Cytotechnology最新文献

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are respiratory conditions associated with high mortality rates, primarily due to intense pulmonary inflammation. Lipopolysaccharide (LPS), an essential constituent of the wall of gram-negative bacteria, has the potential to trigger inflammation, ultimately leading to ALI. This study assessed the protective role of selegiline in mitigating LPS-induced inflammation in A549 cells. A549 cells were divided into seven groups: the negative control (PBS, 100 μM), the positive control (H₂O₂, 100 μM), the LPS group (1 µg/ml), groups treated with selegiline (Sel) (20, 30, and 40 μM) + LPS, and a group receiving Sel (40 μM). Parameters, including cell viability, iNOS activity, levels of SOD, CAT, and GPx, along with the pro-inflammatory cytokines IL-1β and TNF-α, as well as the protein expressions of NF-kB, NLRP3, and Caspase-1, were analyzed. Findings revealed that exposure to LPS increased oxidation and inflammation in A549 cells. Conversely, Sel markedly enhanced antioxidant capacity and diminished the iNOS activity, IL-1β, and TNF-α levels, as well as reduced the expression of inflammatory markers, including NF-kB, NLRP3, and Caspase-1 proteins. These results indicate that Sel may ameliorate LPS-induced pulmonary inflammation through its beneficial properties, including antioxidant and anti-inflammatory effects.

Sevoflurane, a widely utilized inhalational anesthetic, has been shown in previous studies to inhibit the proliferation and invasion of non-small cell lung cancer (NSCLC) cells. However, whether sevoflurane affects the metastasis of small cell lung cancer (SCLC) cells remains unexplored. This study's primary objective was to investigate the mechanism by which sevoflurane induces calcium overload and EFHD1 upregulation, thereby contributing to pyroptosis in SCLC cells. NCI-H446 cells were treated with sevoflurane at concentrations of 0%, 0.6%, 3% and 15%. Cell proliferation was assessed using the CCK8 assay, while cell migration and invasion were analyzed using the Transwell assay. EFHD1 expression was determined by q-PCR and Western blot. Calcium content, ROS levels, and CCO content were measured using respective assay kits, and mitochondrial morphology was observed using transmission electron microscopy. Western blotting was performed to detect the expression of caspase-1 and GSDMD. All procedures were repeated under conditions of EFHD1 overexpression or silencing. Sevoflurane inhibited the proliferation, migration, and invasion of NCI-H446 cells, concurrently exerting a significant pro-pyroptotic effect. This was mediated by mitochondrial calcium overload, EFHD1 upregulation, and subsequent mitochondrial damage-induced pyroptosis. Sevoflurane significantly enhances pyroptosis in NCI-H446 cells, thereby potentially reducing cancer cell dissemination during surgery.

Understanding the molecular mechanisms regulating osteo/cementogenic differentiation is critical for optimizing cell-based strategies in periodontal regeneration. This study employed multivariate statistical modeling to investigate, in vitro, the interaction between bone morphogenetic protein 2 (BMP2) and asporin (ASPN) in human periodontal ligament stem cells (hPDLSCs) and its impact on mineralization potential. Four primary hPDLSC populations-two with high osteo/cementogenic potential (HOP) and two with low potential (LOP)-were cultured under standard (SDM) and osteo/cementogenic (OM) conditions. Mineralization was assessed using Alizarin Red Staining (AR-S) and alkaline phosphatase (ALP) activity, while gene expression of ASPN, BMP2, osteocalcin (OCN), and runt-related transcription factor 2 (RUNX2) was quantified by quantitative real-time polymerase chain reaction (qRT-PCR). BMP2 protein levels were measured using the Luminex system. Data analysis incorporated Principal Component Analysis (PCA), Linear Discriminant Analysis (LDA), Pearson correlations, correlation networks, and multivariate regression models in RStudio. HOP cells exhibited higher mineralization and BMP2 expression compared to LOP cells (p < 0.05). ASPN negatively correlated with BMP2 expression and mineralization, particularly in LOP cells (p < 0.01), reinforcing its inhibitory role. Multivariate modeling identified BMP2 as a key positive regulator of osteo/cementogenesis, while ASPN emerged as a significant inhibitory factor (p < 0.001). This study highlights the potential of multivariate models as powerful tools for uncovering molecular interactions and identifying novel therapeutic targets, paving the way for advancements in periodontal regeneration and cell-based therapies.

Graphical abstract: Created in BioRender. Sacramento, C. (2025).

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00810-x.

[This corrects the article DOI: 10.1007/s10616-021-00496-x.].

Understanding the impact of critical process parameters on CHO cell cultivation and critical quality attributes (CQAs) of monoclonal antibody is an essential requirement for robust process scale-up and technology transfer. The effect of process parameters on lactate metabolism is of particular interest as it has been shown to be associated with cell growth and productivity. Here, a comprehensive study is presented to investigate the impact of potentially critical process parameters (aeration rate, agitation speed, dissolved oxygen level and copper concentration) on process performance and CQAs. Results demonstrated that dissolved oxygen level has a dominate positive effect on lactate consumption. Scale-relevant parameters (aeration rate and agitation speed) and copper concentration have smaller but also significant positive influence on lactate consumption. Based on these results, it can be summarized that low dissolved oxygen, agitation and aeration rates plus low copper concentration, would lead to insufficient oxygen availability for cell respiration (hypoxia). Under this condition, cells would rely on anaerobic glycolysis to generate energy and hence promote lactate production. This study also revealed a linkage between hypoxic condition and antibody purity and charge heterogeneity. This work provides critical insights for lactate metabolic and product quality control through optimizing critical process parameters and medium component.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00816-5.

Endometrial cancer (EC) is one of the common malignancies of the female reproductive system, and metastasis is critical to the progression and prognosis of EC. As a transcription factor, ELF3 is widely involved in the regulation of cancer cell growth and metastasis, but its role in EC is unclear. The purpose of this study was to explore the role and mechanism of ELF3 in regulating EC progression. In this study, we collected 8 pairs of EC tissue and normal paracancerous endometrial tissue, and injected human endometrial cancer cells (Ishikawa) into the left axilla of nude mice to construct a model of subcutaneous tumorigenesis in nude mice. The growth of Ishikawa cells and tumor tissues of EC nude mice was evaluated by CCK-8, clone formation assay, immunohistochemistry, etc., and the expression of related proteins and genes was detected by Western blot and RT-qPCR. In this study, it was found that the expression of ELF3 was up-regulated in EC, and knockdown of ELF3 could inhibit the proliferative activity and colony formation of EC cells, promote the expression of E-cadherin, inhibit the expression of N-cadherin and Vimentin, and reduce the epithelial-mesenchymal transition (EMT) of EC cells, thereby inhibiting the migration of EC cells to a certain extent and alleviating the development of EC in vivo. Mechanistically, ELF3 inhibits TPM1 transcription by binding to the promoter region of TPM1 gene, thereby inhibiting TPM1 expression, promoting EMT, proliferation and migration of EC cells, and accelerating the occurrence and development of EC in vivo. ELF3 promotes the development of EC through TPM1 regulation of EMT, and inhibition of ELF3 may become a potential new target for the treatment of EC.

Mitochondrial medicine has shown great promise as a therapeutic approach for treating currently incurable diseases. Preclinical studies highlight its safety and efficacy, but significant challenges remain in translating these therapies from bench to bedside. Key unresolved issues include understanding the mechanisms behind the reparative potential of transplanted mitochondria, such as their viability and functionality in an extracellular environment, especially under elevated calcium ion concentrations. Additionally, challenges related to mitochondrial sourcing, delivery methods, and ethical considerations need to be addressed for broader clinical adoption. This review analyses these challenges and explores strategies to overcome them, including refining mitochondrial sourcing, delivery techniques, and storage solutions. We also emphasise the need for rigorous ethical guidelines and regulatory frameworks to ensure safe and global implementation, paving the way for mitochondrial medicine's broader clinical use.

The development of novel treatment strategies is essential to treat Cervical Cancer (CC) as it is the fourth-leading cancer among women. Gloriosa superba is a medicinal plant that retains various pharmacological activities. It possesses significant anticancer properties that have been previously studied. However, the anticancer efficacy of the nanocombination of G. superba tuber and seed has not yet been studied in CC. This study aimed to evaluate the anticancer efficacy of chitosan-encapsulated G. superba tuber nanoparticles and chitosan-encapsulated G. superba seed nanoparticles (CEGSTNs and CEGSSNs) via targeting the SKA3-mediated PI3K/AKT/mTOR pathway in CC. The CEGSTNs and CEGSSNs were synthesized and characterized by UV, DLS, zeta potential, FTIR, and TEM analysis. The anticancer efficacy on cell viability, proliferation, and apoptosis was investigated, and RT-PCR was used to measure the expression of the SKA3-mediated PI3K/AKT/mTOR pathway in HeLa cell lines. Furthermore, the acute toxicity assessment was conducted in Wistar rats, and body weights, haematological, and biomedical parameters, as well as histopathological studies, were performed. Characterisation techniques confirmed the synthesis of CEGSTNs and CEGSSNs. Both exhibited significant anticancer activity, induced apoptosis, and downregulated SKA3 expression, which inactivated the PI3K/AKT/mTOR pathway in HeLa cells. Acute toxicity analysis showed no toxicity or adverse effects in the treatment group. Overall, these results suggested that CEGSTNs have exhibited more anticancer efficacy than CEGSSNs. Moreover, CEGSTNs induced apoptosis and suppressed the proliferation of cells via the downregulation of the SKA3-mediated PI3K/AK/mTOR pathway.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00809-4.

[This corrects the article DOI: 10.1007/s10616-022-00528-0.].

Previously, there have been reports of decreased expression of GRIM-19 and increased expression of MDM2 in lung adenocarcinoma. However, the relationship between GRIM-19 and MDM2 in lung adenocarcinoma has not been reported yet. In this study, we demonstrated that GRIM-19mRNA expression was reduced in lung adenocarcinoma tissue while MDM2mRNA expression was increased. Pearson correlation coefficient analysis showed a significant correlation between GRIM-19mRNA and MDM2mRNA (r = -0.970, p < 0.001). In this study, we further detected by experiments that the overexpression of GRIM-19 lowered the expression level of MDM2 protein, suggesting that GRIM-19 may affect the occurrence and development of lung adenocarcinoma by inhibiting the expression of MDM2. Cellular functional and subcutaneous tumour formation experiments in nude mice have shown that GRIM-19 inhibits the proliferation, invasion, and migration ability of lung adenocarcinoma cells, promotes cell apoptosis, and ultimately inhibits lung adenocarcinoma development. In addition, overexpression of GRIM-19 in A549 cells resulted in significant changes in the expression of EMT-related proteins Snail, N-Cadherin, E-Cadherin, and Vimentin, indicating that GRIM-19 may inhibit the metastasis of lung adenocarcinoma cells by regulating the EMT pathway.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-025-00797-5.