatment, the patient reported dramatically reduced pain/tingling, and no longer needed medication, which resulted in the resolution of her mental clouding. At 12 months post-treatment, some symptoms recurred, but the patient maintained substantial cognitive improvements and required a reduced dose of medication.

We have demonstrated, for the first time, CT-guided MSC injection into the foramen ovale to result in significant improvement in trigeminal neuralgia without side effects.

With the passage of time, the incidence rate of diabetes continues to rise. As a common and serious complication of diabetes, the economic burden of diabetes ulcers is also increasing. However, there is currently no unified clinical treatment strategy for its repair and research, and the treatment effect is not satisfactory. Exosomes derived from MSCs play a crucial role in improving diseases.

This study used the explant culture method to isolate human umbilical cord mesenchymal stem cells (hucMSCs) from Wharton’s jelly, which were identified by flow cytometry and differentiation potential and evaluated tumorigenic potential. The supernatant of P3-P5 cell culture medium was collected and high-concentration human umbilical cord mesenchymal stem cell exosomes (hucMSCs-Exos) were isolated through ultracentrifugation. Qualitatively identified were finished by transmission electron microscopy (TEM), nanosight tracking analysis (NTA) and Western blot (WB). CCK-8 assay and cell scratch experiment were used to evaluate the effects of hucMSCs-Exos on proliferation and migration ability of cells; chicken embryo chorioallantoic membrane experiment (CAM) was used to evaluate the angiogenic effect of hucMSCs-Exos. In order to study the effect of hucMSCs-Exos on diabetes wound healing, the diabetes mouse model was constructed by a high-fat feeding and a low-dose STZ injection. Fasting blood glucose and blood lipids were measured and oral glucose tolerance test (OGTT) was conducted; HucMSCs-Exos were subcutaneously injected into the T2DM wound model mice which were established by a high-fat diet and by a low-dose streptozotocin (STZ), and the wound healing was evaluations.

Cells were isolated by explant culture method, and flow cytometry analyses showed that these isolated cells were positive for cells markers CD29, CD90, and CD105, while negative for CD34 and CD45; moreover, these cells could be induced to osteoblasts, adipocytes, and islet-like cells. The tumorigenic experiments showed that these cells have no tumorigenicity. HucMSCs-Exos, separated by ultracentrifugation, displayed spherical or ellipsoidal vesicles with a diameter of around 120nm, positive for CD9, CD63, and CD81. And they could promote cell proliferation and migration, as well as promote vascular growth (p < 0.01). In vivo experiments showed that hucMSCs-Exos had a promoting effect on wound repair in diabetes mellitus type 2 (T2DM) mice (p < 0.01).

This study provides a new scheme for the treatment of T2DM ulcer.