Autologous bone transplantation is an important method for cranial bone repair and reconstruction. This study systematically investigated the effects of different cryopreservation strategies and disinfection methods on autologous bone cells. Bone marrow mesenchymal stem cells (BMSCs) and collagen were added to the basic cryopreservation solution to evaluate the preservation effect. Peracetic acid-UV (PX-UV), supercritical CO₂ (SC-CO₂) and low-temperature plasma (cold atmospheric plasma, CAP) were used to treat skull specimens. Cell viability was detected by CCK-8, cell apoptosis was evaluated by TUNEL staining. RT-qPCR was performed to measure the expression of osteogenic-related genes, including Runt-related transcription factor 2 (Runx2), osteocalcin (OCN), type I collagen (COL1), osteopontin (OPN), and alkaline phosphatase (ALP). Adding BMSCs and collagen to the basic cryoprotectant solution significantly increased the survival rate of cryopreserved bone cells (P < 0.05), inhibited cell apoptosis, and upregulated the expression of osteogenic-related genes. In the disinfection experiment, there were no significant differences between the PX-UV disinfection group and the control group in terms of cell viability, apoptosis rate, and expression of osteogenic genes (P > 0.05). Adding BMSCs and collagen to the basic cryoprotectant solution is expected to improve the preservation effect of autologous bone tissue cells. The PX-UV disinfection has a relatively small impact on the cell survival rate and osteogenic properties of autologous bone cells.
Hypoxia promotes metabolic adaptation and aggressive behavior in prostate cancer, but the contribution of lysine crotonylation to this response is incompletely defined. This study examined whether enoyl-CoA hydratase 1 (ECHS1) is associated with hypoxia-responsive phosphoglycerate kinase 1 (PGK1) crotonylation and malignant phenotypes. ECHS1 expression and prognosis were evaluated using The Cancer Genome Atlas. PC-3 and LNCaP cells were exposed to 1
Cisplatin (CDDP) resistance reduces the treatment effect in non-small cell lung cancer. Lung cancer stem-like cells are closely linked to drug tolerance. Mitochondrial dysfunction may affect their stemness and response to treatment. This study examined whether peptide nucleic acid conjugated with triphenylphosphonium (PNA-TPP) could alter the response of A549-derived stem-like spheres to cisplatin. We also explored whether this effect was associated with Drp1 (dynamin-related protein 1)-related mitochondrial changes. A549-derived stem-like spheres were established by single-cell cloning and serum-free suspension culture. PNA-TPP was designed to target the mitochondrial DNA D-loop region and was used at 10 nmol/mL for 24 h. Cell viability, colony formation, sphere formation, apoptosis, migration, and invasion were assessed using CCK-8, colony formation, sphere formation, Annexin V-FITC/PI flow cytometry, wound healing, and Transwell assays. Mitochondrial membrane potential, reactive oxygen species (ROS), and ATP levels were detected using JC-1 staining, DCFH-DA flow cytometry combined with DHE fluorescence staining, and ELISA, respectively. Drp1 mRNA expression was examined by qRT-PCR, and Drp1 protein levels in the mitochondrial fraction were assessed by Western blotting. Drp1 overexpression and shRNA-mediated knockdown were used to evaluate its involvement. PNA-TPP increased Drp1 mRNA expression and mitochondrial Drp1 levels. PNA-TPP reduced mitochondrial membrane potential and ATP levels, increased ROS levels, and decreased sphere-forming ability and octamer-binding transcription factor 4 (OCT4), SRY-box transcription factor 2 (SOX2), and Nanog homeobox (NANOG) expression. Compared with CDDP treatment alone, PNA-TPP pretreatment plus CDDP resulted in lower cell viability, colony formation, migration, and invasion, together with increased apoptosis and a higher cleaved caspase-9/total caspase-9 ratio. Drp1 overexpression partly mimicked these effects, whereas Drp1 knockdown partially restored mitochondrial membrane potential and stemness-related phenotypes after PNA-TPP treatment. PNA-TPP treatment was associated with Drp1-related mitochondrial dysfunction, weakened stem-like phenotypes, and an enhanced response to cisplatin treatment in A549-derived stem-like spheres. Not applicable.
Although therapeutic advancements have dramatically improved outcomes for cancer patients in recent decades, advanced cancer remains a significant challenge. There is still an urgent need for new and more effective strategies that minimize toxicity and improve survival outcomes in cancer patients. Natural products from microorganisms are a rich and promising source of bioactive compounds with substantial therapeutic potential in cancer treatment. Recently, B. proteolyticus strain OSUB18, isolated from switchgrass, has been demonstrated to promote plant growth and regulate reactive oxygen species (ROS) production. Here, we investigated the anticancer potential of this plant-associated microbe and demonstrated that its crude extracts exhibited broad-spectrum cytostatic activity across diverse solid tumor cell lines. Importantly, OSUB18 extracts displayed limited cytotoxicity toward normal human mammary epithelial cells and mouse fibroblasts. This potentially selective cytostatic activity was retained within a water soluble, heat-labile, and protease-resistant fraction. The extracts primarily suppressed cell proliferation, accompanied by increased lipid peroxidation, with preliminary evidence suggesting the potential involvement of ferroptosis. These findings suggest that the bioactive component may be a selective cytostatic agent with multifaceted mechanisms of action. Not applicable.
This study investigated whether L-NAME, a nitric oxide synthase (NOS) inhibitor commonly used to model hypertension-associated endothelial dysfunction, can exert direct pro-atherogenic effects on vascular smooth muscle cells (VSMCs) independent of systemic hemodynamic influences. Mouse aortic VSMCs (MOVAS) were treated with non-cytotoxic concentrations of L-NAME (50 µM) or oxidized low-density lipoprotein (ox-LDL, 100 µg/mL), which served as a positive atherogenic control. Lipid accumulation was assessed by Oil Red O staining. The expression of contractile phenotype markers (ACTA2, MYH11, and α-SMA), the synthetic phenotype marker SPP1, and matrix metalloproteinases (MMP-2 and MMP-9) was quantified by qRT-PCR and immunofluorescence. Both L-NAME and ox-LDL treatments caused marked intracellular lipid accumulation in MOVAS cells, consistent with VSMC-derived foam cell formation. L-NAME and ox-LDL reduced the mRNA expression of contractile phenotype markers (ACTA2 and MYH11, p < 0.05) and decreased α-SMA protein expression (p < 0.001). Conversely, both treatments increased the mRNA expression of the synthetic phenotype marker SPP1 (p < 0.05) and matrix-remodeling genes MMP-2 and MMP-9 (p < 0.05). These effects were directionally similar to those induced by ox-LDL. L-NAME directly promotes lipid accumulation and synthetic phenotype-associated marker changes in MOVAS cells under the present in vitro conditions. Because eNOS/NOS activity, intracellular NO levels, rescue pathways, and primary VSMC validation were not assessed, these data should be interpreted as phenotypic evidence that NOS/NO pathway disruption may contribute to VSMC dysfunction rather than definitive proof of a single eNOS-dependent mechanism. Not applicable.
