Spina bifida (SB) is associated with progressive spinal cord injury and limited therapeutic options for neural protection. Amniotic fluid stem cells (AFSCs) have shown regenerative potential, but their effects on SB-related neuronal injury and the underlying mechanisms remain unclear. In this study, we investigated whether AFSCs alleviate neuronal injury-related changes in SB rats through the modulation of autophagy via the FOXO1/STAT3 pathway. SB-like lesions were induced in neonatal rats, followed by the intramedullary injection of AFSCs or vehicle on postnatal day 1. Behavioral assessment was performed using Basso, Beattie, and Bresnahan locomotor scoring on days 2, 7, and 14 after injection, and lesion-centered lumbosacral spinal cord tissues were collected on day 15. AFSC treatment partially improved hindlimb locomotor scores and reduced neuronal injury-related alterations in SB rats. AFSCs also increased autophagy-associated markers, including LC3II/I and Beclin1, and reduced p62 mRNA expression in vivo and in neural stem cells isolated from SB rats. Mechanistically, AFSC treatment was associated with reduced FOXO1 and enhanced STAT3 signaling. FOXO1 overexpression or STAT3 inhibition attenuated AFSC-mediated effects, whereas siRNA-mediated FOXO1 knockdown enhanced these effects; combined STAT3 knockdown partially reversed the effects of FOXO1 silencing. These findings suggest that AFSCs alleviate SB-related neuronal injury-associated changes by regulating autophagy through the FOXO1/STAT3 axis. This study provides preclinical evidence for an experimental neuroprotective mechanism of AFSCs, although further validation is required before translational applicability can be considered.
Background Periodontitis is a chronic inflammatory condition characterized by destruction of periodontal support tissues and dysregulated immune responses, particularly involving cytokines like tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β). Identifying molecules that regulate both inflammation and tissue regeneration is essential for targeted therapies. Following preliminary transcriptomic evidence on periodontal tissues, this study aimed to define the specific immunomodulatory and osteogenic regulatory effects of TRAB domain-containing protein 2A (TRABD2A).Methods RNA sequencing was performed on periodontal tissues from patients with periodontitis and periodontally healthy individuals to identify differentially expressed genes and conduct functional enrichment analysis. TRABD2A expression and its effects on cell proliferation, inflammation, osteogenic markers expression, and mineralization were assessed in human periodontal ligament stem cells (hPDLSCs) treated with recombinant TRABD2A, Porphyromonas gingivalis lipopolysaccharide (LPS), or a TRABD2A inhibitor (o-Phenanthroline).Results RNA sequencing revealed significant TRABD2A upregulation in periodontitis tissues, along with elevated bone-related genes runt-related transcription factor 2 (Runx2) and receptor activator of nuclear factor-κB Ligand (RANKL) expression. In vitro experiments demonstrated that TRABD2A reduced the secretion of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) and promoted the proliferation of hPDLSCs. Furthermore, TRABD2A upregulated Runx2 expression and suppressed RANKL expression, while Alizarin Red staining results further confirmed its role in promoting osteogenic differentiation. The underlying mechanism may be associated with the enhancement of β-catenin expression.Conclusion TRABD2A exerts dual functions by promoting osteogenic differentiation and modulating inflammatory responses, highlighting its potential as an immunoregulatory and pro-regenerative target for periodontitis treatment.
Background While secreted phosphoprotein 1 (SPP1) drives macrophage M2 polarization and cancer progression in multiple malignancies, its role in thyroid cancer remains poorly understand. This study investigated the roles and mechanisms of thyroid cancer-derived exosomal SPP1 in regulating macrophage M2 polarization and malignant progression in thyroid cancer.Methods SPP1 protein expression in thyroid cancer tissues and cell lines was assessed. Exosomes from TPC1 cells were isolated and characterized for SPP1 expression. THP-1 cell-differentiated M0 macrophages were treated with unmodified or SPP1-knockdown exosomes, followed by detection of M2 phenotype markers and analysis of cytokine secretion profiling in response to LPS stimulation. The conditioned macrophages were then co-cultured with thyroid cancer cells, and cell proliferation, migration and invasion were examined. Mechanistic studies focused on CD44/JAK2/STAT3 signaling using an anti-CD44 monoclonal antibody. Additionally, tumor growth and macrophage infiltration were assessed in a xenograft tumor mouse model following exosomal SPP1 knockdown.Results SPP1 was upregulated in thyroid cancer and enriched in TPC1-derived exosomes. These exosomes induced macrophage M2 polarization, thereby promoting cancer cell proliferation, migration and invasion. However, SPP1 knockdown in exosomes abolished these effects. Exosomal SPP1 directly interacted with CD44 receptors and activated the JAK2/STAT3 signaling pathway to drive M2 polarization. CD44 blockade reversed M2 polarization and pro-tumorigenic effects. SPP1-deficient exosomes reversed tumor growth and attenuated M2 macrophage infiltration in xenograft model.Conclusions Thyroid cancer-derived exosomal SPP1 promotes tumor progression by driving macrophage M2 polarization via the CD44/JAK2/STAT3 signaling pathway. The exosomal SPP1-CD44 axis represents a promising therapeutic target for thyroid cancer.
Background This study aimed to determine the potential mechanism by which pancreatic stem cell-derived beta cells (PSCs-β) assist in the body's glucose-lowering capacity in type 1 diabetes (T1D) rats.Methods We screened the transcriptomic changes in the pancreatic islets of T1D mice to extract key genes from the GSE169275 dataset. Cell proliferation, cell cycle distribution, apoptosis, PSC-β differentiation ability, and insulin production levels were analyzed after MLPH overexpression/knockdown in PSCs. PSC-β-overexpressing MLPH were transplanted into T1D rats, and the changes in fasting blood glucose level, glucose tolerance and insulin, glucagon and C-peptide contents were examined. After the target genes of MLPH were analysed using the database, immunoprecipitation was introduced for validation. Whether RAB3A is involved in the regulatory effects of MLPH on the proliferation and differentiation of PSCs was further verified.Results MLPH overexpression further enhanced the proliferation of PSCs, inhibited apoptosis and accelerated the differentiation of PSCs to PSC-β cells and insulin secretion. After the transplantation of MLPH-overexpressing PSC-β cells, the pancreatic islet tissue damage was restored, the insulin expression was substantially elevated and the glucagon content decreased. RAB3A knockdown counteracted the effects of MLPH on the proliferation, differentiation and insulin secretion of PSCs.Conclusion MLPH overexpression is favourable for the differentiation of PSCs to insulin β-cells. Transplantation of MLPH-overexpressing PSC-β cells restored the ability of T1D rats to manage a glycemic load by promoting RAB3A expression.
Objective To explore the synergistic effect of nano-pearl powder (NPP) and adipose-derived stem cell exosomes (ADSC-Exos) on the osteogenic potential of MC3T3-E1 cells.Methods The water-soluble matrix of NPP (NPP-WSM) was extracted via freeze-drying, and ADSC-Exos were isolated by ultracentrifugation. NPP-WSM was incorporated into ADSC-Exos through co-incubation to generate NPP-WSM-Exos. MC3T3-E1 cells were treated with NPP-WSM or NPP-WSM-Exos. Cell proliferation and migration were evaluated using CCK-8 and wound-healing assays, respectively. Osteogenic differentiation was assessed by Alizarin Red S staining and alkaline phosphatase (ALP) activity. The expression of osteogenesis-related genes (COL1A1, RUNX2, OCN, and OPN) was measured by qPCR and Western blotting. Transcriptome sequencing (RNA-seq) was conducted to identify signaling pathways activated by NPP-WSM-Exos.Results NPP-WSM-Exos displayed distinct exosome morphology and biomarkers, confirming their successful preparation. Significantly, NPP-WSM-Exos enhanced the viability of MC3T3-E1 cells compared to NPP-WSM alone and upregulated the expression of osteogenic genes, including COL1A1, RUNX2, OCN, and OPN, at both the transcriptional and translational levels. Additionally, NPP-WSM-Exos strongly promoted mineralization, as evidenced by the increased calcification observed through Alizarin Red S staining, and elevated alkaline phosphatase (ALP) activity, indicating excellent potential for osteogenic differentiation. Transcriptome sequencing showed that NPP-WSM-Exos significantly enhanced the PI3K/AKT pathway in MC3T3-E1 cells, while protein level detection indicated that NPP-WSM-Exos could increase AKT phosphorylation levels and inhibit GSK3β activity to improve osteogenic efficiency.Conclusion The use of adipose-derived stem cell exosomes to encapsulate NPP-WSM can increase the utilization of WSM, promote the proliferation of MC3T3-E1, and enhance the osteogenic differentiation ability.
