Organogenesis最新文献

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.

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.

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 evaluates the effectiveness of optimized individualized nursing interventions on clinical outcomes in intensive care unit (ICU) patients with severe pneumonia.

Methods: 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.

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 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.

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.