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.
Animal experimental studies involving the Igaki-Tamai stent (ITS), a bioresorbable poly-l-lactic acid scaffold, in peripheral arteries are limited, and existing studies evaluated only short-term (3-month) outcomes. This study compared arterial responses associated with the ITS and bare metal stent (BMS) over 6 months using intravascular ultrasound (IVUS) analysis and evaluated feasibility in porcine iliac arteries. Four miniature pigs underwent stent implantation with the ITS in the right iliac artery and the BMS in the left iliac artery. Follow-up evaluations at 6, 12, and 24 weeks included angiographic and IVUS analyses to assess neointimal hyperplasia, percent area stenosis (%AS), and percent in-stent volume obstruction (%VO). Histological analysis was performed to evaluate tissue injury and inflammation scores. At 6 weeks, the neointimal area did not differ significantly between the ITS and BMS groups (8.49 ± 2.10 mm² vs 13.47 ± 6.67 mm², P = .205). However, the ITS group exhibited a significantly smaller neointimal area at 12 weeks (6.87 ± 1.15 mm² vs 20.65 ± 10.99 mm², P = .050) and 24 weeks (5.20 ± 0.85 mm² vs 22.32 ± 12.03 mm², P = .042). %AS and %VO were significantly lower in the ITS group at all follow-ups. The ITS group showed reduced tissue damage (injury score: 0.80 ± 0.430 vs 1.74 ± 0.908, P < .001) and inflammation (inflammation score: 1.25 ± 0.516 vs 1.67 ± 0.832, P < .001) compared with the BMS group. The ITS was associated with reduced vessel injury, lower inflammatory response, and favorable luminal remodeling over 6 months in healthy porcine iliac arteries.
Decellularized blood vessels with low immunogenicity and excellent biocompatibility are promising for tissue engineering and clinical applications. However, current decellularization methods face limitations in cell removal efficiency, matrix preservation, and biosafety. This study optimized the Triton X-100/SDS (TX-100/SDS) decellularization method using ultrasound technology by systematically evaluating the effects of ultrasound power, temperature, and processing time on decellularization efficiency. The optimized method achieved a 72% reduction in nucleic acid residues at 100 W power while preserving matrix integrity and significantly reducing chemical reagent residues. Structural and biosafety evaluations confirmed that the optimized scaffolds met biological safety standards and demonstrated excellent stability, providing a strong foundation for developing high-performance decellularized vascular materials for clinical applications.
Endovascular aneurysm repair (EVAR) is a widely accepted treatment for aortic pathologies owing to its minimally invasive nature. However, long-term complications, such as stent graft migration and infection, remain unresolved, primarily due to the persistent presence of synthetic materials and limited tissue integration. This pilot study evaluated the feasibility of a novel tissue-engineered stent graft (TESG) combining a bioresorbable poly-L-lactic acid (PLLA) stent with decellularized porcine veins. The veins were processed using a sodium dodecyl sulfate and the Triton X-100 decellularization protocol. Histological and ultrastructural analyses confirmed effective cell removal while preserving extracellular matrix components. Quantitative deoxyribonucleic acid (DNA) analysis showed a > 97% reduction in DNA content. The TESGs were assembled by suturing the decellularized veins into bioresorbable PLLA stents and implanted into porcine iliac arteries (n = 3). Commercially available prosthetic grafts were used as control implants to evaluate differences in tissue responses. Graft patency and morphology were assessed at implantation and on postoperative day 14 using angiography and intravascular ultrasonography. All TESGs remained patent, with no evidence of thrombosis or aneurysmal changes. Histological analysis revealed early endothelialization and smooth muscle cell infiltration within the TESG wall, in contrast to the prosthetic graft controls, which lacked comparable cellular integration. This study demonstrated the short-term feasibility and biological compatibility of a fully bioresorbable TESG. Although long-term outcomes remain to be established, these results support further development of TESG to reduce late complications through improved tissue integration and avoidance of permanent synthetic materials.
OBJECTIVE:Periodontal ligament stem cells (PDLSCs), undifferentiated mesenchymal cells with multipotent differentiation and self-renewal capacities, constitute the optimal MSC population for periodontal regeneration. This study sought to elucidate the effects of concentrated growth factor (CGF) on the osteogenic differentiation of PDLSCs and analyze the underlying mechanisms. METHODS:PDLSCs were isolated from the molars of patients with malocclusion and characterized by flow cytometry, osteogenic induction, lipogenic induction, ARS staining and ORO staining. PDLSCs were treated with osteogenic induction medium containing different concentrations of CGF. The osteogenic ability of CGF in PDLSCs was analyzed via ALP staining, ARS staining, and ALP activity assays. WNK1, RUNX2 and OPN were detected by RT-qPCR. WNK1, RUNX2, OPN, β-catenin, GSK3β and p-GSK3β were detected by WB. The role of CGF in PDLSC osteogenic differentiation through the Wnt pathway was verified. RESULTS:PDLSCs were successfully isolated and cultured in vitro. After CGF treatment, ALP activity, mineralization nodule formation, and the expression of RUNX2 and OPN in PDLSCs were increased, with 0.1 mg/mL CGF showing the best osteogenic differentiation ability. WNK1, β-catenin, and p-GSK3β/GSK3β were elevated. CGF activated the Wnt pathway through WNK1. The promoting effects of CGF on osteogenic differentiation of PDLSCs were partially reversed after inhibition of WNK1 and the Wnt pathway. CONCLUSION:CGF promotes PDLSC osteogenic differentiation by activating the Wnt pathway through WNK1.
MicroRNAs (miRNAs) can be transported to tumor cells through exosomes secreted by bone marrow mesenchymal stem cells (BMSC-Exos) and exert regulatory functions within cells. Here, we aim to investigate the functional mechanism of miR-29a-3p carried by BMSC-Exos in the treatment of NSCLC. Based on the miRNA/mRNA gene expression data in the UCSC dataset (1029 NSCLC and 110 normal samples), bioinformatics analysis predicted the expression levels of miR-29a-3p and DNMT3A in NSCLC samples and their association with prognosis. Exosomes were isolated from BMSCs and characterized. BMSC-Exos with expressed or knocked out miR-29a-3p were treated A549 cells, and their biological effects on cells were evaluated, including proliferation, migration, and apoptosis. Western blotting was employed to explore the involvement of DNMT3A and JAK2/STAT3 signaling pathways. Furthermore, the binding between miR-29a-3p and DNMT3A was verified through dual-luciferase reporter assay. Following transfection with miR-29a-3p mimic and DNMT3A overexpression vectors, their roles in the biological processes of NSCLC were analyzed. In NSCLC, decreased expression of miR-29a-3p or increased expression of DNMT3A was closely associated with poor prognosis. miR-29a-3p can be transferred from BMSCs to A549 cells via exosomes, thereby inhibiting cell proliferation and migration while promoting apoptosis. DNMT3A was identified as a target gene of miR-29a-3p. Mechanistically, miR-29a-3p upregulation led to decreased DNMT3A expression and impaired JAK2/STAT3 signaling pathway. Overall, this study demonstrated that BMSC-derived exosomal miR-29a-3p restrained NSCLC by reducing DNMT3A/JAK2/STAT3 axis. These findings may provide new insights for the development of NSCLC treatment strategies.
This study delves into the rejuvenating effects of SS-31 on aged human Bone Marrow-Derived Mesenchymal Stem Cells (BM-MSCs), focusing on its potential to restore their diminished osteogenic differentiation capacity, a critical issue in geriatric medicine and bone tissue engineering. SS-31 significantly improved mitochondrial function, increasing ATP production by 35% and reducing ROS levels by 40% in aged BM-MSCs. Osteogenic differentiation was enhanced, as evidenced by a 2.8-fold increase in ALP activity and a 3.5-fold increase in Alizarin Red S staining intensity. Additionally, SS-31 reduced NOS2 expression by 50%, highlighting its therapeutic potential in age-related bone loss. SS-31 intervention not only normalizes mitochondrial structure and function, reducing ROS levels and enhancing oxygen consumption rates, but also targets the NOS2 gene, a potential drug target, which upon knockdown, leads to a substantial upregulation of osteogenic markers and an improvement in mitochondrial function. In conclusion, the findings of this study highlight the therapeutic potential of SS-31 in reversing the age-related decline in BM-MSC function by specifically inhibiting NOS2 expression and restoring mitochondrial function. This research provides a scientific basis for the development of new treatments for osteoporosis and other age-related bone diseases, emphasizing the importance of targeting mitochondrial function and cellular senescence in regenerative therapies.
BACKGROUND:Epilepsy is a common disease of the nervous system. Recent advances in epigenetics have revealed DNA methylation as a key mechanism in epilepsy pathogenesis, particularly through dysregulation of GABAergic signaling. Baicalein has been shown to have anticonvulsant and neuroprotective effects. However, its epigenetic regulatory effects on GABA receptor function remain unexplored. METHODS:The status epilepticus (SE) model was induced by lithium chloride-pilocarpine (LiCl-PILO) in Sprague-Dawley (SD) rats. The rats were divided into control group, epileptic SE group and baicalein intervention group. Morris water maze (MWM) test, Nissl staining, immunofluorescence and enzyme-linked immunosorbent assay (ELISA) were used to detect cognitive functions and neuronal damage. Online sites, chromatin immunoprecipitation (ChIP) and western blotting were used to identify DNA methyltransferase 1 (DNMT1)-mediated methylation of gamma-aminobutyric acid type A receptor subunit delta (GABRD) promoter region. RESULTS:Baicalein treatment significantly prolonged the latency of SE onset and seizure onset, and improved the development of epilepsy. Meanwhile, baicalein improved the cognitive impairment in rats induced by LiCl-PILO. After treatment with baicalein, a sustained elevation in the number of neurons and NeuN levels was observed, along with a decrease in the contents of tumor necrosis factor -alpha (TNF-α), interleukin-1β (IL-1β), and ionized calcium-binding adapter molecule 1 (Iba-1) in the hippocampus. Mechanistically, baicalein interacted with DNMT1 to suppress GABRD promoter region methylation, thus increasing GABRD protein level in the hippocampus of rats induced by LiCl-PILO. CONCLUSION:This study identifies DNMT1/GABRD axis as a novel epigenetic target for epilepsy intervention. Baicalein's ability to enhance tonic inhibition through demethylation of GABRD provides a groundbreaking strategy for drug-resistant epilepsy.
Podocyte damage is a central feature of lupus nephritis (LN), making the identification of potential therapeutic targets to prevent podocyte injury and improve treatment outcomes essential. ORM1 has been suggested as a significant candidate gene in LN. In this study, mouse podocytes were induced using Immunoglobulin G (IgG) extracted from lupus patients. To investigate the role of ORM1, ORM1 knockdown was performed, and the effects on podocyte viability and apoptosis were assessed using the cell counting kit-8 (CCK-8) assay and flow cytometry. Additionally, autophagy markers LC3II/I and p62 were measured by western blotting and immunofluorescence, and the expression of the AMPK/mTOR signaling pathway was evaluated using western blotting. The results showed an upregulation of ORM1 in the LN model. Upon stimulation with IgG from LN patients, ORM1 knockdown reversed the reduction in podocyte viability, decreased the apoptosis rate, and reduced the elevated levels of autophagy, followed by an increase in AMPK phosphorylation and a decrease in mTOR phosphorylation. In conclusion, these results suggest that ORM1 modulates the expression of autophagy-related components in podocytes through the AMPK/mTOR signaling pathway, thereby influencing podocyte damage in the LN model in vitro.
Neural stem cell (NSC) possess the essential properties of pluripotency and self-renewal, making them promising candidates for the treatment of neurological disorders such as Alzheimer’s disease (AD), Parkinson’s disease (PD), and spinal cord injuries. While previous studies have identified the long non-coding RNAs (lncRNAs) Pnky as a regulator of NSC differentiation into neurons via RNA splicing, its role in NSC differentiation and proliferation through the Wnt/β-catenin pathway remains unclear. In this study, we investigated the mechanism by which Pnky influences the Wnt/β-catenin pathway to promote NSC differentiation into neurons. Using cck8 assays, western blot analysis, and quantitative polymerase chain reaction (qPCR), we found that Pnky knockdown significantly enhanced NSC proliferation and promoted their differentiation into neurons. Additionally, Pnky knockdown resulted in the downregulation of the neural stem cell marker Nestin and upregulation of the neuronal marker β3-Tubulin, through activation of the β-catenin signaling pathway. Conversely, inhibiting the β-catenin pathway hindered both NSC differentiation and proliferation. These findings suggest that targeting the Pnky-mediated Wnt/β-catenin pathway may offer novel strategies for the treatment, diagnosis, and drug development of central nervous system diseases.
Objective This study probed the effect of targeted regulation of CD151 by microRNA-214-3p (miR-214-3p) delivered by bone marrow mesenchymal stem cells-secreted exosomes (BMSCs-exo) on oxidative stress and apoptosis of neurons in Alzheimer’s disease (AD).Methods Rat BMSCs were isolated, from which MSCs-exo were extracted and identified. The AD rat model was established and injected with MSC-exo suspension. Meanwhile, miR-214-3p and CD151 interfering lentivirus were transfected in MSCs. After injection, learning and cognitive ability of the rats were assessed, as well as neuronal apoptosis and oxidative stress injury. miR-214-3p and CD151 levels were determined, and their relationship was explored.Results AD rats had prolonged escape latency, weakened learning and cognitive ability, increased neuronal apoptosis in the hippocampal CA3 region, and aggravated oxidative stress. After MSC-exo injection, these changes in AD rats were partially rescued. CD151 was targeted by miR-214-3p, and MSC-exo improved AD in rats through the miR-214-3p/CD151 axis.Conclusion MSC-exo down-regulates CD151 by targeting miR-214-3p to enhance antioxidant capacity, thereby improving the pathological injury of AD rats.
Objective This study evaluates the effectiveness of optimized individualized nursing interventions on clinical outcomes in intensive care unit (ICU) patients with severe pneumoniaMethods In this randomized controlled trial, 76 patients with severe pneumonia were randomized into a control group and an experimental group. Both groups received routine nursing care. On this basis, the experimental group received optimized individualized nursing. After the nursing intervention, clinical outcomes, respiratory function, coagulation function, Acute Physiology and Chronic Health Evaluation II (APACHE II) score, and St. George’s Respiratory Problems Questionnaire (SGRQ) score were assessed, and the complication and mortality rates were counted.Results After the intervention, compared with the control group, the experimental group exhibited shorter times of fever reduction, white blood cell count recovery, and off-boarding and ICU stay, higher oxygenation index, lower rapid shallow breathing index, respiratory rate, activated partial thromboplastin time, prothrombin time, fibrinogen, and D-Dimer levels, lower APACHE II scores and SGRQ scores (p < 0.05). Additionally, the experimental group possessed a lower complication rate and mortality rate than the control group (p < 0.05).Conclusion Implementing optimized individualized nursing can significantly enhance recovery and reduce complications in ICU patients with severe pneumonia.
Fiber-photometric is a novel optogenetic method for recording neural activity in vivo, which allows the use of calcium indicators to observe and study the relationship between neural activity and behavior in free-ranging animals. Calcium indicators also convert changes in calcium concentration in cells or tissues into recordable fluorescent signals, which can then be observed using the system of fiber-photometric. To date, there is a paucity of relevant literature on the proper selection and application of fiber-photometric indicators. Therefore, this paper will detail how to correctly select and apply fiber-photometer indicators in four sections: the basic principle of optical fiber photometry, the selection of calcium fluorescent probes and viral vector systems, and the measurement of specific expression of fluorescent proteins in specific tissues. Therefore, the correct use of suitable fiber optic recording indicators will greatly assist researchers in exploring the link between neuronal activity and neuropsychiatric disorders.
Objective This trial probed the correlation between miR-31 expression and endometrial receptivity (ER) in patients with repeated implantation failure (RIF) of in vitro fertilization and embryo transfer (IVF-ET).Methods A retrospective study of 80 infertility patients who underwent IVF-ET assisted conception treatment were divided into RIF group and normal pregnancy group (control group) according to the pregnancy outcome after embryo transfer. General information of both groups was collected. Endometrial tissues were collected in the middle luteal phase of the menstrual cycle before IVF-ET. miR-31 levels in endometrial tissues were measured, and endometrial tolerance indicator pulsatility index (PI), resistance index (RI), and endometrial thickness (Em) were detected. The correlation between endometrial miR-31 levels and ER indices was evaluated by Pearson method. ROC curves were utilized to analyze the efficacy of miR-31 in predicting RIF occurrence. The influencing factors of RIF were analyzed by binary Logistic regression.Results RIF patients had increased miR-31 expression level and endometrial tolerance indicator PI, and RI while decreased Em (p < 0.05). miR-31 in RIF patients was positively correlated with PI and RI, and negatively correlated with Em (p < 0.05). The area under the curve for miR-31 to predict the occurrence of RIF was 0.899, with a sensitivity of 0.750 and a specificity of 0.950. PI, RI, and miR-31 were risk factors for developing RIF in IVF-ET women, and Em was a protective factor (p < 0.05).Conclusion miR-31 in RIF patients is positively correlated with PI and RI, and negatively correlated with Em.
Tetramethylpyrazine (TMP) has been confirmed to suppress inflammation in endometriosis (EMs). Herein, this study investigated whether and how TMP affected NLRP3 inflammasomes and oxidative stress in EMs. After establishment of an EMs rat model, rats were treated with different concentrations of TMP. The size of endometriotic lesions and the latency and frequency of torsion in rats were recorded, followed by the measurement of relevant indicators (TNF-α, IL-6, IL-2, IL-10, MDA, SOD, GSH, CAT, ROS, NLRP3, ASC, GSDMD, caspase-1, Nrf2, and HO-1). The study experimentally determined that TMP treatment markedly decreased the size of endometriotic lesions and improved torsion in rats with EMs. The levels of inflammatory proteins, oxidative stress markers, NLRP3 inflammasome, and pyroptotic proteins were elevated in rats with EMs, all of which were reversed upon TMP treatment. Additionally, the activities of SOD, GSH, and CAT were lowered in rats with EMs, which were partly abrogated by TMP treatment. Furthermore, the downregulation of Nrf2 and HO-1 was counteracted by TMP treatment. To sum up, TMP represses excessive oxidative stress, NLRP3 inflammasome activation, and pyroptosis in rats with EMs. Additionally, TMP may activate the Nrf2/HO-1 pathway.
Zinc ions play a pivotal role in facilitating the development of cartilage in mice. Nevertheless, the precise underlying mechanism remains elusive. Our investigation was centered on elucidating the impact of zinc deficiency on cartilage maturation by modulating SUMO1 and UBC9 at both the protein and mRNA levels. We administered a regimen inducing zinc deficiency to gravid mice from E0.5 until euthanasia. Subsequently, we subjected the embryos to scrutiny employing HE, Safranin O staining and IHC. Primary chondrocytes were isolated from fetal mouse femoral condyles and utilized for Western blot analysis to discern the expression profiles of SUMO1, SUMO2/3, UBC9, SOX9, MMP13, Collagen II, RUNX2, and aggrecan. Furthermore, ATDC5 murine chondrocytes were subjected to treatment with ZnCl2, followed by RT-PCR assessment to scrutinize the expression levels of MMP13, Collagen II, RUNX2, and aggrecan. Additionally, we conducted Co-IP assays on ZnCl2-treated ATDC5 cells to explore the interaction between SOX9 and SUMO1. Our investigation unveiled that zinc deficiency led to a reduction in cartilage development, as evidenced by the HE results in fetal murine femur. Moreover, diminished expression levels of SUMO1 and UBC9 were observed in the IHC and Western blot results. Furthermore, Western blot and Co-IP assays revealed an augmented interaction between SOX9 and SUMO1, which was potentiated by ZnCl2 treatment. Significantly, mutations at the SUMOylation site of SOX9 resulted in alterations in the expression patterns of crucial chondrogenesis factors. This research underscores how zinc ions promote cartilage development through the modification of SOX9 by SUMO1.
With the rapid development of the field of life sciences, traditional 2D cell culture and animal models have long been unable to meet the urgent needs of modern biomedical research and new drug development. Establishing a new generation of experimental models and research models is of great significance for deeply understanding human health and disease processes, and developing effective treatment measures. As is well known, long research and development cycles, high risks, and high costs are the “three mountains” facing the development of new drugs today. Organoids and organ-on-chips technology can highly simulate and reproduce the human physiological environment and complex reactions in vitro, greatly improving the accuracy of drug clinical efficacy prediction, reducing drug development costs, and avoiding the defects of drug testing animal models. Therefore, organ-on-chips have enormous potential in medical diagnosis and treatment.
