Neural stem cells (NSCs) are vulnerable to oxidative stress, which triggers aging and subsequently leads to a reduced regenerative capacity of the central nervous system (CNS). Due to the challenges in acquiring aged human NSCs and the lack of an oxidative stressinduced aging model specifically designed for human NSCs, research related to the aging mechanisms and the screening of anti-aging drugs have been limited. Here, we aimed to establish an oxidative stress-induced senescence model of NSCs by using D-galactose (D-gal). Human embryonic stem cells (hESC) were differentiated into hESC-NSCs using a type I collagen method. hESC-NSCs were characterized by flow cytometry combined with immunofluorescence. A senescence model of hESC-NSCs was established using D-gal and characterized by CCK-8 assay, neurosphere formation, crystal violet staining, DNA damage assay, SA-β-gal staining and ROS levels measurement. To further explore the profile of gene expression in D-gal-induced hESCNSCs senescence model, transcriptome sequencing was performed and analysed by bioinformatics method, following verified by qPCR. The hESC-derived NSCs senescence model demonstrated reduced proliferation and elevated β-galactosidase activity, accompanied by DNA damage and increased levels of reactive oxygen species. Furthermore, transcriptome analysis unveiled the potential central role of the MAPK signaling pathway in D-gal-induced senescence, which involves the key genes including DDIT3, ATF3, CEBPB, JUN, and CCND1. We presented an oxidative stress-induced senescence model of hESC-NSCs and identified key pathways and genes related to D-gal-induced senescence. Our study might offer an alternative approach for investigating human NSC aging and provide valuable data for understanding the underlining mechanisms of oxidative stress-induced aging.

This study aimed to assess the knowledge, perception, and willingness of the Saudi population towards stem cell treatment and banking, the associated factors, and their predictive abilities. A cross-sectional study was conducted from September to December 2022 in Saudi Arabia using a structured bilingual, self-administered online survey to collect sociodemographic information and determine the knowledge and understanding, perception, and willingness of the general population. Bloom's cut-off points were used to distribute the scores into three categories, namely strong (80-100%), moderate (60-79%), and weak (<59%). Descriptive statistics were used to assess each domain, while t-tests, ANOVA, and binary logistic regression were used to assess factors influencing each domain and their predictive abilities. The study conscripted 440 respondents, mainly females (70%) aged 18-24 years (56.1%), mostly single (44.3%), Saudi nationals (89%), and college graduates (56.6%). Social media (53.4%) was the primary source of information. 77.95% of respondents exhibited a low level of knowledge. Females, Saudi nationals, respondents with Islamic beliefs, college graduates, and those who received information from family physicians and social media had significantly better knowledge. 50.68% of respondents showed a strong perception. Non-Saudi participants, those who received information from family and friends, and respondents with a high prevalence of hypertension and diabetes in their families had a better perception. However, only 21.59% showed strong willingness toward stem cell treatment and banking, including Saudi nationals and college graduates. Gender was found to be a significant predictor for better knowledge and perception, while no sociodemographic variables significantly predicted willingness. This study emphasizes the need for increased awareness, educational campaigns, and targeted strategies considering various socio-demographic factors to improve the knowledge, perception, and willingness of the general population toward stem cell treatments and banking in Saudi Arabia.

To investigate the protective mechanism of Qishen Yiqi Dropping Pills (Qishen) and its key active ingredients in combination with exosomes from bone marrow mesenchymal stem cells (BMSCs) against myocardial ischemia/reperfusion (I/R) injury. Infarct area was assessed by Evan's Blue/TTC double staining, myocardial apoptosis was analyzed by TUNEL staining, ATP content and NAD+/NADH ratio were detected biochemically, macrophage phenotype of myocardial tissues was detected by flow cytometry, activation of the mTOR/PI3K/Akt pathway was detected by Western blotting, and miR-155-5p expression was detected by qRT-PCR. 155-5p expression. Exo were given to identify the M1/M2 phenotypic transition by immunofluorescence, and the molecular mechanism was verified as in the in vivo experiments. Compared with the model group, the Qishen, ginsenoside, and Exo group significantly reduced the infarcted area of the heart and promoted M2 and M2 phenotypic conversion, promoted M2- type macrophage infiltration, up-regulated the p-Akt/Akt ratio, and inhibited the expression of miR- 155-5p, but the combination therapy group did not show a synergistic effect, but the above protective effects were significantly weakened by the removal of macrophages. Ginsenoside and Exo synergistically promoted M2 polarization, activated the mTOR/PI3K/Akt pathway and upregulated miR-155-5p expression. Qishen, particularly its active component ginsenoside, synergizes with BMSC-Exo to alleviate myocardial I/R injury by modulating macrophage polarization via the miR-155- 5p/mTOR/PI3K/Akt signaling axis. Qishen synergistically regulates the miR-155-5p/mTOR/PI3K/Akt signaling axis through ginsenoside components in BMSCs exosomes, promoting macrophage polarization toward M2-type, improving myocardial energy metabolism and attenuating I/R injury, and this protective effect is macrophage-dependent.

Acute Spinal Cord Injury (SCI) often causes motor and sensory deficits. SDF-1 promotes stem cell survival and proliferation, while FBN1 may impact repair mechanisms. This study investigates how SDF-1 promotes SCI treatment by inducing BMSC maturation through BMP-8-mediated FBN1 inhibition. Bone marrow mesenchymal stem cells were induced to differentiate with BMP-8 and transfected with related plasmids (oe-NC, oe-SDF-1, oe-FBN1, si-BMP-8). CCK-8 and alizarin red staining were used to assess cell growth and differentiation. Western blotting was used to detect the levels of SDF-1, FBN1, and BMP-8. In a rat SCI model, cells with plasmids were injected, and motor recovery was assessed using BBB scoring. Immunofluorescence assay detected SDF-1 expression, while Western blotting was used to detect SDF-1, FBN1, and BMP-8. In cell experiments, BMP-8 induced successful differentiation of BMSCs. After overexpression of SDF-1, the proliferation and differentiation of BMSCs were increased. In animal experiments, the BBB score increased after overexpression of SDF-1. These findings suggest a potential therapeutic mechanism in which SDF-1 promotes spinal cord repair by modulating the BMP-8/FBN1 axis. The suppression of FBN1 appears to be a key step in enhancing BMSC function. Targeting this pathway could offer new strategies for regenerative treatment following SCI. In acute spinal cord injury, SDF-1 enhances the differentiation of bone marrow mesenchymal stem cells induced by BMP-8 through the suppression of FBN1.

Acute myeloid leukemia (AML) arises from the aberrant proliferation of white blood cells, red blood cells, or platelets. Various therapy modalities are available for individuals diagnosed with AML. While Cytarabine remains a standard treatment, exosomes, especially those derived from cord blood, have emerged as promising adjuncts. Exosomes are a specific kind of extracellular vesicles that have been identified as candidate biomolecules for the treatment of AML. The current study looked at how cord blood-derived exosomes affect the U937 cell lines compared to Cytarabine as a fundamental component of standard AML treatment. The first stage involved processing umbilical cord blood and isolating the mononuclear cells, CB-MNCs. The exosomes were isolated and verified using transmission electron microscopy, western blotting, and dynamic light scattering. Subsequently, for 72 hours, the U937 cells were cultivated and exposed to the exosomes, Cytarabine, and a combination of both. Afterward, apoptosis was evaluated using flow cytometry. The activity of Caspase 3/7 was assessed using a special kit. The real-time PCR technique was used to evaluate the gene expression in the proliferation and apoptosis pathways. Finally, the activity of NF-κB and AMPK was assessed using western blotting. The flowcytometry analysis showed that the apoptosis rate of U937 cells after being exposed to CB-MNC exosomes, Cytarabine, and a mix of them was 19.60 (p < 0.05) %, 44.05 % (P < 0.0001), and 47.45 % (P < 0.0001), respectively. The activity of Caspase 3-7 was 0.32 mU/mL and 0.45 mU/mL (P < 0.001) Cytarabine and Mix groups. The qRT-PCR study revealed a notable upregulation of apoptotic genes and a downregulation of anti-apoptotic gene expression in the Mix group. Western blot analysis revealed a decrease in NF-κB phosphorylation in all treatment groups. In addition, the phosphorylation of AMPK increased in all treatment groups. The study evaluated the effects of CB-MNC-derived exosomes on U937 cells, both alone and in combination with Cytarabine. Results demonstrated that while exosomes induced moderate apoptosis, their combination with Cytarabine significantly enhanced cell death, increased Caspase 3/7 activity, and altered gene expression in favor of apoptosis. The treatment also inhibited NF-κB and activated AMPK signaling pathways. In conclusion, our findings indicated that CB-MNCs-derived exosomes together with Cytarabine have a cooperative effect on U937 cells through increasing apoptosis rate and reducing the proliferation rate, and may serve as a promising adjunct in AML therapy.

Diabetic foot ulcer (DFU) is a severe complication of diabetes mellitus, affecting up to 15% of diabetic patients and leading to high rates of hospitalization, morbidity, and lower limb amputation. This case study details the management of a 67-year-old diabetic male patient with a DFU complicated by osteomyelitis. The patient, with a long history of diabetes and multiple comorbidities, was treated with a comprehensive regimen that included antibiotics, debridement, and platelet-rich plasma (PRP) therapy. PRP was administered weekly for 15 weeks. Clinical, laboratory, and imaging data were employed for the evaluation of the disease improvement. As a result, a marked improvement in wound healing was observed, characterized by reduced wound size, accelerated closure of the wound, and enhanced tissue regeneration. However, the patient developed deep vein thrombosis, which was successfully managed with anticoagulants. The study highlights PRP's potential in DFU treatment due to its regenerative properties despite the risk of adverse effects. The efficacy of PRP aligns with previous studies, showing improved healing rates and infection control. Future research should focus on large-scale trials to optimize PRP protocols and confirm the safety and efficacy of this therapeutic method in DFU.

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.

Hematopoietic stem cells (HSCs) represent the most primitive cell population endowed with the ability for self-renewal and differentiation. They possess the capacity to differentiate into all types of blood cells, each serving unique functions. Traditional theories have established a clear hierarchical relationship between HSCs, their progenitors, and mature blood cells. The identification of distinct cell populations within the hematopoietic system forms the foundation of the hematopoietic differentiation model. However, recent research has led to a constant evolution of our understanding of the hierarchical structure of hematopoietic differentiation, particularly in the context of megakaryocyte differentiation pathways. Megakaryocytes are essential for platelet production, a critical process in hemostasis and thrombosis. Understanding the mechanisms underlying megakaryocyte-biased HSCs differentiation holds significance for both basic research and clinical applications. In this review, we consolidate the latest research progress concerning the evidence supporting these nonclassical pathways of megakaryocytic differentiation. Furthermore, we delve into the alterations observed in these pathways under conditions of steady state, transplantation, stress, and aging.

Fetal Bovine Serum (FBS), the conventional supplement for Mesenchymal Stromal Cell (MSC) culture, presents ethical issues, batch variability, and risks of pathogen transmission. This study aimed to evaluate human-derived Umbilical Cord Serum (UCS) and Human Platelet Lysate (HPL) as xeno-free alternatives to FBS and to assess the synergistic effects of nano-curcumin and crocin as supplements to enhance the proliferation and survival of human umbilical cord-derived MSCs. Human umbilical cord-derived MSCs were cultured in media supplemented with 10% FBS (control), UCS, or HPL. These groups were further treated with nano-curcumin (0.3 μM) or crocin (2.5 μM), either individually or in combination. Cell proliferation was measured using the MTT assay, apoptosis was assessed by Annexin V/PI flow cytometry, and pluripotency gene expression (Sox2, Nanog, Oct4) was analyzed by RT-qPCR. UCS and HPL supplements significantly increased MSC proliferation compared to the FBS control (p < 0.001). Specifically, UCS reduced the population doubling time by approximately 50%. Supplementation with crocin reduced apoptosis by up to 30% (p = 0.04) and significantly enhanced the expression of the pluripotency genes Sox2 and Nanog, particularly in cultures supplemented with HPL. In contrast, nano-curcumin inhibited MSC proliferation and increased apoptosis across all tested conditions. The results demonstrate that UCS and HPL are effective, viable alternatives to FBS, promoting superior MSC expansion. The anti-apoptotic and stemness-enhancing properties of crocin highlight its potential as a valuable additive for improving culture quality and cell survival. The cytotoxic effects observed with nano-curcumin underscore a critical need for dose-optimization studies. The primary limitation of this study is the use of fixed concentrations for the supplements, which warrants further investigation across a range of doses. UCS and HPL are robust, ethically sound replacements for FBS in MSC biomanufacturing. Crocin can further enhance culture outcomes by improving cell survival and maintaining stem cell properties. These findings support the development of optimized, xeno-free culture systems for scalable MSC production, which is crucial for advancing regenerative medicine therapies.

Intra-articular injection of autologous microfragmented adipose-derived mesenchymal stem cells (AMSCs) has shown potential for symptom relief and cartilage regeneration in osteoarthritis (OA). However, histological evidence in humans remains limited. We present a 46-year-old female patient who had symptomatic left hip OA and underwent a single injection of autologous microfragmented AMSCs under ultrasound guidance after unsuccessful hip arthroscopy. At the 12-month follow-up, the patient was pain-free, fully mobile, and had returned to normal daily activities without limitations, indicating an excellent clinical outcome. Eighteen months after the treatment, due to symptom recurrence, the patient later underwent total hip arthroplasty, allowing histological analysis of the joint. Examination revealed areas of hyaline-like cartilage in regions previously affected by degeneration. This case provides clinical and histological evidence of cartilage regeneration following intra-articular autologous microfragmented AMSCs therapy for hip OA. Although symptom recurrence occurred at 18 months, findings suggest this treatment may offer a regenerative option warranting further study.