Iron overload-associated osteoporosis presents a significant challenge to bone health. This study examines the effects of arecoline (ACL), an alkaloid found in areca nut, on bone metabolism under iron overload conditions induced by ferric ammonium citrate (FAC) treatment. The results indicate that ACL mitigates the FAC-induced inhibition of osteogenesis in zebrafish larvae, as demonstrated by increased skeletal mineralization and upregulation of osteogenic genes. ACL attenuates FAC-mediated suppression of osteoblast differentiation and mineralization in MC3T3-E1 cells. RNA sequencing analysis suggests that the protective effects of ACL are related to the regulation of ferroptosis. We demonstrate that ACL inhibits ferroptosis, including oxidative stress, lipid peroxidation, mitochondrial damage, and cell death under FAC exposure. In this study, we have identified heme oxygenase-1 (HO-1) as a critical mediator of ACL inhibiting ferroptosis and promoting osteogenesis, which was validated by HO-1 knockdown and knockout experiments. The study links ACL to HO-1 activation and ferroptosis regulation in the context of bone metabolism. These findings provide new insights into the mechanisms underlying the modulation of osteogenesis by ACL. Targeting the HO-1/ferroptosis axis is a promising therapeutic approach for treating iron overload-induced bone diseases.

Cobalt alloys have numerous applications, especially as critical components in orthopedic biomedical implants. However, recent investigations have revealed potential hazards associated with the release of nanoparticles from cobalt-based implants during implantation. This can lead to their accumulation and migration within the body, resulting in adverse reactions such as organ toxicity. Despite being a primary interface for cobalt nanoparticle (CoNP) exposure, skeletal muscle lacks comprehensive long-term impact studies. This study evaluated whether selenium nanoparticles (SeNPs) could mitigate CoNP toxicity in muscle cells and zebrafish models. CoNPs dose-dependently reduced C2C12 viability while elevating reactive oxygen species (ROS) and apoptosis. However, low-dose SeNPs attenuated these adverse effects. CoNPs downregulated myogenic genes and α-smooth muscle actin (α-SMA) expression in C2C12 cells; this effect was attenuated by SeNP cotreatment. Zebrafish studies confirmed CoNP toxicity, as it decreased locomotor performance while inducing muscle injury, ROS generation, malformations, and mortality. However, SeNPs alleviated these detrimental effects. Overall, SeNPs mitigated CoNP-mediated cytotoxicity in muscle cells and tissue through antioxidative and antiapoptotic mechanisms. This suggests that SeNP-coated implants could be developed to eliminate cobalt nanoparticle toxicity and enhance the safety of metallic implants.

Ginsenosides have been used in traditional Chinese medicine to treat fractures. Ginsenoside Re, a key component of P. ginseng, however, the molecular mechanisms underlying these effects remain inadequately understood. This study aims to investigate the effects of Ginsenoside Re on bone formation in fractures and to explore the underlying molecular mechanisms involved. Cell proliferation and osteogenic differentiation were evaluated using the Cell Counting Kit-8 (CCK-8) assay and alkaline phosphatase (ALP) staining, respectively. Quantitative PCR (qPCR) and western blotting were employed to quantify mRNA and protein expression levels. To assess bone formation in vivo, a zebrafish caudal fin regeneration model was utilized, with calcein staining applied to visualize mineralized tissue. Additionally, a bone morphogenetic protein 2a (bmp2a) knockout zebrafish model was generated using the CRISPR-Cas9 system to further investigate the molecular mechanisms underlying ginsenoside Re-mediated bone formation. Ginsenoside Re promotes normal development and survival in zebrafish. In a zebrafish model, ginsenoside Re enhances bone regeneration, accompanied by upregulated expression of osteogenic markers. Following a 10-day treatment with 20 μM ginsenoside Re, mRNA sequencing (mRNA-Seq) analysis identified bmp2a as a critical regulator of bone formation. In vitro, ginsenoside Re stimulates osteogenic differentiation in MC3T3-E1 cells, elevating both mRNA and protein levels of bone morphogenetic protein 2 (BMP2). Co-treatment with noggin or SB203580 reverses ginsenoside Re-induced osteogenesis: noggin suppresses BMP2 expression and p38 phosphorylation, whereas SB203580 specifically inhibits p38 activity. In a bmp2a knockout zebrafish model, the pro-osteogenic effects of ginsenoside Re are completely abolished. Similarly, in wild-type zebrafish, both noggin and SB203580 attenuate the bone-promoting effects of ginsenoside Re. Noggin reduces the expression of bmp2a and p38a, while SB203580 specifically targets p38a. In conclusion, our findings demonstrate, for the first time, that ginsenoside Re at an optimal concentration of 20 μM enhances bone formation by modulating the osteogenic system. Mechanistically, we reveal that this pro-osteogenic effect is mediated through the BMP2/p38 signaling pathway.

Introduction: The escalating global surge in Rifampicin-resistant strains poses a formidable challenge to the worldwide campaign against tuberculosis (TB), particularly in developing countries. The frequent reports of suboptimal treatment outcomes, complications, and the absence of definitive treatment guidelines for Rifampicin-resistant spinal TB (DSTB) contribute significantly to the obstacles in its effective management. Consequently, there is an urgent need for innovative and efficacious drugs to address Rifampicin-resistant spinal tuberculosis, minimizing the duration of therapy sessions. This study aims to investigate potential targets for DSTB through comprehensive proteomic and pharmaco-transcriptomic analyses.Methods: Mass spectrometry-based proteomics analysis was employed to validate potential DSTB-related targets. PPI analysis confirmed by Immunohistochemistry (IHC) and Western blot analysis.Results: The proteomics analysis revealed 373 differentially expressed proteins (DEPs), with 137 upregulated and 236 downregulated proteins. Subsequent Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses delved into the DSTB-related pathways associated with these DEPs. In the context of network pharmacology analysis, five key targets—human leukocyte antigen A chain (HLAA), human leukocyte antigen C chain (HLA-C), HLA Class II Histocompatibility Antigen, DRB1 Beta Chain (HLA-DRB1), metalloproteinase 9 (MMP9), and Phospholipase C-like 1 (PLCL1)—were identified as pivotal players in pathways such as “Antigen processing and presentation” and “Phagosome,” which are crucially enriched in DSTB. Moreover, pharmaco-transcriptomic analysis can confirm that 58 drug compounds can regulate the expression of the key targets.Discussion: This research confirms the presence of protein alterations during the Rifampicin-resistant process in DSTB patients, offering novel insights into the molecular mechanisms underpinning DSTB. The findings suggest a promising avenue for the development of targeted drugs to enhance the management of Rifampicin-resistant spinal tuberculosis.