Stem cells from human exfoliated deciduous teeth are widely used in the field of tissue repair and regeneration, but the tissue regeneration effect has limitations in practical applications. Using miRNA to intervene in the directional differentiation of stem cells from human exfoliated deciduous teeth is an important development direction for tissue repair and regeneration in the future.

To investigate the effect of miR-26b on the neurogenic and angiogenic differentiation of stem cells from human exfoliated deciduous teeth.

Dental pulp stem cells were isolated and extracted from human exfoliated deciduous teeth, and induced to differentiate into nerves and blood vessels. The expressions of neurogenic markers Nestin, NSE, βIII-Tubulin, and angiogenic markers CD31, VEGFR2, ANG-1 and miR-26b were detected. Dental pulp stem cells were divided into blank control group, miR-26 overexpression group, and negative control group. RT-qPCR, cell immunofluorescence staining, and western blot assay were used to detect the expression changes of related markers of neurogenic and angiogenic induction of stem cells from human exfoliated deciduous teeth in each group.

(1) Stem cells from human exfoliated deciduous teeth had multi-directional differentiation potential and could differentiate into osteogenesis, adipogenesis, neurogenesis, and vasculogenesis. (2) The expression level of miR-26b increased during the neurogenic and angiogenic differentiation of stem cells from human exfoliated deciduous teeth. (3) Compared with the blank control group and negative control group, the mRNA expression of neuroblast-related genes βIII-Tubulin, Nestin, NSE and angiogenesis-related genes CD31, VEGFR2, and ANG-1 in stem cells from human exfoliated deciduous teeth in the miR-26b overexpression group was significantly increased (P < 0.01), βIII-Tubulin, Nestin, CD31, and VEGFR2 protein expression was significantly increased (P < 0.01). The above results show that overexpression of miR-26b can promote the neurogenic and angiogenic differentiation of stem cells from human exfoliated deciduous teeth.

Hepatocyte-like cells induced by mesenchymal stem cells are promising seed cells for liver regeneration or liver tissue engineering. The efficiency of traditional two-dimensional culture for hepatocyte induction is low, and more and more research is focused on three-dimensional culture for inducing hepatocyte differentiation.

To summarize three-dimensional culture models for the hepatic induction of mesenchymal stem cells, focus on research progress on the epigenetic regulation mechanisms of mesenchymal stem cell hepatogenic differentiation, providing a theoretical basis for improving the differentiation efficiency of mesenchymal stem cells.

Relevant articles in the PubMed and other databases such as CNKI were searched, using Chinese and English search terms “mesenchymal stem cell, 3D culture, hepatogenic differentiation, hepatocyte-like cells, epigenetics.” Additionally, the literature tracing method was employed to find some of the literature for a comprehensive review and analysis.

(1) Common three-dimensional culture models for the hepatogenic differentiation of mesenchymal stem cells currently include spheroids, biological scaffolds, bioprinting, and microfluidic chips. Each of these models has its own advantages and disadvantages in the process of inducing hepatogenic differentiation. (2) During the differentiation of mesenchymal stem cells into hepatocyte-like cells, epigenetic regulation plays a key role, primarily involving histone modification, DNA methylation, and the regulation of non-coding RNAs. (3) Under three-dimensional culture conditions, epigenetic modifications, especially histone acetylation, play an important role in promoting the hepatogenic differentiation of mesenchymal stem cells.

Previous studies have found that growth arrest and DNA damage-inducible protein 45β (Gadd45b) is beneficial to the repair of acute cerebral ischemia, but the action mechanism is still unclear.

To investigate the effect and mechanism of Gadd45b on white matter demyelinating lesions in rats with chronic cerebral ischemia.

SD rats were randomly divided into four groups: sham operation group, model group, empty vector group, and Gadd45b overexpression group, with 15 rats in each group. Gadd45b-overexpressing lentivirus and no-load lentivirus were injected into the bilateral hippocampus and bilateral ventricles of rats. One week after lentivirus transfection, the rat model of chronic hypoperfusion cerebral ischemia was established by bilateral common carotid artery ligation. Three weeks after the bilateral common carotid artery ligation, the learning and cognitive functions of rats were evaluated by novel object recognition test. Luxol fast blue staining was used to observe the changes of myelin structure in the corpus callosum of rats. Hematoxylin-eosin staining and Nissl staining were used to observe the damage of the rat corpus callosum. Immunofluorescence staining was used to detect the expression of myelin basic protein and neurofilament protein 200 in the corpus callosum of rats. Immunofluorescence double staining was used to detect the expression of astrocyte markers GFAP/C3d and GFAP/S100A10 in rat brain tissue. ELISA was used to detect the levels of tumor necrosis factor-α and interleukin-6 in the supernatant of brain tissue.

(1) Gadd45b overexpression could significantly improve the learning and cognitive function of rats with chronic cerebral ischemia, and improve demyelination and pathological damage in the rat corpus callosum. (2) The results of immunofluorescence showed that Gadd45b overexpression significantly increased the expression levels of myelin basic protein and neurofilament protein 200 in the brain tissue of rats with chronic cerebral ischemia. (3) Gadd45b overexpression reduced GFAP/C3d double positive cells and increased GFAP/S100A10 double positive cells in the brain tissue of rats with chronic cerebral ischemia. (4) Gadd45b overexpression reduced the levels of tumor necrosis factor-α and interleukin-6 in the brain tissue of rats with chronic cerebral ischemia. It is concluded that Gadd45b overexpression improves cognitive dysfunction by promoting the A2 phenotype transformation of astrocytes, alleviating white matter myelin structure damage and neuroinflammation in rats with chronic cerebral ischemia.

Long noncoding RNA TP53TG1 (lncRNA TP53TG1) is involved in regulating the proliferation, migration, invasion, and apoptosis of various cancer cells, but there are few reports on its role in other cells.

To investigate the effects and pathways of lncRNA TP53TG1 on the proliferation and differentiation of human stem cells from the apical papilla.

Human stem cells from the apical papilla were isolated and cultured, and then transfected with lncRNA TP53TG1 overexpression lentivirus. RT-qPCR was used to detect the overexpression efficiency of lncRNA TP53TG1. Western blot assay was used to detect the relative expression levels of PI3K, AKT, ERK, P38, Smad3, and their phosphorylated proteins. Human stem cells from the apical papilla were divided into the empty lentiviral vector transfection group and the lncRNA TP53TG1 overexpression group. CCK-8 assay was used to measure the cell proliferation. Alkaline phosphatase activity was detected by alkaline phosphatase staining on day 5 of osteogenic induction. Formation of mineralized nodules was detected by alizarin red staining on day 21 of osteogenic induction. RT-qPCR was used to detect the mRNA expression levels of dentin sialophosphoprotein, Runt-related transcription factor 2, dentin matrix protein 1, and bone sialoprotein on days 3, 7, and 14 of osteogenic induction. Western blot assay was used to detect the protein expression levels of dentin sialophosphoprotein and Runt-related transcription factor 2 on days 3, 7, and 14 of osteogenic induction.

(1) RT-qPCR results showed that the lentivirus was successfully integrated into the genome of stem cells from the apical papilla. Western blot assay results showed that overexpression of lncRNA TP53TG1 up-regulated the protein levels of p-PI3K and p-AKT without affecting the expression of phosphorylated proteins in other pathways. (2) Starting from day 3 of cell culture, overexpression of lncRNA TP53TG1 significantly promoted the proliferation of stem cells from the apical papilla. (3) In the process of inducing odontogenic differentiation of stem cells from the apical papilla, overexpression of lncRNA TP53TG1 promoted the expression of odontogenic and osteogenic differentiation-related genes and proteins, significantly increased alkaline phosphatase activity and mineralized nodule formation. (4) The results show that lncRNA TP53TG1 may promote the odontogenic and osteogenic differentiation of stem cells from the apical papilla by activating the PI3K/AKT signaling pathway.

Enhancing the differentiation of induced pluripotent stem cells into lung stem cells is crucial for repairing lung injuries. NKX2.1 is the earliest marker of lung epithelial differentiation and plays a significant regulatory role in lung development. However, the impact of its expression on the differentiation of induced pluripotent stem cells into lung stem cells remains inadequately understood.

To investigate the effect of NKX2.1 on the differentiation of induced pluripotent stem cells into lung stem cells.

Induced pluripotent stem cells were cultured in vitro. The expression of specific pluripotent stem cell genes was assessed using real-time fluorescence quantitative PCR. NKX2.1 was overexpressed in induced pluripotent stem cells, which were then induced to differentiate into lung stem cells. The expression of FoxA2, SOX9, and P63 was determined via quantitative PCR and immunofluorescence on day 7 of induction of differentiation. The expression of the alveolar marker SPB and SPC was evaluated through immunofluorescence staining on day 7 of induction of differentiation.

(1) Induced pluripotent stem cells in vitro were tightly packed and showed typical clonoid growth and significantly expressed stem cell-specific genes OCT-4, SOX2, and NANOG. (2) Compared with the non-transfected control group, the expression of NKX2.1 in human induced pluripotent stem cells was significantly increased in the NKX2.1 overexpression group (P < 0.000 1). (3) Seven days after induction of differentiation, compared with the non-transfected control group, the expression of lung stem cell-related markers FoxA2, SOX9, and P63 was significantly increased in the NKX2.1 overexpression group (P < 0.000 1). (4) Thirteen days after induction of differentiation, compared with the non-transfected control group, the fluorescence intensity of alveolar cell marker molecules SPB and SPC increased significantly in the overexpression NKX2.1 group. The results show that NKX2.1 can promote the differentiation of induced pluripotent stem cells into lung stem cells.

Premature ovarian failure has manifested a trend of younger, and stem cell therapy has been progressively implemented in clinical practice in recent years. Nevertheless, given the extensive range of sources and variegated existence of stem cells in diverse tissues, certain disparities prevail in their biological characteristics and functions. In this paper, the therapeutic efficacies of dissimilar sources of mesenchymal stem cells on animal models of premature ovarian failure were contrasted, with the aim of providing a basis for the clinical application of stem cells.

The animal model experiments of mesenchymal stem cell therapy for premature ovarian failure were retrieved from PubMed, The Cochrane Library, and EMbase, as well as Chinese databases such as CNKI, WanFang, VIP, and China Biomedical Literature Service. The search period extended from the inception to December 31, 2023. Two researchers independently screened the literature, extracted and analyzed the data. The quality of the included studies was evaluated by means of the SYRCLE animal experiment bias risk assessment table. Main outcome measures: Follicle stimulating hormone, estradiol, luteinizing hormone, the quantity of follicles at all levels. Secondary outcome measure: Pregnancy rate. Network meta-analysis, mapping, and tabulation were executed using Stata 17.0 software after assessing the risk of bias in the included studies.

Totally 24 animal experiment studies were incorporated, and the overall quality of the literature was mediocre, encompassing 7 distinct sources of mesenchymal stem cells. They were umbilical cord-derived mesenchymal stem cells, menstrual blood-derived mesenchymal stem cells, placenta-derived mesenchymal stem cells, human cord blood-derived mesenchymal stem cells, bone marrow-derived mesenchymal stem cells, adipose-derived mesenchymal stem cells, and amnio-derived mesenchymal stem cells. The network meta-analysis demonstrated that (1) in contrast to the blank group, mesenchymal stem cells from various sources were effective in enhancing the pregnancy rate and estradiol, reducing follicle-stimulating hormone and luteinizing hormone, augmenting the number of follicles at all levels, and diminishing the number of atretic follicles. (2) According to the area map under the cumulative sequencing curve, the three stem cells with the most prominent efficacy in improving estradiol levels were umbilical cord-derived mesenchymal stem cells (72.7%) > adipose-derived mesenchymal stem cells (72.6%) > menstrual blood-derived mesenchymal stem cells (71.7%). (3) The three kinds of stem cells with the highest efficacy in reducing follicle-stimulating hormone levels were the adipose-derived mesenchymal stem cells (96.3%) > human cord blood-derived mesenchymal stem cells (65.4%) > umbilical cord-derived mesenchymal stem cells (63.9%). (4) The three kinds of stem cells with the highest efficacy in reducing luteinizing hormone levels were adipose-derived mesenchymal stem cells (100.0%) > umbilical cord-derived mesenchymal stem cells (51.6%) > human cord blood-derived mesenchymal stem cells (46.8%). (5) The top three kinds of stem cells for increasing the number of primordial follicles were human cord blood-derived mesenchymal stem cells (76.3%) > umbilical cord-derived mesenchymal stem cells (75.5%) > menstrual blood-derived mesenchymal stem cells (57.5%). (6) The top three kinds of stem cells for increasing the number of primary follicles were umbilical cord-derived mesenchymal stem cells (75.3%) > adipose-derived mesenchymal stem cells (53.0%) > the placenta-derived mesenchymal stem cells (51.7%). (7) The top three kinds of stem cells for increasing the number of secondary follicles were adipose-derived mesenchymal stem cells (76.1%) > menstrual blood-derived mesenchymal stem cells (66.8%) > umbilical cord-derived mesenchymal stem cells (66.5%). (8) The top three kinds of stem cells in reducing the number of atretic follicles were adipose-derived mesenchymal stem cells (99.9%) > bone marrow-derived mesenchymal stem cells (68.1%) > umbilical cord-derived mesenchymal stem cells (53.4%).

(1) For animal models of premature ovarian failure, the results of the network meta-analysis disclosed that various stem cell transplantation treatments were preponderant over the blank or placebo group to varying extents, and the efficacies were comparable. (2) The results indicated that umbilical cord-derived mesenchymal stem cells were the most frequently utilized and adipose-derived mesenchymal stem cells were the most potent. More high-quality experimental study data are requisite in the future for further validation.

With aging, the regenerative capacity and differentiation function of bone marrow mesenchymal stem cells progressively decline, reducing bone tissue repair efficacy. Thus, identifying bone marrow mesenchymal stem cell subpopulations with enhanced osteogenic potential is of significant importance for advancing bone tissue engineering.

To evaluate the osteogenic differentiation potential differences between STRO-1 positive and negative bone marrow mesenchymal stem cells under osteogenic induction conditions.

SD rat bone marrow mesenchymal stem cells were isolated and cultured. The expression of CD29, CD45, CD90, and STRO-1 was identified via flow cytometry and immunofluorescence. Immunomagnetic cell sorting was used to separate STRO-1 positive and negative bone marrow mesenchymal stem cells. The cells of two groups were subjected to osteogenic induction for 7 and 14 days. qRT-PCR and western blotting were performed to analyze differences in osteogenesis-related gene expression (Collagen I, Runt-related transcription factor 2, osteoprotegerin, and osteocalcin) and protein levels. Alizarin red staining and alkaline phosphatase staining were used to observe calcium nodule formation.

Flow cytometry showed high expression levels of CD29 and CD90 and low expression of CD45, with a positive STRO-1 expression rate of 12.8%. Immunofluorescence results were consistent with those of flow cytometry. After magnetic cell sorting, STRO-1 positive cells demonstrated a higher colony formation rate than STRO-1 negative cells. On day 14, STRO-1 positive cells showed significantly higher osteogenic differentiation potential than on day 7, with significantly elevated osteogenesis-related marker levels compared to STRO-1 negative cells (P < 0.01). The findings indicate that STRO-1 positive bone marrow mesenchymal stem cells exhibit significant advantages in osteogenic potential, providing a theoretical basis for their selection as ideal seed cells in bone tissue engineering. In future applications, they may represent a promising therapeutic approach for bone defect repair.

Human pluripotent stem cell-derived cardiomyocytes offer an ideal cellular resource for studying heart diseases, conducting drug screening, developing in vitro heart models, and exploring potential cell therapies. However, human pluripotent stem cell-derived cardiomyocytes are characterized by immaturity with limited specific gene expression, low Ca²⁺ processing levels, and underdeveloped structural, metabolic, and electrophysiological features. These limitations significantly impede the application of human pluripotent stem cell-derived cardiomyocytes.

To review the academic progress and clinical application of promoting the maturation of human pluripotent stem cell-derived cardiomyocytes by in vitro synthetic microenvironment.

CNKI, WanFang, VIP, PubMed, Web of Science, and Medline databases were searched, with “human pluripotent stem cells, human myocardial cells, hPSC-CMs, mature, OA, human pluripotent stem cell-derived cardiomyocytes, hPSC-CMs” as English search terms and “human pluripotent stem cells, cardiomyocytes, mature, OA, hPSC-CMs” as Chinese search terms. All relevant literature published from January 2002 to July 2024 was retrieved and 82 articles were included in the review.

(1) In recent years, in vitro synthetic microenvironments have attracted extensive attention due to their excellent intrinsic properties such as stiffness, plasticity, nanoscale morphology, and chemical functionality. (2) Human pluripotent stem cell-derived cardiomyocytes can be used as an effective platform for the treatment of cardiovascular diseases. (3) Mechanical stimulation, electrical stimulation, addition of biochemical molecules, and three-dimensional culture methods are effective methods to promote the maturation of human pluripotent stem cell-derived cardiomyocytes, which can further promote the clinical application of human pluripotent stem cell-derived cardiomyocytes.

Exosomes have been confirmed to be closely related to cartilage degeneration in osteoarthritis. However, the role and mechanism of exosome-derived genes in cartilage degeneration of osteoarthritis have not been fully elucidated.

Bioinformatics analyses were used to screen the genes related to cartilage degeneration in the synovial exosomes of patients with osteoarthritis, and to determine their biological functions and signaling pathways in order to provide new therapeutic targets for delaying cartilage degeneration in osteoarthritis.

Firstly, osteoarthritis-related exosome dataset GSE185059 and cartilage degeneration dataset GSE114007 were downloaded from Gene Expression Omnibus (GEO) database to screen exosome-derived cartilage degeneration related genes. GO functional and KEGG pathway enrichment analyses were performed based on the screened exosome-derived cartilage degeneration related genes. Protein-protein interaction network was drawn and Ingenuity Pathway Analysis (IPA) was conducted to screen and obtain key exosome-derived cartilage degeneration-related genes. Finally, qRT-PCR was used to verify the expression of key genes in osteoarthritis cartilage tissue and interleukin-1β stimulated chondrocyte models.

(1) There were 831 differentially expressed genes in the GSE185059 dataset and 5 323 differentially expressed genes in the GSE114007 dataset. A total of 94 exosome-derived cartilage degeneration related genes were screened after the intersection of these differentially expressed genes, of which 51 genes were down-regulated and 43 genes were up-regulated. (2) GO functional enrichment analysis showed that the up-regulated genes were mainly involved in the positive regulation of cell-cell adhesion, the positive regulation of T cell activation, and chronic inflammatory response, while the down-regulated genes were mainly involved in biological processes such as cell aggregation, cartilage differentiation and development, and skeletal system morphogenesis. (3) KEGG pathway enrichment analysis showed that exosome-derived cartilage degeneration-related genes were mainly involved in tryptophan enrichment metabolism, vitamin B6 metabolism, and leukocyte transendothelial migration. (4) The constructed protein-protein interaction network confirmed the existence of multiple interaction relationships among exosome-derived cartilage degeneration-related genes. Combined with five algorithms in CytoHubba software, four key exosome-derived cartilage degeneration-related genes were further screened, namely THY1, CYP1A1, NFKB2, and COL6A3. (5) The results of qRT-PCR showed that compared with normal cartilage, the expressions of THY1 and COL6A3 in osteoarthritic cartilage were increased, while the expression of CYP1A1 and NFKB2 was decreased. Similarly, compared with the unstimulated group, the expression of THY1 and COL6A3 in the interleukin-1β induced chondrocytes was upregulated, while the expression of CYP1A1 and NFKB2 was downregulated. (6) These results indicate that THY1, CYP1A1, NFKB2, and COL6A3 are genes related to cartilage degeneration in the exosomes of synovial fluid of patients with osteoarthritis, and may participate in the pathogenesis of osteoarthritis by regulating biological processes such as protein tyrosine kinase activity and lipid metabolism, as well as nuclear factor-κB signaling pathway and focal adhesion signaling pathway. However, the specific regulatory roles and molecular mechanisms of these key genes in cartilage degeneration need to be further verified by experiments.

Hematopoietic stem cell transplantation is a widely used treatment method to cure malignant and nonmalignant diseases originated from hematological cells. Mobilization and collection of sufficient hematopoietic stem cells are the preconditions to ensure rapid and sustained hematopoietic reconstitution after hematopoietic stem cell transplantation. However, the most commonly used granulocyte colony-stimulating factor with or without chemotherapy still has a mobilization failure rate of 10% to 40%.

To review the present status of hematopoietic stem cell mobilization in recent years, analyze the advantages and disadvantages of different mobilization plans, and be looking forward for new mobilization methods.

Using “hematopoietic stem cell, hematopoietic stem cell transplantation, hematopoietic stem cell mobilization, cytokines, thrombopoietin, CXCR4 antagonists, integrin antagonist, chemotherapy, mobilization efficiency” as Chinese and English keywords, articles published from 1990 to 2024 were searched on CNKI, PubMed, and WanFang databases. A total of more than 300 articles were retrieved, and 68 articles were finally included.

More and more studies have found that granulocyte colony-stimulating factor combined with other agents including plerixafor, interleukins, thrombopoietin, and integrin antagonist could improve hematopoietic stem cell mobilization. Combined use can reduce the dose of granulocyte colony-stimulating factor and related adverse reactions. Some new drugs, such as soluble recombinant FLT3-ligand (CDX-301) and dual α9β1/α4β1 integrin inhibitor BOP, can be combined with granulocyte colony-stimulating factor to promote hematopoietic stem cell mobilization. In addition, some potential mobilization targets, such as prostaglandin E2 receptor and sphingosine 1-phosphate, are still in the research stage. In addition to inherent patient characteristics and treatment options, incorporating biomarkers into the factors affecting mobilization and developing new predictive models will help to effectively predict the failure of hematopoietic stem cell mobilization and improve stem cell mobilization technology.